Compositions and methods for the diagnosis and treatment of body weight disorders, including obesity

This application is a divisional of U.S. application Ser. No. 09/245,041, filed Feb. 5, 1999, now U.S. Pat. No. 6,274,339 which claims benefit to Provisional Application No. 60/093,630, filed on Jul. 21, 1998 and of Provisional Application No. 60/104,978, filed Oct. 20, 1998, the contents of each of which are incorporated by reference in their entirety.

1. INTRODUCTION

The present invention relates to mammalian mahogany genes, including the human mahogany gene, which are novel genes involved in the control of mammalian body weight. The invention encompasses nucleotide sequences of the mahogany gene, host cell expression systems of the mahogany gene, and hosts which have been transformed by these expression systems, including transgenic animals. The invention also encompasses novel mahogany gene products, including mahogany proteins, polypeptides and peptides containing amino acid sequences mahogany proteins, fusion proteins of mahogany proteins polypeptides and peptides, and antibodies directed against such mahogany gene products.

The present invention also relates to methods and compositions for the diagnosis and treatment of mammalian body weight disorders, including obesity, cachexia, and anorexia, and for the identification of subjects susceptible to such disorders. Further, the invention relates to methods of using the mahogany gene and gene products of the invention for the identification of compounds which modulate the expression of the mahogany gene and/or the activity of the mahogany gene product. Such compounds can be useful as therapeutic agents in the treatment of mammalian body weight disorders, including obesity, cachexia, and anorexia.

2. BACKGROUND OF THE INVENTION

Obesity represents the most prevalent of body weight disorders, and it is the most important nutritional disorder in the western world, with estimates of its prevalence ranging from 30% to 50% within the middle-aged population. Other body weight disorders, such as anorexia nervosa and bulimia nervosa, which together affect approximately 0.2% of the female population of the western world, also pose serious health threats. Further, such disorders as anorexia and cachexia (wasting) are also prominent features of other diseases such as cancer, cystic fibrosis, and AIDS.

Obesity, defined as an excess of body fat relative to lean body mass, also contributes to other diseases. For example, this disorder is responsible for increased incidence of diseases such as coronary artery disease, hypertension, stroke, diabetes, hyperlipidemia, and some cancers (See, e.g., Nishina, P. M. et al., 1994, Metab. 43: 554-558; Grundy, S. M. & Barnett, J. P., 1990, Dis. Mon. 36: 641-731). Obesity is not merely a behavioral problem, i.e., the result of voluntary hyperphagia. Rather, the differential body composition observed between obese and normal subjects results from differences in both metabolism and neurologic/metabolic interactions. These differences seem to be, to some extent, due to differences in gene expression, and/or level of gene products or activity (Friedman, J. M. et al., 1991, Mammalian Gene 1: 130-144).

The epidemiology of obesity strongly shows that the disorder exhibits inherited characteristics (Stunkard, 1990, N. Eng. J. Med. 322: 1438). Moll et al. have reported that, in many populations, obesity seems to be controlled by a few genetic loci (Moll et al., 1991, Am. J. Hum. Gen. 49: 1243). In addition, human twin studies strongly suggest a substantial genetic basis in the control of body weight, with estimates of heritability of 80-90% (Simopoulos, A. P. & Childs, B., eds., 1989, in “Genetic Variation and Nutrition in Obesity”, World Review of Nutrition and Diabetes 63, S. Karger, Basel, Switzerland; Borjeson, M., 1976, Acta. Paediatr. Scand. 65: 279-287).

In other studies, non-obese persons who deliberately attempted to gain weight by systematically over-eating were found to be more resistant to such weight gain and able to maintain an elevated weight only by very high caloric intake. In contrast, spontaneously obese individuals are able to maintain their status with normal or only moderately elevated caloric intake. In addition, it is a commonplace experience in animal husbandry that different strains of swine, cattle, etc., have different predispositions to obesity. Studies of the genetics of human obesity, and of animal models of obesity demonstrate that obesity results from complex defective regulation of both food intake, food induced energy expenditure, and of the balance between lipid and lean body anabolism.

There are a number of genetic diseases in man and other species which feature obesity among their more prominent symptoms, along with, frequently, dysmorphic features and mental retardation. For example, Prader-Willi syndrome (PWS; reviewed in Knoll, J. H. et al., 1993, Am. J. Med. Genet. 46: 2-6) affects approximately 1 in 20,000 live births, and involves poor neonatal muscle tone, facial and genital deformities, and generally obesity.

In addition to PWS, many other pleiotropic syndromes have been characterized which include obesity as a symptom. These syndromes are genetically straightforward, and appear to involve autosomal recessive alleles. Such diseases include, among others, Ahlstroem, Carpenter, Bardet-Biedl, Cohen, and Morgagni-Stewart-Monel Syndromes.

A number of models exists for the study of obesity (see, e.g., Bray, G. A., 1992, Prog. Brain Res. 93: 333-341; and Bray, G. A., 1989, Amer. J. Clin. Nutr. 5: 891-902). For example, animals having mutations which lead to syndromes that include obesity symptoms have also been identified. Attempts have been made to utilize such animals as models for the study of obesity, and the best studied animal models to date for genetic obesity are mice. For reviews, see, e.g., Friedman, J. M. et al., 1991, Mamm. Gen. 1: 130-144; Friedman, J. M. and Liebel, R. L., 1992, Cell 69: 217-220.

Studies utilizing mice have confirmed that obesity is a very complex trait with a high degree of heritability. Mutations at a number of loci have been identified which lead to obese phenotypes. These include the autosomal recessive mutations obese (ob), diabetes (db), fat (fat), and tubby (tub).

The dominant Yellow mutation (Ay) at the agouti locus causes a pleiotropic syndrome which causes moderate adult onset obesity, a yellow coat color, and a high incidence of tumor formation (Herberg, L. and Coleman, D. L., 1977, Metabolism 26:59), and an abnormal anatomic distribution of body fat (Coleman, D. L., 1978, Diabetologia 14:141-148). The mutation causes the widespread expression of a protein which is normally seen only in neonatal skin (Michaud, E. J. et al., 1994, Genes Devel. 8:1463-1472). The agouti protein has been reported to be a competitive antagonist of α-MSH binding to the melanocortin receptors MC1-R and MC4-R in vitro (Lu et al., 1996, Nature 371:799-802), and the authors speculated that de-regulated ubiquitous expression of agouti may lead to obesity by antagonism of melanocortin receptors expressed outside the hair follicles.

Mahogany (mg) and mahoganoid (md) are mutations that suppress the phenotypic effects of agouti protein in vivo (Lane and Green, 1960, J. Hered. 51: 228-230). The mahogany and mahoganoid mutation have been mapped to mouse chromosomes 2 and 16, respectively (Green, 1989, “Catalog of mutant genes and polymorphic loci”, pp. 12-403 in Genetic Variants and Strains of the Laboratory Mouse , Lyon, M. F. and Searle, A. G., eds., Oxford University Press, Oxford). Mutations of both mg and md have been shown to suppress the effects of agouti on obesity as well as on coat color (Miller et al., 1997, Genetics 146: 1407-1415).

In summary, therefore, obesity, which poses a major, worldwide health problem, represents a complex, highly heritable trait. Given the severity, prevalence, and potential heterogeneity of such disorders, there exists a great need for the identification of those genes that participate in the control of body weight.

3. SUMMARY OF THE INVENTION

The present invention relates to the identification of novel nucleic acid molecules and proteins encoded by such nucleic acid molecules that are involved in the control of mammalian body weight, and which, further, are associated with mammalian body weight disorders such as obesity, cachexia, and anorexia. The nucleic acid molecules of the present invention represent the genes corresponding to the mammalian mahogany gene, including the human mahogany gene.

In particular, the compositions of the present invention include nucleic acid molecules which comprise the following sequences: (a) nucleotide sequences of the murine mahogany gene (SEQ ID NO.: 1) shown in FIG. 2A, as well as allelic variants and homologs thereof, as shown, e.g., in FIGS. 8A-C and 9 A; (b) nucleotide sequences that encode the mahogany gene product amino acid sequence (SEQ ID NO.: 2) shown in FIG. 2B; (c) nucleotide sequences that encode portions of the mahogany gene product corresponding to its functional domains and individual exons; (d) nucleotide sequences comprising the novel mahogany gene sequences disclosed herein that encode mutants of the mahogany gene product in which all or a part of one or more of the domains is deleted or altered, as shown, e.g., in FIG. 6; (e) nucleotide sequences that encode fusion proteins comprising the mahogany gene product (SEQ ID NO.: 2), or one or more of its domains fused to a heterologous polypeptide; (f) nucleotide sequences within the mahogany gene, as well as chromosome sequences flanking the mahogany gene, see, e.g., FIG. 3, which can be utilized as part of the methods of the present invention for the diagnosis of mammalian body weight disorders, including obesity, cachexia, and anorexia, which are mediated by the mahogany gene, as well as for the identification of subjects susceptible to such disorders; (g) nucleic acid sequences that hybridize to the above described sequences under stringent or moderately stringent conditions, particularly human mg homologues, as shown, e.g., FIG. 10 . The nucleic acid molecules of the invention include, but are not limited to, cDNA and genomic DNA sequences of the mahogany gene.

The present invention also encompasses expression products of the nucleic acid molecules listed above; i.e., proteins and/or polypeptides that are encoded by the above mahogany nucleic acid molecules.

Agonists and antagonists of the mahogany gene and/or gene product are also included in the present invention. Such agonists and antagonists will include, for example, small molecules, large molecules, and antibodies directed against the mahogany gene product. Agonists and antagonists of the invention also include nucleotide sequences, such as antisense and ribozyme molecules, and gene or regulatory sequence replacement constructs, that can be used to inhibit or enhance expression of the mahogany gene.

The present invention further encompasses cloning vectors, including expression vectors, that contain the nucleic acid molecules of the invention and can be used to express those nucleic acid molecules in host organisms. Further, host organisms which have been transformed with these nucleic acid molecules are also encompassed in the present invention. Host organisms of the invention include organisms transformed with the cloning vectors described above, e.g., transgenic animals, particularly non-human transgenic animals, and particularly transgenic non-human mammals.

The transgenic animals of the invention include animals that express a mutant variant or polymorphism of a mahogany gene, particularly a mutant variant or polymorphism of a mahogany gene that is associated with a weight disorder such as obesity, cachexia, or anorexia. The transgenic animals of the invention further include those that express a mahogany transgene at higher or lower levels than normal. The transgenic animals of the invention further include those which express the mahogany gene in all their cells, “mosaic” animals which express the mahogany gene in only some of their cells, and those in which the mahogany gene is selectively introduced into and expressed in a specific cell type(s). The transgenic animals of the invention also include “knock-out” animals. Knock-out animals comprise animals which have been engineered to no longer express the mahogany gene.

The present invention also relates to methods and compositions for the diagnosis of mammalian body weight disorders, including obesity, cachexia, and anorexia, as well as for the identification of subjects susceptible to such disorders. Such methods comprise, for example, measuring expression of the mahogany gene in a patient sample, or detecting a mutation in the mahogany gene in the genome of a mammal, including a human, suspected of exhibiting such a weight disorder. The nucleic acid molecules of the invention can also be used as diagnostic hybridization probes, or as primers for diagnostic PCR analysis to identify of mahogany gene mutations, allelic variations, or regulatory defects, such as defects in the expression of the mahogany gene. Such diagnostic PCR analyses can be used to diagnose individuals with a body weight disorder associated with a particular mahogany gene mutation, allelic variation, or regulatory defect. Such diagnostic PCR analyses can also be used to identify individuals susceptible to such body weight disorders and hyperphagia.

Methods and compositions, including pharmaceutical compositions, for the treatment of body weight disorders such as obesity, cachexia, and anorexia are also included in the invention. Such methods and compositions are capable of modulating the level of mahogany gene expression and/or the level of activity of the mahogany gene product. Such methods include, for example, modulating the expression of the mahogany gene and/or the activity of the mahogany gene product for the treatment of a body weight disorder which is mediated by some other gene, for example by the agouti gene.

The invention still further relates to methods for identifying compounds which modulate the expression of the mammalian mahogany gene and/or the synthesis or activity of mammalian mahogany gene products. Such compounds include therapeutic compounds which can be used as pharmaceutical compositions to reduce or eliminate the symptoms of mammalian body weight disorders such as obesity, cachexia, and anorexia. Cellular and non-cellular assays are described that can be used to identify compounds that interact with the mahogany gene and/or gene product, e.g., modulate the activity of the mahogany gene and/or bind to the mahogany gene product. Such cell-based assays of the invention utilize cells, cell lines, or engineered cells or cell lines that express the mahogany gene product.

In one embodiment, such methods comprise contacting a compound to a cell that expresses a mahogany gene, measuring the level of mahogany gene expression, gene product expression, or gene product activity, and comparing this level to the level of mahogany gene expression, gene product expression, or gene product activity produced by the cell in the absence of the compound, such that if the level obtained in the presence of the compound differs from that obtained in its absence, a compound that modulates the expression of the mammalian mahogany gene and/or the synthesis or activity of mammalian mahogany gene products has been identified.

In an alternative embodiment, such methods comprise administering a compound to a host, e.g., a transgenic animal that expresses a mahogany transgene or a mutant mahogany transgene, and measuring the level of mahogany gene expression, gene product expression, or gene product activity. The measured level is compared to the level of mahogany gene expression, gene product expression, or gene product activity in a host that is not exposed to the compound, such that if the level obtained when the host is exposed to the compound differs from that obtained when the host is not exposed to the compound, a compound that modulates the expression of the mammalian mahogany gene and/or the synthesis or activity of mammalian mahogany gene products, and/or the symptoms of a mammalian body weight disorder, such as obesity, cachexia, or anorexia, has been identified.

The Example presented in Section 6, below, describes the genetic and physical mapping of the mahogany gene to a specific 700 kb interval of mouse chromosome 2. The example presented in Section 7, below, describes the identification of a transcription unit within this chromosome interval, referred to herein as the MG gene, which represents the mahogany gene. The expression and sequence analysis of this candidate mahogany gene is described in the example presented in Section 8, below. These experiments prove that the candidate gene MG is indeed the mahogany gene.

As used herein, the following terms shall have the abbreviations indicated.

BAC, bacterial artificial chromosomes

bp, base pair(s)

EST, expressed sequence tag

mg, mahogany gene

RFLP, restriction fragment length polymorphism

RT-PCR, reverse transcriptase PCR

SSCP, single-stranded conformational polymorphism

SSLP, simple sequence length polymorphisms

STS, short tag sequence

YAC, yeast artificial chromosome

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 . Physical map of the mahogany interval of mouse chromosome 2.

FIG. 2 . Panel A(1)-A(9): cDNA nucleotide sequence of the wild-type (C57BL/6J) murine mahogany gene (SEQ ID NO: 1), including the 5′ and 3′ untranslated regions, and Panel B: the derived amino acid sequence (SEQ ID NO: 2) of the mahogany gene product.

FIG. 3 . Genomic structure and nucleotide sequences derived from the wild-type (C57BL/6J) mouse genomic regions containing the mg gene. Panel A, genomic structure; Panel B(1)-B(17), genomic sequence c56 (SEQ ID NO: 3); Panel C(1)-C(6), genomic sequence c96 (SEQ ID NO: 4); Panel D(1)-D(86), genomic sequence of c110/111 (SEQ ID NO: 5).

FIG. 4 . Structural depiction of MG cDNA without introns. CUB=CUB domain, metal=metallothionin domain; T-transmembrane domain.

FIGS. 5 ( 1 )- 5 ( 4 ). Nucleotide sequence of primers used to amplify each of the exons in the mg gene.

FIG. 6 . Nucleotide sequence of the wild-type (SEQ ID NO: 6) and mahogany mutant (SEQ ID NO: 7) sequences in exon 15 of the MG gene. Bases shown in bold are deleted in Mg3J mutant mg.

FIG. 7 . Differential 5′ start sequences in the murine mahogany gene showing splice forms akm1003 and akm1004.

FIG. 8 . Panel A-C, cDNA sequence (SEQ ID NO: 8) from one form of the differential 5′ start site found in the murine (akm1003), Panel D, amino acid sequence (SEQ ID NO: 9) encoded by the cDNA of Panel A-C; Panel E, hydropathy plot of the akm1003 amino acid sequence.

FIG. 9 . Panel A, cDNA sequence (SEQ ID NO: 10) from one form of the differential 5′ start site found in the murine (akm1004); Panel B, amino acid sequence (SEQ ID NO: 11) encoded by the cDNA of Panel A; Panel C, hydropathy plot of the akm1004 amino acid sequence.

FIG. 10 . Nucleotide sequence (SEQ ID NO: 12) of a contig containing a portion of the human MG cDNA, panel A(1)-A(7) and the translated amino acid sequence (SEQ ID NO: 13), panel B.

FIG. 11 . Effect of mg on MC4r−/− induced weight gain in females (FIG. 11A) and males (FIG. 11 B); values depicted are the mean+/−SD within a designated time interval.

FIG. 12 . Effect of mg on monogenic obese mutants Lepr db (FIG. 12 A), tub (FIG. 12 B), Cpe fat (FIG. 12 C), and on high fat diet induced obesity (FIG. 12 D); the values indicated are the mean+/−SD of the weight length ratio for each animal.

FIG. 13 . Genetic and physical map of the region surrounding the mg locus; all MIT markers are presented with shortened names, e.g., D2MIT77 is indicated as D2M77; locations of loci which also mapped on the human cytogenetic map are indicated in parentheses after the gene symbol.

FIG. 13 A. The genetic map of the mg gene region on the Millennium BSB mapping panel (Misumi, D. J. et al., 1997, Science 278:135-138);

FIG. 13 B. The genetic map obtained from crosses segregating mg mutant alleles;

FIG. 13 C. The ˜1 Mb BAC contig across the mg gene region of mouse Chromosome 2;

FIG. 13 D. The transcriptional units identified in the mg region; the filled box indicates the mg gene, whereas the hatched box is a member of the High Mobility Group (HMG) gene family which sits between coding exons 21 and 22 of the mg gene.

FIG. 14 . Northern blot analysis with C3H/HeJ (lane 1), and three mutant alleles of mg: C3HeB/FeJ-mg 3J (Lane 2), LDJ/Le-mg (Lane 3), and C3H/HeJ-mg J (Lane 4); the size marker is shown on the left, and hybridization with actin is shown below for loading comparisons.

FIG. 15 . In situ hybridization data: FIG. 15A demonstrates widespread expression of mg throughout the mouse brain is seen in an antisense autoradiographic image of a C3H/HeJ brain at the level of the 3rd ventricle; decreased expression in mg mutants is documented in selected antisense darkfield images of 10 μm whole mount cross sections of the ventromedial hypothalamic nucleic (VMH) of C3H/HeJ (FIG. 15 B), LDJ/Le-mg (FIG. 15 C), and C3HeB/FeJ-mg 3J (FIG. 15 D).

FIG. 16 . Alignment of the MG protein sequence with its family members showing the transmembrane region (indicated in brackets) and cytoplasmic tail (FIG. 16 A); and a schematic of the molecular modular architecture of MG (FIG. 16 B).

FIGS. 17A-D. Sequence alignment of the predicted MG protein sequence (top) with the Attractin protein sequence. Characteristic MG domains are as indicated. See Section 10.2 for details.

FIGS. 18A-D. Panel A1-A8: cDNA nucleotide sequence (SEQ ID NO: 14) of the long splice variant of the human ortholog of the mahogany gene, and Panel B-D: the derived amino acid sequence (SEQ ID NO: 15) of the mahogany gene product which it encodes.

FIGS. 19A-D. Panel A-C: cDNA nucleotide sequence (SEQ ID NO: 16) of a shorter splice variant of the human ortholog of the mahogany gene, and Panel D: the derived amino acid sequence (SEQ ID NO: 17) of the mahogany gene product which it encodes.

FIGS. 20A-C. Panel A-B: cDNA nucleotide sequence (SEQ ID NO: 18) of a second shorter splice variant of the human ortholog of the mahogany gene, and Panel C: the derived amino acid sequence (SEQ ID NO: 19) of the mahogany gene product which it encodes.

5. DETAILED DESCRIPTION OF THE INVENTION

Described herein is the identification of the novel mammalian mahogany (mg) gene, including the human mahogany gene, which is involved in the control of mammalian body weight. Also described are recombinant mammalian, including human mahogany DNA molecules, cloned genes, and degenerate variants thereof. The compositions of the present invention further include mg gene products (e.g., proteins) that are encoded by the mg DNA molecules of the invention, and the modulation of mg gene expression and/or mg gene product activity in the treatment of mammalian body weight disorders, including obesity, cachexia, and anorexia. Also described herein are antibodies against mg gene products (e.g., proteins), or conserved variants or fragments thereof, and nucleic acid probes useful for the identification of mg gene mutations, and the use of such nucleic acid probes in diagnosing mammalian body weight disorders, including obesity, cachexia, and anorexia. Further described are methods for the use of the mg gene and/or mg gene products in the identification of compounds which modulate the activity of the mg gene product.

5.1. The Mahogany Gene

The mahogany genes are novel mammalian genes involved in the control of body weight. The nucleic acid sequences of the mahogany genes, including the murine mahogany gene (SEQ ID NO: 1) shown in FIG. 2A, (with splice variants shown in FIGS. 8A-C and 9 A) are described herein. The genomic sequence and structure, i.e., the intron/exon structure, of the mahogany genes have also been elucidated, FIG. 3 .

The mahogany gene nucleic acid molecules of the present invention comprise: (a) the DNA sequence shown in FIG. 2 (SEQ ID NO: 1), or any DNA sequence that encodes the amino acid sequence of the mahogany gene product shown in FIG. 2, (SEQ ID NO: 2); (b) nucleotide sequences comprising the novel mahogany sequences disclosed herein that encode mutants of the mahogany gene product in which all or a part of one or more of the domains is deleted or altered, FIG. 6; (c) nucleotide sequences that encode fusion proteins comprising a mahogany gene product, or one of its domains fused to a heterologous polypeptide; and (d) nucleotide sequences within a mahogany gene, nucleotide sequences on the chromosome flanking the mahogany gene, FIG. 3, which can be utilized as part of the methods of the invention for identifying and diagnosing individuals who exhibit or are susceptible to weight disorders, including obesity, cachexia, and anorexia.

The mahogany nucleotide sequences of the invention further comprise: (a) any nucleotide sequence that hybridizes to the complement of a nucleic acid molecule that encodes a mahogany gene product under highly stringent conditions, e.g., hybridization to filter-bound DNA in 0.5 M NaHPO 4 , 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C., and washing in 0.1×SSC/0.1% SDS at 68° C. (Ausubel F. M. et al., eds., 1989, Current Protocols in Molecular Biology, Vol. I, Green Publishing Associates, Inc., and John Wiley & Sons, Inc., New York, at p. 2.10.3) particularly human mg sequences, FIG. 10; and (b) any nucleotide sequence that hybridizes to the complement of a nucleic acid molecule that encodes a mahogany gene product under less stringent conditions, such as moderately stringent conditions, e.g., washing in 0.2×SSC/0.1% SDS at 42° C. (Ausubel et al., 1989, supra), yet which still encodes a functionally equivalent mahogany gene product.

“Functionally equivalent”, as utilized herein, refers to a gene product (e.g., a protein) capable of exhibiting a substantially similar in vivo activity as the endogenous mg gene products encoded by the mg gene sequences described above. The in vivo activity of the mg gene product, as used herein, refers to the ability of the mg gene product, when present in an appropriate cell type, to ameliorate, prevent, or delay the appearance of the mahogany phenotype relative to its appearance when that cell type lacks a functional mahogany gene product.

The invention also includes nucleic acid molecules, preferably DNA molecules, that are the complements of the nucleotide sequences described above. Among the nucleic acid molecules of the invention are deoxyoligonucleotides (“oligos”) which hybridize under highly stringent or moderately stringent conditions to the mahogany nucleic acid molecules described above. Exemplary highly stringent conditions may refer, e.g., to washing in 6×SSC/0.05% sodium pyrophosphate at 37° C. (for 14-base oligos), 48° C. (for 17-base oligos), 55° C. (for 20-base oligos), and 60° C. (for 23-base oligos). These nucleic acid molecules may encode or act as antisense molecules, useful, for example, in mahogany gene regulation, and/or as antisense primers in amplification reactions of mahogany gene nucleic acid sequences. With respect to mahogany gene regulation, such techniques can be used to regulate, for example, weight disorders such as obesity, cachexia, or anorexia. Such sequences may also be used as part of ribozyme and/or triple helix sequences, which are also useful for mahogany gene regulation. Still further, such molecules may be used as components of diagnostic methods whereby, for example, the presence of a particular mahogany allele associated with a weight disorder, such as obesity, cachexia, or anorexia, may be detected. Among the molecules which can be used for diagnostic methods, such as those which involve amplification of genomic mahogany sequences, are primers or probes that can routinely be obtained using the genomic and cDNA sequences disclosed herein.

The nucleotide sequences of the present invention also include (a) DNA vectors that contain any of the foregoing mahogany coding sequences and/or their complements; (b) DNA expression vectors that contain any of the foregoing mahogany coding sequences operatively associated with a regulatory element that directs the expression of the coding sequences; and (c) genetically engineered host cells and organisms that contain any of the foregoing mahogany coding sequences operatively associated with a regulatory element that directs the expression of the coding sequence in the host cell. As used herein, regulatory elements include, but are not limited to inducible and non-inducible promoters, enhancers, operators, and other elements known to those skilled in the art that drive and regulate gene expression. Such regulatory elements include, but are not limited to, the cytomegalovirus hCMV immediate early gene, the early or late promoters of SV40 adenovirus, the lac system, the trp system, the TAC system, the TRC system, the major operator and promoter regions of phage A, the control regions of fd coat protein, the promoter for 3′-phosphoglycerate kinase, the promoters of acid phosphatase, and the promoters of the yeast alpha-mating factors.

In addition to the mahogany gene sequences described above, homologs of such sequences, exhibiting extensive homology to one or more domains of the mahogany gene product can be present in other species. In a preferred embodiment, the mahogany gene homologue maps to a chromosomal region that is syntenic to the chromosomal region of the mahogany gene. In a particularly preferred embodiment, a human mahogany gene homologue sequence maps to a human chromosome region that is syntenic to the region of mouse chromosome 2 to which the murine mahogany gene maps, namely 20p15, and comprises the contiged human MG cDNA provided herein. Further, there can also exist homologue genes at other genetic loci within the genome of the same species which encode proteins having extensive homology to one or more domains of the mahogany gene product. Such mahogany homologs can include, for example, secreted forms of the mahogany sequences, see, e.g., Duke-Cohan, J. S. et al. (1998, Proc. Natl. Acad. Sci. U.S.A. 95:11336-11341). Such sequences, can be used, for example, in the screening assays, described in Section 5.4.2 below, for compounds that interact with the mahogany gene and/or its gene product and that may therefore be useful in treating and ameliorating body weight disorders.

Other mahogany homologs can be identified and readily isolated, without undue experimentation, by molecular biological techniques well known in the art, and are therefore within the scope of the present invention. As an example, in order to clone a human mahogany gene homologue using isolated murine mahogany gene sequences, such murine mahogany gene sequences may be labeled and used to screen a cDNA library constructed from mRNA obtained from appropriate cells or tissues derived from the organism (in this case, human) of interest. With respect to the cloning of such a human mahogany homologue, a human cDNA library may, for example be used for screening, such as a cDNA library obtained from mRNA isolated from brain tissues, particularly containing hypothalamic regions.

The hybridization washing conditions used should be of a lower stringency when the cDNA library is derived from an organism different from the type of organism from which the labeled sequence was derived. With respect to the cloning of a human mahogany homologue, for example, hybridization can be performed for 4 hours at 65° C. using Amersham Rapid Hyb™ buffer (Cat. #RPN1639) according to manufacturer's protocol, followed by washing, with a final washing stringency of 1.0×SSC/0.1% SDS at 50° C. for 20 minutes being preferred.

Low stringency conditions are well known to those of skill in the art, and will vary predictably depending on the specific organisms from which the library and the labeled sequences are derived. For guidance regarding such conditions see, for example, Sambrook et al., 1989, Molecular Cloning, A Laboratory Manual, Cold Springs Harbor Press, N.Y.; and Ausubel et al., 1989, Current Protocols in Molecular Biology, Green Publishing Associates and Wiley Interscience, N.Y.

Alternatively, the labeled fragment may be used to screen a genomic library derived from the organism of interest, again, using appropriately stringent conditions.

Further, a mahogany gene homologue may be isolated from nucleic acid of the organism of interest by performing PCR using two degenerate oligonucleotide primer pools designed on the basis of amino acid sequences within the mahogany gene product disclosed herein. The template for the reaction may be cDNA obtained by reverse transcription of mRNA prepared from, for example, human or non-human cell lines or tissue known or suspected to express a mahogany gene allele.

The PCR product may be subcloned and sequenced to ensure that the amplified sequences represent the sequences of a mahogany gene nucleic acid sequence. The PCR fragment may then be used to isolate a full length cDNA clone by a variety of methods. For example, the amplified fragment may be labeled and used to screen a cDNA library, such as a bacteriophage cDNA library. Alternatively, the labeled fragment may be used to isolate genomic clones via the screening of a genomic library. This method has been used to isolate sequences encoding each of the murine MG gene exons as well as to isolate contigs containing the human MG sequences provided herein, FIG. 10 .

PCR technology may also be utilized to isolate full length cDNA sequences. For example, RNA may be isolated, following standard procedures, from an appropriate cellular or tissue source (i.e., one known, or suspected, to express the mahogany gene). A reverse transcription reaction may be performed on the RNA using an oligonucleotide primer specific for the most 5′ end of the amplified fragment for the priming of the first strand synthesis. The resulting RNA/DNA hybrid may then be “tailed” with guanines using a standard terminal transferase reaction, they hybrid may be digested with RNAase H, and second strand synthesis may then be primed with a poly-C primer. Thus, cDNA sequences upstream of the amplified fragment may easily be isolated. For a review of cloning strategies which may be used, see e.g., Sambrook et al., 1989 supra.

Mahogany gene sequences may additionally be used to isolate mutant mahogany alleles. Such mutant alleles may be isolated from individuals either known or proposed to have a phenotype which contributes to the symptoms of body weight disorders such as obesity, cachexia, or anorexia or disorders associated with hyperphagia. Mutant alleles and mutant allele products may then be utilized in the therapeutic and diagnostic systems described below. Additionally, such mahogany gene sequences can be used to detect mahogany gene regulatory (e.g. promoter) defects which can affect body weight.

A cDNA of a mutant mahogany gene may be isolated, for example, by using PCR, a technique which is well known to those of skill in the art. In this case, the first cDNA strand may be synthesized by hybridizing an oligo-dT oligonucleotide to mRNA isolated from tissue known or suspected to be expressed in an individual putatively carrying the mutant mahogany allele, and by extending the new strand with reverse transcriptase. The second strand of the cDNA is then synthesized using an oligonucleotide that hybridizes specifically to the 5′ end of the normal gene. Using these two primers, the product is then amplified via PCR, cloned into a suitable vector, and subjected to DNA sequence analysis through methods well known to those of skill in the art. By comparing the DNA sequence of the mutant mahogany allele to that of the normal mahogany allele, the mutation(s) responsible for the loss of alteration of activity of the mutant mahogany gene product can be ascertained.

Alternatively, a genomic library can be constructed using DNA obtained from an individual suspected of or known to carry the mutant mahogany allele, or a cDNA library can be constructed using RNA from a tissue known, or suspected to express the mutant mahogany allele. The normal mahogany gene or any suitable fragment thereof may then be labeled and used as a probe to identify the corresponding mutant mahogany allele is such libraries. Clones containing the mutant mahogany gene sequences may then be purified and subjected to sequence analysis according to methods well known to those of skill in the art.

Additionally, an expression library can be constructed utilizing cDNA synthesized from, for example, RNA isolated from a tissue known, or suspected to express a mutant mahogany allele in an individual suspected of or known to carry such a mutant allele. In this manner, gene products made by the putatively mutant tissue may be expressed and screened using standard antibody screening techniques in conjunction with antibodies raised against the normal mahogany gene product as described, below, in Section 5.3. For screening techniques, see, for example, Harlow, E. and Lane, eds., 1988, “Antibodies: A Laboratory Manual”, Cold Spring Harbor Press, Cold Spring Harbor. In cases where a mahogany mutation results in an expressed gene product with altered function (e.g., as a result of a missense or a frameshift mutation) a polyclonal set of anti-mahogany gene product antibodies are likely to cross-react with the mutant mahogany gene product. Library clones detected via their reaction with such labeled antibodies can be purified and subjected to sequence analysis according to methods well known to those of skill in the art.

5.2. Protein Products of the Mahogany Gene

Mahogany gene products (e.g., proteins), polypeptides and peptide fragments, mutant, truncated, or deleted forms of the mahogany gene product, and/or fusion proteins of the mahogany gene product can be prepared for a variety of uses. For example, such gene products, or peptide fragments thereof, can be used for the generation of antibodies in diagnostic assays, or for the identification of other cellular or extracellular products involved in the regulation of mammalian body weight.

FIG. 2 shows the amino acid sequence (SEQ ID NO: 2) of the murine form of a mahogany gene product. Mahogany gene products, also referred to herein as mahogany proteins, of the present invention additionally include those gene products encoded by the mahogany gene sequences described in Section 5.1, above, particularly the human homologue of the murine mg gene, a fragment of which is shown in FIG. 10 .

In addition, mahogany gene products may include proteins that represent functionally equivalent gene products. Such an equivalent mahogany gene product may contain deletions, including internal deletions, additions, including additions yielding fusion proteins, or substitutions of amino acid residues within and/or adjacent to the amino acid sequence encoded by the mahogany gene sequences described, in Section 5.1, above, but that result in a “silent” change, in that the change produces a functionally equivalent mahogany gene product. Such amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues involved. For example, nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; positively charged (basic) amino acids include arginine, lysine, and histidine; and negatively charged (acidic) amino acids include aspartic acid and glutamic acid.

“Functionally equivalent”, as utilized herein, refers to a gene product (e.g., a protein) capable of exhibiting a substantially similar in vivo activity as the endogenous mg gene products encoded by the mg gene sequences described in Section 5.1, above. The in vivo activity of the mg gene product, as used herein, refers to the ability of the mg gene product, when present in an appropriate cell type, to ameliorate, prevent, or delay the appearance of the mahogany phenotype relative to its appearance when that cell type lacks a functional mahogany gene product.

Alternatively, where alteration of function is desired, deletion or non-conservative alterations can produce altered, including reduced-activity, mahogany gene products. Such alterations can, for example, alter one or more of the biological functions of the mahogany gene product. Further, such alterations can be selected so as to generate mahogany gene products that are better suited for expression, scale up, etc. in the host cells chosen. For example, cysteine residues can be deleted or substituted with another amino acid residue in order to eliminate disulfide bridges.

As another example, altered mahogany gene products can be engineered that correspond to mutants or variants of the mahogany gene product associated with mammalian weight disorders, such as obesity, cachexia, or anorexia. Altered mahogany gene products can also be engineered that correspond to mutants or variants of the mahogany gene product known to neutralize or ameliorate the symptoms of body weight disorders, such as obesity, cachexia, or anorexia, which are mediated by some other gene, including, but not limited to, body weight disorders mediated by the agouti gene.

Also within the scope of the present invention are peptides and/or proteins corresponding to one or more domains of the mahogany protein or any one of the individual exon encoded regions of the MG protein, as well as fusion proteins in which the full length mahogany protein, a mahogany peptide, or a truncated mahogany protein or peptide is fused to an unrelated heterologous protein. Such proteins and peptides can be designed on the basis of the mahogany nucleotide sequence disclosed in Section 5.1, above, and/or on the basis of the mahogany amino acid sequence disclosed in the Section.

Fusion proteins of the invention include, but are not limited to, IgFc fusions which stabilize the mahogany protein or peptide and prolong half life in vivo; or fusions to any amino acid sequence that allows the fusion protein to be anchored to the cell membrane; or fusions to an enzyme, fluorescent protein, or luminescent protein which provides a marker function.

The mahogany gene products, peptide fragments thereof and fusion proteins thereof, may be produced by recombinant DNA technology using techniques well known in the art. Thus, methods for preparing the mahogany gene products, polypeptides, peptides, fusion peptide and fusion polypeptides of the invention by expressing nucleic acid containing mahogany gene sequences are described herein. Methods that are well known to those skilled in the art can be used to construct expression vectors containing mahogany gene product coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. See, for example, the techniques described in Sambrook, et al., 1989, supra, and Ausubel, et al., 1989, supra. Alternatively, RNA capable of encoding mahogany gene product sequences may be chemically synthesized using, for example, synthesizers. See, for example, the techniques described in “Oligonucleotide Synthesis”, 1984, Gait, ed., IRL Press, Oxford.

A variety of host-expression vector systems may be utilized to express the mahogany gene product coding sequences of the invention. Such host-expression systems represent vehicles by which the coding sequences of interest may be produced and subsequently purified, but also represent cells that may, when transformed or transfected with the appropriate nucleotide coding sequences, exhibit the mahogany gene product of the invention in situ. These include but are not limited to microorganisms such as bacteria (e.g., E. coli, B. subtilis ) transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing mahogany gene product coding sequences; yeast (e.g., Saccharomyces, Pichia) transformed with recombinant yeast expression vectors containing the mahogany gene product coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the mahogany gene product coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing mahogany gene product coding sequences; or mammalian cell systems (e.g., COS, CHO, BHK, 293, 3T3) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter).

In bacterial systems, a number of expression vectors may be advantageously selected depending upon the use intended for the mahogany gene product being expressed. For example, when a large quantity of such a protein is to be produced, for the generation of pharmaceutical compositions of mahogany gene product or for raising antibodies to mahogany gene product, for example, vectors that direct the expression of high levels of fusion protein products that are readily purified may be desirable. Such vectors include, but are not limited, to the E. coli expression vector pUR278 (Ruther et al., 1983, EMBO J. 2, 1791), in which the mahogany gene product coding sequence may be ligated individually into the vector in frame with the lac Z coding region so that a fusion protein is produced; pIN vectors (Inouye and Inouye, 1985, Nucleic Acids Res. 13, 3101-3109; Van Heeke and Schuster, 1989, J. Biol. Chem. 264, 5503-5509); and the like. pGEX vectors may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. The pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned target gene product can be released from the GST moiety.

In an insect system, Autographa californica , nuclear polyhidrosis virus (AcNPV) is used as a vector to express foreign genes. The virus grows in Spodoptera frugiperda cells. The mahogany gene product coding sequence may be cloned individually into non-essential regions (for example the polyhedrin gene) of the virus and placed under control of an AcNPV promoter (for example the polyhedrin promoter). Successful insertion of mahogany gene product coding sequence will result in inactivation of the polyhedrin gene and production of non-occluded recombinant virus (i.e., virus lacking the proteinaceous coat coded for by the polyhedrin gene). These recombinant viruses are then used to infect Spodoptera frugiperda cells in which the inserted gene is expressed. (e.g., see Smith, et al., 1983, J. Virol. 46, 584; Smith, U.S. Pat. No. 4,215,051).

In mammalian host cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, the mahogany gene product coding sequence of interest may be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence. This chimeric gene may then be inserted in the adenovirus genome by in vitro or in vivo recombination. Insertion in a non-essential region of the viral genome (e.g., region E1 or E3) will result in a recombinant virus that is viable and capable of expressing mahogany gene product in infected hosts. (e.g., See Logan and Shenk, 1984, Proc. Natl. Acad. Sci. USA 81, 3655-3659). Specific initiation signals may also be required for efficient translation of inserted mahogany gene product coding sequences. These signals include the ATG initiation codon and adjacent sequences. In cases where an entire mahogany gene, including its own initiation codon and adjacent sequences, is inserted into the appropriate expression vector, no additional translational control signals may be needed. However, in cases where only a portion of the mahogany gene coding sequence is inserted, exogenous translational control signals, including, perhaps, the ATG initiation codon, must be provided. Furthermore, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of appropriate transcription enhancer elements, transcription terminators, etc. (see Bittner, et al., 1987, Methods in Enzymol. 153, 516-544).

In addition, a host cell strain may be chosen that modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may be important for the function of the protein. Different host cells have characteristic and specific mechanisms for the post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed. To this end, eukaryotic host cells that possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product may be used. Such mammalian host cells include but are not limited to CHO, VERO, BHK, HeLa, COS, MDCK, 293, 3T3, and WI38.

For long-term, high-yield production of recombinant proteins, stable expression is preferred. For example, cell lines that stably express the mahogany gene product may be engineered. Rather than using expression vectors that contain viral origins of replication, host cells can be transformed with DNA controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA, engineered cells may be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci that in turn can be cloned and expanded into cell lines. This method may advantageously be used to engineer cell lines that express the mahogany gene product. Such engineered cell lines may be particularly useful in screening and evaluation of compounds that affect the endogenous activity of the mahogany gene product.

A number of selection systems may be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler, et al., 1977, Cell 11, 223), hypoxanthine-guanine phosphoribosyltransferase (Szybalska and Szybalski, 1962, Proc. Natl. Acad. Sci. USA 48, 2026), and adenine phosphoribosyltransferase (Lowy, et al., 1980, Cell 22, 817) genes can be employed in tk − , hgprt − or aprt − cells, respectively. Also, antimetabolite resistance can be used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler, et al., 1980, Natl. Acad. Sci. USA 77, 3567; O'Hare, et al., 1981, Proc. Natl. Acad. Sci. USA 78, 1527); gpt, which confers resistance to mycophenolic acid (Mulligan and Berg, 1981, Proc. Natl. Acad. Sci. USA 78, 2072); neo, which confers resistance to the aminoglycoside G-418 (Colberre-Garapin, et al., 1981, J. Mol. Biol. 150, 1); and hygro, which confers resistance to hygromycin (Santerre, et al., 1984, Gene 30, 147).

Alternatively, the expression characteristic of an endogenous mahogany gene within a cell line or microorganism my be modified by inserting a heterologous DNA regulatory element into the genome of a stable cell line or cloned microorganism such that the inserted regulatory element is operatively linked with the endogenous mahogany gene. For example, an endogenous mahogany gene which is normally “transcriptionally silent”, i.e., a mahogany gene which is normally not expressed, or is expressed only a very low levels in a cell line or microorganism, may be activated by inserting a regulatory element which is capable of promoting the expression of a normally expressed gene product in that cell line or microorganism. Alternatively, a transcriptionally silent, endogenous mahogany bene may be activated by insertion of a promiscuous regulatory element that works across cell types.

A heterologous regulatory element may be inserted into a stable cell line or cloned microorganism, such it is operatively linked with an endogenous mahogany gene, using techniques, such as targeted homologous recombination, which are well known to those of skill in the art, and described e.g., in Chappel, U.S. Pat. Nos. 4,215,051; 5,578,461 to Sherwin et al.; International Application No. PCT/US92/09627 (WO93/09222) by Selden et al.; and International Application No. PCT/US90/06436 (WO91/06667) by Skoultchi et al., each of which is incorporated by reference herein in its entirety.

Alternatively, any fusion protein may be readily purified by utilizing an antibody specific for the fusion protein being expressed. For example, a system described by Janknecht, et al. allows for the ready purification of non-denatured fusion proteins expressed in human cell lines (Janknecht, et al., 1991, Proc. Natl. Acad. Sci. USA 88, 8972-8976). In this system, the gene of interest is subcloned into a vaccinia recombination plasmid such that the gene's open reading frame is translationally fused to an amino-terminal tag consisting of six histidine residues. Extracts from cells infected with recombinant vaccinia virus are loaded onto Ni 2+ .nitriloacetic acid-agarose columns and histidine-tagged proteins are selectively eluted with imidazole-containing buffers.

The mahogany gene products can also be expressed in transgenic animals. Animals of any species, including, but not limited to, mice, rats, rabbits, guinea pigs, pigs, micro-pigs, goats, sheep, and non-human primates, e.g., baboons, monkeys, and chimpanzees may be used to generate mahogany transgenic animals. The term “transgenic,” as used herein, refers to animals expressing mahogany gene sequences from a different species (e.g., mice expressing human mahogany gene sequences), as well as animals that have been genetically engineered to over express endogenous (i.e., same species) mahogany sequences or animals that have been genetically engineered to no longer express endogenous mahogany gene sequences (i.e., “knock-out” animals), and their progeny.

Any technique known in the art may be used to introduce a mahogany gene transgene into animals to produce the founder lines of transgenic animals. Such techniques include, but are not limited to pronuclear microinjection (Hoppe and Wagner, 1989, U.S. Pat. No. 4,873,191); retrovirus mediated gene transfer into germ lines (Van der Putten, et al., 1985, Proc. Natl. Acad. Sci., USA 82, 6148-6152); gene targeting in embryonic stem cells (Thompson, et al., 1989, Cell 56, 313-321); electroporation of embryos (Lo, 1983, Mol. Cell. Biol. 3, 1803-1814); and sperm-mediated gene transfer (Lavitrano et al., 1989, Cell 57, 717-723) (For a review of such techniques, see Gordon, 1989, Transgenic Animals, Intl. Rev. Cytol. 115, 171-229).

Any technique known in the art may be used to produce transgenic animal clones containing a mahogany transgene, for example, nuclear transfer into enucleated oocytes of nuclei from cultured embryonic, fetal or adult cells induced to quiescence (Campbell, et al., 1996, Nature 380, 64-66; Wilmut, et al., Nature 385, 810-813).

The present invention provides for transgenic animals that carry a mahogany transgene in all their cells, as well as animals that carry the transgene in some, but not all their cells, i.e., mosaic animals. The transgene may be integrated as a single transgene or in concatamers, e.g., head-to-head tandems or head-to-tail tandems. The transgene may also be selectively introduced into and activated in a particular cell type by following, for example, the teaching of Lasko et al. (Lasko, et al., 1992, Proc. Natl. Acad. Sci. USA 89, 6232-6236). The regulatory sequences required for such a cell-type specific activation will depend upon the particular cell type of interest, and will be apparent to those of skill in the art. When it is desired that the mahogany transgene be integrated into the chromosomal site of the endogenous mahogany gene, gene targeting is preferred. Briefly, when such a technique is to be utilized, vectors containing some nucleotide sequences homologous to the endogenous mahogany gene are designed for the purpose of integrating, via homologous recombination with chromosomal sequences, into and disrupting the function of the nucleotide sequence of the endogenous mahogany gene. The transgene may also be selectively introduced into a particular cell type, thus inactivating the endogenous mahogany gene in only that cell type, by following, for example, the teaching of Gu, et al. (Gu, et al., 1994, Science 265, 103-106). The regulatory sequences required for such a cell-type specific inactivation will depend upon the particular cell type of interest, and will be apparent to those of skill in the art.

Once transgenic animals have been generated, the expression of the recombinant mahogany gene may be assayed utilizing standard techniques. Initial screening may be accomplished by Southern blot analysis or PCR techniques to analyze animal tissues to assay whether integration of the transgene has taken place. The level of mRNA expression of the transgene in the tissues of the transgenic animals may also be assessed using techniques that include but are not limited to Northern blot analysis of tissue samples obtained from the animal, in situ hybridization analysis, and RT-PCR (reverse transcriptase PCR). Samples of mahogany gene-expressing tissue, may also be evaluated immunocytochemically using antibodies specific for the mahogany transgene product.

5.3. Antibodies to Mahogany Gene Products

Described herein are methods for the production of antibodies capable of specifically recognizing one or more mg gene product epitopes, or epitopes of conserved variants, or peptide fragments of the mg gene products. Further, antibodies that specifically recognize mutant forms of mg gene products, are encompassed by the invention.

Such antibodies may include, but are not limited to, polyclonal antibodies, monoclonal antibodies (mAbs), humanized or chimeric antibodies, single chain antibodies, Fab fragments, F(ab′) 2 fragments, fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above. Such antibodies may be used, for example, in the detection of a mg gene product in an biological sample and may, therefore, be utilized as part of a diagnostic or prognostic technique whereby patients may be tested for abnormal levels of mg gene products, and/or for the presence of abnormal forms of such gene products. Such antibodies may also be utilized in conjunction with, for example, compound screening schemes, as described, below, in Section 5.4.2, for the evaluation of the effect of test compounds on mg gene product levels and/or activity. Additionally, such antibodies can be used in conjunction with the gene therapy techniques described, below, in Section 5.4.3.2, to, for example, evaluate the normal and/or engineered mahogany-expressing cells prior to their introduction into the patient.

Anti-mg gene product antibodies may additionally be used in methods for inhibiting abnormal mg gene product activity. Thus, such antibodies may, therefore, be utilized as part of weight disorder treatment methods.

For the production of antibodies against a mg gene product, various host animals may be immunized by injection with a mg gene product, or a portion thereof. Such host animals may include, but are not limited to rabbits, mice, and rats, to name but a few. Various adjuvants may be used to increase the immunological response, depending on the host species, including but not limited to Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, dinitrophenol, and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and Corynebacterium parvum.

Polyclonal antibodies are heterogeneous populations of antibody molecules derived from the sera of animals immunized with an antigen, such as a mg gene product, or an antigenic functional derivative thereof. For the production of polyclonal antibodies, host animals such as those described above, may be immunized by injection with mg gene product supplemented with adjuvants as also described above.

Monoclonal antibodies, which are homogeneous populations of antibodies to a particular antigen, may be obtained by any technique that provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique of Kohler and Milstein, (1975, Nature 256, 495-497; and U.S. Pat. No. 4,376,110), the human B-cell hybridoma technique (Kosbor et al., 1983, Immunology Today 4, 72; Cole et al., 1983, Proc. Natl. Acad. Sci. USA 80, 2026-2030), and the EBV-hybridoma technique (Cole et al., 1985, Monoclonal Antibodies And Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Such antibodies may be of any immunoglobulin class including IgG, IgM, IgE, IgA, IgD and any subclass thereof. The hybridoma producing the mAb of this invention may be cultivated in vitro or in vivo. Production of high titers of mAbs in vivo makes this the presently preferred method of production.

In addition, techniques developed for the production of “chimeric antibodies” (Morrison, et al., 1984, Proc. Natl. Acad. Sci., 81, 6851-6855; Neuberger, et al., 1984, Nature 312, 604-608; Takeda, et al., 1985, Nature, 314, 452-454) by splicing the genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used. A chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine mAb and a human immunoglobulin constant region. (See, e.g., Cabilly et al., U.S. Pat. No. 4,816,567; and Boss et al., U.S. Pat. No. 4,816,397, which are incorporated herein by reference in their entirety.)

In addition, techniques have been developed for the production of humanized antibodies. (See, e.g., Queen, U.S. Pat. No. 5,585,089, which is incorporated herein by reference in its entirety.) An immunoglobulin light or heavy chain variable region consists of a “framework” region interrupted by three hypervariable regions, referred to as complementarily determining regions (CDRs). The extent of the framework region and CDRs have been precisely defined (see, “Sequences of Proteins of Immunological Interest”, Kabat, E. et al., U.S. Department of Health and Human Services (1983). Briefly, humanized antibodies are antibody molecules from non-human species having one or more CDRs from the non-human species and a framework region from a human immunoglobulin molecule.

Alternatively, techniques described for the production of single chain antibodies (U.S. Pat. No. 4,946,778; Bird, 1988, Science 242, 423-426; Huston, et al., 1988, Proc. Natl. Acad. Sci. USA 85, 5879-5883; and Ward, et al., 1989, Nature 334, 544-546) can be adapted to produce single chain antibodies against mahogany gene products. Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain polypeptide.

Antibody fragments that recognize specific epitopes may be generated by known techniques. For example, such fragments include but are not limited to: the F(ab′) 2 fragments, which can be produced by pepsin digestion of the antibody molecule and the Fab fragments, which can be generated by reducing the disulfide bridges of the F(ab′) 2 fragments. Alternatively, Fab expression libraries may be constructed (Huse, et al., 1989, Science, 246, 1275-1281) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity.

5.4. Uses of the Mahogany Genes, Gene Products, and Antibodies

Described herein are various applications of the mahogany genes, of the mahogany gene products, including peptide fragments thereof, and of antibodies directed against mahogany gene products and peptide fragments thereof. Such applications include, for example, prognostic and diagnostic evaluation of body weight disorders and the identification of subjects with a predisposition to such disorders, as described below, in Section 5.4.1. Additionally, such applications include methods for the treatment of body weight and body weight disorders, as described, below, in Section 5.4.2, and for the identification of compounds which modulate the expression of the mahogany gene and/or the activity of the mahogany gene product, as described in Section 5.4.3, below. Such compounds can include, for example, other cellular products which are involved in body weight regulation. These compounds can be used, for example, in the amelioration of body weight disorders, including obesity, cachexia, and anorexia.

While, for clarity, the uses described in this section are primarily uses related to body weight disorder abnormalities, it is to be noted that each of the diagnostic and therapeutic treatments described herein can additionally be utilized in connection with other defects associated with the mahogany gene, such as hyperpigmentation, hyperphagia and other disorders resulting in increased metabolic rates.

5.4.1. Diagnosis of Body Weight Disorder Abnormalities

A variety of methods can be employed for the diagnostic and prognostic evaluation of body weight disorders, including obesity, cachexia, and anorexia, and for the identification of subjects having a predisposition to such disorders.

Such methods may, for example, utilize reagents such as the mahogany gene nucleotide sequences described in Section 5.1, and antibodies directed against mahogany gene products, including peptide fragments thereof, as described, above, in Section 5.3. Specifically, such reagents may be used, for example, for:

(1) the detection of the presence of mahogany gene mutations, or the detection of either over- or under-expression of mahogany gene relative to levels of mahogany expression in a wild-type, non-body weight disorder state which correlates with certain body weight disorders or susceptibility toward such body weight disorders;

(2) the detection of over- or under-abundance of mahogany gene product relative to the abundance of mahogany gene product in a wild-type non-body weight disorder state which correlates with certain body weight disorders or susceptibility toward such body weight disorders; and

(3) the detection of an aberrant level of mahogany gene product activity relative to mahogany gene product activity levels in a wild-type, non-body weight disorder state which correlates with certain body weight disorders or susceptibility toward such body weight disorders.

Mahogany gene nucleotide sequences can, for example, be used to diagnose a body weight disorder using, for example, the techniques for detecting mutations in the mahogany gene described above in Section 5.1, above.

The methods described herein may be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one specific mahogany gene nucleic acid or anti-mahogany gene product antibody reagent described herein, which may be conveniently used, e.g., in clinical settings, to screen and diagnose patients exhibiting body weight disorder abnormalities, and to screen those individuals exhibiting a predisposition to developing a body weight disorder abnormality.

For the detection of mahogany gene mutations, any nucleated cell can be used as a starting source for genomic nucleic acid. For the detection of mahogany gene expression or mahogany gene products, any cell type or tissue in which the mahogany gene is expressed may be utilized, such as, for example, tissues or cells shown herein to express the MG gene.

Nucleic acid-based detection techniques are described, below, in Section 5.4.1.1. Peptide detection techniques are described, below, in Section 5.4.1.2.

5.4.1.1. Detection of Mahogany Gene Nucleic Acid Molecules

Mutations or polymorphisms within the mahogany gene can be detected by utilizing a number of techniques. Nucleic acid from any nucleated cell can be used as the starting point for such assay techniques, and may be isolated according to standard nucleic acid preparation procedures which are well known to those of skill in the art.

Genomic DNA may be used in hybridization or amplification assays of biological samples to detect abnormalities involving mahogany gene structure, including point mutations, insertions, deletions and chromosomal rearrangements. Such assays may include, but are not limited to, Southern analyses, single stranded conformation polymorphism analyses (SSCP), and PCR analyses.

Diagnostic methods for the detection of mahogany gene-specific mutations can involve for example, contacting and incubating nucleic acids obtained from a sample, e.g., derived from a patient sample or other appropriate cellular source with one or more labeled nucleic acid reagents including recombinant DNA molecules, cloned genes or degenerate variants thereof, such as described in Section 5.1, above, under conditions favorable for the specific annealing of these reagents to their complementary sequences within or flanking the mahogany gene. Preferably, the lengths of these nucleic acid reagents are at least 15 to 30 nucleotides.

After incubation, all non-annealed nucleic acids are removed from the nucleic acid:mahogany molecule hybrid. The presence of nucleic acids that have hybridized, if any such molecules exist, is then detected. Using such a detection scheme, the nucleic acid from the cell type or tissue of interest can be immobilized, for example, to a solid support such as a membrane, or a plastic surface such as that on a microtiter plate or polystyrene beads. In this case, after incubation, non-annealed, labeled nucleic acid reagents of the type described in Section 5.1 are easily removed. Detection of the remaining, annealed, labeled mahogany nucleic acid reagents is accomplished using standard techniques well-known to those in the art. The mahogany gene sequences to which the nucleic acid reagents have annealed can be compared to the annealing pattern expected from a normal mahogany gene sequence in order to determine whether a mahogany gene mutation is present.

In a preferred embodiment, mahogany gene mutations or polymorphisms can be detected by using a microassay of mahogany nucleic acid sequences immobilized to a substrate or “gene chip” (see, e.g. Cronin, et al., 1996, Human Mutation 7:244-255).

Alternative diagnostic methods for the detection of mahogany gene specific nucleic acid molecules, in patient samples or other appropriate cell sources, may involve their amplification, e.g., by PCR (the experimental embodiment set forth in Mullis, 1987, U.S. Pat. No. 4,683,202), followed by the analysis of the amplified molecules using techniques well known to those of skill in the art, such as, for example, those listed above. The resulting amplified sequences can be compared to those that would be expected if the nucleic acid being amplified contained only normal copies of the mahogany gene in order to determine whether a mahogany gene mutation exists.

Among those mahogany nucleic acid sequences which are preferred for such amplification-related diagnostic screening analyses are oligonucleotide primers which amplify mahogany exon sequences. The sequences of such oligonucleotide primers are, therefore, preferably derived from mahogany intron sequences so that the entire exon, or coding region, can be analyzed as discussed below. Primer pairs useful for amplification of mahogany exons are preferably derived from adjacent introns. Appropriate primer pairs can be chosen such that each of the 25 mahogany exons are amplified. Primers for the amplification of mahogany exons can be routinely designed by one of ordinary skill in the art by utilizing the exon and intron sequences of mahogany shown in Figures, particularly FIGS. 3 and 5 (1)- 5 (4).

Additional mahogany nucleic acid sequences which are preferred for such amplification-related analyses are those which will detect the presence of a mahogany polymorphism which doffers from the consensus mahogany sequence depicted in Figures, particularly those that detect the polymorphism identified in exon 15 (FIG. 7 ). Such polymorphisms include ones which represent mutations associated with body weight disorders such as obesity, cachexia, or anorexia.

Further, well-known genotyping techniques can be performed to type polymorphisms that are in close proximity to mutations in the mahogany gene itself, including mutations associated with weight disorders such as obesity, cachexia, or anorexia. Such polymorphisms can be used to identify individuals in families likely to carry mutations in the mahogany gene. If a polymorphism exhibits linkage disequilibrium with mutations in the mahogany gene, the polymorphism can also be used to identify individuals in the general population who are likely to carry such mutations. Polymorphisms that can be used in this way include restriction fragment length polymorphisms (RFLPs), which involve sequence variations in restriction enzyme target sequences, single-base polymorphisms, and simple sequence length polymorphisms (SSLPs).

For example, Weber (U.S. Pat. No. 5,075,217) describes a DNA marker based on length polymorphisms in blocks of (dC-dA)n-(dG-dT)n short tandem repeats. The average separation of (dC-dA)n-(dG-dT)n blocks is estimated to be 30,000-60,000 bp. Markers that are so closely spaced exhibit a high frequency co-inheritance, and are extremely useful in the identification of genetic mutations, such as, for example, mutations within the mahogany gene, and the diagnosis of diseases and disorders related to mutations in the mahogany gene.

Also, Caskey et al. (U.S. Pat. No. 5,364,759) describe a DNA profiling assay for detecting short tri and tetra nucleotide repeat sequences. The process includes extracting the DNA of interest, such as the mahogany gene, amplifying the extracted DNA, and labelling the repeat sequences to form a genotypic map of the individual's DNA.

A mahogany probe could additionally be used to directly identify RFLPs. Further, a mahogany probe or primers derived from the mahogany sequence could be used to isolate genomic clones such as YACs, BACs, PACs, cosmids, phage, or plasmids. The DNA contained in these clones can be screened for single-base polymorphisms or SSLPs using standard hybridization or sequencing procedures.

The level of mahogany gene expression can also be assayed. For example, RNA from a cell type or tissue known, or suspected, to express the mahogany gene, such as muscle, brain, kidney, testes, heart, liver, lung, skin, hypothalamus, spleen, and adipose tissue may be isolated and tested utilizing hybridization or PCR techniques such as are described, above. The isolated cells can be derived from cell culture or from a patient. The analysis of cells taken from culture may be a necessary step in the assessment of cells to be used as part of a cell-based gene therapy technique or, alternatively, to test the effect of compounds on the expression of the mahogany gene. Such analyses may reveal both quantitative and qualitative aspects of the expression pattern of the mahogany gene, including activation or inactivation of mahogany gene expression.

In one embodiment of such a detection scheme, a cDNA molecule is synthesized from an RNA molecule of interest (e.g., by reverse transcription of the RNA molecule into cDNA). All or part of the resulting cDNA is then used as the template for a nucleic acid amplification reaction, such as a PCR amplification reaction, or the like. The nucleic acid reagents used as synthesis initiation reagents (e.g., primers) in the reverse transcription and nucleic acid amplification steps of this method are chosen from among the mahogany gene nucleic acid reagents described in Section 5.1. The preferred lengths of such nucleic acid reagents are at least 9-30 nucleotides.

For detection of the amplified product, the nucleic acid amplification may be performed using radioactively or non-radioactively labeled nucleotides. Alternatively, enough amplified product may be made such that the product may be visualized by standard ethidium bromide staining or by utilizing any other suitable nucleic acid staining method.

As an alternative to amplification techniques, standard Northern analyses can be performed to determine the level of mRNA expression of the mahogany gene, if a sufficient quantity of the appropriate cells can be obtained.

Additionally, it is possible to perform such mahogany gene expression assays “in situ”, i.e., directly upon tissue sections (fixed and/or frozen) of patient tissue obtained from biopsies or resections, such that no nucleic acid purification is necessary. Nucleic acid reagents such as those described in Section 5.1 may be used as probes and/or primers for such in situ procedures (see, for example, Nuovo, G. J., 1992, “PCR In Situ Hybridization: Protocols And Applications”, Raven Press, NY).

5.4.1.2. Detection of Mahogany Gene Products

Mahogany gene products, including both wild-type and mutant mahogany gene products, conserved variants, and polypeptide fragments thereof, which are discussed, above, in Section 5.2, may be detected using antibodies which are directed against such mahogany gene products. Such antibodies, which are discussed in Section 5.3, below, may thereby be used as diagnostics and prognostics for a body eight disorder. Such methods may be used to detect abnormalities in the level of mahogany gene expression or of mahogany gene product synthesis, or abnormalities in the structure, temporal expression, and/or physical location of mahogany gene product. The antibodies and immunoassay methods described herein have, for example, important in vitro applications in assessing the efficacy of treatments for body weight disorders such as obesity, cachexia, and anorexia. Antibodies, or fragments of antibodies, such as those described below, may be used to screen potentially therapeutic compounds in vitro to determine their effects on mahogany gene expression and mahogany gene product production. The compounds that have beneficial effects on body weight disorders, such as obesity, cachexia, and anorexia, can thereby be identified, and a therapeutically effective dose determined.

In vitro immunoassays may also be used, for example, to assess the efficacy of cell-based gene therapy for a body weight disorders, including obesity, cachexia, and anorexia. Antibodies directed against mahogany gene products may be used in vitro to determine, for example, the level of mahogany gene expression achieved in cells genetically engineered to produce mahogany gene product. In the case of intracellular mahogany gene products, such an assessment is done, preferably, using cell lysates or extracts. Such analysis will allow for a determination of the number of transformed cells necessary to achieve therapeutic efficacy in vivo, as well as optimization of the gene replacement protocol.

The tissue or cell type to be analyzed will generally include those that are known, or suspected, to express the mahogany gene. The protein isolation methods employed herein may, for example, be such as those described in Harlow and Lane (1988, “Antibodies: A Laboratory Manual”, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). The isolated cells can be derived from cell culture or from a patient. The analysis of cells taken from culture may be a necessary step in the assessment of cells to be used as part of a cell-based gene therapy technique or, alternatively, to test the effect of compounds on the expression of the mahogany gene.

Preferred diagnostic methods for the detection of mahogany gene products, conserved variants or peptide fragments thereof, may involve, for example, immunoassays wherein the mahogany gene products or conserved variants or peptide fragments are detected by their interaction with an anti-mahogany gene product-specific antibody.

For example, antibodies, or fragments of antibodies, such as those described, above, in Section 5.3, may be used to quantitatively or qualitatively detect the presence of mahogany gene products or conserved variants or peptide fragments thereof. This can be accomplished, for example, by immunofluorescence techniques employing a fluorescently labeled antibody (see below, this Section) coupled with light microscopic, flow cytometric, or fluorimetric detection. Such techniques are especially preferred for mahogany gene products that are expressed on the cell surface.

The antibodies (or fragments thereof) useful in the present invention may, additionally, be employed histologically, as in immunofluorescence or immunoelectron microscopy, for in situ detection of mahogany gene products, conserved variants or peptide fragments thereof. In situ detection may be accomplished by removing a histological specimen from a patient, and applying thereto a labeled antibody that binds to a mahogany polypeptide. The antibody (or fragment) is preferably applied by overlaying the labeled antibody (or fragment) onto a biological sample. Through the use of such a procedure, it is possible to determine not only the presence of the mahogany gene product, conserved variants or peptide fragments, but also its distribution in the examined tissue. Using the present invention, those of ordinary skill will readily recognize that any of a wide variety of histological methods (such as staining procedures) can be modified in order to achieve in situ detection of a mahogany gene product.

Immunoassays for mahogany gene products, conserved variants, or peptide fragments thereof will typically comprise: (1) incubating a sample, such as a biological fluid, a tissue extract, freshly harvested cells, or lysates of cells in the presence of a detectably labeled antibody capable of identifying mahogany gene products, conserved variants or peptide fragments thereof; and (2) detecting the bound antibody by any of a number of techniques well-known in the art.

The biological sample may be brought in contact with and immobilized onto a solid phase support or carrier, such as nitrocellulose, that is capable of immobilizing cells, cell particles or soluble proteins. The support may then be washed with suitable buffers followed by treatment with the detectably labeled mahogany gene product specific antibody. The solid phase support may then be washed with the buffer a second time to remove unbound antibody. The amount of bound label on the solid support may then be detected by conventional means.

By “solid phase support or carrier” is intended any support capable of binding an antigen or an antibody. Well-known supports or carriers include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylases, natural and modified celluloses, polyacrylamides, gabbros, and magnetite. The nature of the carrier can be either soluble to some extent or insoluble for the purposes of the present invention. The support material may have virtually any possible structural configuration so long as the coupled molecule is capable of binding to an antigen or antibody. Thus, the support configuration may be spherical, as in a bead, or cylindrical, as in the inside surface of a test tube, or the external surface of a rod. Alternatively, the surface may be flat such as a sheet, test strip, etc. Preferred supports include polystyrene beads. Those skilled in the art will know many other suitable carriers for binding antibody or antigen, or will be able to ascertain the same by use of routine experimentation.

One of the ways in which the mahogany gene product-specific antibody can be detectably labeled is by linking the same to an enzyme, such as for use in an enzyme immunoassay (EIA) (Voller, A., “The Enzyme Linked Immunosorbent Assay (ELISA)”, 1978, Diagnostic Horizons 2, 1-7, Microbiological Associates Quarterly Publication, Walkersville, Md.); Voller, A. et al., 1978, J. Clin. Pathol. 31, 507-520; Butler, J. E., 1981, Meth. Enzymol. 73, 482-523; Maggio, E. (ed.), 1980, Enzyme Immunoassay, CRC Press, Boca Raton, Fla.,; Ishikawa, E. et al., (eds.), 1981, Enzyme Immunoassay, Kgaku Shoin, Tokyo). The enzyme which is bound to the antibody will react with an appropriate substrate, preferably a chromogenic substrate, in such a manner as to produce a chemical moiety that can be detected, for example, by spectrophotometric, fluorimetric or by visual means. Enzymes that can be used to detectably label the antibody include, but are not limited to, malate dehydrogenase, staphylococcal nuclease, delta-5-steroid isomerase, yeast alcohol dehydrogenase, α-glycerophosphate, dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, β-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase and acetylcholinesterase. The detection can be accomplished by colorimetric methods that employ a chromogenic substrate for the enzyme. Detection may also be accomplished by visual comparison of the extent of enzymatic reaction of a substrate in comparison with similarly prepared standards.

Detection may also be accomplished using any of a variety of other immunoassays. For example, by radioactively labeling the antibodies or antibody fragments, it is possible to detect mahogany gene products through the use of a radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March, 1986). The radioactive isotope can be detected by such means as the use of a gamma counter or a scintillation counter or by autoradiography.

It is also possible to label the antibody with a fluorescent compound. When the fluorescently labeled antibody is exposed to light of the proper wave length, its presence can then be detected due to fluorescence. Among the most commonly used fluorescent labeling compounds are fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine.

The antibody can also be detectably labeled using fluorescence emitting metals such as 152 Eu, or others of the lanthanide series. These metals can be attached to the antibody using such metal chelating groups as diethylenetriaminepentacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA).

The antibody also can be detectably labeled by coupling it to a chemiluminescent compound. The presence of the chemiluminescent-tagged antibody is then determined by detecting the presence of luminescence that arises during the course of a chemical reaction. Examples of particularly useful chemiluminescent labeling compounds are luminol, isoluminol, theromatic acridinium ester, imidazole, acridinium salt and oxalate ester.

Likewise, a bioluminescent compound may be used to label the antibody of the present invention. Bioluminescence is a type of chemiluminescence found in biological systems in which a catalytic protein increases the efficiency of the chemiluminescent reaction. The presence of a bioluminescent protein is determined by detecting the presence of luminescence. Important bioluminescent compounds for purposes of labeling are luciferin, luciferase and aequorin.

5.4.2. Screening Assays for Compounds that Interact with the Mahogany Gene or Gene Product

The following assays are designed to identify compounds that bind to a mahogany gene product, compounds that bind to proteins, or portions of proteins that interact with a mahogany gene product, compounds that interfere with the interaction of a mahogany gene product with proteins and compounds that modulate the activity of the mahogany gene (i.e., modulate the level of mahogany gene expression and/or modulate the level of mahogany gene product activity). Assays may additionally be utilized that identify compounds that bind to mahogany gene regulatory sequences (e.g., promoter sequences; see e.g., Platt, 1994, J. Biol. Chem. 269, 28558-28562), which is incorporated herein by reference in its entirety, and that can modulate the level of mahogany gene expression. Such compounds may include, but are not limited to, small organic molecules, such as ones that are able to cross the blood-brain barrier, gain to and/or entry into an appropriate cell and affect expression of the mahogany gene or some other gene involved in the body weight regulatory pathway, or intracellular proteins.

Methods for the identification of such proteins are described, below, in Section 5.4.2.2. Such proteins may be involved in the control and/or regulation of body weight. Further, among these compounds are compounds that affect the level of mahogany gene expression and/or mahogany gene product activity and that can be used in the therapeutic treatment of body weight disorders, including obesity, cachexia, and anorexia, as described, below, in Section 5.9.

Compounds may include, but are not limited to, peptides such as, for example, soluble peptides, including but not limited to, Ig-tailed fusion peptides, and members of random peptide libraries; (see, e.g., Lam, et al., 1991, Nature 354, 82-84; Houghten, et al., 1991, Nature 354, 84-86), and combinatorial chemistry-derived molecular library made of D- and/or L-configuration amino acids, phosphopeptides (including, but not limited to members of random or partially degenerate, directed phosphopeptide libraries; see, e.g., Songyang, et al., 1993, Cell 72, 767-778), antibodies (including, but not limited to, polyclonal, monoclonal, humanized, anti-idiotypic, chimeric or single chain antibodies, and FAb, F(ab′) 2 and FAb expression library fragments, and epitope-binding fragments thereof), and small organic or inorganic molecules.

Compounds identified via assays such as those described herein may be useful, for example, in elaborating the biological function of the mahogany gene product and for ameliorating body weight disorders, such as obesity, cachexia, or anorexia. Assays for testing the effectiveness of compounds identified by, for example, techniques such as those described in Sections 5.4.2.1-5.4.2.3, are discussed, below, in Section 5.4.2.4.

5.4.2.1. In Vitro Screening Assays for Compounds that Bind to the Mahogany Gene Product

In vitro systems may be designed to identify compounds capable of binding the mahogany gene products of the invention. Compounds identified may be useful, for example, in modulating the activity of unimpaired and/or mutant mahogany gene products, may be useful in elaborating the biological function of the mahogany gene product, may be utilized in screens for identifying compounds that disrupt normal mahogany gene product interactions, or may in themselves disrupt such interactions.

The principle of the assays used to identify compounds that bind to the mahogany gene product involves preparing a reaction mixture of the mahogany gene product and the test compound under conditions and for a time sufficient to allow the two components to interact and bind, thus forming a complex that can be removed and/or detected in the reaction mixture. These assays can be conducted in a variety of ways. For example, one method to conduct such an assay involves anchoring a mahogany gene product or a test substance onto a solid support and detecting mahogany gene product/test compound complexes formed on the solid support at the end of the reaction. In one embodiment of such a method, the mahogany gene product may be anchored onto a solid support, and the test compound, which is not anchored, may be labeled, either directly or indirectly.

In practice, microtiter plates are conveniently utilized as the solid support. The anchored component may be immobilized by non-covalent or covalent attachments. Non-covalent attachment may be accomplished by simply coating the solid surface with a solution of the protein and drying. Alternatively, an immobilized antibody, preferably a monoclonal antibody, specific for the protein to be immobilized may be used to anchor the protein to the solid surface. The surfaces may be prepared in advance and stored.

In order to conduct the assay, the non-immobilized component is added to the coated surface containing the anchored component. After the reaction is complete, unreacted components are removed (e.g., by washing) under conditions such that any complexes formed will remain immobilized on the solid surface. The detection of complexes anchored on the solid surface can be accomplished in a number of ways. Where the previously non-immobilized component is pre-labeled, the detection of label immobilized on the surface indicates that complexes were formed. Where the previously non-immobilized component is not pre-labeled, an indirect label can be used to detect complexes anchored on the surface; e.g., using a labeled antibody specific for the previously non-immobilized component (the antibody, in turn, may be directly labeled or indirectly labeled with a labeled anti-Ig antibody).

Alternatively, a reaction can be conducted in a liquid phase, the reaction products separated from unreacted components, and complexes detected; e.g., using an immobilized antibody specific for mahogany gene product or the test compound to anchor any complexes formed in solution, and a labeled antibody specific for the other component of the possible complex to detect anchored complexes.

5.4.2.2. Assays for Proteins that Interact with the Mahogany Gene Product

Any method suitable for detecting protein-protein interactions may be employed for identifying mahogany gene product-protein interactions.

Among the traditional methods that may be employed are co-immunoprecipitation, cross-linking and co-purification through gradients or chromatographic columns. Utilizing procedures such as these allows for the identification of proteins that interact with mahogany gene products. Such proteins can include, but are not limited, the mahoganoid gene product.

Once isolated, such a protein can be identified and can be used in conjunction with standard techniques, to identify proteins it interacts with. For example, at least a portion of the amino acid sequence of a protein that interacts with the mahogany gene product can be ascertained using techniques well known to those of skill in the art, such as via the Edman degradation technique (see, e.g., Creighton, 1983, “Proteins: Structures and Molecular Principles,” W. H. Freeman & Co., N.Y., pp. 34-49). The amino acid sequence obtained may be used as a guide for the generation of oligonucleotide mixtures that can be used to screen for gene sequences encoding such proteins. Screening may be accomplished, for example, by standard hybridization or PCR techniques. Techniques for the generation of oligonucleotide mixtures and the screening are well-known. (See, e.g., Ausubel, supra, and 1990, “PCR Protocols: A Guide to Methods and Applications,” Innis, et al., eds. Academic Press, Inc., New York).

Additionally, methods may be employed that result in the simultaneous identification of genes that encode a protein which interacts with a mahogany gene product. These methods include, for example, probing expression libraries with labeled mahogany gene product, using mahogany gene product in a manner similar to the well known technique of antibody probing of λgt11 libraries.

One method that detects protein interactions in vivo, the two-hybrid system, is described in detail for illustration only and not by way of limitation. One version of this system has been described (Chien, et al., 1991, Proc. Natl. Acad. Sci. USA, 88, 9578-9582) and is commercially available from Clontech (Palo Alto, Calif.).

Briefly, utilizing such a system, plasmids are constructed that encode two hybrid proteins: one consists of the DNA-binding domain of a transcription activator protein fused to the mahogany gene product and the other consists of the transcription activator protein's activation domain fused to an unknown protein that is encoded by a cDNA that has been recombined into this plasmid as part of a cDNA library. The DNA-binding domain fusion plasmid and the cDNA library are transformed into a strain of the yeast Saccharomyces cerevisiae that contains a reporter gene (e.g., HBS or lacZ) whose regulatory region contains the transcription activator's binding site. Either hybrid protein alone cannot activate transcription of the reporter gene: the DNA-binding domain hybrid cannot because it does not provide activation function and the activation domain hybrid cannot because it cannot localize to the activator's binding sites. Interaction of the two hybrid proteins reconstitutes the functional activator protein and results in expression of the reporter gene, which is detected by an assay for the reporter gene product.

The two-hybrid system or related methodologies may be used to screen activation domain libraries for proteins that interact with the “bait” gene product. By way of example, and not by way of limitation, mahogany gene products may be used as the bait gene product. Total genomic or cDNA sequences are fused to the DNA encoding an activation domain. This library and a plasmid encoding a hybrid of a bait mahogany gene product fused to the DNA-binding domain are co-transformed into a yeast reporter strain, and the resulting transformants are screened for those that express the reporter gene. For example, a bait mahogany gene sequence, such as the open reading frame of the mahogany gene, can be cloned into a vector such that it is translationally fused to the DNA encoding the DNA-binding domain of the GAL4 protein. These colonies are purified and the library plasmids responsible for reporter gene expression are isolated. DNA sequencing is then used to identify the proteins encoded by the library plasmids.

A cDNA library of the cell line from which proteins that interact with bait mahogany gene product are to be detected can be made using methods routinely practiced in the art. According to the particular system described herein, for example, the cDNA fragments can be inserted into a vector such that they are translationally fused to the transcriptional activation domain of GAL4. Such a library can be co-transformed along with the bait mahogany gene-GAL4 fusion plasmid into a yeast strain that contains a lacZ gene driven by a promoter that contains GAL4 activation sequence. A cDNA encoded protein, fused to a GAL4 transcriptional activation domain that interacts with bait mahogany gene product will reconstitute an active GAL4 protein and thereby drive expression of the HIS3 gene. Colonies that express HIS3 can be detected by their growth on petri dishes containing semi-solid agar based media lacking histidine. The cDNA can then be purified from these strains, and used to produce and isolate the bait mahogany gene product-interacting protein using techniques routinely practiced in the art.

5.4.2.3. Assays for Compounds that Interfere with Mahogany Gene Product Macromolecule Interaction

The mahogany gene products may, in vivo, interact with one or more macromolecules, such as proteins. For example, the mahogany gene products may, in vivo, interact with the mahoganoid gene products. Other macromolecules which interact with the mahogany gene products may include, but are not limited to, nucleic acid molecules and those proteins identified via methods such as those described, above, in Sections 5.4.2.1-5.4.2.2. For purposes of this discussion, the macromolecules are referred to herein as “binding partners”. Compounds that disrupt mahogany gene product binding to a binding partner may be useful in regulating the activity of the mahogany gene product, especially mutant mahogany gene products. Such compounds may include, but are not limited to molecules such as peptides, and the like, as described, for example, in Section 5.4.2.1 above.

The basic principle of an assay system used to identify compounds that interfere with the interaction between the mahogany gene product and a binding partner or partners involves preparing a reaction mixture containing the mahogany gene product and the binding partner under conditions and for a time sufficient to allow the two to interact and bind, thus forming a complex. In order to test a compound for inhibitory activity, the reaction mixture is prepared in the presence and absence of the test compound. The test compound may be initially included in the reaction mixture, or may be added at a time subsequent to the addition of mahogany gene product and its binding partner. Control reaction mixtures are incubated without the test compound or with a compound which is known not to block complex formation. The formation of any complexes between the mahogany gene product and the binding partner is then detected. The formation of a complex in the control reaction, but not in the reaction mixture containing the test compound, indicates that the compound interferes with the interaction of the mahogany gene product and the binding partner. Additionally, complex formation within reaction mixtures containing the test compound and normal mahogany gene product may also be compared to complex formation within reaction mixtures containing the test compound and a mutant mahogany gene product. This comparison may be important in those cases wherein it is desirable to identify compounds that disrupt interactions of mutant but not normal mahogany gene product.

The assay for compounds that interfere with the interaction of the mahogany gene products and binding partners can be conducted in a heterogeneous or homogeneous format. Heterogeneous assays involve anchoring either the mahogany gene product or the binding partner onto a solid support and detecting complexes formed on the solid support at the end of the reaction. In homogeneous assays, the entire reaction is carried out in a liquid phase. In either approach, the order of addition of reactants can be varied to obtain different information about the compounds being tested. For example, test compounds that interfere with the interaction between the mahogany gene products and the binding partners, e.g., by competition, can be identified by conducting the reaction in the presence of the test substance; i.e., by adding the test substance to the reaction mixture prior to or simultaneously with the mahogany gene product and interactive intracellular binding partner. Alternatively, test compounds that disrupt preformed complexes, e.g., compounds with higher binding constants that displace one of the components from the complex, can be tested by adding the test compound to the reaction mixture after complexes have been formed. The various formats are described briefly below.

In a heterogeneous assay system, either the mahogany gene product or the interactive binding partner, is anchored onto a solid surface, while the non-anchored species is labeled, either directly or indirectly. In practice, microtiter plates are conveniently utilized. The anchored species may be immobilized by non-covalent or covalent attachments. Non-covalent attachment may be accomplished simply by coating the solid surface with a solution of the mahogany gene product or binding partner and drying. Alternatively, an immobilized antibody specific for the species to be anchored may be used to anchor the species to the solid surface. The surfaces may be prepared in advance and stored.

In order to conduct the assay, the partner of the immobilized species is exposed to the coated surface with or without the test compound. After the reaction is complete, unreacted components are removed (e.g., by washing) and any complexes formed will remain immobilized on the solid surface. The detection of complexes anchored on the solid surface can be accomplished in a number of ways. Where the non-immobilized species is pre-labeled, the detection of label immobilized on the surface indicates that complexes were formed. Where the non-immobilized species is not pre-labeled, an indirect label can be used to detect complexes anchored on the surface; e.g., using a labeled antibody specific for the initially non-immobilized species (the antibody, in turn, may be directly labeled or indirectly labeled with a labeled anti-Ig antibody). Depending upon the order of addition of reaction components, test compounds that inhibit complex formation or that disrupt preformed complexes can be detected.

Alternatively, the reaction can be conducted in a liquid phase in the presence or absence of the test compound, the reaction products separated from unreacted components, and complexes detected; e.g., using an immobilized antibody specific for one of the binding components to anchor any complexes formed in solution, and a labeled antibody specific for the other partner to detect anchored complexes. Again, depending upon the order of addition of reactants to the liquid phase, test compounds that inhibit complex formation or that disrupt preformed complexes can be identified.

In an alternate embodiment of the invention, a homogeneous assay can be used. In this approach, a preformed complex of the mahogany gene product and the interactive binding partner is prepared in which either the mahogany gene product or its binding partners is labeled, but the signal generated by the label is quenched due to complex formation (see, e.g., U.S. Pat. No. 4,109,496 by Rubenstein which utilizes this approach for immunoassays). The addition of a test substance that competes with and displaces one of the species from the preformed complex will result in the generation of a signal above background. In this way, test substances that disrupt mahogany gene product/binding partner interaction can be identified.

In another embodiment of the invention, these same techniques can be employed using peptide fragments that correspond to the binding domains of the mahogany gene product and/or the binding partner (in cases where the binding partner is a protein), in place of one or both of the full length proteins. Any number of methods routinely practiced in the art can be used to identify and isolate the binding sites. These methods include, but are not limited to, mutagenesis of the gene encoding one of the proteins and screening for disruption of binding in a co-immunoprecipitation assay. Compensating mutations in the gene encoding the second species in the complex can then be selected. Sequence analysis of the genes encoding the respective proteins will reveal the mutations that correspond to the region of the protein involved in interactive binding. Alternatively, one protein can be anchored to a solid surface using methods described in this Section above, and allowed to interact with and bind to its labeled binding partner, which has been treated with a proteolytic enzyme, such as trypsin. After washing, a short, labeled peptide comprising the binding domain may remain associated with the solid material, which can be isolated and identified by amino acid sequencing. Also, once the gene coding for the segments is engineered to express peptide fragments of the protein, it can then be tested for binding activity and purified or synthesized.

For example, and not by way of limitation, a mahogany gene product can be anchored to a solid material as described, above, in this Section by making a GST-1 fusion protein and allowing it to bind to glutathione agarose beads. The binding partner can be labeled with a radioactive isotope, such as 35 S, and cleaved with a proteolytic enzyme such as trypsin. Cleavage products can then be added to the anchored GST-1 fusion protein and allowed to bind. After washing away unbound peptides, labeled bound material, representing the binding partner binding domain, can be eluted, purified, and analyzed for amino acid sequence by well-known methods. Peptides so identified can be produced synthetically or produced using recombinant DNA technology.

5.4.2.4. Assays for the Identification of Compounds that Ameliorate Body Weight Disorders

Compounds, including but not limited to binding compounds identified via assay techniques such as those described, above, in Sections 5.4.2.1-5.4.2.3, can be tested for the ability to ameliorate body weight disorder symptoms, including obesity, cachexia, and anorexia. It should be noted that the assays described herein can identify compounds that affect mahogany activity by either affecting mahogany gene expression or by affecting the level of mahogany gene product activity. For example, compounds may be identified that are involved in another step in the pathway in which the mahogany gene and/or mahogany gene product is involved, such as, for example, a step which is either “upfield” or “downfield” of the step in the pathway mediated by the mahogany gene. Such compounds may, by affecting this same pathway, modulate the effect of mahogany on the development of body weight disorders. Such compounds can be used as part of a therapeutic method for the treatment of the disorder.

Described below are cell-based and animal model-based assays for the identification of compounds exhibiting such an ability to ameliorate body weight disorder symptoms.

First, cell-based systems can be used to identify compounds that may act to ameliorate body weight disorder symptoms. Such cell systems can include, for example, recombinant or non-recombinant cell, such as cell lines, that express the mahogany gene.

In utilizing such cell systems, cells that express mahogany may be exposed to a compound suspected of exhibiting an ability to ameliorate body weight disorder symptoms, at a sufficient concentration and for a sufficient time to elicit such an amelioration of such symptoms in the exposed cells. After exposure, the cells can be assayed to measure alterations in the expression of the mahogany gene, e.g., by assaying cell lysates for mahogany mRNA transcripts (e.g., by Northern analysis) or for mahogany gene products expressed by the cell; compounds that modulate expression of the mahogany gene are good candidates as therapeutics.

In addition, animal-based systems or models for a mammalian body weight disorder, for example, transgenic mice containing a human or altered form of mahogany gene, may be used to identify compounds capable of ameliorating symptoms of the disorder. Such animal models may be used as test substrates for the identification of drugs, pharmaceuticals, therapies and interventions. For example, animal models may be exposed to a compound suspected of exhibiting an ability to ameliorate symptoms, at a sufficient concentration and for a sufficient time to elicit such an amelioration of body weight disorder symptoms. The response of the animals to the exposure may be monitored by assessing the reversal of the symptoms of the disorder.

With regard to intervention, any treatments that reverse any aspect of body weight disorder-like symptoms should be considered as candidates for human therapeutic intervention in such a disorder. Dosages of test agents may be determined by deriving dose-response curves, as discussed in Section 5.5.1, below.

5.4.3. Compounds and Methods for the Treatment of Body Weight Disorders

Described below are methods and compositions whereby body weight disorders, including obesity, cachexia, and anorexia, may be treated. Such methods can comprise, for example administering compounds which modulate the expression of a mammalian mahogany gene and/or the synthesis or activity of a mammalian mahogany gene product, so that symptoms of the body weight disorder are ameliorated. Alternatively, in those instances whereby the mammalian body weight disorder results from mahogany gene mutations, such methods can comprise supplying the mammal with a nucleic acid molecule encoding an unimpaired mahogany gene product such that an unimpaired mahogany gene product is expressed and symptoms of the disorder are ameliorated.

In another embodiment of methods for the treatment of mammalian body weight disorders resulting from mahogany gene mutations, such methods can comprise supplying the mammal with a cell comprising a nucleic acid molecule that encodes an unimpaired mahogany gene product such that the cell expresses the unimpaired mahogany gene product, and symptoms of the disorder are ameliorated.

Because a loss of normal mahogany gene function results in the restoration of a non-obese phenotype in individuals exhibiting an agouti mutation (e.g. individuals that ectopically express the agouti gene in all tissues) a decrease or elimination of normal mahogany gene product would facilitate progress towards a normal body weight state in such individuals. Methods for inhibiting or reducing the level of mahogany gene product synthesis or expression can include, for example, methods such as those described in Section 5.4.3.1.

Alternatively, symptoms of certain body weight disorders such as, for example, cachexia and anorexia, which involve a lower than normal body weight phenotype, may be ameliorated by increasing the level of mahogany gene expression and/or mahogany gene product activity. Methods for enhancing the expression or synthesis of mahogany can include, for example, methods such as those described below, in Section 5.4.3.2.

5.4.3.1. Inhibitory Antisense, Ribozyme and Triple Helix Approaches

In another embodiment, symptoms of body weight disorders may be ameliorated by decreasing the level of mahogany gene expression and/or mahogany gene product activity by using mahogany gene sequences in conjunction with well-known antisense, gene “knock-out,” ribozyme and/or triple helix methods to decrease the level of mahogany gene expression. Among the compounds that may exhibit the ability to modulate the activity, expression or synthesis of the mahogany gene, including the ability to ameliorate the symptoms of a mammalian body weight disorder, are antisense, ribozyme, and triple helix molecules. Such molecules may be designed to reduce or inhibit either unimpaired, or if appropriate, mutant target gene activity. Techniques for the production and use of such molecules are well known to those of skill in the art.

Antisense RNA and DNA molecules act to directly block the translation of mRNA by hybridizing to targeted mRNA and preventing protein translation. Antisense approaches involve the design of oligonucleotides that are complementary to a target gene mRNA. The antisense oligonucleotides will bind to the complementary target gene mRNA transcripts and prevent translation. Absolute complementarily, although preferred, is not required.

A sequence “complementary” to a portion of an RNA, as referred to herein, means a sequence having sufficient complementarily to be able to hybridize with the RNA, forming a stable duplex; in the case of double-stranded antisense nucleic acids, a single strand of the duplex DNA may thus be tested, or triplex formation may be assayed. The ability to hybridize will depend on both the degree of complementarily and the length of the antisense nucleic acid. Generally, the longer the hybridizing nucleic acid, the more base mismatches with an RNA it may contain and still form a stable duplex (or triplex, as the case may be). One skilled in the art can ascertain a tolerable degree of mismatch by use of standard procedures to determine the melting point of the hybridized complex.

In one embodiment, oligonucleotides complementary to non-coding regions of the mahogany gene could be used in an antisense approach to inhibit translation of endogenous mahogany mRNA. Antisense nucleic acids should be at least six nucleotides in length, and are preferably oligonucleotides ranging from 6 to about 50 nucleotides in length. In specific aspects the oligonucleotide is at least 10 nucleotides, at least 17 nucleotides, at least 25 nucleotides or at least 50 nucleotides.

Regardless of the choice of target sequence, it is preferred that in vitro studies are first performed to quantitate the ability of the antisense oligonucleotide to inhibit gene expression. It is preferred that these studies utilize controls that distinguish between antisense gene inhibition and nonspecific biological effects of oligonucleotides. It is also preferred that these studies compare levels of the target RNA or protein with that of an internal control RNA or protein. Additionally, it is envisioned that results obtained using the antisense oligonucleotide are compared with those obtained using a control oligonucleotide. It is preferred that the control oligonucleotide is of approximately the same length as the test oligonucleotide and that the nucleotide sequence of the oligonucleotide differs from the antisense sequence no more than is necessary to prevent specific hybridization to the target sequence.

The oligonucleotides can be DNA or RNA or chimeric mixtures or derivatives or modified versions thereof, single-stranded or double-stranded. The oligonucleotide can be modified at the base moiety, sugar moiety, or phosphate backbone, for example, to improve stability of the molecule, hybridization, etc. The oligonucleotide may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger, et al., 1989, Proc. Natl. Acad. Sci. U.S.A. 86, 6553-6556; Lemaitre, et al., 1987, Proc. Natl. Acad. Sci. U.S.A. 84, 648-652; PCT Publication No. WO88/09810, published Dec. 15, 1988) or the blood-brain barrier (see, e.g., PCT Publication No. WO89/10134, published Apr. 25, 1988), hybridization-triggered cleavage agents (see, e.g., Krol et al., 1988, BioTechniques 6, 958-976) or intercalating agents (see, e.g., Zon, 1988, Pharm. Res. 5, 539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

The antisense oligonucleotide may comprise at least one modified base moiety which is selected from the group including but not limited to 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine.

The antisense oligonucleotide may also comprise at least one modified sugar moiety selected from the group including but not limited to arabinose, 2-fluoroarabinose, xylulose, and hexose.

In yet another embodiment, the antisense oligonucleotide comprises at least one modified phosphate backbone selected from the group consisting of a phosphorothioate, a phosphorodithioate, a phosphoramidothioate, a phosphoramidate, a phosphordiamidate, a methylphosphonate, an alkyl phosphotriester, and a formacetal or analog thereof.

In yet another embodiment, the antisense oligonucleotide is an α-anomeric oligonucleotide. An α-anomeric oligonucleotide forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other (Gautier, et al., 1987, Nucl. Acids Res. 15, 6625-6641). The oligonucleotide is a 2′-0-methylribonucleotide (Inoue, et al., 1987, Nucl. Acids Res. 15, 6131-6148), or a chimeric RNA-DNA analogue (Inoue, et al., 1987, FEBS Lett. 215, 327-330).

Oligonucleotides of the invention may be synthesized by standard methods known in the art, e.g., by use of an automated DNA synthesizer (such as are commercially available from Biosearch, Applied Biosystems, etc.). As examples, phosphorothioate oligonucleotides may be synthesized by the method of Stein, et al. (1988, Nucl. Acids Res. 16, 3209), methylphosphonate oligonucleotides can be prepared by use of controlled pore glass polymer supports (Sarin, et al., 1988, Proc. Natl. Acad. Sci. U.S.A. 85, 7448-7451), etc.

While antisense nucleotides complementary to the target gene coding region sequence could be used, those complementary to the transcribed, untranslated region are most preferred.

Antisense molecules should be delivered to cells that express the target gene in vivo. A number of methods have been developed for delivering antisense DNA or RNA to cells; e.g., antisense molecules can be injected directly into the tissue site, or modified antisense molecules, designed to target the desired cells (e.g., antisense linked to peptides or antibodies that specifically bind receptors or antigens expressed on the target cell surface) can be administered systemically.

However, it is often difficult to achieve intracellular concentrations of the antisense sufficient to suppress translation of endogenous mRNAs. Therefore a preferred approach utilizes a recombinant DNA construct in which the antisense oligonucleotide is placed under the control of a strong pol III or pol II promoter. The use of such a construct to transfect target cells in the patient will result in the transcription of sufficient amounts of single stranded RNAs that will form complementary base pairs with the endogenous target gene transcripts and thereby prevent translation of the target gene mRNA. For example, a vector can be introduced e.g., such that it is taken up by a cell and directs the transcription of an antisense RNA. Such a vector can remain episomal or become chromosomally integrated, as long as it can be transcribed to produce the desired antisense RNA. Such vectors can be constructed by recombinant DNA technology methods standard in the art. Vectors can be plasmid, viral, or others known in the art, used for replication and expression in mammalian cells. Expression of the sequence encoding the antisense RNA can be by any promoter known in the art to act in mammalian, preferably human cells. Such promoters can be inducible or constitutive. Such promoters include but are not limited to: the SV40 early promoter region (Bernoist and Chambon, 1981, Nature 290, 304-310), the promoter contained in the 3′ long terminal repeat of Rous sarcoma virus (Yamamoto, et al., 1980, Cell 22, 787-797), the herpes thymidine kinase promoter (Wagner, et al., 1981, Proc. Natl. Acad. Sci. U.S.A. 78, 1441-1445), the regulatory sequences of the metallothionein gene (Brinster, et al., 1982, Nature 296, 39-42), etc. Any type of plasmid, cosmid, YAC or viral vector can be used to prepare the recombinant DNA construct which can be introduced directly into the tissue site. Alternatively, viral vectors can be used that selectively infect the desired tissue, in which case administration may be accomplished by another route (e.g., systemically).

Ribozyme molecules designed to catalytically cleave target gene mRNA transcripts can also be used to prevent translation of target gene mRNA and, therefore, expression of target gene product. (See, e.g., PCT International Publication WO90/11364, published Oct. 4, 1990; Sarver, et al., 1990, Science 247, 1222-1225).

Ribozymes are enzymatic RNA molecules capable of catalyzing the specific cleavage of RNA. (For a review, see Rossi, 1994, Current Biology 4, 469-471). The mechanism of ribozyme action involves sequence specific hybridization of the ribozyme molecule to complementary target RNA, followed by an endonucleolytic cleavage event. The composition of ribozyme molecules must include one or more sequences complementary to the target gene mRNA, and must include the well known catalytic sequence responsible for mRNA cleavage. For this sequence, see, e.g., U.S. Pat. No. 5,093,246, which is incorporated herein by reference in its entirety.

While ribozymes that cleave mRNA at site specific recognition sequences can be used to destroy target gene mRNAs, the use of hammerhead ribozymes is preferred. Hammerhead ribozymes cleave mRNAs at locations dictated by flanking regions that form complementary base pairs with the target mRNA. The sole requirement is that the target mRNA have the following sequence of two bases: 5′-UG-3′. The construction and production of hammerhead ribozymes is well known in the art and is described more fully in Myers, 1995, Molecular Biology and Biotechnology: A Comprehensive Desk Reference , VCH Publishers, New York, (see especially FIG. 4, page 833) and in Haseloff and Gerlach, 1988, Nature, 334, 585-591, which is incorporated herein by reference in its entirety.

Preferably the ribozyme is engineered so that the cleavage recognition site is located near the 5′ end of the target gene mRNA, i.e., to increase efficiency and minimize the intracellular accumulation of non-functional mRNA transcripts.

The ribozymes of the present invention also include RNA endoribonucleases (hereinafter “Cech-type ribozymes”) such as the one that occurs naturally in Tetrahymena thermophila (known as the IVS, or L-19 IVS RNA) and that has been extensively described by Thomas Cech and collaborators (Zaug, et al., 1984, Science, 224, 574-578; Zaug and Cech, 1986, Science, 231, 470-475; Zaug, et al., 1986, Nature, 324, 429-433; published International patent application No. WO 88/04300 by University Patents Inc.; Been and Cech, 1986, Cell, 47, 207-216). The Cech-type ribozymes have an eight base pair active site which hybridizes to a target RNA sequence whereafter cleavage of the target RNA takes place. The invention encompasses those Cech-type ribozymes which target eight base-pair active site sequences that are present in the target gene.

As in the antisense approach, the ribozymes can be composed of modified oligonucleotides (e.g., for improved stability, targeting, etc.) and should be delivered to cells that express the target gene in vivo. A preferred method of delivery involves using a DNA construct “encoding” the ribozyme under the control of a strong constitutive pol III or pol II promoter, so that transfected cells will produce sufficient quantities of the ribozyme to destroy endogenous target gene messages and inhibit translation. Because ribozymes unlike antisense molecules, are catalytic, a lower intracellular concentration is required for efficiency.

Endogenous target gene expression can also be reduced by inactivating or “knocking out” the target gene or its promoter using targeted homologous recombination (e.g., see Smithies, et al., 1985, Nature 317, 230-234; Thomas and Capecchi, 1987, Cell 51, 503-512; Thompson, et al., 1989, Cell 5, 313-321; each of which is incorporated by reference herein in its entirety). For example, a mutant, non-functional target gene (or a completely unrelated DNA sequence) flanked by DNA homologous to the endogenous target gene (either the coding regions or regulatory regions of the target gene) can be used, with or without a selectable marker and/or a negative selectable marker, to transfect cells that express the target gene in vivo. Insertion of the DNA construct, via targeted homologous recombination, results in inactivation of the target gene. Such approaches are particularly suited in the agricultural field where modifications to ES (embryonic stem) cells can be used to generate animal offspring with an inactive target gene (e.g., see Thomas and Capecchi, 1987 and Thompson, 1989, supra). However this approach can be adapted for use in humans provided the recombinant DNA constructs are directly administered or targeted to the required site in vivo using appropriate viral vectors.

Alternatively, endogenous target gene expression can be reduced by targeting deoxyribonucleotide sequences complementary to the regulatory region of the target gene (i.e., the target gene promoter and/or enhancers) to form triple helical structures that prevent transcription of the target gene in target cells in the body. (See generally, Helene, 1991, Anticancer Drug Des., 6(6), 569-584; Helene, et al., 1992, Ann. N.Y. Acad. Sci., 660, 27-36; and Maher, 1992, Bioassays 14(12), 807-815).

Nucleic acid molecules to be used in triplex helix formation for the inhibition of transcription should be single stranded and composed of deoxynucleotides. The base composition of these oligonucleotides must be designed to promote triple helix formation via Hoogsteen base pairing rules, which generally require sizeable stretches of either purines or pyrimidines to be present on one strand of a duplex. Nucleotide sequences may be pyrimidine-based, which will result in TAT and CGC + triplets across the three associated strands of the resulting triple helix. The pyrimidine-rich molecules provide base complementarily to a purine-rich region of a single strand of the duplex in a parallel orientation to that strand. In addition, nucleic acid molecules may be chosen that are purine-rich, for example, contain a stretch of G residues. These molecules will form a triple helix with a DNA duplex that is rich in GC pairs, in which the majority of the purine residues are located on a single strand of the targeted duplex, resulting in GGC triplets across the three strands in the triplex.

Alternatively, the potential sequences that can be targeted for triple helix formation may be increased by creating a so called “switchback” nucleic acid molecule. Switchback molecules are synthesized in an alternating 5′-3′, 3′-5′ manner, such that they base pair with first one strand of a duplex and then the other, eliminating the necessity for a sizeable stretch of either purines or pyrimidines to be present on one strand of a duplex.

In instances wherein the antisense, ribozyme, and/or triple helix molecules described herein are utilized to inhibit mutant gene expression, it is possible that the technique may so efficiently reduce or inhibit the transcription (triple helix) and/or translation (antisense, ribozyme) of mRNA produced by normal target gene alleles that the possibility may arise wherein the concentration of normal target gene product present may be lower than is necessary for a normal phenotype. In such cases, to ensure that substantially normal levels of target gene activity are maintained, therefore, nucleic acid molecules that encode and express target gene polypeptides exhibiting normal target gene activity may, be introduced into cells via gene therapy methods such as those described, below, in Section 5.9.2 that do not contain sequences susceptible to whatever antisense, ribozyme, or triple helix treatments are being utilized. Alternatively, in instances whereby the target gene encodes an extracellular protein, it may be preferable to co-administer normal target gene protein in order to maintain the requisite level of target gene activity.

Anti-sense RNA and DNA, ribozyme, and triple helix molecules of the invention may be prepared by any method known in the art for the synthesis of DNA and RNA molecules, as discussed above. These include techniques for chemically synthesizing oligodeoxyribonucleotides and oligoribonucleotides well known in the art such as for example solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding the antisense RNA molecule. Such DNA sequences may be incorporated into a wide variety of vectors that incorporate suitable RNA polymerase promoters such as the T7 or SP6 polymerase promoters. Alternatively, antisense cDNA constructs that synthesize antisense RNA constitutively or inducibly, depending on the promoter used, can be introduced stably into cell lines.

5.4.3.2. Gene Replacement Therapy

Mahogany gene nucleic acid sequences, described above in Section 5.1, can be utilized for the treatment of a mammalian body weight disorders, including obesity, cachexia, and anorexia. Such treatment can be in the form of gene replacement therapy. Specifically, one or more copies of a normal mahogany gene or a portion of the mahogany gene that directs the production of a mahogany gene product exhibiting normal mahogany gene function, may be inserted into the appropriate cells within a patient, using vectors that include, but are not limited to adenovirus, adeno-associated virus, and retrovirus vectors, in addition to other particles that introduce DNA into cells, such as liposomes.

Because the mahogany gene is expressed in the brain, such gene replacement therapy techniques should be capable delivering mahogany gene sequences to these cell types within patients. Thus, in one embodiment, techniques that are well known to those of skill in the art (see, e.g., PCT Publication No. WO89/10134, published Apr. 25, 1988) can be used to enable mahogany gene sequences to cross the blood-brain barrier readily and to deliver the sequences to cells in the brain. With respect to delivery that is capable of crossing the blood-brain barrier, viral vectors such as, for example, those described above, are preferable.

In another embodiment, techniques for delivery involve direct administration of such mahogany gene sequences to the site of the cells in which the mahogany gene sequences are to be expressed.

Additional methods that may be utilized to increase the overall level of mahogany gene expression and/or mahogany gene product activity include using target homologous recombination methods, discussed in Section 5.2, above, to modify the expression characteristic of an endogenous mahogany gene in a cell or microorganism by inserting a heterologous DNA regulatory element such that the inserted regulatory element is operatively linked with the endogenous mahogany gene in question. Targeted homologous recombination can be thus used to activated transcription of an endogenous mahogany gene that is “transcriptionally silent”, i.e., is not normally expressed, or to enhance the expression of an endogenous mahogany gene that is normally expressed.

Further, the overall level of mahogany gene expression and/or mahogany gene product activity may be increased by the introduction of appropriate mahogany-expressing cells, preferably autologous cells, into a patient at positions and in numbers that are sufficient to ameliorate body weight disorder symptoms. Such cells may be either recombinant or non-recombinant.

Among the cells that can be administered to increase the overall level of mahogany gene expression in a patient are normal cells, preferably brain cells, that express the mahogany gene. Alternatively, cells, preferably autologous cells, can be engineered to express mahogany gene sequences, and may then be introduced into a patient in positions appropriate for the amelioration of the body weight disorder symptoms. Alternately, cells that express an unimpaired mahogany gene and that are from a MHC matched individual can be utilized, and may include, for example, brain cells. The expression of the mahogany gene sequences is controlled by the appropriate gene regulatory sequences to allow such expression in the necessary cell types. Such gene regulatory sequences are well known to the skilled artisan. Such cell-based gene therapy techniques are well known to those skilled in the art, see, e.g., Anderson, U.S. Pat. No. 5,399,349.

When the cells to be administered are non-autologous cells, they can be administered using well known techniques that prevent a host immune response against the introduced cells from developing. For example, the cells may be introduced in an encapsulated form which, while allowing for an exchange of components with the immediate extracellular environment, does not allow the introduced cells to be recognized by the host immune system.

Additionally, compounds, such as those identified via techniques such as those described, above, in Section 5.4.2, that are capable of modulating mahogany gene product activity can be administered using standard techniques that are well known to those of skill in the art. In instances in which the compounds to be administered are to involve an interaction with brain cells, the administration techniques should include well known ones that allow for a crossing of the blood-brain barrier.

5.5. Pharmaceutical Preparations and Methods of Administration

The compounds that are determined to affect mahogany gene expression or gene product activity can be administered to a patient at therapeutically effective doses to treat or ameliorate body weight disorders, such as obesity, anorexia, or cachexia. A therapeutically effective dose refers to that amount of the compound sufficient to result in amelioration of symptoms of such a disorder.

5.5.1. Effective Dose

Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD 50 (the dose lethal to 50% of the population) and the ED 50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD 50 /ED 50 . Compounds that exhibit large therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED 50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC 50 (i.e., the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.

5.5.2. Formulations and Use

Pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more physiologically acceptable carriers or excipients.

Thus, the compounds and their physiologically acceptable salts and solvates may be formulated for administration by inhalation or insufflation (either through the mouth or the nose) or oral, buccal, parenteral or rectal administration.

For oral administration, the pharmaceutical compositions may take the form of, for example, tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g., pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulphate). The tablets may be coated by methods well known in the art. Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid). The preparations may also contain buffer salts, flavoring, coloring and sweetening agents as appropriate.

Preparations for oral administration may be suitably formulated to give controlled release of the active compound.

For buccal administration the compositions may take the form of tablets or lozenges formulated in conventional manner.

For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

The compositions may, if desired, be presented in a pack or dispenser device that may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration.

6. EXAMPLE

Genetic and Physical Mapping of the Mahogany Locus

In the Example presented herein, studies are described which, first, define the genetic interval on mouse chromosome 2 within which the mahogany gene lies, and second, successfully narrow the interval to approximately 0.29 cM. Further, the physical mapping of this interval is described.

Mouse crosses were performed to obtain homozygous mg/mg mice. First, LDJ-Le-mg mice were crossed with CAST/Ei mice. The F1s were back-crossed with LDJ-Le-mg mice and the resulting litters scored for coat color. Mice showing coat color of mg/mg homozygotes were genotyped to using D2/NDS3 and D2/MIT19 markers to identify meiotic events. Mice showing recombinant events were fine structure mapped using various markers shown in FIG. 1 . All genotyping was performed using PCR-SSLP and then analyzed using PAGE.

After 2300 meiosesis, the mahogany gene was mapped to a 0.99 cM interval FIG. 1 . This corresponded to an interval width of 700 kb.

Physical Mapping of the Genetic Interval: The 700 kb mahogany region on mouse chromosome 2 is shown in FIG. 1 . Genetic markers, clones spanning the region and open reading frames in the interval are shown in the figure.

7. EXAMPLE

Identification of a Candidate Mahogany Gene

In the Example presented herein, a gene is identified within the cloned DNA described in the Example in Section 6, above, which corresponds to a candidate mahogany gene.

Clones spanning the 700 kb region were sequenced and open reading frames were identified and analyzed through this interval. Nucleic acid sequencing was performed using ABI sequencers and the manufactures recommended procedures. Many novel sequences encoding proteins are located in this integral, see the bottom of FIG. 1 . With each open reading frame identified, mutational analysis, primarily via SSCP analysis, was used with the three alleles of the mahogany phenotype mice to identify which of the open reading frames within this interval contain a mutation in an mg mouse.

A mutation was found in one of the genomic/cDNA sequences found in the integral in mg3J mice. FIGS. 3 and 2 provide the genomic and cDNA sequences surrounding the mutation, FIG. 6 shows the mutation in mg3J, and FIGS. 8 and 9 show splice variants in the 5′ end of the murine mg gene. The mutation in mg3J mice is a deletion of a GCTGC sequence which results in the creation of a frameshift. Based on the chromosomal location and mutation identification, the cDNA provided in FIG. 2 and the corresponding genomic DNA which contains the contigs provided in FIG. 3 represent the mg gene/locus.

Further analysis of cDNA clones identified two distinct splice variants in the 5′ end of the mg gene. FIG. 7 provides an analysis of the structure of the two splice variants, denoted akm1003 and akm1004. FIGS. 8 and 9 provide the nucleic acid and amino acid sequence of the 5′ ends of these splice variants and structural analysis of the protein encoded by the 5′ regions.

Analysis of libraries of human cDNA sequences led to the identification of three forms of the human ortholog of the mg gene: a long form (FIGS. 18A-D) and two shorter splice forms, each of which is shown in FIGS. 19A-D and 20 A-C.

8. EXAMPLE

Characterization of the Mahogany Gene

In the example presented herein, the nucleic acid sequence of the mahogany gene transcript identified in the example presented in Section 7, above, is used to generate Northern analysis data which characterize the expression of the mahogany transcript in a number of tissues both of wild type mice, and of mice exhibiting the mahogany phenotype. The results presented in this example are consistent with the mg gene being the mahogany gene.

For Northern analysis, polyA RNA was isolated from wild-type and the original mg mutant, mg3J and mg-Lester mice and utilized from the Northern analysis following standard protocols. Northern blots prepared from this mRNA was hybridized with a probe obtained from sequences common to the akm1003 and akm1004 sequences. Specifically PCR primers TTCCTCACTGG and GGACACACAG were used to amplify cDNA from the akm1003 sequence which had been radiolabelled by random priming using a Gibco-BRL kit according to the manufacturer's recommended protocol.

An mg transcript was found in all mice examined in mRNA isolated from brain (minus the hypothalamus), kidney, heart, testes, liver, skin, and hypothalamus. No expression was seen in muscle.

In a Northern blot run on RNA samples from mahogany mice, the mg transcript was found to be expressed at a reduced level in all tissues in mRNA isolated from mg3J mice, as a varied size fragment in mg-Lester derived mRNA, and at different levels and sizes in original mg mutant mice derived mRNA.

These results are consistent with the mg gene disclosed herein as being the mahogany gene.

9. EXAMPLE

Effects of the Mahogany Gene on Genetic and Dietary Obesity

This section describes experiments which examine whether the mg gene acts specifically within the agouti pathway. Specifically, these experiments test whether mg can suppress the obesity of other monogenic obese mutants as well as whether it can suppress diet-induced obesity. The results show that mg does not suppress obesity in any of the monogenic obese mutants. However, mg can suppress diet-induced obesity. Thus, the mg gene and its corresponding gene product and compounds that modulate mg expression and/or activity have implications in the treatment of diet-induced obesity disorders, as well as in the treatment of disorders related directly to the mg or agouti gene.

9.1. Materials and Methods

Genetic crosses: The crosses, and the number of animals for each (n) were (LDJ/Le-mg/mg×CAST/Ei)×LDJ/Le-mg/mg (n=1588), (C3HeB/FeJ-mg 3J /mg 3J ×CAST/Ei)×C3HeB/FeJ-mg 3J /mg 3J (n=324), (C3HeB/FeJ-mg 3J /mg 3J ×MOLF/Ei)×C3HeB/FeJ-mg 3J /mg 3J (n=216) and (C3HeB/FeJ-mg 3J /mg 3J ×C57BL6/J)×C3HeB/FeJ-mg 3J /mg 3J (n=309). The 2437 N 2 mice were analysed by coat colour to determine their genotype at the mg locus. As mice change color slightly at each hair molt and because the phenotype of mg/mg vs. mg/+ can be subtle, all mice were phenotyped at the same age by a single person. Genomic DNA was made from a tail biopsy of each mouse and analysed for multiple simple sequence length repeat polymorphism (SSLP) markers. The first ˜100 mice were typed for a series of polymorphic Mit genetic markers (Deitrich, W. F. et al., 1996, Nature 380:149-152) from distal mouse chromosome 2 in order to accurately delimit the position of mg. With the first ˜100 mice it was determined that mg mapped approximately 15 cM proximal of Agouti between markers D2Mit19 and D2Nds3 (FIG. 13 ). All remaining animals were genotyped for D2Mit19 and D2Nds3. Animals recombinant in that interval were typed with all available Mit markers between and for the ever growing number of markers developed during the project which, finally totaled 265 markers.

9.2. Result

The murine mahogany (mg) gene is known to act in a dosage dependent manner within the agouti pathway, to compensate for the agouti overexpression and for lack of signaling from the nul allele McIr (Miller, K. A. et al., 1997, Genetics 146:1407-1415; Dinulescu, D. M. et al., Proc. Natl. Acad. Sci. , in press; Robbins, L. S. et al., 1993, Cell 72:827-834). The phenotype of mice homozygous for both mg and a null allele of McIr (recessive yellow, McIr e ) is yellow, the same as the phenotype of McIr e /McIr e mice, indicating that mg is not acting downstream of McIr. A similar experiment was performed with obese Mcr4 knock out mice (FIG. 11 ). For both sexes, all the animals homozygous for Mc4r−/− were approximately equally obese and were heavier than the mice wild-type at Mc4r independent of the genotype for mg. This data strengthens and confirms the McIr data previously published, strongly suggesting that mg acts at or upstream of both melanocortin receptors.

To test whether mg acts specifically within the agouti pathway, experiments were performed to determine whether mg can suppress the obesity of other monogenic obese mutants of the mouse and whether it could suppress diet-induced obesity. Appropriate genetic crosses were set up to product mice segregating mg and one of the mouse obesity mutations Cpe fat , tub, or Lepr db such that all combinations of homozygous and heterozygous animals were on the same mix of genetic background. No suppression of obesity was seen for any of the monogenic obese mutants (FIG. 12) lending credence to the assumed specificity of action within the agouti pathway. To ask whether mg can suppress diet induced obesity C3HeB/FeJ-mg 3J and C3H/HeJ mice were placed, at weaning, either on normal chow having a physiological fuel value (PFV) of 3.63 kcal/gm with 9% fat, or onto a high fat diet having a PFV of 4.53 kcal/gm with 42.2% fat. Food consumption and body weight were measured weekly. Converting the grams of food consumed to calories indicated that C3H/HeJ mice on normal chow and high fat diet consumed ˜97 kCal/week and ˜96 kCal/week, respectively. C3HeB/FeJ-mg 3J mice on normal chow and high fat diet consumed ˜83 kCal/week and ˜81 kCal/week, respectively. Despite the equal calorie intake, the C3H/HeJ mice on the high fat diet readily gained more weight than the C3H/HeJ mice on normal chow (p=0.0004). In stark contrast, the C3HeB/FeJ-mg 3J mice on either diet showed no statistically significant difference in weight (FIG. 12 D). Female data showed the same trends, although there was no statistical significance between any of the mice on either diet.

10. Experiment: Mapping and Sequencing of the Mahogany Gene

This section describes experiments wherein the murine mahogany gene was genetically and physically mapped to an approximately 0.6 cM interval, and then sequenced. The murine mg sequence obtained was then used to isolate and sequence the human mg gene. Northern and in situ analyses of mg expression in mouse tissue are also described, and sequence motifs of the predicted MG polypeptide are discussed.

10.1. Materials and Methods

Physical Mapping: More than 36,000 individual sequences from the region were compared by BLAST (Altschul, S. F. et al., 1990, J. Mol. Biol. 215:403-410) to publicly available sequence databases and analyzed using GRAIL (Guan, X. et al., 1992, Proc. Eighth IEEE Conference on AI Applications: 9-13) to identify potential coding sequence. In addition, sequences from overlapping BACs were assembled using phrap (Sing, C. F. et al., 1998, Genome Res. 8:175-185; Ewing B. and Green, P., 1998, Genome Res. 8:186-194; Gordon, D. et al., 1998, Genome Res. 8:195-202), and the resulting contigs were also analyzed using BLAST and GRAIL to aid in gene prediction. This data was displayed in ACEdb (Durbin, Richard and Mieg, Jean Thierry, 1991, A C. elegans Database, Documentation, code, and data available from anonymous FTP servers at lirmm.lirmm.fr, cele, mrc-lmb.cam.ac.uk, and ncbi.nlm.nih.gov) to further visualize predicted exons and their relationships to each other.

Northern Blot Analysis: PolyA+ RNA was extracted from the tissues indicated from wild-type, C3H/HeJ and the three mutant alleles of mg, C3HeB/FeJ-mg 3J , LDJ/Le-mg, and C3H/HeJ-mg L , according to the manufacturer's instructions. RNA STAT-60 (Tel-Test, Inc., 1511 Sounty Rd. 129, Friendswood, Tex. 77546) was used to isolate total RNA. PolyA+ was isolated using Poly(A)Pure™ mRNA purification kit (Ambion, Inc., 2130 Woodward St. #200, Austin, Tex. 78744). 2 μg of each mRNA was separated on a 1% agarose-formaldehyde gel, transferred to nylon, and hybridized with a probe for mg corresponding to nt 990-1406 of the murine cDNA sequence with Rapid-hyb Buffer (Amersham LIFE SCIENCE, Gaithersberg, Md.). Filters were washed with 0.11×SSC, 0.1% SDS and exposed to KODAK X-omat film overnight.

10.2. Results

A positional cloning strategy was undertaken to identify the mg gene. Multiple genetic crosses were set up to produce second generation mice (n-2437) segregating mg which were used to genetically localise the mg locus (FIG. 13 B). When the genetic map critical interval for mg was resolved to ˜0.6 cM physical mapping was initiated. Approximately 1 Mb was contiged with 30 BACs (FIG. 13 C), most of which were made into random sheared libraries for shot gun sequencing. At completion of the project it was estimated that 85% sequence coverage across the interval had been achieved and that all genes within the region had been found. Twenty-nine genes were identified, 15 of which are novel genes. Within the final minimal interval for mg, indicated by the arrows in FIG. 13, there were eleven genes of which nine were unknown. All of these genes were tested as candidates for mg by examining the three mutant alleles of the mahogany locus, the original allele, mg, that arose in a stock of Swiss×C3H mice, and two alleles that have independently arisen on the C3H background, C3HeB/FeJ-mg 3J /mg 3J and C3H/He-mg L /mg L . Each gene was examined by Northern blot analysis and RT-PCR analysis of RNA from tissues from wild-type and mg mutant mice, by Southern blot analysis of DNA from wild-type and mg mutant mice, and by SSCP analysis of genomic PCR products designed to cover the intron-exon boundaries of much of each of the genes. In all, 20 genes were analyzed in this manner, one of which showed a northern blot difference between the wild type and mutant alleles (FIG. 14 ).

The wild type expression pattern of this gene gives three bands of size ˜9 kb, 4.5 kb, and 3.8 kb, of which the larges message is the most prominent (FIG. 14 ). The smaller two bands can be seen in all tissues but, depending upon tissue, may require extended exposure. Each of the different mg alleles gave a different expression pattern. C3HeB/FeJ-mg 3J /mg 3J has extremely low expression, the 9 kb message only being very faint in brain, hypothalamus, and fat on northerns. C3H/He-mg L /mg L expresses a single aberrant band of approximately 9.5-10 kb in kidney, heart, muscle, fat, and, most prominently, brain and hypothalamus. The LDJ/Le-mg/mg shows an altered ratio of the three wild type messages: the 9 kb message is reduced, while the two smaller messages are more highly expressed, in particular being very abundant in fat and hypothalamus. In situ analysis was used to look more closely at mg expression in the brain and specifically the hypothalamus. Overall hybridization in LDJ/Le-mg/mg looks equivalent to that of wild type, and the C3HeB/FeJ-mg 3J /mg 3J shows an overall reduction of expression. Close examination of the hypothalamic region in both wild type and mutant alleles revealed differences in the ventromedial hypothalamic nucleus (VMH). Both C3HeB/FeJ-mg 3J /mg 3J and the LDJ/Le-mg/mg have reduced VMH expression (FIG. 15) which is particularly interesting as many neuropeptides and receptors known to be involved in body weight regulation are expressed in the VMH, including Mc4r.

Initially, two overlapping mouse cDNAs of 1051 bps and 2419 bps were identified. Using these cDNAs as a starting point it was possible to build over 7990 bps of human sequences, using both the public EST database and an in house database, as well as identifying one cDNA clone from a human liver library. The 23 ESTs used in the contiging are listed in Table I below. Using the derived human sequence, it was then possible to estimate the intron-exon boundaries within the mouse genomic sequence. These were verified by PCR amplification and sequencing. In total, 4079 bps of mouse sequence was obtained, of which 4011 bp are coding sequence. The mouse genomic locus spans over 160 kb, and has 31 identified exons, at least one of which is differentially spliced.

The mutant mahogany alleles were also sequenced, checking all intron-exon boundaries. A 5 bp deletion at 2809 nt was found in the coding sequence of the mg gene from C3HeB/FeJ-mg 3J /mg 3J which introduces a stop codon a position 937, two codons 3′ of the deletion. This mutation will result in a seriously truncated protein lacking many interesting domains, as discussed below. The mg 3J allele is the same allele that showed extremely low expression levels. The combined Northern blot analysis, in situ hybridization analysis, and sequence analysis of the mutant mg 3J allele strongly suggest that this gene is the mouse mahogany gene.

The 4011 bp of open reading frame (ORF) of mouse MG predicts a 1336 amino acid polypeptide with molecular mass of 148,706 D (FIG. 17, top sequence). BLAST searches of the NCBI and SwissProt protein databases identified two human paralogues with a similar modular architecture (KIAA0534, Genbank accession no. 3043592; and MEGF8, Genbank accession no. AB011541), as well as a C. elegans homologue (YC81_CAEEL, Genbank accession no. Q19981).

Another human protein, Attractin or DPPT-L (Duke-Cohen, J. S. et al., 1998, Proc. Natl. Acad. Sci. U.S.A. 95:11336-11341), appears to be a 1198 amino acid residue, approximately 134,000 D, secreted splice variant of the MG polypeptide. An alignment of the predicted MG (top) and Attractin (bottom) amino acid sequences is shown in FIG. 17 . Attractin has not identified as being involved in the regulation of body weight. Rather, the protein is reported to mediate an interaction between T lymphocytes and monocytes that leads to the adherence and spreading of monocytes that become foci for T lymphocyte clustering (see Duke-Cohen et al., supra).

Searching the MG polypeptide with the SMART domain tool (Schultz, J. et al., 1998, Proc. Natl. Acad. Sci. U.S.A. 95:5857-5864) revealed sequence motifs that may provide further clues to its biological function (FIG. 16B, FIG. 17 ). The single transmembrane spanning MG protein has a large extracellular sequence of 1289 amino acids containing three EGF domains (Nakayama, M. et al., 1998, Genomics 51:27-34), two laminin-like EGF repeats, a CUB domain (Bork, P. and Beckmann, G., 1993, Mol. Biol. 231:539-545), a C-type lectin domain (Drickamer, K., 1995, Nat. Struct. Biol. 6:437-439; Weis W. I., and Drickamer, K., 1996, Ann. Rev. Biochem. 65:441-473), two plexin-like repeats (Maestrini, E. et al., 1996, Proc. Natl. Acad. Sci. U.S.A. 93:674-678), and six consecutive kelch repeats (Bork, P. and Doolittle, R. F., 1994, J. Mol. Biol. 236:1277-1282). Multiple EGF domains are commonly found in Type-1 membrane proteins involved in cell adhesion and receptor-ligand interactions (Schultz, J. et al, 1998, Proc. Natl. Acad. Sci. USA 95:5857-5864). Laminin-EGF-like modules are found in a variety of proteoglycans such as perlecan and heparin sulphate proteoglycan. As CUB domains also frequently occur in glycosylated proteins and c-type lectins are known to be carbohydrate binders, it is likely that MG is heavily glycosylated and that carbohydrate interactions are essential for its function. Many kelch motif containing proteins have been found that, like MG, have multiple consecutive domains. Such consecutive four-stranded β-sheet Kelch motifs form a bladed beta “propeller fold” that is common in many sialidases and other enzymes (Maestrini, E. et al., supra). Unlike the other well recognized domains, the “plexin” repeat is less well defined. It was first recognized as a triple repeat in the Xenopus gene plexin that has similarity to MET (Bork, P. and Beckmann, G., 1993, Mol. Biol. 231:539-545). Since then, this cysteine rich repeat has been found in 6 MET gene family members, three of which signal via tyrosine kinase and three of which are hypothesized to have putative signaling function via a novel conserved cytoplasmic domain. However, it is fascinating that there is an eight amino acid stretch that is 100% conserved in the four proteins shown in FIG. 16A from human, mouse, and C. elegans . The conservation of sequence across such widely evolutionary divergent species strongly indicates a functional domain, possible a putative signaling motif.

The multi-domain structure of MG is complex, but draws many similarities from receptor and receptor-like proteins. The full-length MG polypeptide is predicted to be a large membrane-spanning protein with multiple extracellular domains that may have a binding or gathering function as well as a highly conserved putative signaling motif in the cytoplasmic tail.

11. References Cited

The present invention is not to be limited in scope by the specific embodiments described herein, which are intended as single illustrations of individual aspects of the invention. Functionally equivalent methods and components are within the scope of the present invention. Indeed, various modifications of the invention, in addition to those shown and described herein, will become apparent to those skilled in the art from the foregoing description and accompanying drawings.

All publications and patent applications mentioned in the specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

129

1

8827

Mus musculus

1

gaattccggg cgaaggggag ccggcgtgcg gggtgtgtat gtgttcgctg ggcgccggct 60

cagccccagg aagatggtgg cggtggcggc ggcggcggcg actgaggcgc ggctgagggg 120

gagcacgagg acgacagcag cgcctgcggg caggaagggc aggcagcacc gaccctgcac 180

cgcgacaggg gcctggaggc cgggaccgcg cgcccggctg tgtctcccgc gggtgctgtc 240

gcgggcgctg cccccgccgc cgctgctgcc gctgctcttt tcgctgctgc tgctgccgct 300

gccccgggag gccgaggccg ctgcggtggc ggcggcggtg tccggctcgg ccgcagccga 360

ggccaaggaa tgtgaccggc cgtgtgtcaa cggcggccgc tgcaaccctg gcaccggcca 420

gtgcgtctgc cccacgggct gggtgggcga gcaatgccag cactgcgggg gccgcttcag 480

gacatctgtc tcacgcctat aatcacagct gttcggaagg tgaggctgga ggaacagttc 540

gaggcaagct tcggctacag aataagttca agagtaacct ggggcaactt gggcttgtct 600

ccaaaaccaa aatgagcgaa aaggagcaag ctagagtctt ttgggaaaat tttagctgac 660

taatttttca ccgagaacta actggctctt ctggatttgt aacagatgga cctgggaatt 720

ataaatataa gacgaagtgc acatggctca ttgaaggaca gccaaataga ataatgagac 780

ttcgcttcaa ccattttgct acagaatgta gctgggacca tttatatgtt tatgatgggg 840

actcaatcta cgcacctctg attgctgcct ttagtggcct cattgttcct gaaagagatg 900

gcaatgagac ggctcctgag gtcactgtca cttcaggtta tgcactgctg cattttttca 960

gtgatgctgc ttataatctg actggattta atatcactta caattttgac atgtgtccga 1020

ataattgctc aggccgagga gagtgtaaga gcagtaacag cagcagcgct gttgagtgtg 1080

aatgttctga aaactggaaa ggggagtcgt gtgacattcc tcactgtaca gacaactgtg 1140

gctttcctca ccgaggcatc tgtaatgcaa gcgataccag agggtgctcc tgctttcctc 1200

actggcaggg tcctggatgt tcaattcctg tgccagctaa ccagtctttt tggactcgag 1260

aagaatattc tgatttaaag cttcccagag cctctcataa agctgtggtc aatggaaata 1320

taatgtgggt tgttggcgga tatatgttca accattcaga ttacagcatg gttttagcgt 1380

atgacctgac ttctagggaa tggcttccac taaaccattc tgtgaacagt gtggttgtaa 1440

gatatggtca ttctttggca ttacataagg ataaaatcta catgtatgga ggaaaaattg 1500

attcaacagg gaacgtgacc aatgagctga gagtatttca tattcataat gaatcatggg 1560

tattgttaac tccgaaagct aaggatcagt atgcagtggt tggacactca gcacacattg 1620

ttacactggc atctggccgt gtggtcatgt tggtcatctt cggtcattgc ccactctatg 1680

gatatataag cgttgtgcag gaatatgact tggaaaagaa cacatggagt atattacata 1740

ctcagggtgc tcttgtgcaa gggggttatg gccacagtag tgtttatgat gacaggacca 1800

aggctctgta cgttcatggt ggctacaagg ctttcagcgc caacaaatac cggcttgcag 1860

atgacctcta cagatacgat gtggatactc agatgtggac cattcttaag gacagccgat 1920

ttttccgtta cttgcataca gctgtgatag tgagtggaac catgctggtg tttggaggga 1980

acacacacaa tgacacttcc atgagccacg gtgccaaatg cttctcctcg gacttcatgg 2040

cttatgacat tgcttgtgac cgatggtcag tgcttcccag acctgagctc catcatgatg 2100

tcaacagatt tggccattca gcagtcttgt acaacagcac catgtatgtg ttcggcggct 2160

tcaacagcct cctcctcagt gacgtcttgg tctttacctc ggagcagtgc gatgcacacc 2220

gcagtgaagc tgcttgtgtg gcagcaggac ctggtatccg gtgtctgtgg gacacacagt 2280

cgtctcgatg tacctcctgg gagttggcaa ctgaagaaca agcagaaaag ttaaaatcag 2340

agtgtttttc taaaagaacc cttgaccatg acagatgtga ccagcacaca gattgttaca 2400

gctgcacagc caataccaat gactgccact ggtgcaatga tcactgtgtc cctgtgaacc 2460

acagctgcac agaaggccag atctccattg ccaagtatga gagttgcccc aaggataacc 2520

ccatgtacta ctgcaataag aaaaccagct gcaggagctg tgccctagac cagaactgcc 2580

agtgggagcc ccggaatcaa gagtgcatcg ccctgccgga aaatatctgt ggcaatggct 2640

ggcatttggt tggaaactcg tgtctgaaaa tcactactgc taaggagaat tatgacaatg 2700

ctaaattgtc ctgtaggaac cacaatgcct ttttggcttc cctcacatcc cagaagaagg 2760

tggagtttgt ccttaagcag cttcgattaa tgcaatcatc tcaaagtatg tccaagctca 2820

ctctgactcc atgggttggt cttcggaaga tcaatgtgtc ttactggtgc tgggaggata 2880

tgtctccatt cacaaatagt ttgctgcagt ggatgccatc tgagcccagt gatgctggct 2940

tctgtgggat cttgtcagag cctagtactc ggggattaaa ggctgcaacc tgcatcaacc 3000

ctctcaatgg cagcgtctgt gaaaggcctg caaaccacag tgccaagcag tgccggacac 3060

catgtgccct gcggacagcg tgtggcgagt gcactagcag cagctcggag tgcatgtggt 3120

gcagtaacat gaagcagtgt gtggactcca atgcctacgt ggcctccttc ccttttggcc 3180

agtgtatgga atggtatacg atgagcagct gcccacctga aaattgctct ggctactgta 3240

cctgcagcca ttgcttggag cagccaggct gtggttggtg tactgatcct agcaatactg 3300

ggaaaggaaa atgtattgag ggcagctata aaggacctgt gaagatgccg tcacaggcct 3360

ctgcaggaaa tgtgtatcca cagccccttc tgaactccag catgtgtcta gaggacagca 3420

gatacaactg gtctttcatt cactgtccag cttgccagtg caacggacac agcaaatgca 3480

tcaaccagag tatctgtgag aagtgtgagg acctgaccac gggcaagcac tgcgagacct 3540

gcatatctgg cttctatggt gacccgacta atggaggcaa atgtcagcca tgcaagtgca 3600

atgggcacgc atcactgtgc aacaccaaca ccggcaagtg cttctgtacc accaaaggtg 3660

tcaaggggga cgagtgccag ctatgtgagg tagaaaatcg ataccaagga aaccctctca 3720

aaggaacatg ctactatacc cttctcattg actatcagtt cacctttagc ctgtcccagg 3780

aagacgaccg ctactacaca gccatcaact ttgtggctac tcctgatgaa caaaacaggg 3840

atttggacat gttcatcaat gcctccaaaa acttcaacct caacatcacc tgggccacca 3900

gcttcccagc cggaacccag actggagaag aggtgcctgt tgtttcaaaa accaacatca 3960

aggaatacaa agatagcttc tctaatgaga aatttgattt tcgcaaccat ccaaacatca 4020

ctttctttgt ttatgtcagt aatttcactt ggcccatcaa aattcagatt gccttctccc 4080

agcacagcaa cttcatggac ctggtacagt tcttcgtgac tttcttcagt tgtttcctct 4140

cgctgcttct ggtggctgca gtggtctgga agatcaagca gagctgttgg gcatccaggc 4200

ggagagagca acttcttcgg gagatgcaac agatggccag ccgccccttt gcttctgtaa 4260

acgttgcctt ggaaacagat gaggagcctc ctgatcttat tggggggagt ataaagactg 4320

ttcccaaacc cattgcactg gagccgtgtt ttggcaacaa agccgctgtc ctctctgtgt 4380

ttgtgaggct ccctcgaggc ctgggtggca tccctcctcc tgggcagtca ggtcttgctg 4440

tggccagcgc cctggtggac atttctcagc agatgccgat agtgtacaag gagaagtcag 4500

gagccgtgag aaaccggaag cagcagcccc ctgcacagcc tgggacctgc atctgatgct 4560

ggggccaggg actctcccac gcacgagcta gtgagtggca caccagagcc atctgcaggg 4620

aagggcgtgg cggggaaatg gctgtgcggt gcgggacgga agactggaaa ccctcaaagc 4680

atctgactca cctgcatgat cacaagcttt ctttgacggt ttctcccatc cgtgttccag 4740

catctaacct tttacttttg cataggaaat acttgattta attacaggtc cagggatgag 4800

ctgatggttg ctggaggagg ccagtgtaga gccagtgaga gaactaggaa tgacactcag 4860

gttcactgtg gaaaactgtt cttgggactg tctcaactgt gcaaaaaaca aaagatggag 4920

tgtttacaag tagacattcg tcatcagttg ttcttgaaca tggtctttta aaaactagtc 4980

agatgaatta acttgttttc atctgaagcc tgctatcttt tttaaaagat gtgctattta 5040

ttcttgcacg atttaggcaa ttatctctct tccagggagt accttttttt ctagttgaga 5100

attaataatg gtccatctct tttgatcata tcaagctagg atagaagggg ggctatttta 5160

aatgtcaagg tcagcagtgt tactttgaat gtaaactggt ataataggta gttttctata 5220

gtaacttgat taatttagtc ttaatccatt tgaaactctc tcttcctttc tctctgcctg 5280

tccctctcct tctccatctc accctccctc tctcacacat acacacacaa acacatacac 5340

acaacactaa gtgcctagac tttaaataga tctagcaatt ggaaagttag taagcctaag 5400

tttttacata attgcattcc tacattcttg taaaatttaa atagctacca ttggcaatct 5460

gctttttttc taaaatctga tttgcagcca ggaaagaatt ttctcaccca aggaacattt 5520

gatctagcag cagggatgag aggaaagcag aaatgaatga actgtgaaag ctcctgtttt 5580

tattatcaaa aaggacactg tcaagaaggc gccccctgcc cccacccccg tgtcacccta 5640

ggcctgataa gcgatcagag gaaaggactc attcatgtca cgcttccttg agcagaaaag 5700

agcactgaga gcacttggga cccctggatc agagagcatc tgtgtgtcct gcagcctcct 5760

ctgaacttgt ggttcattct caggctgggg tggactcaga tgccaggaaa gggacagcct 5820

cccattgtca ggcagaagct gcccaaagcc tggagaagga cttgtttgcc ctctttcccc 5880

caggaggggc tcgacccacc caccctccct ctcagaccaa ggtggtggct gtgaggaggg 5940

cagcaaatgc tgacaaggat gaaaagcaca tggaaaaaaa tggacgagga gggaaaactc 6000

tgccaaatgg aaaatgacca aatttaagag ggtgggacag tcccctgctc ctctcccaga 6060

gggcactgct tggaaattgt gttttcccca tttatggtgc tctgtattct ggcattatgc 6120

agcagcctcc cagaagctct cttctgcttc aaaacctggg atctctggca ttaccctatt 6180

gggatggacc gctggacagc aatgctcgag tttgtgaatt tggagagata ctcaaaagag 6240

ctaaaactgc agcattttac ctttaaatgc agtgcctaga gagagagtat tgtctcttcc 6300

ccaacactaa ccccactccc atgaagaatt gcctggaaag atgttttcaa ggaatttgaa 6360

ccataaaaca ctatctgatg cacagaacac ctctactttg agactcacct ctcataaagc 6420

ttctttttca cattactgtt aaagaccaga cgttctagaa aagacccctc ctctcatgag 6480

ctcccccatc cctgctacag aacacagcac ccatggcgcc tgcagtggac tggcccctta 6540

attcccacag gcccccccag caaggccaaa gggaggcccc tgggtattgt cctcctacaa 6600

ggaagatcct ctttgtttgt tcaaaggacc agttttccta ggccaaagaa gtctcttccc 6660

catgttagtc ctatgccttg aaatatcatg caccatgacc cacagccatc tggttatgtc 6720

ttattttttt cctaaaagat aatgtttatt tttaaaaagg aaggaagaag caagtgaagt 6780

ttcattctgc tccagcggtg gggaagccgc tgaatccacc tgcttctcct ttgcaaccga 6840

cagcaaacag ctttctccgg cctcagggca gaaaaaggga atggcaggga gtaagaggcg 6900

ctgggctcgg agcctgtttc caagaaggaa ttggttgtca tctggcagtg ttgcgcgtca 6960

caagagagcc tgtatataaa ttaaaatagt caagacaaca ctgaccttgc acttgtacat 7020

aactatacag tagtgtccag aatgttcaga cattcggagt gtacataaaa cagaaaaaat 7080

cttcatgtat ttttattaaa tataacaatg tctgagtttc acctaagatg tttttgtgcc 7140

atatgctgga tatccaggtt ctcgccaggc cccgatacat gaataacaaa cccaagaaac 7200

gcatccccat tgtgtgatgt gttcagatgc atctggcacc aattaggtat ttcttaaaac 7260

aggactcatc tgtcagagtg cacatgaaaa atcaggcagg gaatcgaaac gacagcgctg 7320

gaggagactc aggaagcaga ggcgtccctg ccgctgccct tggccctgca agcacatcat 7380

gaccctttct ggcagcctct tggtgctctg ggtagtgagg gatgaccagt cttgtcctga 7440

gaaatgtttc tcttagtctt taagttcaaa gactaacctg tagcaatcag actttccaaa 7500

agggggttct ccattttttg tagttttgtc taaattttta atgaccattt cctggaatca 7560

gtttattata ctgaaaactg ggggtgggag tagggagcta gtttgttgat aaatagttcc 7620

catttccccg tggagaattt gacataccct ggactcctgt gtgcctcctg ccatccctgc 7680

acacagcctg gggagaagcc tgtgcctccc cgtgtggaga gaaggcaacc ccagatcccc 7740

tgagctaacc cggaggaaag gcagtcctgg acagaagact gtcagcagaa ggaaagtact 7800

ggactacccg tgggtaagtc ctgccattca agactggaga cacctgggaa ataaaaagag 7860

cagggcactg ctggtgggaa gaggcatttt accttccagt gcaaatcctg ctcctttgat 7920

ttaatggggt gtactggggc caggggctga ttcacttcct tgggagatgg tggtgttttc 7980

atgaacatct ttgatccttc catttcattt attcatccat ccattcaaca agtatttgct 8040

aaacactaac ttaagctaat gctagggtag tgactgagat gtaaaaatag attttagaat 8100

taaaacaaaa tccaagtcct cacacccctg tcatcccagg agatctttcc ttgtggtggt 8160

ttctgtgaga attggccatc ctgaggacac agccaggacg gcagaggcct cctggcctca 8220

gggcatgccc tgcctacctt ctgaaatgtt taccccattg accaaacttg gctccagcca 8280

ttgcggtggt ttctagatag ccaggcccac caagagatat tgccccttga tgagagtcaa 8340

acaccctgcc tacaaggaga tgttttgaaa tggagaggaa aattggcacc tcatctttta 8400

aaggcagtaa tggaattgat tttcagtaac tgaatttgtg cacaaaacat tctaaacact 8460

agtgaagcct gtttcgttga actaattctg gctctggaaa tgtttttgtt ttatagttat 8520

ttacgatttc gtttgtttgg attcaagctt agtttgttaa tatgtataat ttagcatcta 8580

ttacactcat gtaaatatgg agtaagtatt gtaaactatt tcattgcggg gattgtgggt 8640

gttatacata catttaggac tgcaattttt tggtattttt tgtattgtaa aataacagct 8700

aatttaagca ggaacaagag aactaaggga ggtctgtgca ttttaaacac aaatgtgaag 8760

aacttgtata taaacaaaag taaatactat aatacaaact tccttctgaa ataaaagtag 8820

atctggt 8827

2

1260

Mus musculus

2

Met Arg Leu Arg Phe Asn His Phe Ala Thr Glu Cys Ser Trp Asp His

1 5 10 15

Leu Tyr Val Tyr Asp Gly Asp Ser Ile Tyr Ala Pro Leu Ile Ala Ala

20 25 30

Phe Ser Gly Leu Ile Val Pro Glu Arg Asp Gly Asn Glu Thr Ala Pro

35 40 45

Glu Val Thr Val Thr Ser Gly Tyr Ala Leu Leu His Phe Phe Ser Asp

50 55 60

Ala Ala Tyr Asn Leu Thr Gly Phe Asn Ile Thr Tyr Asn Phe Asp Met

65 70 75 80

Cys Pro Asn Asn Cys Ser Gly Arg Gly Glu Cys Lys Ser Ser Asn Ser

85 90 95

Ser Ser Ala Val Glu Cys Glu Cys Ser Glu Asn Trp Lys Gly Glu Ser

100 105 110

Cys Asp Ile Pro His Cys Thr Asp Asn Cys Gly Phe Pro His Arg Gly

115 120 125

Ile Cys Asn Ala Ser Asp Thr Arg Gly Cys Ser Cys Phe Pro His Trp

130 135 140

Gln Gly Pro Gly Cys Ser Ile Pro Val Pro Ala Asn Gln Ser Phe Trp

145 150 155 160

Thr Arg Glu Glu Tyr Ser Asp Leu Lys Leu Pro Arg Ala Ser His Lys

165 170 175

Ala Val Val Asn Gly Asn Ile Met Trp Val Val Gly Gly Tyr Met Phe

180 185 190

Asn His Ser Asp Tyr Ser Met Val Leu Ala Tyr Asp Leu Thr Ser Arg

195 200 205

Glu Trp Leu Pro Leu Asn His Ser Val Asn Ser Val Val Val Arg Tyr

210 215 220

Gly His Ser Leu Ala Leu His Lys Asp Lys Ile Tyr Met Tyr Gly Gly

225 230 235 240

Lys Ile Asp Ser Thr Gly Asn Val Thr Asn Glu Leu Arg Val Phe His

245 250 255

Ile His Asn Glu Ser Trp Val Leu Leu Thr Pro Lys Ala Lys Asp Gln

260 265 270

Tyr Ala Val Val Gly His Ser Ala His Ile Val Thr Leu Ala Ser Gly

275 280 285

Arg Val Val Met Leu Val Ile Phe Gly His Cys Pro Leu Tyr Gly Tyr

290 295 300

Ile Ser Val Val Gln Glu Tyr Asp Leu Glu Lys Asn Thr Trp Ser Ile

305 310 315 320

Leu His Thr Gln Gly Ala Leu Val Gln Gly Gly Tyr Gly His Ser Ser

325 330 335

Val Tyr Asp Asp Arg Thr Lys Ala Leu Tyr Val His Gly Gly Tyr Lys

340 345 350

Ala Phe Ser Ala Asn Lys Tyr Arg Leu Ala Asp Asp Leu Tyr Arg Tyr

355 360 365

Asp Val Asp Thr Gln Met Trp Thr Ile Leu Lys Asp Ser Arg Phe Phe

370 375 380

Arg Tyr Leu His Thr Ala Val Ile Val Ser Gly Thr Met Leu Val Phe

385 390 395 400

Gly Gly Asn Thr His Asn Asp Thr Ser Met Ser His Gly Ala Lys Cys

405 410 415

Phe Ser Ser Asp Phe Met Ala Tyr Asp Ile Ala Cys Asp Arg Trp Ser

420 425 430

Val Leu Pro Arg Pro Glu Leu His His Asp Val Asn Arg Phe Gly His

435 440 445

Ser Ala Val Leu Tyr Asn Ser Thr Met Tyr Val Phe Gly Gly Phe Asn

450 455 460

Ser Leu Leu Leu Ser Asp Val Leu Val Phe Thr Ser Glu Gln Cys Asp

465 470 475 480

Ala His Arg Ser Glu Ala Ala Cys Val Ala Ala Gly Pro Gly Ile Arg

485 490 495

Cys Leu Trp Asp Thr Gln Ser Ser Arg Cys Thr Ser Trp Glu Leu Ala

500 505 510

Thr Glu Glu Gln Ala Glu Lys Leu Lys Ser Glu Cys Phe Ser Lys Arg

515 520 525

Thr Leu Asp His Asp Arg Cys Asp Gln His Thr Asp Cys Tyr Ser Cys

530 535 540

Thr Ala Asn Thr Asn Asp Cys His Trp Cys Asn Asp His Cys Val Pro

545 550 555 560

Val Asn His Ser Cys Thr Glu Gly Gln Ile Ser Ile Ala Lys Tyr Glu

565 570 575

Ser Cys Pro Lys Asp Asn Pro Met Tyr Tyr Cys Asn Lys Lys Thr Ser

580 585 590

Cys Arg Ser Cys Ala Leu Asp Gln Asn Cys Gln Trp Glu Pro Arg Asn

595 600 605

Gln Glu Cys Ile Ala Leu Pro Glu Asn Ile Cys Gly Asn Gly Trp His

610 615 620

Leu Val Gly Asn Ser Cys Leu Lys Ile Thr Thr Ala Lys Glu Asn Tyr

625 630 635 640

Asp Asn Ala Lys Leu Ser Cys Arg Asn His Asn Ala Phe Leu Ala Ser

645 650 655

Leu Thr Ser Gln Lys Lys Val Glu Phe Val Leu Lys Gln Leu Arg Leu

660 665 670

Met Gln Ser Ser Gln Ser Met Ser Lys Leu Thr Leu Thr Pro Trp Val

675 680 685

Gly Leu Arg Lys Ile Asn Val Ser Tyr Trp Cys Trp Glu Asp Met Ser

690 695 700

Pro Phe Thr Asn Ser Leu Leu Gln Trp Met Pro Ser Glu Pro Ser Asp

705 710 715 720

Ala Gly Phe Cys Gly Ile Leu Ser Glu Pro Ser Thr Arg Gly Leu Lys

725 730 735

Ala Ala Thr Cys Ile Asn Pro Leu Asn Gly Ser Val Cys Glu Arg Pro

740 745 750

Ala Asn His Ser Ala Lys Gln Cys Arg Thr Pro Cys Ala Leu Arg Thr

755 760 765

Ala Cys Gly Glu Cys Thr Ser Ser Ser Ser Glu Cys Met Trp Cys Ser

770 775 780

Asn Met Lys Gln Cys Val Asp Ser Asn Ala Tyr Val Ala Ser Phe Pro

785 790 795 800

Phe Gly Gln Cys Met Glu Trp Tyr Thr Met Ser Ser Cys Pro Pro Glu

805 810 815

Asn Cys Ser Gly Tyr Cys Thr Cys Ser His Cys Leu Glu Gln Pro Gly

820 825 830

Cys Gly Trp Cys Thr Asp Pro Ser Asn Thr Gly Lys Gly Lys Cys Ile

835 840 845

Glu Gly Ser Tyr Lys Gly Pro Val Lys Met Pro Ser Gln Ala Ser Ala

850 855 860

Gly Asn Val Tyr Pro Gln Pro Leu Leu Asn Ser Ser Met Cys Leu Glu

865 870 875 880

Asp Ser Arg Tyr Asn Trp Ser Phe Ile His Cys Pro Ala Cys Gln Cys

885 890 895

Asn Gly His Ser Lys Cys Ile Asn Gln Ser Ile Cys Glu Lys Cys Glu

900 905 910

Asp Leu Thr Thr Gly Lys His Cys Glu Thr Cys Ile Ser Gly Phe Tyr

915 920 925

Gly Asp Pro Thr Asn Gly Gly Lys Cys Gln Pro Cys Lys Cys Asn Gly

930 935 940

His Ala Ser Leu Cys Asn Thr Asn Thr Gly Lys Cys Phe Cys Thr Thr

945 950 955 960

Lys Gly Val Lys Gly Asp Glu Cys Gln Leu Cys Glu Val Glu Asn Arg

965 970 975

Tyr Gln Gly Asn Pro Leu Lys Gly Thr Cys Tyr Tyr Thr Leu Leu Ile

980 985 990

Asp Tyr Gln Phe Thr Phe Ser Leu Ser Gln Glu Asp Asp Arg Tyr Tyr

995 1000 1005

Thr Ala Ile Asn Phe Val Ala Thr Pro Asp Glu Gln Asn Arg Asp Leu

1010 1015 1020

Asp Met Phe Ile Asn Ala Ser Lys Asn Phe Asn Leu Asn Ile Thr Trp

1025 1030 1035 1040

Ala Thr Ser Phe Pro Ala Gly Thr Gln Thr Gly Glu Glu Val Pro Val

1045 1050 1055

Val Ser Lys Thr Asn Ile Lys Glu Tyr Lys Asp Ser Phe Ser Asn Glu

1060 1065 1070

Lys Phe Asp Phe Arg Asn His Pro Asn Ile Thr Phe Phe Val Tyr Val

1075 1080 1085

Ser Asn Phe Thr Trp Pro Ile Lys Ile Gln Ile Ala Phe Ser Gln His

1090 1095 1100

Ser Asn Phe Met Asp Leu Val Gln Phe Phe Val Thr Phe Phe Ser Cys

1105 1110 1115 1120

Phe Leu Ser Leu Leu Leu Val Ala Ala Val Val Trp Lys Ile Lys Gln

1125 1130 1135

Ser Cys Trp Ala Ser Arg Arg Arg Glu Gln Leu Leu Arg Glu Met Gln

1140 1145 1150

Gln Met Ala Ser Arg Pro Phe Ala Ser Val Asn Val Ala Leu Glu Thr

1155 1160 1165

Asp Glu Glu Pro Pro Asp Leu Ile Gly Gly Ser Ile Lys Thr Val Pro

1170 1175 1180

Lys Pro Ile Ala Leu Glu Pro Cys Phe Gly Asn Lys Ala Ala Val Leu

1185 1190 1195 1200

Ser Val Phe Val Arg Leu Pro Arg Gly Leu Gly Gly Ile Pro Pro Pro

1205 1210 1215

Gly Gln Ser Gly Leu Ala Val Ala Ser Ala Leu Val Asp Ile Ser Gln

1220 1225 1230

Gln Met Pro Ile Val Tyr Lys Glu Lys Ser Gly Ala Val Arg Asn Arg

1235 1240 1245

Lys Gln Gln Pro Pro Ala Gln Pro Gly Thr Cys Ile

1250 1255 1260

3

17056

Mus musculus

3

agattttatg ccttcgtaca cgcctcccat aagatggaca aggtgtacta attactgcca 60

ttactgttgc tgaccccaga ggtcaatgtc ctcacatggc ctctactggc actgtctggg 120

cagaaactgt atatccaact ggtgaacctg aaagccctat gactacttgg tgtctctggt 180

gctaacccta gtcgttgggg catcttactg tatcctggta aggaaagaca tccaggctcc 240

ccacttaymk wwacyrgywm rgmycakgsy mgrgcyaawk tkctgtrrmr tctggctggc 300

atagagacat tactattgaa agttttgtct ttctaaatcc ttggactaaa gagagcacaa 360

gattttctgg aagatcttgc tttaaatttt ttttttattc ttttgagatg ctacatataa 420

ttagaggccc tgcacatgga ggcgagaacc ccacctctgg gctacatcct acgtcttttc 480

cttagggtat ttttttttct ttcttgtacc tatcagtatt actaagttgc aaatgtgctc 540

agcagtaaat ttaacataca taggcaaaaa gaaaagtctc aggacaccct gcctcacact 600

gtttactgtg ctcaggagta ctgagccata ctgttttctt gctgctgctt tttttctctt 660

ggttgtttac acacagtgtt caaggtgtgt taatcatagt tagtatttca attttttctt 720

aggtcagcaa gaaagctcac agaggaagag tgctttgctg ccagcctgat gacctgggtg 780

acccaagtga tctcacctac agggtgggag cacagcacag cattccaagt ctttttctga 840

ccacacaggc actatggcac acaaacacac aggatacata aatgttaaaa aaaaaaaaag 900

acttttatat ttttctccat ataatttaaa agattcctct ttcaacattc cttttgcaaa 960

gcagtatcat tgtgtttgta tatgtgtgtc cttccacatt ttgtcttcaa ttctaaattt 1020

ttagaattgt tagcctggtc ctctcatttc tactactttc tctagtaaac tgtcctttca 1080

tattacacat cgctctcctg tcacctgttt tagagctgtc atccatttta taaggttact 1140

tcactgttct acactacttt gtgtctttta attactatgc ctggggtgat tcaaaaactg 1200

tctgtgatgg gttggttgag gatggctcca ataggttcaa cacttggtcc tgattggtgg 1260

aactgttttg ggaaggatta ggaggtgtga ccttgtgggg gagtgtgtca ctgggagtga 1320

gtgacctttg aggtttcaaa agcccatgct aggcccagtg tctgtctgcc tgtctgtctg 1380

tctcctccct ttgctctttc ttccctccca cttgcttgca gatcagattc gagctcttag 1440

ctactgctcc cgtgctgtgc cttgctgcta ccatgcttct tgccatgatg ttcatagact 1500

tactctctga aactgtaaat aagcccccta ataaaatgct ttcttttaaa actgccttga 1560

tcatggtgtc tcttcaaaga aatagaacat taacaaaaac actataccaa actgcctaat 1620

agtcctacta attttatgat gagtgctagt gctttataat cactagaaga aaaaatttcc 1680

aggccataaa attaacatgg ttttaagtat gtataaatct tgtcttgaaa tctgttttct 1740

ataactaact ctaatatgat aatgtatatt ctaccttcaa aaaagcacaa ataagacttc 1800

aaaccctggg aattgttaga caaaggccat ttaatactaa taagctataa actgaaacca 1860

tctgatatat gaaaactatt aataaaatca agataaaata acccctattt atataactta 1920

ctatatacct aaagcaaaat atcaaagaaa gtaccttaaa aagataaatt attcttattt 1980

tgacaatgaa ttctttgggg cgttaaattg tagaatatca acacatatca agaaagttta 2040

gaagaaaact accaaagttt aaacagactt tcctcggtaa ttactggtga tttcttggct 2100

tttttttttt acactgcagt ttttcagggt ggaaacttaa gctttgtaca gaagcactta 2160

ccaccactct cagagctgga aatggctcaa agggcaaagc attacaagcc tggcaacctg 2220

aaccaaatac ccaaaacact tgcaaaggtg aaaggagaaa actaactcca ggaagttgtc 2280

cttcgagctc ctcttgcaca ccactgtata caccccctta tatacactca gttaccataa 2340

ataaaatgtt tcattataaa gacacttacg ctaaaaccat gctgtaatct gaatggttga 2400

acatatatcc gccaacaacc cacattatat ttccattgac cacagcttta tgagaggctc 2460

tgggaagctt taaatcagaa tattcttctc gagtccaaaa agactggtta gctggcacag 2520

gaattgagca tccaggacct aataaaaaaa aaaaaaacaa caacaacaac aaatagcttc 2580

acaaaatgca gcctgaaagt ttatagtatt ccaagttcca atctaagtgc aaagaatatt 2640

taaagacttg tggggctaga gagatggctc agtggttaag aaaactgact gctcttcttg 2700

gaggtcctga gttcaaatcc cagcaactac atggtggctc acaaccatat gtaatgggga 2760

tctgatgccc tcttctggtg tgtctgaaga cagcaacaat gtactcacat gaaataaata 2820

aattaatttt ttaaaaaaca gaccagaaaa aaaaaaaaaa aaaagacttg tgtttccttt 2880

agcacttaag cgcaaacatc tttaacttgt ggggttttaa aggtttttac atgtacaggt 2940

attttgttta catgtatgcc tatataccac ttgcttgctt ggtacccaat gatgtcagga 3000

aaaggcattg aatcccctgg aactagagtt acagatctta tgagctactt tgtggatgct 3060

aggatcaaac ctgagtcctc tggaagagca accagtactc ttaaccaaga agccatctgc 3120

ttagcaccta acatgagttt ttaacttact caagatacag accaaaacca atcactccct 3180

tataaaattt aatactacac actttctgat aatttggcaa tttctgataa tcaggttaaa 3240

cttttttaga ggtaaaaatc ttgctgaagc aacatttagt agaaagggta gaccaagggg 3300

ttattatatt aactcatgtg gaaaaggcat tagggttgaa atataatgac agatcaaaat 3360

cgatcttctg gcaagtccag gcgctgaata gatgaaagag acaaagggag aattggacaa 3420

actaaaaaca tttacatgaa cacttacttt ctgaggacct aagcatagaa ggaaaatcac 3480

taaaccaacg atgactgctt cctcaatacc ccagggaatt ccctacagta ccttagtacc 3540

cggttgtgtt gggtaatggc actagatgac agcactgaga ctctaaggaa cgcttgtcct 3600

cctctcagct tgagtctctg cttctctatc accagaccat gttccctaat tcccacgaat 3660

gagttgcaaa ggatttgtca aacctttcca caattctaag cacatagata acaaccacat 3720

atatgtaaat tcaaagaatc tgaataaatg gagatgaatg cttaaatgcc acctgataca 3780

tgattaacat aaggcgtatg gctgctaaaa taaactccct acagttcact aactcagaac 3840

tttctgtgag ggaaaggact ttgaagggca gctcctaccc tgccagtgag gaaagcagga 3900

gcaccctctg gtatcgcttg cattacagat gcctcggtga ggaaagccac agttgtctgt 3960

acagtgagga atgtcacacg actccccttt ccagttttca gaacattcac actcaacagc 4020

gctgctgctg ttactgctct tacactctcc tcggcctgag caattattcg gacacatgtc 4080

aaaactacaa agacaggaga aaacgaagtc aacaatttca actaagcaac attgcaacta 4140

atgcagacct tcctccttca gtttaagttc agttcatttg caagtgtgac tgcaggactt 4200

accagttagc ccaagtgtgc tcacagagct ctgtgtagct agagccccag gctcaagtaa 4260

tgaaatcaaa tcaaccttgc tgcattcaca tatgaagaag gaagaataaa taactcacaa 4320

agttagagaa attacaaaac aatagacatt tgtgcaaaat cacttagact tagctcaaga 4380

ctggcaacca ggatcctact ctttctggta gctcattagt aaagagttct acaaaagcag 4440

caaggtcatg ctaggaagtg gaggaaggag aggaagccaa tgagctgcca acattcacgg 4500

tatacatttc tctgtaaaga ttctgagaat taacagaatt taagattatt ttccagtgat 4560

gtagttaaag gtctttagta acttttatca gcttagaagg agaagagcag ttaacttcat 4620

gtatgagttt aagtgtctca tgacttaaga taacagtttt gctacaattt gaaatgccat 4680

acttcagact ttttaaaggg gtgcattagt ggactattac aatagcttaa aaatatagat 4740

ttctcctact gatgattatt actgagacac tactagtctt tattaaattc acttagcaaa 4800

actcctgaca ttttcttcca gcagcggaag aatgtctctc tcttctagga gatcctcagt 4860

gacaagatct agaaagacca agaactgtgg tcccaaccag tggggctgat atttgtttaa 4920

ccttttagct cctgtttctt caattatgaa aaaaaaaaaa aagaagaaga agaaaatcca 4980

tgttaaaatt tagcaaggag cctgactagc tagaagcctc cctccaatat attagtgtta 5040

ttaagtcatt tgagtagtat cacaaatatt aaatctaaat atcttacttg taagtgatat 5100

taaatccagt cagattataa gcagcatcac tgaaaaaatg cagcagtgca taacctgaag 5160

tgacagtgac ctcaggagcc gtctcattgc catctctttc aggaacaatg aggccactga 5220

aatgtaaaca cagaccagat tacagcaact tcaacagaaa ctgtctatat gttactattt 5280

gatcctgctg ctcctgttcc aacacacact gtaaatgtga ctctagctgg cctcaaattc 5340

acagacccac ctgcttccac ctcctgggtt ataggcatgc gctactatgc ccaacatcta 5400

aaaggatttg aaatctatga ctttgattga atttttggtt ttttgttttt gctataaact 5460

ttttattata atactctcaa gtctctacaa taacattatt aacaaacttt atgaattgac 5520

aactgtcaaa tatatactgt tgaaagaaaa tactttacat atttttgtaa tatgtatcat 5580

ataatctttt taatgtattt tatagatgtc ttatataagt aaaaatagaa aagtttactg 5640

atttataatc cttatactat tagctttcag acgtattttt gttgttaaac tggtaacaca 5700

ttttatgttt ataattcaca ataagcactg ccactgaagg tgccaaaggc tccctagaat 5760

ctcagtaaga acctagtggg taatatttga agttttggat gccagtaaat tcatgtgtaa 5820

agatttattg agtaagtgac taccagcggg acagtggtgg tgcacgcctt tagtcccagc 5880

acttgggagg cagaggcagg cgaatttctg agttcgaggc cagcctggtc tacagagtga 5940

gttcccagga taaccagggc tacacagaga aaccctgtca ccctgtctca aaaaaaaaaa 6000

aaaaaaaaaa aagaatatac catttttaag gcatttgatc cacaaaatca taccaccttg 6060

ttttacaaaa gatatatatt aacttgaagg ctggaaatgg tggcacatgt ctttagtccc 6120

agtattggga agacagaccc agatggatct ctgagttcaa gaccagcatg gtctacatag 6180

tgaattccat gtaagtttgt ccgtgtgtgt aacttgaaac ctcattatag aatggaagtg 6240

tctaccccac cccacttacc aacagtaagg aatattatgt tggtcccgct catttaatac 6300

atggtgtact cccaaggtaa atcattttca tgtttagtcg ctcctattat tttttccatt 6360

atcaattcac tacaactact accaccaatc acatttagcc actagaaaag ccatgtgatt 6420

tgctccacac atacaacttc actcaataaa taaacatctt atcagtacta ctctctcttt 6480

cactcactca atccctagtc ccctaagttt ttggacgatt acaccaggta aattcctact 6540

tcagggttat gaccatctta aaaactacga cctagcaatt ctctttgtat aagaaatact 6600

tccccgtata tacacagaaa aacaaagaac actactacag cactattcag atgacaactg 6660

actaaaagtc acctaattgc ttatttatgg gagttgatta aattagtcat tacaaatctg 6720

taggtctgca agactaacca agagcttcgt gaggacaata ggtagggcta cccagagaaa 6780

ccctgtcacc ctgtctcgaa aaaaaaaaaa aaaaagggag gcacagagaa aaaacaacag 6840

gcccggggta cctgtacatc tatgtaagcg taggtacatg cacataaaag tgactacaag 6900

agaacataaa cagagagcgc cgatgagaag aggatgggat ttttcattta atttgcgtgt 6960

atgagagcac ctatatgtgc atgttatccg caccaaagtg tgtagggtac attatgtgag 7020

tgtgcctgca gacgtcactg tcaggtgtct tcaatcactc ccctcctttt tcttctggag 7080

ataagagttt catgaagtag tactggctgg actagaactc actatgcaaa ccaggctggc 7140

cttgaattct cagagagcct cttgagtgct ggaattatat gcatgtgccg caacacagcc 7200

cacctcattt tggggggtag gatctttcac tgaacctgag ctcactgatt ggttagaccg 7260

gactggccag taagttccag gacctctctt gtctccgcct cttcagcact gtgatcacag 7320

gctcacaacc acacctggac ttttacttga gtcctggaga tctaaactca gctctccatg 7380

cctgtgcaga aggaattaaa ctgagccagc tgtctcagta tcaagagaga acataggaac 7440

tgtaagattc tgacagtact ctagggctta cagaacaccg acacattttc tactatgtat 7500

tcagttaata aaagaataaa tacaaacaaa aaaacatgag aaacatatag aggcagagac 7560

agacagacac acacacacac acacacacac acgcacacac acacacacac acacacgcac 7620

ttagacgggt gtgggggaag aaagagcaag gccacctaga aacaggtacg ttccatgcaa 7680

atgatcacag gaaaggattg gggattttta accacttgtg ggaaatgctg tactctccta 7740

ttctagcaca gatttgagga aaaagtagac cagagagtct gtccttccac atatcctgga 7800

aagtcactga catgtccaag ttttgatttc ttcataggga caatgagaga aacccagact 7860

atctcacagc agcacagcaa ggaccaacca gcagagcagg agaagtgctt acagcagtgt 7920

gctgctagaa ggtgcaacag tcttcttaca gagggcattt aaatatgcag gatggataag 7980

tttgccaact acaactacag aggctggaca aggtaggaca gcttcttcac tgtcaaagac 8040

gtttgggcag ttgcttctat ttaccttaaa atcaaactgt gacagctgtg gcatatatag 8100

atttctccca gaatgaaaac acattaactc acttatgtca ataatatgga gtaaacacaa 8160

acatagtcta tctagctcag catgcaagac atgtgaggaa gaggagctac tgtgagtccc 8220

tatccctgtc cctaaggaaa ccaatatatg taaatgtagt ctaagctgca ggcagttctt 8280

caactgccta ccccaggctg ctcaccactt cacattctaa gcacagacta gaaagtatga 8340

tcaacctctg aacactgtgc tataatgtta ccatcaatct cacacacaaa tttcataaca 8400

ttttaagtaa gtctatgatg attctatgtt gtgtcccagt tatataagat ccataggtca 8460

cagggtagac attcaaggac accaacattt ggaattttgg gtttttttgg tgtactgtat 8520

atacttgcta gtgcaggtac ccatgctcat gtgtgtagaa gttgggcgtc tttcttctat 8580

cactgtctac tttatatttt ctttattgtt tcatttgata tgtataggtg ttttgcctgc 8640

atatatgtgt atgtttgttg ccagaagagg gtattgaatt ccctgggact agagttacag 8700

gtggttgtga ggcaccatta tgggtactgg gactcaatcc tgggttctct ggaagggcag 8760

ccagtacttt taatcactga gccatctctt tagcttcctt cgttcattcg ttcgttcatt 8820

ccttcattcc ttcattcctt cattcagagg attgagatac cttcctcagt taggctggct 8880

agccaatgga ctctgggaat ctatctgttc agctattctc tccttcccca tccaagtgct 8940

ggggatacag gcaggtccta ctgggttcat tttgaaaaat tacagaacta tgtattttct 9000

tcataaatct gaaactcagc ataactgtct caggctaaca tggaatccct aaatatatat 9060

gaggcacaac ctgactttac caactgtact atgtaaattt gctagtatat tagtcaacac 9120

ttaatggaaa aacatctgat aaaaacaact tacaggccaa taggcaagga gacacttggg 9180

gaggtggatt caaggcagtc actggattct tgaatttaag tccagcctag gctacatgag 9240

attctgcttc aaaaaataaa caattaaatt tatgggggaa agaatgatgt attttggttt 9300

cagaaattcc atcctatcat ccaagggaga tattgtataa cagcgaagtt cctcagctca 9360

cagcagtcag tagcatatag acaatcctgg ctccaagcct atgaaaacac agcctgtact 9420

aaaggtgtgt tcctgtgttt tgagtgagat gtgcccccta agtcttgtgt atttgaatac 9480

ttggcactca cttggtggcg atttgggagg aattaggagg tgtggccttg gtggaaaagg 9540

agcatcacta gggtcaaggt ttcaaaatcc tcctgccatc atccccaata tgtcctctct 9600

gcctcctgct tgcagttcaa gctatgagct cttagttact acttccacca cctacccctg 9660

ctatctctgc tccatcatca tggactccta ttctggtgga actgttagtc caaaaaagtc 9720

ctttcttcta caacttgatt tgatgccaga tctagccccc cagcctagct agcaatatac 9780

caaggtatac catcttgaac tctaggtgtc tctcaatcca atcaagctac ataagattaa 9840

ccatcatacc tagtcatccc caaatcagtg tatctctctc ctcccaagac tataagctcc 9900

tcaagggtca aaatatgtag aaaggaagaa agattctcaa aggtcaagga tcagaccttg 9960

gtgaggattg agcactgtct acactttgcc tggtaaagaa gggtccacaa tgtaaaagag 10020

aactgacctg aacagttttc aattaggtgc taacaaatgt ctcatacgta ttgagtttct 10080

tataaataaa taaataaata aataaataaa taagcaagca agcaagcaag cacttaagag 10140

cactagctgc tttcttcctg aagacctggt ttcaattacc cagcacttat acagaggctc 10200

ataccaattg taactccagt ttgatgatat ccaacatctt cttctagcct tcagacacca 10260

agcaccaagc atgtaatggt ataacacatg tataccaaac acccatacaa accaattttt 10320

aaaaaaatat tcgagccggc gtggtggcgc acgcctttaa tcccagcact cgggagacag 10380

aggcaggtgg atttctgagt tcgaggccag cgtggtctac agagtgagtt ccaggacagc 10440

cagggctgca cagagaaacc ctgtctcgaa aaaccaaaaa aaaaaaaaaa aaaaaaaaat 10500

agtcatttta gggctggaga gatggctcag gggttaagag cactgactgt tcttccagag 10560

gtccttagtt caatatccag caaccacatg gtggctcaca gccatttgta atggggatcc 10620

aatatcccat tctggtgtgt ctgaagacag ctatagtgta aataaaataa agaaatcata 10680

taaataaaat aaataaatct ttttaaaaat attaattaac ccaggctgaa cctaaactta 10740

caaacttccc acattaggct ctttaatgcg ggtgttatag gtctgaatac cagcttaaga 10800

ataatattct tctgaagaat gtgccctggt caatcaccat gaccacacct gccaacaggt 10860

ccttcataaa atacttggta tatgttgaat gttccataaa attatggagc tagaaaaggt 10920

agtgagctag aaggatatta aagatataaa ccattgcccc agtggtcctc acatttgtct 10980

agtaatagaa cgttgttaaa ctgtttttat ttagaatttc aatatataaa agacaaatat 11040

gaaatagtcc ggaagcaaat taagctacag cttgcagcaa agccagatag aatgcagatt 11100

aaactaacac agtacctttg tcttatgttt tagatgctaa agtctagtct acaaccccag 11160

ctgcccttga actcttagca gtcctcttgc ttcagcctct catgctgcta gggttaaaag 11220

tatgtgcgac cacacacagt tttgaagttt agagcactta aatgatctat tcagcaactc 11280

aggcaggatt tacactgaaa gtaaattatc ttatgaatcc tttttggttt tccttttatt 11340

catttcattc atgcacctta catgaactat ctattgctag gctgtctcta tactggatgc 11400

tcagcacatc accaacatgc cgattcttct actggtacaa tggcaatgct gagaaaacca 11460

cacaacctaa gacagtaggg aggtggtgct ctgattgttg gtgttgttgt tgttgttgtt 11520

ttggtttttc gagacagggt ttctctgtgt agccctggct gtcctagaac tcactctgta 11580

gaccgggctg gcctcaaact cagaaatccg cctgcctctg cctcccaagt gctgggatta 11640

aaggcgtgtg ccaccacgcc gggctctggt gctctgattt ttaaatacaa caattttcag 11700

ctagcaatgt aactcagtag taaaatgcct gcccagcatg cacaaggctc cagactggac 11760

cctgagcacc acaacacttt ttaaaagatg tgtttatttt attttatgtg catgagtgtt 11820

ttgcttacat gaatgtctgc actgtgttta cctggtgcct gtgaaggtta gaaggcaatg 11880

gagctatgga gagttgtaaa ctaccatgtg gaaatggagc tatggagagt tgtaaactac 11940

catgtgggta ctaggaattg aatcagggca ctcctctgca agaacaacaa aggctcttaa 12000

cagctaaaat attactacaa acccacacca caaaatttta aattgataga cattatcacc 12060

ttagttctag atagagaatg tgcttggcat tgtaagtact aaaaaggttt tggggtggat 12120

cttttatatt atctcactat aattttataa aattaatact caaatatgtt ataagttaag 12180

gtttttattt ttgtttttca tttctgtatt ttgtctatgt agctctgcct ggcctgaaac 12240

tcatgggaac ttgactggcc tcaaactcag agagacctga acggccctgc ctccaaagag 12300

ctgggactaa ccatgcccaa cagtaggtag ctttaatacc taaccagtgt attagttcat 12360

gctctcaatt aaccaacatt ctctacatac agaaattttt atgcctattt aatcaaatac 12420

acagtctaag taaactctaa gtacaactgc ttggctcata ttcttacaat ggctatggct 12480

agctaattca aaggccagtc acataaaagg gtctctatga attctgatta acaaatgcag 12540

ttaaatagat gaattcctaa aaagtagtat cataataata tcatatttag tttttgtgct 12600

tccattatag tttgaggtgc ctcctcccat aatgcaaggt atatttcaaa taatagatat 12660

atacatggtt aacacatggc aaatgccatt ttaaatgctt agcacagcct gctctttggc 12720

tccattaagt gaaactctta agttctcagt taaaataatt gttggagagc tataggagca 12780

atgggtggag aactagtctt ctaatttgtc ctttgcctcc ttgcgtacta agtagtccct 12840

ccctcactat gtggcattcc agcagactac caccaagaga agaacagaaa agtgttgatt 12900

tctttctaaa gtaaagaaat aaggggccag tgagatacct cagcaggtca aagccatttg 12960

cctagaaacc aaagttcaat ccttggaagc cctgtaaagg tggaattaga aaacagactc 13020

cacaaaactg tcctctaacc tccactcggg cacacatgtg ccaacccctc cattctccct 13080

cccccacata caaagtaaca ataaactttc agaaaattta agttgctacg catggtgatt 13140

gatgaatgtc tttaattcta gctcttggga agcagaagtg ggtggatctc tgtcagttca 13200

agaccaacct ggtctatata gtgtgttcca ggcatccagg actacacaca cacacacaaa 13260

attacgtgaa ggaagtagaa tgtttgaagg aaagaagtct ggaaatgggg atggagagag 13320

acctcagcaa ttaagaaaag gtcttgcacc ggacgtggtg gtgcatgcct ttaatcccag 13380

cactcgggag gcagaggcag gcggatttct gagttcgagg ccagcctggt ctacaaagtg 13440

agttccagga cagccagggc tacacagaga aacccagtct cgaaaaaacc aaaaccaaaa 13500

acagaaaacc agtatgatag gtcaggcaat tggatcgaga caggacactc aagatagcta 13560

gcctgtgcaa tatagaaaga agtctcatgg aagagagagg gaagggaagg aggggggaga 13620

gagagagaga gagagagaga gagagagaga gagagagaga gagaatgaga gcgagagagc 13680

gagcgcacct cagttgatac aagattgggg ccctgagttc catccccagc atcccataaa 13740

ttgggtgtag cagcacacac ctgtatccca gcagagaggc aaaagacaag ttcaaagtcc 13800

tatatggaaa aagtgtgaga tcagcctgga gacctggtgt gtggcagtgg ggtgaggggt 13860

gtcatcaagg agaaggctta gtaagtaaag gacctgcgtt ggttcttgag ttcaagtctc 13920

cagcaatcag agaaagccag aaccattgca caaacttgta agccaagtgt tggactggac 13980

agagacaggc aaatgtttga ggtccaggtt cagtaagaga ccctatctca aaaaatctga 14040

tggagagtaa cactggaaga actcagagtg agtcacacat gcacacacag gtgaatgtgt 14100

atacaaaggg ggcagggagg gagaatgaga ggagactggg agatatctgt agttcatgtc 14160

tgtaattcta gcacttcaga ggcagctgga gctacacagc aagaccccgt ctcaaaaaca 14220

aacccaagcc tgacagtggt gaggtacacc tttaagccca gaggcaggag aatctctgag 14280

ttcaagggca gcctgagtga gttccaggac aaccagggct ccacaaagaa acactgtctt 14340

gaaaaaaacc aaaaccaacc aaacaaaaaa gaatcaaaaa caaccaccac cactacaaca 14400

aagcaaacaa gggagaaggt ataaaatgct taggagagtc ttcctttagt ctccatcctt 14460

tgggtactcc ttccccacag aaagccacta ctaccaattt cttacataag ctgctgtttt 14520

agacacaggt tttttttttt tttaaatata gtaacatatt catgtgtagc tcatttttct 14580

agtgagtggt tggtccttct tttaacagtt taaaggacct ctatgtttaa aggcgattgg 14640

cccttgtctg gagtatgggt tgtattttcc caatttgtga gttttaccca acctattgcc 14700

tattacctat ggccatttat tcttgtcgat aagtagtttc caattgtatg actatggtca 14760

cagtgttcca tggactcttc tgccgctaga cagcccctgg gtctgaattt gagatggtta 14820

caagggtgat tggctctgct ccctgggtgc tgggattaaa ggcgtgcacc tccacaccca 14880

atttgttctg ttttgtaaga aatgaggttt tattgtgttg ctcaggctga tctcagtctc 14940

ctggcctcaa ggtatcctcc catgtcgata cacagcacaa ggcgtaggaa aagtggcaga 15000

tttttttaaa ttaagttttc tttccaaaat atagattcag aaatgtgaga ttttcacaaa 15060

gtgaacctgc tcacttccct ggctctmgaa tctccattgt ggctcccgcc catccctttt 15120

gcccaccagt ggctgttgta ttgacttcta tcccattcct taactatacc tgtccttggt 15180

cttcgctgtg aacttgcttg ggctgagaat caccttgttc cgggcacatc aggtcagtga 15240

gggtgtttcc agagagtttt aacagagacc agaagaccca ctccaaatgt gggtggcaat 15300

acctgatgtt ctgtcatcct ggactgggta ggaagaggaa agtaagaagc aaacggcacc 15360

cccacctctc tgtctgcttc ctcgccgaca caaagtgacc agggcctccc actcctgccc 15420

cctcagctag agacacttgc tgccatcttt ccaaccactc tgagactgtg cctactaacc 15480

gtgacccaaa ataaatgttt ccttccttaa ggttgccttt gttagctcct ttaatagagc 15540

ggtaggacat gtaactgcca caggcagcca tcgctgccag cccctcccac tgaccgtctg 15600

agaaccacac tcagctgtag gcacagctct catagctgtg tgggcgtagc tctgtctact 15660

cggtcattcc cctgctgccg agcatttatt gttttcagtt cctggctgat gggtagcact 15720

gttatgaaca tcctagtaca aatctcaggg tgacacgcgc cttcattttt cctgagaaaa 15780

tgcccaagga taaaatgcta gggccaaggg aagaatattt caccattaag agacactggt 15840

caggactgga aagatggccc agtggttaag agcactgact actcttccag aggtcctgag 15900

ttcaattctc agcaaccaca tggtggctca caaccatctg taatgggatc caatgtcctc 15960

ttctggtgtg tctgaagaca gtgacagtgt acctacatac atgaaataaa taaataaata 16020

tctgagagag acagacagac agacactggc tagtcatctc acaatgttct catgtttaaa 16080

atatgatacc atttgtataa agcagaaaca caggaaaaat aaaatctgtg gtattatatt 16140

tgatttttaa attaacttga ttagtgaagt tagcagctac actgggcagg ggttgggagt 16200

ggggtactct gaagtgctgg tatttctggt tttgtttttt gtttgtttgt ttttttatct 16260

tatttatatt acatagaaag ccattttgct aatacactta ccatgtgtat atattgtgct 16320

tgaattacag ctaagtaatt atttctgagg ggctttagac tactgaagat tgggcccaat 16380

gagccccacc ccaagtagtc tccaacatcc ctcttggaag tacttgagag caaagattca 16440

agtcacatgt ccccaaaccc tcagcagcca ccacccttta ggtgtggctt ttgctctcgg 16500

tcatcctgga acatcttgcc atctttggtt tgttctctcc ctgtcttgcc tctggtagag 16560

ctgggtttct gtgcttctat tcaaccatgt acaagaacca tgtgccacct gccatgtgcc 16620

aagcctgtgc cagtccctgt gagcgagcag cccaccccgt gagttatcat gtgaggagct 16680

atgaggagca ggaaggggcc cggatgactt cagcagacag tatgaagcaa gcactgtgcg 16740

atttatgctc cctggccaca tgcccacaga tggtgtctga gacactagcg tttaatattt 16800

gaattctcca cattctagcc tagacatttt ggttgcaaga agaaaattga ctccagttgt 16860

atcctggaat gaaatttatt ggaggaaaat actggacagg ctcccagaga aaatacgata 16920

ttcaggcaca aaaagaaatg gggactgagg atctgaagtt caaggtcatc tgtaatgaga 16980

ttgaagtcag tttgggctac atgggacctg gtctaggggg aatggggaag agaagggaag 17040

ggatcgagat agggat 17056

4

5973

Mus musculus

4

caatgtgctc tgacgattaa tgggctagaa atgtgtggct gttgattagt gaaaagatgt 60

catggttcag gagattggta gtctctgtgg gaagacaact cactgaaagg gaggaaatag 120

cctggaagag ataaagagac agtgatcagc taggaagctt aaaatttaaa ttttgttgga 180

agtactgtta ggaatactag cagaggccag atgaatgtat ggttaagtta tagcaaagga 240

aaagattgtt aatggtgagg ttaggaatgc agggtgacac caacctgtaa tgtcagcatt 300

agcgagatag aagcaggtgt ttaaggccat tctctgctac ttagcaagtt gaggccaatc 360

tggaccacat gagacctttt ttcaaaaata aatctcctta aacaaaagag gctgggtttt 420

ttgatagatt cttcaagatg ttaatgtaaa taaatggaag accaaggatg gcatgctaat 480

atcctcagtg tctgaagaag gactatgtag tgttggctgc tgactctgaa gtaagtgctc 540

attactgaca gatagtgtat cttagagcct ggcagatggg atggaagtga ggaagcaagt 600

agcacctttg tatattatgt tctaagtagc cagagatact tgacacaaaa caaagttgag 660

aaaatgtatc ttctagaaaa tacagacatg gaaaggtgtc ctttctataa aagaggtatt 720

aaacattaac ctgaaaaaaa agttagcaaa ttgggctttg gcaaatgaat atagtcaagt 780

ttcattttta ttttgttttt tgtatatgac tgtttggctt gttgtaccat gtgtgttcct 840

ggtgcctagg aagtcagctg gagttacaga tggtgtgagt tgccatgtgg gtgctgagag 900

atgaacctag gtcctctgga agagcagtta gtgctcttaa ccactgagcc atctctctag 960

ttccttctgt agaattttca ttaatttaca aaggagaaag tataaatgat aaaaccatga 1020

gaagatagac cggcactaga attagtggag tcaaaatgtt aatgatatgt cagatacgcc 1080

ttatatgagg aagttgcaaa attatgaaaa tccaggcact ccactgagtt agaaatctag 1140

gctctgatgc atactgctat ggtaaggtag caagtggcca ttgagtgcag aagtgagtct 1200

ggatgggtct tctggtgttg tggagcacac agactgctgt cttctgcatt gcagtttcac 1260

ctgtatttcc ttggaactac ttagctttgc aactaggcgt taaaaaaaac tttatattta 1320

tggttttaag ttatttattt gttttatttt attttatgag acatagtctc actctctaac 1380

ctaggctggc ctggaactgc ctaggtaact tgagctggtg attctcttgc catagccttc 1440

taaaatttta gattgcaggc ataagccaga ccactcctga cttttgtagc catttttctg 1500

acatgaagtg taactttgct ttcataacta aaatgattta gttgttttgt tattgtttaa 1560

tcccttttgc tttgaatgta tccttttgtg tgggtggcag atatataacc acagactttt 1620

ccacaggcat cctaccctag gtccagaaat gactctgaga cgtcttatat atgaatgaat 1680

gcctaggcca atagctttgg ctgatttcca cgggttcata gctcagttat cccatttaaa 1740

ctagtctaag tcatgccatg aggctacata cccctccttc agtttcaggc gactgtcttc 1800

tcagttgtgt aatgtcctat cctctgttgc tgctccccaa cccccatcct tgcgtcatag 1860

tccgtctgtc ctcgtctccc cccatttact tgcacaacgg actctactct agaagtcctc 1920

tctgtgctgg agcttgcacc tccgctctcc ccgtctaagc taataggcaa cagcattgta 1980

cagacaggtg atgcttccat acatcgcaca ggagattctc cctacacaga tacttattca 2040

tccagcgtga atgcaaccgt ccaggcgtgt tctcctagtt gtagtacatg ctgttgtatc 2100

agtctgatga atttctttgt ctttacaacc aagaaagata atactgtaag aaattttgac 2160

taacattttt tctttattta aattacagac taactggctc ttctggattt gtaacagatg 2220

gacctgggaa ttataaatat aagacgaagt gcacatggct cattgaagga cagtaagtta 2280

taatggctga ctttatttta atttattata agagcacagt atagcacaaa atacttccat 2340

gtgtgttatt gctatttctt gagacaggac ctttctgact gagtaactca ggctgacctt 2400

gaattttgct atgctacctc tgcttcccaa gtgctagggt ggtaggtgtg gaccaccatg 2460

ccctgctgct aaaataccgt tcattgatgc ttttcatttg gatagtgttc ttgcttttta 2520

aaatttactt tttgggggac cagagagatg gctcagtggg taaagtgctt gctgaacaag 2580

tctggttatg tgagttaatc cctggctccc acagtggaag agtgactcct gaaagttgtc 2640

ttctgactcc cacgcttgtg catgcacgca cacacacaaa taataaaata aaaaattaaa 2700

aggaaatttt cttttttggg tgatagggat tgaacctatg acttcactaa gcaagtgctc 2760

tattgttaaa taattccttt aatttgtggg tttttttttt ttaggttcca agttgactta 2820

atgttataaa tgaaagatac ataccagaaa tttgcatatt tctaatagtt taaaaaactt 2880

agttaaaatc tttttaatag tttgcttaaa tctttatata ataatgctat tatatcattt 2940

ttctaaatat tgattttatt atcagcaaaa cagtaaatga gccatcagaa taaccactgt 3000

agcctgtttc cctggccctc tgtccttcca tctgtctatc ttctcttttt tttccttttt 3060

tgtgcctgtc atttagggca aagcatttta gtctctgaac aaaactttga aatttccaag 3120

taactcttgt ttatttgttg tgtctcatat tcaacccaag aaatattatt tactaactca 3180

tttaaaagca acaattataa cccactacat gttagcagaa aaacctattt gtttttattg 3240

agacgggatc acactagtaa gcactacatg gcatggcgtt cactgtgtag atcaggcagg 3300

ctggcttcgt gctcttgaca gtcctcctgt gtttgtctct cacttctgag tgctgggatt 3360

atagacatta ccaacacacc gatttggggg gttggggtac tgggatcagt ccagagttgc 3420

atggatgcta ggcaagcact ccaccaactt agctatatcc ctggtcataa atgtcataag 3480

gaaaaaaatt ccttatattt aaagaaattt taagaattgc attgtttaag atttcacaga 3540

tctctttgct atctggcaat cttttttgat attttgtttt gtttttaaaa atatgtggta 3600

tgtaaacaaa cttaaatatg aatgggacag ttccagatga gagtgaaaag ttaaatattt 3660

gggagaaaaa ttgataggtt tatctattat ggaaaatttc agagatttta gtaaaatttg 3720

aaaatggagc tgggaggtct gaggtagtca tctaaagctg ccagttgtag agcgtgttgg 3780

agtgtggagt cagagggagt tactgataca cttgttgaaa ttgcccaggc ttcatgggaa 3840

gtgatgaggg gctgttactg tgactctggg cagggcttgt tagtttcctt tggatttagt 3900

ctcagtcaga gttgatacat agtttcctga ggacgtggct ttttggtaca gtgctgtgaa 3960

aaggcagaga agcaggtaaa cttagaaaat gtgtgttttt aaagtgatgt gttatgaaat 4020

cttacgtaag atgaataaag aaagaagtgg ggacactgag ggctcctgtt tctaaatgtt 4080

aaaagcaagg ctggaaacat tctttgaagg cccctgaagt cagagcccgt gtctcttttg 4140

gttcccagga catttttgat attcccttac acatagcaaa tactaactag atctctgaca 4200

aatgcaggaa agctgtttat atttatatat atttatattg tatatttttc tccttataaa 4260

ttctttaaaa gtctgtttta gtagttaatg ttatgattat tataaattac ttaattattt 4320

ttctaggcca aatagaataa tgagacttcg cttcaaccat tttgctacag aatgtagctg 4380

ggaccattta tatgtttatg atggggactc aatctacgca cctctgattg ctgcctttag 4440

gtaagccctg ctgcatttca tctcaggaag taagtgtgtc tccaggatgg agtccgtgct 4500

gcatttactt tattctgcag tcacactcat ctcatggaat tagttctgtt ctggtgagct 4560

acagttcact tggtttttat gtactgggtc gttttccatg tatactagta tgtagccacg 4620

gttagtcttg aacttctggt tctcctgcct ccaccttcca agtgctagga gtataggctt 4680

gtgccactgt gcctgactca tttcacattc ttgaactgtg aagttttgat aacactatta 4740

aatttacctg ctatttgtga ttttgttaaa gtttgcatta aaaagttttt gactatattg 4800

ataatatttt tgtgacaaat ttaaatcaga aaccatacct ttcttgttct tgtatgtatt 4860

tcattccata ggcccttagg aataactttt ttcaatagta tatagttctc tcagtttgta 4920

tatatgtatt attagggata ggaggagctt tctggaagac tatttataaa ttggacaatg 4980

gctagctgtt gagagtgagg aatttgctag ttttgttttg taaatccctc cccaatgcat 5040

ctgtattagt gatttaataa aataatgcaa ttttgtcagt tatatgggtt gcactgaatt 5100

tttgctattt tattttaaga aagatttttg tgtgtctaca gtgtatatga gtgtatgata 5160

tgtgtgcgtg tgcatgtgtg tgtgtacttc tatgcaggta ctcacatgct atggtgtgca 5220

cgtaaggttg gagtgcagcc tcacatgttg atcattatat tccaccttgt tagagatagg 5280

ttgtctttgt tgtttgctgc ggcctggagc tggagctgga gctaacgagt ctcagccacc 5340

tgacatgggt actgggaacc aagagcagca agacctcttc ttcttcttct tcttttttca 5400

ttttcggttt ttcaagacag ggtttctctg tatagccctg gctgtcctgg aactcactct 5460

gtagaccagg ctggcctcga actcagaaat ctgcctgcct ctgcctccca aatgctggga 5520

ttaaaggtgt gtgccaccac cacccagcct aaaagatttt cttactaaaa tatatttcta 5580

aattaattag ttggaatctg gttcatactt ctttttgaaa caaaaccagc attttttttc 5640

attctacata cagagacatt gacactagac actggttatg agtagttact ataagaatgg 5700

gaaattattc cacccttgta aaacttaata caactcctta tcaggctctg aagacttttt 5760

aaaagcaaga attgtatata acacacagaa atgatttaga ctatttagat ctttattgca 5820

tgggatttta aaattattat tgtatttcgt gggcatgttt tgtctatgta gcatatgtgc 5880

ctgtagaggc aaccaccaag taggtcctgg gaatcaaacc tggtacccct gctcttaggt 5940

gttcttaact gctgagccat ctctccagtc ctc 5973

5

90050

Mus musculus

5

aggcaagaaa gagccagcga gcctccagac agaccattag aaattccaca gtcagcacaa 60

tagggagaac agtaaatctt acattaaaag aaggccaggg cctggtagca aaaggtttta 120

atttaagcac ttgagaggga gaggaggcaa atctctctga ttgggggttg gggttaatgg 180

tgaatgccat gacaccctgc tcagagttag ccttctcccc taaaaaaatt ttaaattcat 240

tttcaatgct gacacagtta atcatagaca ttgtatctca gacacctcaa catactccag 300

actgcagcac cagcccactg ctgaggctgt cgttcagttg gtagaaggca tgctcagcat 360

tcgcgaagca ccagacttca tccttagcac tacataaaac tgggtgtggt catgcacact 420

tataacttca gcaccatgga ggcagaggca ggatgatgag aacttgagga tcattctcag 480

ttacataggg agtttgaggt taagcagggg tacaggaggc ctgtctcaaa caaacagaca 540

aacagacaaa caaacaaact tcaaaaaact cttgaagtac taggcctagt acgtgctgag 600

attgtaggta tatgtcatca tgcctgttgt agaatgagtg agagcggact ccataggctt 660

atagatttga atcttggtgt ctgtctatgt catgtcatcc ctgcacaaaa gcccacacta 720

ggcccacaca ttctctctgt ctgctgcatg tgggttagat gtgagctctc agctgctgct 780

ccagtgccat gcctgcctgc tgccaggctc cagccatgac ggtcagggac taactctctg 840

aaactgtaac caagttccca atgaaatgct ttctttcata agttgccttg gtcatggtgt 900

ctgcttcaca gcaataacac ggtgactaag atacctggct cctcccctcc ccaccccacc 960

attatttacc ataaagtaaa caatacacag ttggataaca tgatactgaa gttattttcc 1020

tgtttcctga tgtaacccaa ttttggacaa gattaagcct taaatagcaa gctgtgaggc 1080

aggataaaga aaaagctcgc aggccaatgt ctgctttacc aaattctgtt cagcagtcta 1140

aagctgccgt cacctcgact cctgtgatgg catttccatc actatcttag atattccctg 1200

ggtcacaacc ttttagtaca cagattgcaa ctctgatgga atggctgact gcttggctaa 1260

ttaaagcaag ctagagtttg tctggcttcc ttgtctgaat ggggaggtgg tatttacaaa 1320

attttgtaaa taaactacta tatttgcatg atgtatataa atttgatgtg gctgctttta 1380

aatcatttaa cctaaactgt cccacagaat catctgtttg attggaaaga ttgtagcttc 1440

aagagaattt ctgctgaacc tgaaatgatt cataatgatg tgtctgaaga atgtgtgcta 1500

tcacctacgg tttttgtttt agttgatatt tgtactttaa gatttccttt atgtatgtgt 1560

gtgtgtgtgt atttatgtga atgtatacct catgtatgtg gtgctcaagg acacctgaag 1620

aagggctctg gagctggagt tacagggagt tgtgagtgct aggaaagaaa gctgggtaca 1680

ctgggaaatc aaaaggtgct tctaaccact gagaaatcct gccagcccct tggtttatta 1740

aaaatatcaa acaaaaccaa cactagttac ataagtatct ctctctcttt ctttctctct 1800

ttctttctct ctctctttct ctctctctct ctctgtcaca cacacacaca cacacacaca 1860

cacacacaca cacacaaagg atccataata gttcttctgt atcccggtta aatataagtt 1920

cttaggggct agagagatgg ctcagcagtt aagagtgctt gttgttcttc cagaggaccc 1980

aagttcagat cctagtacac acatcaggca gctcacagct acccatatct ccagctccag 2040

gaagaaccaa tcaatgccta tggcccatgc aagcaccagc acacatatgc tccacaaaca 2100

tccatatata tagctaaaag taataaaaat aaatcttcaa aaaattaatt ctggttgaac 2160

tgaaaaagat cacctaacat ttagaaaaag cagtttacta gtgaatagga cataaatcat 2220

ggtatcaaat attctgttgt taaaggaagc aactagaaaa agcatgtgtt tgaaataacc 2280

aatggataca aaacaaatga ggcaacccca acatctgtca gtaccttgca aaccaacaca 2340

ataaatttga ttttatttaa atcgtagtta tttttcatgc tagtagtttt gaaacacaat 2400

aaatttgatt ttatttaaat cgtagttatt tttcatgcta gtagttttga aaccaagatc 2460

tagattttgt atagccacat aaatacacat tagaattgca aactgatacg agcttcatct 2520

tcatcagtct ctcttcatga aaagcagtta cagggactga gacatgactc agcagttacg 2580

gcatgggctg ttcttccata cgacatggat tcaattctca gtgcccaaat gttggctcac 2640

aaccatttgt aactctggtc ccaggggatc tgacactctt cttggcttct atggccactg 2700

tattcatacg gtacacagac acatatgcag gcaaaactca acaaaaaaaa taaggtttaa 2760

aaaaaagaat tagaacttaa aggcacttca ttccgtcagc actaaatcag cctctctgga 2820

gtcttcccac ttcatgagaa aatcgtcagc tctccactgc tgtctgtggc tgaggagcag 2880

gacctggaca acgttcagag attgtcagtg catctctttt cttctttggt ttgctgtcat 2940

caggttcact gtcacattcc ctttgtacca tccttccttt aacagccttt tgaaaatgca 3000

gaaatgttgg atgctgcctt cagttcacac aggctgtctt tttagctcct catctatcta 3060

tgcttaattt gttagtggtg ctcacccatg tatgtgttta tgtcatgaag ccacaagatg 3120

agccttgatt gagtcttgct gtcagtgtgg atcacagaaa tgacacccta tcatctttgc 3180

ttcctgcttg ttagaagtca ttgattctgc ttatactcaa ggcccacagt attatacttg 3240

ggtgtgaacc ccaggaagca gggaggtggg gggtgtcatg gatactactc agatatctga 3300

ctgttgtgat atttcatcag ttctcattgg tcctatcttt aaaatctgcc ctacatctag 3360

agctggctgt ggtggtgtgt gtggtggcat cagtatcaga acttggatta cagaggcagg 3420

aagattgtga tttttgaggc cagaataggt gcatacaaag atcctgtctg caaaagaaac 3480

aaatgtgcaa ataattataa ctactttact aatagcctaa ctaataacca ctgctagtgc 3540

tgtgtccacg aaaaggtgaa gtaaactgtg aaaatgactt ccccttctgt gtgacacacg 3600

ccgtcatgtg attttacttg tgtctcatca ttgtttttcc ttctgtttgc atgtgtgaat 3660

gttcacatgt ggaagccaga agtcagtgtt gagtgtcttc ataattgatc tctattctct 3720

ttgttttgag acagggtttt gagactaagc ccagtgctca gtgattcatc cagtaaactg 3780

tagggagctt cctgtctctg cctccacagt gttgggatta caagcatgat ccaaattatg 3840

tgacaagcgc tttactaact tagccatgtc ctcagctccc cactcccctt ttcttttctt 3900

cttctttttt ttagacttac ttgtttattt ttatgaatgt cttgcctgca tgcatacaca 3960

cacacacaca cacacacaca cacacacaca ccccacatgc aagcaattcc agaagagggc 4020

attgaatccc tgaaactgga gttccagtta actgtgagcc tgtcatgtgc gtactgggag 4080

ctaaatccgg gttctctgga aggtcagcaa ggtcttacct gggagccgtc tctttagctc 4140

atgtgtttct ctcttgaagc aagaaaccta ggaatcattt tgaaacttcc ttcacagcct 4200

ttatcataac ttcacgtcaa tttttaccta ctctttcaac aaatacatgt tatatttact 4260

tatttttatg tttagcctgc tattggtttc tacttagcct cttgcagtag agttctgtca 4320

gatttatgtt tctattgctt ttaatttatt tgtaaaggtg aatgggaaaa tatttaaaaa 4380

ttacagatcc catcatttac tatattctta aaagccatgg ctagccaggc ttggttgtgc 4440

atgcttgtac tcccaggact ctgacaactc agtaaggagg agagtgaatc agaaaatagc 4500

gccagcctgt gctgcttagc aagaaacaga aacaagtaca atcacacaca tagaaaatcc 4560

cccattaata ccatcccatt agatataatg gtcctgtatg accattcaac cactgtttgt 4620

cctctgtact gcagtaacag tcttctgccc ttgcccgtga agcacgtgcg caccccgcct 4680

ccaagtgctt ttgcactggt gtcctccgtc tagatgtcct gttactatat gtaaggactg 4740

gtttctcctc ctctttacag ttcaatctaa ttgtctcatg aaaagatctt tcctgaccat 4800

ctggttcaga caggttctcc ctgttgttgt tttgtttttt gttttatagt tctaaattcc 4860

tttcaggaac ttttgcttat tttaaattcc ctgagtgcat acgtgtgctt gttgttgctc 4920

atgctcgttg tttgggctta ctttactatc agctctggat gtggttcaca gaaggtgctc 4980

aggggagcac tctcagccac tcatctcaca cgggttatag atatatgtat tgatgctacg 5040

tttgcttgtg agccatgttt taaagattag aatatctttt ctatgtgtac tctatcaaaa 5100

cacatgttag ggctttatct attttataca gatattggtg ttcttgcttt actaattttc 5160

atggaatttc ggtgaatatt agtattttag ataggaagac ttgtctcaaa atgtagctca 5220

gctggttgag tgcctgcctg catgtagaaa gccctgtatt cactctccag cacctcagaa 5280

gtgggccatg gtgcatatgc tgtcatctca gcactccgga gggagagaaa agagaatctg 5340

gagttcaagg ttatccttgg ctatataaca agtccaagat cagcctgggc tacatggcat 5400

cctgcctcaa aatcaaacac caaatcaaaa agctcacatc ttgatccaaa agaaggtaga 5460

gagaatacac tgggaaagtc tttgaaacct caaagctaac tccaagtgac agtgacacct 5520

ccttagcagg gccataaatt ctaatccttc cccaaagccc accaactgga gaccaagtat 5580

tcaaagataa gaatctatgc agtccattct ccttcaaact accacagtag gttttcttaa 5640

aaaaagaaaa aagaatattt taattgattg tgattattca gtattattca tgaataatca 5700

tgaactacat ggcaggacta taaactatta ttttttttaa agatttattt atttatttta 5760

tgtatgtgag tacactgtag ctgtcttcag acacaccaga agagagcatc aaatcccatt 5820

acagatggtt gtgagccacc aagtggttgc tgggaattga actcaggacc tctggaagaa 5880

cagtcagttc tcttaaccac taagccatct ctccagcccc tataaactat tattatattt 5940

ataaaatata aatccgtgag tctgtgcacc cctgtgtgca catggatggg acatctttga 6000

actggattat atcatactta gaagaataca agatactctg ttttgtcatt tgggtgaaaa 6060

tatggtctgt ttattttgca ggtatgacct gacttctagg gaatggcttc cactaaacca 6120

ttctgtgaac agtgtggttg taagatatgg tcattctttg gcattacata aggtaaacta 6180

tctcaactct tcaccaagca agaagttcaa ctcttcctgt tgctttatgt cattgaatac 6240

tatcgagctt tggttttagt tggtataagc tttgttttga tgtcatggag gtatataatt 6300

caccaagttg tcaccaagtt gtaattggaa attgaagtta gaacgatttt aatccatggt 6360

gtcttgcatt tggatactct gatcacagtt aacaatgaag attaaatagt gtcagcaagc 6420

ctatgcccat tatcaagtct agcatactgc atgcgtgtga ctgagtagcc attgttatct 6480

ccttgttttg agcgtatatt gtagaatgag gcaactgtat tttccacacc attttcgttc 6540

tgtaacacgt ttcatgtaga gaaggtgatt tagagagggg aagaatgtga ttgtattggt 6600

tggttctttc tctatgctat tcctagcaag tcaccgaaga gctcatgtta ctcacacttc 6660

ttaagctggg atcacaatga gattgtgaac cactcattgt tgttttccaa tataattttt 6720

aaaaagatgt atttattttt attttatgtg tgtgggtgtt ttgcctgcat gtatgcctgt 6780

gtatactgtt cctccagagg tcagaagagg atggcatcag aactggtggc tgttagctgc 6840

catgtgggta ctaggaacta aacccgggtc ctctgcaaga gcagcaagtg ttcataaact 6900

ctctctccag ccctagagtt gatttcttaa tggttttaaa aatcctgttt acatctttct 6960

tataggataa aatctacatg tatggaggaa aaattgattc aacagggaac gtgaccaatg 7020

agctgagagt atttcatatt cataatgaat catgggtatt gttaactccg aaagctaagg 7080

atcagtatgc agtggttgga cactcagcac acattgttac actggcatct ggccgtgtgg 7140

tcatgttggt catcttcggt cattgcccac tctatggata tataagcgtt gtgcaggaat 7200

atgacttggg tatgtatttt ttccagtgga ggcatcttga atatcatact gagaacccct 7260

gcccttatta ttaggacacc gtaacaaaat tcagcatgat cttgatccag taccttgtct 7320

tgaaatagta tcagtagata actggtgaga ttgaggttgt tgaagtccct gtgcaacagc 7380

tgtttcttac ttgtcaaggt ctagtcttgg cttgggaggg gttctgagga aaggggtgtc 7440

aaaaaaccca aaaagtccaa ttgtaggtcc aagctggcag ctgtatattg cattaaggaa 7500

agctgaggga aatttgggat atttatttca tctattagtc tacatcaagc aagtcaagcg 7560

ctcacagtca acgtttgcac cctcaaatta gtaacaaaag agggggaact gaggagtcca 7620

gcatggtcct ggttgggaca gaatgacatg gttccagccc tgagacaggg gcagcaggtc 7680

cgggcctcca tggatgtcac actatggaca taaacctgtt tgtataataa tgtacatatt 7740

tcatgctcct cttctgagta atgtccttct gttaatgtga atgacttcat gataatcaga 7800

gccagtgtga gtctgggaag taaatggtgg gaccttcagg acagctctta aggctgtgga 7860

aaagaacatg agttcaaaac catatacttc ctcaactata caaaaataga aggatgcaat 7920

atgaattgta tgaggggctt cacagatcta aaggaacaaa agcagcttcg ctgtgagcca 7980

acttgtcaga aagatattga gtaagcagtt aaagagattt agggagtgct gattgctaga 8040

ggaggccacc cagctaagtt tgtgcttaca aaggcagaca aagtcctgag ttcagggtgg 8100

gcctggaaca gagcaaggtt agttagacct tggtgtggta gaaatggtaa tttccagaca 8160

ggatacccaa ctagtttttg tgcttaacag aggcaggtag atctctgaat tcttttgtaa 8220

tgttaaaagg aaatgtgtgc ttgttgtctc ccaaggggcc tgagtcccag gatgctgatt 8280

tataggaaac ctggagtaac tgggtttatg acctgcagga gacgagctat ccagaatgtt 8340

ttttgcaata gcaagagaga actgcctgga gaactgcctt cagcaaagaa tagcaagaga 8400

aagctgtcta gagagagagc tgtctgtaga gaaagccggt cagagagaaa gtagactgga 8460

aaactgtctc cagcttggac ccacaatttg actttttgtt tttgttgaca agttgccctc 8520

ccccagaaac accttcctca ggacccctcc caagccaagg cagggccttg gcccttcttg 8580

tcagcttgca aggagccaaa gatagcatta aatgctttgg atatcaaaat aagcaaaatg 8640

caaaacagta aacactctaa aataattctg gctagtcctt taaatattag gccagtgcac 8700

tgttatttta ccttaatgta taatcttgtg ttacatttta ttgtttttat tgtataatag 8760

gaatgtcaga attataattt tgtaacattt gtttgacatt cctgtgaaaa tgcatctaaa 8820

gatcattaaa gtgcatctga agatcataag gactcactga ggagcacagg gaattaagtg 8880

tctgcttaag agaactttga atctttaatc tttagaattt gttttaaaaa tttgaatctt 8940

gccagtgtgg tggcgcatcc ctttggtccc agcactcaag gggcagaggc aggtgtatct 9000

ccattagtgt gaggccagcc tggtctacag agcaagttcc aggccaggca gggtacacag 9060

agaaacccta gcttaacaaa acaaaacaaa atatgaatct ttaaaaactt gttctgtgaa 9120

aatttcatac atgtatacaa tatagcttgt tcatatccac cgccattcct tccagctcct 9180

ctaggctttc ccagtgcatc tccttcctag ccttatggcc tccctttcag ggtgaaggtt 9240

agcacactga gtccagttag tgctgatccg atgcagtctt gtctagatgg tcttctttat 9300

aataaggtga aagtatatcc taaacttccg tcttttgctc taaggtgttt agactttaaa 9360

ctaatgttta aatcgtttaa ataatttatt atttcataag aagaggagcc tgcaacattg 9420

actttaacta ttgtctctta tccagaaaag aacacatgga gtatattaca tactcagggt 9480

gctcttgtgc aagggggtta tggccacagt agtgtttatg atgacaggac caaggctctg 9540

tacgttcatg gtggctacaa ggctttcagc gccaacaaat accggcttgc agatgacctc 9600

tacagatacg atgtggatac tcagatgtgg tgggtgtttt cctagagctt tcccttggta 9660

gtctagaatc tgcagaggca attgattaaa aatactgtgc tatggtttga cttttgttca 9720

gcattgtatg taacaaagtt aggagatcaa tacagtaata gagttaaggt actaatggtg 9780

ctgttgctgt ctgttagtgc ttagtgcttt agacctgatt cactgaactc tagcaaggtt 9840

tcctctcttc agaattctca gcaataaaag ctgtgctgat tttatccata cttaaaaagc 9900

atatccttcc ttttctcttt ttggtgttgg ggatcaaacc ttgtacatga ataggctata 9960

ccatctttat ccatttacat caccaaacag gatgctctcg tgcctatttg atagggtttt 10020

cactcacttc gaactgaaac ttgggttgta agagtatggt acttttagca aatggaaata 10080

aatttgagtt atgatgcaat tataaagcac tggtctctct gtatttccct cctccttcta 10140

ctccctccct cttcctttct gaccccctct ctcaacatac attagagacc atgctttgac 10200

tgtcaattta tgctgtgctg aagatcaggt ctttagtggc tgtgaaccac ggagcctatg 10260

cagtggaagt tctggtctct ggcttttgcc ttactaataa aacactgagc ataaattttg 10320

atttgtattt cacaattctt acctggaatt cttaagtgga attatggagc catagagaat 10380

gaacatttta gggcttttaa tatagtttcc cgaaatttta acagattttc atgattgtta 10440

aaggaagtgg cttacgtata gggggaaatc aagtattgca catttgaatc taaagttata 10500

aagtaattac atttaaattg gcaaataagt attcttttaa aactaacctt atatttatta 10560

tttctaaata aactcaaaag gaccattctt aaggacagcc gatttttccg ttacttgcat 10620

acagctgtga tagtgagtgg aaccatgctg gtgtttggag ggaacacaca caatgacact 10680

tccatgagcc acggtgccaa atgcttctcc tcagacttca tggcttatga cattggtaag 10740

ctttccaaag atgttttagc ttcaggaata ttttctttgc tgatggaaag atcactatgt 10800

taaaataatt gcaccattta aaagaagtcc aggtggtaga atttgcattt aatttgagta 10860

gggttacaca tctattgaaa agcattattt tggattaaac tacattaaat tctttgtgaa 10920

atcactcttc ttaattgctt taattctttt tttaggttga gttaattggt atcttctttc 10980

ttataagtgc cttacatagt agtggtggta gttgtaacca ccagtgttat gttaagtttg 11040

atgggatatg ctgtttccta gaaacctggt tttacacatg ctgttgatgt caatatacat 11100

gtggccagaa gagggcagtg tctgtttatt cctggaaaat aaacatcagc tgctctgttg 11160

tgtaaatatc acccatgtga tgttctttct gtttatttgt ctttgcattt tgagacagcc 11220

tcactatgta gtctaattgg ctgaagctca gtatatagat caaggtgacc ttgaacttag 11280

agaaatcctc ctgcctcttc tgagtgctaa gattaaagat gtgtactacg aatgaaaaaa 11340

aaaaatgtgt actaccacac ctgactagag attcatttta aaaattattc ttattgtgat 11400

aaaatgctca gaataacact caccatctta atgttttaag tagtttagat ttaaatatat 11460

tcctagtgtt attcatgtta taataccatc tgcttgccga cttcttgtaa aactgaaact 11520

ctgcccttaa acaatagttc ctctcttcat ccctcactcc agcctcttga aatcattttc 11580

tatatctcta tgattttgac tagtctaaat taggcatttt ttaaaaaaaa tattttgttt 11640

acttgtatgt gtatgagtgt tttgcatgca tgtatgttaa gcacaccatg tatattcagt 11700

gcccatagaa gccaaaagta ggcatagatt ccccagagct ggaattacag acttttgtga 11760

gccaccatgt gggtgctgga tactgtgccc aaatcctttg gaggaatagt gagtcttctt 11820

agctgttgag ccatcttgtc agccctagat gtttgttttt aacaaacgtg tttttgccag 11880

ccattgagtt tttaaattga gaatgggggg tacactatag ttagtcctta gcttcaagct 11940

tgtggaagca gaaatgagaa gacaatataa tcttaactca ggaggattct tgctggctga 12000

aacaaagatg tgaaattacc tccgagcact cctaagccac tggggtgagc agggtggtct 12060

ggagaggcct tgaagagaag ctgtctgagc ttgttcctgg ggacactggg agtcaaatag 12120

acctcctggg cagggggatt tagtgcagac aagaggcagg aaagtacatg tcaaatattt 12180

aggacttttg aaccgctacc tttcttttgt catggtaaca cagaaggtag caggtgactg 12240

ttagactaga atgttcagat ctgattcaga gtgccaggga tcgttggttg gtcttgtgta 12300

aagtctcaca agtgatagaa tcatatgtgt gtcttagact ttttttgttg taggtatttt 12360

agatttttct tgtttttcct ttttgtaagt ctggccctca cactatggtc caggcaggct 12420

taagacttat ggtaaccatc ctactctgcc tttatgggcc accatgacca atttaagaag 12480

ctctcttggg tggcattgtg ataagtgatc tggaaggggc atattgacag ttagcaggct 12540

gctactgcag aagtcctaat taggtttgta tcaaggccat ggaaggagca gtgacttcta 12600

gtacctggct gttgtgtgtc ttgacaaaaa tataactgcc ctttcttccc aagtgtctac 12660

tatggaccac ctttgccaaa actaaaagca gattcagaga aaaacatatc atgattgcac 12720

atggctataa tccctgaact taggaggatg agaaatatgg caagattgag accagtctga 12780

actatctagt aagaccgtgt ctttaataaa aatagtaaaa attataaaat cagggagtag 12840

gatctgggaa gaagagaatg aagtaagtgt ggggcatatc caattggaga tgtctttagg 12900

acagagctga ttgctgagag gtggttgtag gagaggtgag ttattgtggg gcataaaaga 12960

tgagcaagag tcagagacag ttggagaaca gagtctgaac aagagtagag actaaagaga 13020

gtgtcagaga agcagggaga aaataggtga gattgatgac ctgtgagata tgttaatggc 13080

cagaagagtg gctaaaaatg actggagaat ccttcagact tgtcaacaaa gaaatccttt 13140

agcctaattt agggtgcagg cggctgagga aggacatagg tgaaatatgt gctctgtgtg 13200

ttcattttta ttaaagctta tctgcaaagg cctcagattt gctgtgtact tgtagctgag 13260

gctcttttga actcctggtt ctcctggctc caccttccca agtgctagga ttacagatgt 13320

gtgccctagt taaaatagct gtatacctag cattaaaaat tttaagttag aaaatactgt 13380

ggtgctccgg ggatgcatct cagcagtaga gtgcttgcct gctatacaca aggccctggg 13440

actgatccct agcaccacaa atactaaagc agacattctg gtagggaaaa ctggtagaca 13500

gcagagtggt gaccatcagg aggggggttg tgggtgatga atgactacat taattagaag 13560

ttctgtgcag tatatttatt tcatgccctg aaacattgct gctgctgttg ctgctttcct 13620

ttacacataa taacataact aaaagacaga caagcatgtg gtatgaggct gtggatgagg 13680

cattctttgt tttccttttt tttttttttt tgagacaggg tttctctgac ctggctgtct 13740

gaaactcagt aggtagaaca ggctggcctt gaatgcacag agaccctcct gcttctgcct 13800

tctgagtgct gggttcaaaa tttatgtttt tttctataaa gactgagagt tcacatggac 13860

tatatatgac aacctactct gaaatgtgtt tttctccccc ttagcttgtg accgatggtc 13920

agtgcttccc agacctgagc tccatcatga tgtcaacaga tttggccatt cagcagtctt 13980

gtacaacagg taattggaaa gcaaaggctc tattactgtc ttacatctta tattcatttt 14040

taatatcaac ttcctaacag ttgtatctga atggtaagag gtttggggag aaaaaaggag 14100

agaaggcagt tctaagtgca cgataaggta aggggaatag gactgggagg ttatggggtc 14160

aaagagcaag tctgaagtct gcactatatc caggtgtgtg ctcaggaata cttttctgac 14220

cagcagagct ctttttccat ttgctccagg aaccttagtc ctgtaaagga catgcaaagg 14280

actagggttg tgggccagca atagagtgtt tatctagctt gcacaagatc ctgagttctg 14340

acctcagcat tttgccttct gcaaacacag catttgccat aagggacatg cagaatggcc 14400

attttaccta gtcacttgaa agtgtgcttt aagattgaga aacttaacag cctgctgatg 14460

ctgacttttc ttattttgct tctgttactg ctttctgctt ctttctttaa tactctaatg 14520

cttacattat atagtcctac aggtattcaa attttctgtt ggagtttcct aatacaagta 14580

atttaacttg cattaggaaa aggataaaag tgccattctg gagttgtgaa gaatgaccgt 14640

ttagaagcta gatagtgggg aaagatgata tctttaatca tgtgattatt tagtgtttta 14700

caagtatata ggggattgtg gcaagaccat tgtatgatta gagactaaag tggaaagatt 14760

ttttaaatat cttgttaact tgagtgttat cttaaattac aatctgatgc tttccttcag 14820

aaaaagccct aaatgcctct tgaggttttc atctggcaag tatcatgtca cctggccttg 14880

ctggtggaat ctgccccagc tcatgtgtgt tcttagtgtt ctcctagcac agagttaggc 14940

acgtgtgggc atttgcatac taatgtatag taatagtaac aattgaatga attgtctatt 15000

aaaacattct taagttttac ccaaacacag agaggtcgac aatttgtcat aaaatgtagt 15060

ttatccatga atcaaaatca ggaatgactg tctgaacagt gtttttattt tttattttat 15120

tttattttgt gtaatttctg tgatgtgttt gaatatctca gttttaggca ggattggaaa 15180

tgttagaggt tggtaagagg tcatggttgc agtttgatca tgagagaaat cgatggctct 15240

cccttcattg cagtgttgtc agtcagcagt gtgggatcac ctatgtctaa cagttgttct 15300

aattgagaga ggattacagg agggaaagca gtgagattgt gaggtgctag atgaggagat 15360

ggcatttacc tagcagcctt ctctcccgcc ctcccatcat gtgacctgag agattcacaa 15420

tttctgaaga tatcagctgt gcttagttta agcaatagtt ttattaacta aatccaactt 15480

gattcatgtt attcccaggg aaccagtggt aggattaaaa atgaatccta gtgttctttt 15540

tggttattgg aatgtcaagt tttcagacac tgtaacgaat acagagccat acaatcacta 15600

tatttatttg gtcctttgtt gacttagaaa aattgaagcc cagtttaggt gagctaccaa 15660

atttctcatt gtggattagt attaaacttg cgtggagttg tgggatcttg gaagtggggg 15720

ctaagcatcc gtgtttgtca cagcccagaa ggaacagatg aggttccttt tgaggagtct 15780

tatgtcttta tgaacttgga cttagaaata tttgatgtgt ttaattctgc tgtagttttt 15840

taaactctag ctagtgagca tcttttcaca ggagcgcttg agtctgacct acagccattg 15900

tctgtctctg gtgtgcatat tacaaatgca ctgggagcgt ttcttgaccc aaacatataa 15960

ttagattttt cttctaaaaa ggtctagttt gggaaggaat gaaagggatt agagaaatgt 16020

tgtgggtttg gtatttattt atttatttat ttatttattt aatgtatatg aatgatctat 16080

cttcatgtat acctgcatgc caaaagagga catcagactc atgatggtga tgaaccatca 16140

tgtggttgct gggaattgaa ctcaagacct ctggaaaaac agctggtgat cttaactgct 16200

gaggcatctc tccagcccaa ttgttctgtt ttagtttgag gatgaacatc taatttagag 16260

atgccctgct tttccaaaag tgagttttaa acactaattt ccattgtcag tggattggtc 16320

ttttaagaat ataggtagtg gtggcacacg cctttaatcc cagcacttgg gaggcagagg 16380

caggtggatt tctgagttcg agaccagcct ggtttacaga gtgagttcca ggacagccag 16440

ggatacacag agaaaccctg tctcgaaaag caaacaaaca aaaacaaaac aaacaaacaa 16500

aaacaaacaa aaagaatata ggttggaata ggttggaagc agccaatgat agtgcatacc 16560

tttaatccca gcacttgaga agcagaggca ggtggaactc tgagtttgag gccagcctag 16620

ttagtctaca gagtattttc ctggagagcc aaggctatat atagaaaccc tatcttgaaa 16680

ggccaaaaaa ggaggaaaaa aaaaaaaaag aaaagaaaaa agaaaaaaag aatgcaggtt 16740

gggcagtcag ggtaagtgtc taaggtaaga ggaattcttc aaggtggaaa gtcatgagtt 16800

ctgcgccagc ctaggctaca gagtactgaa aggggaagag actgtccatg tgtcagaccc 16860

tcatttctcc aaaagtcaca tgactatatt ttttctgtat tgcccactct tccatacatg 16920

cacctaacaa taaatattga agttcactct gtggcactat atctatgtga tagacttcta 16980

gaaaagtgat ttaaagttca aaaggtaaat acgtagtttt gtttcaagtt gccaaaatcc 17040

ctttagtaga ctcctacaat cttacatgcc cagtagcagt atagaagctt gcttgttgcc 17100

ttgaagcctc accaattcaa atattaggta acatttgtta catttttctt tgtcagctgg 17160

ataggtaatg aatgacacaa caatgtgttc ccattttctc tgcattacta attgaagtcc 17220

tatcacccac agcagactga agagttcctt taatatttta tggactttga caaacctagg 17280

attcatagct tccatacaga gaggaatttc acaaatagca aagttgggct gttagaagaa 17340

taaaaagaga attctgagta cagcttctca aagaagagtc ccacgtaggt gtcctctggg 17400

atgtgcctag atgcagggtt attgtacagg agctcttctg tctgctctct gatacttgag 17460

attatagggt tgcagggaaa tgcattagat ggcattacaa actgataaga taaagttagg 17520

agctatcaga gatttaggac atggtttttc tctgtaaatg gggcttctgg tgagattcct 17580

agaaaatgct gtttatagct aggaatgggg ttatagctag gaatggggaa agaccttaag 17640

cagttgtgag ctgtggtgga atgcatgtgt tttcagtttg ctaaggcttc cgggaatact 17700

tttcctgtcg ataattttct ttcactctct ttgtagcctt ctttgtatta aaatcctctc 17760

tgcttgcttt tgtgtgtgaa tgtgtgtatg tgtgtgtttg tgtatgtgtg tgtatgcatg 17820

tgcatgtagg tccctacata ggacagaaca tatttcctgg agttataggt gcttgtgagc 17880

agccttttag ggaaccaaac tctgtcctct ggaagagtag cccctttaac tgctgagtca 17940

tttcagcctc aagaatcttc tcttttccct attagtagaa gatgtcatct tagctctagg 18000

aactacacca cctctggcct cagtggacac ccatttacat atgcacatac agcagacaga 18060

catataacta aagataaaat aaatcttttt aaaatgtcat ttccctgtgt actaattttc 18120

catgtacaca ctcacaggta gatttttaaa ctattctgag tgatcacaaa gcagagcaga 18180

aggtgaaatt tgagagaata gatgatatta gtggattttg agaccttgaa aataatgtct 18240

cagagcatta aattaatcac tcatgtatgt atgtatgtat ataagtatgt atgcatgtat 18300

tatgtggatg ggggtgctgt agcacatgtg tggaagtcag aggacaactt tgtgaagtca 18360

tgtttctcct tccatcttta tatggttcca gtgattgagc tcagattgtc tacctgtgta 18420

gcaagtgcct tacctgctga cctgtcgcac tagccctctc agaggacttt taatatttgg 18480

aatatttcta acgattgaca gtcaaaagtt tattgtgagc caggcactta aaatcctagc 18540

acttgtgaga cacaagatgg aggtcagtcc agtctactga gttctagacc agcaagggct 18600

acacagtgaa acctgtctca aaaatttcaa aagcggagct agagaaatta cccaaggagc 18660

taaagggaac tgcaacccta taggtggaac aacaatatga actaaccagt acctgggagc 18720

tcttgtcttt agctgcatat gtatcaaaag atggcctagt cggccatcac tgcaaagaga 18780

ggcccattgg acttgcaaac tttatatgcc ccagtacagg ggaacgccag ggccaaaaag 18840

ggggagtggg tgggtagggg attggggggg tgggtatggg gaacctttgg gatagcattg 18900

aaaatgtaaa cgaggaaaat acctaataat aaaaaaaaga aatgatatca gaaaaaaata 18960

aaaaaataaa aaataaaata aaataaaatt tcaaaagcaa caactcaaac cagccctacg 19020

tcgtgcctct gagttctcag taaattcctt ctctctctcc tctcagcacc atgtatgtgt 19080

tcggcggctt caacagcctc ctcctcagtg acgtcttggt ctttacctcg gagcagtgcg 19140

atgcacaccg cagtgaagct gcttgtgtgg cagcaggacc tggtatccgg tgtctgtggg 19200

acacacagtc gtctcgatgt acctcctggg agttggcaac tgaagaacaa gcagaaaagt 19260

taaaatcaga gtgtttttct aaaagaagta tgttttttct ctacttagaa tttaaaaatc 19320

taattttatc tgaattgtga aggaacctag tctctgtact ttcctgttca ccttactctc 19380

tagttatttc ttaataaaaa aatacacaag atctttggat gggaggaagc atgtggctcc 19440

tggaagctgt tagcaggtaa taagttgtct ttgaattaca caggctttgt gtaccaactc 19500

ctggtctggc tgcaggtgat ctgaagccat agcacaatga aatttgtttt cattttggtt 19560

ttatgagaca gggtcttgct ctatagctca tactggtcaa gctccttgtc agcctcctcc 19620

ttcagcctct tgaatgctgg ggttataggc atgcatcact ggccctactt gggaaatatt 19680

ttgatgacag acatgctata tatttctttg ttcagtttag tagccactag caatctgtta 19740

ttattagata tttgaaatgt ggctatgtaa ctaaggggct aactgttttc ttttctttag 19800

tgtatgtagt gaggcagatg tagtagcaca cgcctgcaat ccagacactc acgaggctga 19860

ggcaagaggc agttctaggc cagcctgggc tgtgtaatga gaccttgtct caagagccaa 19920

aacatcaaca ataaaagaac agtatgtggc tattggctgt tatgttgatg atgaaggtct 19980

agtgttaagg ataagagcct ctaatggtat gatcacatat agcaaattgc tctggtagac 20040

agcagagagc tgctgttctt gaaaagtatt tccagccccc tttagctgta tatagcaagc 20100

agtacagcat aacagacaaa ctatggtccc ttcttctaga gcccctggcg tgctcttgtt 20160

atttttctct cctttgctac ttgcttagtg gttgctctga gcaccacttc accaactcag 20220

cgaagtaacg tgcaaaaatg tttggaaaat aagaatgcct ccaagatatt tgtccatatc 20280

aatctttaaa gtatgaaact acttccttat ctagttgttg cagttacatg agagttatat 20340

taggcagaga ctacttctgt ttttctggta tgtgttaaat aaagttgtgc agggacataa 20400

agctcctgag gctgtgctgt tgattagaat tttggttcat ttatggaaaa cagcttacca 20460

gaacctggta ggattcataa ttctcccgaa acagttagaa ttggtagaat aaccaaaatt 20520

taaagttaag cttaaatata cagtgcattg gaaataatat tatcttctga ggttcagtat 20580

gagcccatta gtttacctca ctttctgggt agacctaatc ctgtcagagt aaacttggca 20640

agaaaagcag cctacatgaa aactgatcag gcagggaagt ttctgtggcc tctcttcctg 20700

cttgtgtatg tcatattcat gaaatgattt atagatggca acatggcttt tagcttcttg 20760

tttggggatt taatgagaat tatgttaggt ctacaaagag tggaagttgt gaaatccaca 20820

ggtttggagt cacatgagta tatagagttc gagttagcaa gtgcctcctg tggggttgtg 20880

ggtcactggg tatacctgca cccaggtagg ccttgcattt gtaacaagga caaatgtatt 20940

ggtctctcat attgctttct taggcttctg cacagcttct ggtgttaatt ctgttgctag 21000

ttgatgtttg tcgtgggaag aaaagcatcc attacttctt agaagctata aaattaacag 21060

acctttgctt ttcactttct ggacactatg ggaggacagt tataaaacag tgtttctcgg 21120

attgtctgct tatatctgtt ttattttaac ctaaacatgg cactgctttt ttcctttcag 21180

tttgactata cactttgctt cctgactatt gttaggagct ttcctacctc agattataca 21240

taagagaggc tgccgcatag ttgatgggtt tgtcttctct ctgtagccct tgaccatgac 21300

agatgtgacc agcacacaga ttgttacagc tgcacagcca ataccaatga ctgccactgg 21360

tgcaatgatc actgtgtccc tgtgaaccac agctgcacag aaggccaggt cagatgctgt 21420

ttttcacgga ttttagggaa tagaaaaatg ctagatgagt gtgagtgtag ggcaaataat 21480

gagtagagtt ctttttaaaa tgggatatcg atttgaattc tactgttgct caggttttct 21540

cttaggaagg gatgctatat acatcctgat tccaaggatc gctcctgctg ctgaggtctt 21600

tgtgcagtgt ttccgaaagc atgttttaca gaatgccctt ggcccatatc tgactcagca 21660

tgacatctgg gctaatcatg tatgatttgt tataggtgat aataggctat gagtaaggtg 21720

atccagcttt tgctgtcttt gatggcttat gacatttttt tctcaaagtt taatgcattt 21780

cataagaaat aagacttgag attgctatgg tgggcacggg ctgggaggag ctctggaaaa 21840

gcagcaggtt cagctttcac gttttacaga taagcattgg ctgaggcttg gtggtgccag 21900

tggttccgtt gggctgctag cttgccagct aaaagcatgt tagtgagaat acacactgtg 21960

gtattcacat tgcagtgctg cttcctgttc attctaattc tatcattcat ccatctacct 22020

atctctatct atctatctat ctatctatct atctacctac ctacctacct acctacctac 22080

ctacctacct acctaccact tatctaattc tatctgtctg tctgtctgtc tttctgtcta 22140

tctatcctcc atctaattct atctatctgt ccacctatct atcatctaat tttatacatc 22200

catccatcta tccatctatc tgtctgtcta tcatatatgt aattctaacc attcatctat 22260

ctatccactt atctgtctgt catctaattc catccattta tgtatctatc tatatatcta 22320

attctatcta ttcaattctt ttcttttttt ctatctttct ttctgcagtt accattctca 22380

gttaattctc actgagttat ttgtgtgaat aacaaaacac ttctcccctg tgttccagat 22440

ctccattgcc aagtatgaga gttgccccaa ggataacccc atgtactact gcaataagaa 22500

aaccagctgc aggagctgtg ccctagacca gaactgccag tgggagcccc ggaatcaaga 22560

gtgcatcgcc ctgccgggta ggccttgcac agggatgtcc tctataaggt ccaagcttgg 22620

tcctccctcc tcagatcaag gtggacctag gaacaagatt gcttattctg tctatttagc 22680

cctctcacta ttgggggggg gggggggcga tattttgtat gtttttaact taaatgtggt 22740

ttttatgtat gtatttacta gcctttgaaa gaaagtgaag tgtcagctca tgttctggag 22800

aattgggggg tagcttagat ccatgttaca aactgtgtcc cactgtcctt ccttctgctg 22860

tgaaggagaa cctggcacta gagctctgtg gtctcagcag cagtcaggaa cctgcaggaa 22920

gcacttactg acagttgtgt gagaagagat ttctgtacca gcatcatctc ccatgtgacc 22980

ttccttcccg actatttcag cagaggttgt tcagggtatt aacttaggtc ctgaggccag 23040

ctagccctga ctaaatctct atgatgtatt tgcttgatca ggatatccag gaaggggagc 23100

ttctgtgctc tccaacatcg aggtttgagg ggaagttggt ctgactcttt tgaaagcatt 23160

ttatttagtt tgctgaatgg gctttagttt agccagtgtt ctattgctgt gaagagatac 23220

catttccaac gtgtaacttt tatgaaagga aacatttaag tgggggcttg cagtctcaga 23280

agctattatc atcatgacag ggagcataga ggcacaaagg caggcattag agtggtagct 23340

gagagctaca tcctcatctg tgagcagagg cagacaaggt gtgaaaaaga cagaacctgg 23400

cctgggcttt tgagacctca aagtctacca cccccagtga gacacttcct ccaacagctc 23460

ctgcaacaaa gctccatccc ccgatccttc tccagtcctg ccactccctg gtgaatgagc 23520

actcacatat atgagcctat gggggtcatt cttactcaag ccactacagg ctttgttttg 23580

tgtctcagac tttatgtcaa tagaatacct agacaccttg ttacaagaca ggcctggaaa 23640

gcctgcagtg ctgactccct gccagtagca cattctgagg agcaagtccc ttaagtcgct 23700

tacctgctct tacattacgc ctttccctga ccatttagtg agcactgttg gtgtccccaa 23760

cctgaacctg gttctgggga aacacttgct tattcacttc cgtgctaatg gccagggagc 23820

aagcatgctt tcatgcaaca ctgtgagttc agtacaacca caggaggaga ttgcagactt 23880

ccttcgtgta ctgtatcact atgaggtttt ccaaaccagt ctccctttca cctcattttt 23940

tggcatgcct tatgtacttg cttatacttt ctatcttatg acatgaaaac agagtggcat 24000

ttggaggctt aaatttatca cattcccaat tcaattccat tttcagttta ctctttctgt 24060

atatacatca gtgtgcagat aaatatctct ttgtgtgagc attggaggcc agaggttaac 24120

ctctggtata ttcttcctct atcactcttc acagggtcct ttgatgaatg tggagctcac 24180

tgattacata gactagctga ctcaaccctc aggcctcata accctgcctc tagccctcag 24240

atgagattac aagcaagcaa aactacgcct ggccttttat gtgggtgctt ggaatttgaa 24300

ctgggtactt atgcttgaca caagtatttt atccactgaa ccatctccca agcctccatt 24360

tgcagttttt tacctcaccc ttccaatata tatatttatt tgtatgccct ttgttcaaga 24420

ttttagtcac cttttacatt tttcttcaaa aataattgca ccaatttctt aataatggca 24480

cccaaaagta ggaacattag cctagagtat accctgtgag ccaggaaatg tgactggtga 24540

gacttgtaaa agggtctttt tattctggcc ctcagcggag gctcagcagt ggagcatgca 24600

tgctgttcct ctggaggacc cgaggtcccc aggggccagg tcacaaccac ttgtaacttt 24660

aactctgatc taatgccctc tatggctttt gtgctatagt ctcttgcact aacccacact 24720

caaggcacac atacacacat tctttaaaag ataaattatt ttattttcaa aggttttttt 24780

ctgcatatag aagttaataa tttgtctgtt atgctcacca gatcctaaca aagcacctga 24840

aattcaaatc aggatgagtt cagatgttca gtattttgaa ctagtaaacc gaactgcata 24900

attcctaaaa ctttgttttc tttcctcttc ccctttaaaa aagaaaatat ctgtggcaat 24960

ggctggcatt tggttggaaa ctcgtgtctg aaaatcacta ctgctaagga gaattatgac 25020

aatgctaaat tgtcctgtag gaaccacaat gcctttttgg cttccctcac atcccagaag 25080

aaggtggagt ttgtccttaa gcagcttcga ttaatgcaat catctcaaag tatggtgagt 25140

taatgtgttc agaactttgg tttctagggc acaacagcag ctcttatgta gaaggccaca 25200

gttgtatgtt atttgcctgg taagagaaag aattacaata aatgattaat aatatactgt 25260

gggcctctat ttcagaggct cttcttttga tacctttctt cttgtcttaa aaagttcagt 25320

actttgcata ttttattagt tgttattatt aagtaaatta taaggtatga acatatggaa 25380

tgaatggtaa tatgtgtaca tattctggtg acatcagatt attttgtact tgatttatat 25440

ctagattctg cttgggaaaa gggagagtaa aatgttagtt acctaggtgt cattaaagcc 25500

atctacagcc cctggaggta ttattatagc acatagtgta atcgtcagta agaaatgtaa 25560

aatctgccca ggttttatag ccttcttcct aaggcttctg aactcagaaa gttctcttac 25620

tctagagcca aactctcaaa tggcttgtag ttactatata gtctcatttg gtatttttct 25680

tggtaagtct aattctaaga cttgtgattt gactgtgatg cttcagtcaa ttagatattc 25740

acagagcagc ttttctgtct atgctggctg tggtacagag agatgtgagg gacatgtttt 25800

tgtctagcca ggagaagaca gaatgcagct cagcatctct catttggcac caccttcatg 25860

tgatgggatg ccggtatggt gtgggtcctg gttgttaaat ctcaggaagt ccatatatcc 25920

agaaatgacc tcaactatag gtggatttct ggcaattagg taaaagtcag cattccttgg 25980

gcacttggga aactggttac catctgcata aaggagtcat ttcccttcta tctggcagaa 26040

gggacatatg gctatctatt gtgcctgtca gcatggaagc acatgctagt ctccaggtcc 26100

ccccaatatc acaagtacct atagcagtga attagttaaa ctgatttggc tcccaatggg 26160

tcaagtacag ctgcacctgc ccaagagctc tttgggtttg caaatgagag acacatagtt 26220

aatttttata tgctttgact agttcagttg ctggacattt ctaatcctcc ctgcagtagc 26280

atacattaac ccctccaact ttcctgagtc aacttactaa ctcaacattt catctctgac 26340

accccagacc taatggcaga gtggccctta gagccacttt cccaattttt tttttatcag 26400

atattttctt tattttcatt tccaatgtcc cctttcctag tttccctgtc ctctccccct 26460

gctccccaac ccacccactc cctcttcctg gccttggcat tcccctatac tggggcatag 26520

agccttcaca ggaccaagga cctctcctcc cattgatgac cgactaggcc atcctctgct 26580

gaatatacag ctagcaccac gagtcccacc atgtgttttc tttgattggt ggtttagtct 26640

cagggagctc tggggtactg gttagttcat attggtgttc actttcccaa attcttacat 26700

ggctggttta gttctttcct gcagctctta ggtctaatcc ctttccttcc tctgtcatgg 26760

tgattgcctt cctctcctat ctcagttcct tgcctgctca atctaaaagt cccacctcca 26820

tctttctgcc cagccactgg ctgtatgcag ttctttatta tcagttgaag ccagctaggg 26880

gcagagacct tcaggtctgt aagtgctttg gggagcagaa ttaagacaaa gcattagaac 26940

caattcccaa caagtacctg ctatacattt caaagtccat attagtctcc tgggtcttcc 27000

cttccccagc tacttgtcct ccttgtaatc caaatgacaa gctttttcac acatctcttt 27060

atctcacatt tccctagccc tggccatgtc cacttgttct tttactctct gctctgctct 27120

ctttccaatg cctctggata ttttctctct cttattcaca ataaaaacca aaccaaacca 27180

aacaaaaaac cttaccctaa taatggagtg gtcacgcctg aggtttcctt actgctcccc 27240

cttgcacacg tcttgtgtct gacacactgg caggctttta ttagcagcag gctctaggag 27300

ctgagagaag cagcaggcac ctctgaggtg gtagttacta gagtgattag aacagacagt 27360

ggagacgtgg ctggaaatat ggactctggt gtttggagcc aagtatggta ggcggcagaa 27420

gccagcagaa gcatgatcca caccttcacc aggttgcttc cattgggaaa ggctggaccc 27480

cttgggaagg ggtccctttg tgccttccta ggtgttcgga gccaggtgtg tgagggatac 27540

agtaaaggga ctgactgcat gactgctcca ttagggtgaa gggttttgtt gtgaatagga 27600

gaaacaaaat gtgcagaggc atctgggaga gagcagagca gagtgaaaag gaagcagtgt 27660

aggcatggtc agggctaggg acagcggaga cagcaagata gcgagtgggt gataaggtga 27720

gagagagtgt gtgtgtgcgg tgcacacatc acgtgcatta taaggaggct gagtagctag 27780

ctggggggag ggaagggcca gaaaactagc atgcactctg aaacgggtac ttgtgatgct 27840

gagggagctt gggggagaag ggcatgcctc aagaccagaa gagggagttg gagttacagt 27900

ttgtaagatg cctaatttga atgctgagat ccaaactctg atcctttggc tgaacatcat 27960

atctgctgag ccatctctcc agcccctaga aaggtggtga tggtggttgt tcttgttttg 28020

ttttattttg tttaaatggg gagccaggta cagtacatca tgcctttaat cccagcagga 28080

gattcaggag atagagacag gtagatctct ttgagttcaa gggcaccttg gtgtgtatag 28140

gaaattccat ccacccaggg ctacagaagg gtaccttgtc tttaaaaaaa aaaaaaagaa 28200

agaaagaaag aaaaagaaaa aaaaaagaga atgaaatttc agagttatgc aagataggag 28260

ctcagtggta gagtgtgtgc ccaggaagtg ctgggtttga ctcctcagaa caacagcagg 28320

ggcagaaact agtctacagg ttcatgagtg gtgttttgtt ttgttttaca taaaatgtgt 28380

tgaattagat aagtagataa aatgtgactc atacacagat aaatagataa aatgtgatac 28440

atgtacctgt acatagaaga ttatgatctc acctttaaaa aggaggaaat agagagtttt 28500

ggtagttaca ccacaggaaa actggaaaag aaaatgtata tatgaggctg tgccccatgg 28560

ctaaaggaac atgtttttaa gtcatttgaa ttcaccaaac agttttaggt aatgatatat 28620

ggttttgcat acaaccagta ttttataaat attagcaagg tcacatcatt tatgaaccaa 28680

catttaaact aaatttgtaa atcatcattt ctttatagca cttgtcatag aacataagta 28740

gtttaaaatg tgattattgc tttgctcttg atgtctgaaa atcttcatgt attctcttct 28800

ttgagccatt tttatgcttt gcagtactgg atgcatattg aagtgatcac ttattttaat 28860

ctaccttgcc tgagtttggg gaatagatgg tttccacatg tctgtgggtt atgcctaagc 28920

tagtggtttt tatgttagag cttgttttgg ggaaggcact ggttgcattc atagctgtgt 28980

ttcttttgcc tgtagtccaa gctcactctg actccatggg ttggtcttcg gaagatcaat 29040

gtgtcttact ggtgctggga ggatatgtct ccattcacaa atagtttgct gcagtggatg 29100

ccatctgagc ccagtgatgc tggcttctgt gggatcttgt cagagcctag tactcgggga 29160

ttaaaggctg caacctgcat caaccctctc aatggcagcg tctgtgaaag gcctggtaag 29220

gacatgggtg catatagtgc tccaggagga gccaagacag caaaggaggc acagctgaat 29280

gagcgctgag gtgatgaagt acttatggca gcagggagag gagcaccaat ttaggcatat 29340

gtatttcaaa cagaacccga ttccagatag tctttcttgg cctctgactg ctttaagcca 29400

tactgaaaac caaaaataaa attgctgaaa gaacccagtt tatattgagc tgcactgttt 29460

cgttggtctc aaagtgttga gaattgttct agaagattat ttccttggtg ttggcagaga 29520

agtgctatgg aggaaacaac aacctgaaac caaagaaaca tttagaaaag cagcaagtca 29580

ggacactatt cagacactgc tggggtgggg ggagaggggc atggccaaag aagccgacag 29640

agccaacacc aggctgtggc aatgtcctgc gctgaggtta aggttagact ccatgaggcc 29700

aggcccagaa cagccataca caaatgagga ctccaaaaca agaggtgcaa gtgtagtgga 29760

gactccatcc ctgcaggtcc tgtttcagga aatgattgta ctttgcctga gtaatacagc 29820

ctaggagcta ctttctgata gggtttttta aatacttaca aagaattatt tatctttaat 29880

catgtggttt tgtatgtgtg tgcttgcaca tgcagtgctt gtgagagaga gtatgtgtga 29940

gagcatgcat gtatgagagt gtgagaatat atgtgagaga gtgtgagtgc atgtgtgcgt 30000

gtgtgcatct gtgtgtacag gtgtgtgtac atgcatgtgt gtataagagt atgtgagagt 30060

gtgggtgtgt gtgtgagagt atgtgagaat atatgtatga gtgtgtgtga gtatgagtgt 30120

atgtgcgtgc ctgcatgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt 30180

agaagtggcc ttggaaaaca gagttgtcag atctcttaga gatatagttg cagttggttg 30240

tgagccatct catatgagcg ctggaagttg aaattgggtt ctctggaatc ctctgggttc 30300

cttgttgaag cctgaatatt ttgataaata tttatgtcat tatccctcaa aattgtaaat 30360

gtagaattta acaaactcag gtcttgagtc atctttgtcc caaggtttgt ttgtttggtt 30420

ttttgttccc ccaccttttc ttcagtgctt ttaaaaaaga gagtccattt tttcctaaat 30480

gtttaaatac agttgaggaa tagaacatct gactccaatt tcctgggttt ccctccatgt 30540

agtgtagtgc tgacctgatt tcagtgtgca ttgaaaactt tgatcacttg gaaggcagct 30600

atgctcacca ctatactacc aatgtctgca atcctatagg agaaacaaca atatgaacta 30660

actagtaccc cccagagctg tgtctctagt tgcatatgta gcagaggatg gcctagtcag 30720

ccatcattgg gaggagaggc ccttggtatt gcgaagatca tatgccccag tacaggggaa 30780

tgccaggacc aggaagcaag agtgggtggg ttggggagca gtgcgggggg ggggggtata 30840

gggggttttg gggatagcat ttgaaatgta aatgaagaaa ataactaata aaaattgcct 30900

taaaaaaaaa caaaaaagaa aagtttttga tcttagctga ccagtgtctc tttgggtctt 30960

aatttccagc aaaccacagt gccaagcagt gccggacacc atgtgccctg cggacagcgt 31020

gtggcgagtg cactagcagc agctcggagt gcatgtggtg cagtaacatg aagcagtgtg 31080

tggactccaa tgcctacgtg gcctccttcc cttttggcca gtgtatggaa tggtatacga 31140

tgagcagctg cccacgtaag tggaaggagc ttttgaacat ttgcaggcaa gttgggcttg 31200

actttctgct caagtccatg cagaagctgg tcgggccggc ccttccagat taacatgtat 31260

gtatagaatg cagcacagtg ttccatgcag taaatcagtt acatcaagga gaaggcacag 31320

ggtacagaaa taccttttct tcttcagggt aatattataa ttcaatctgt ataatgtttc 31380

tacatcttaa tctaccagta tgtaaagtgc tttctagtag aggcctcccc agctcccttt 31440

ttcatccaac atcctgatat taaaaggttg gaaaagtccc tgttatatat tatgtaaaat 31500

gtggggccct ttaaattatt tcagttcaat aatcactata gggtactatt tttaattcat 31560

ggaagttaaa tcatctgtta aaagaaaagg taataacagt aaattcaaat cttgtgatag 31620

tgaattacaa gttggattgt tttgccttgt tttttaatag ctgaaaattg ctctggctac 31680

tgtacctgca gccattgctt ggagcagcca ggctgtggtt ggtgtactga tcctagcaat 31740

actgggaaag gaaaatgtat tgagggcagc tataaaggac ctgtgaagat gccgtcacag 31800

gcctctgcag gaaatgtgta tccacagccc cttctgaact ccagcatgtg tctagaggac 31860

agcagataca actggtcttt cattcactgt ccaggtaaga tgcctgtgta tcctagttca 31920

aatctcgtac ataaactaga cgcccagatc ccttggctca cttgttttct tgactgtgtt 31980

tgagttcttt ctgtgttctg catcaccttg ttggatcata gctggcaaag gtgctctcct 32040

ttctgtgggc tttttcttta cttgattgat tgtttctttg gttgcacaga agctttttag 32100

ctttctgaag tcccatttgc cagttgtcct taattcctgg gcgagtagaa gcctcataaa 32160

aaaaagttcc ttcctacaca tgtatcatgt agggcactgc ctatgtttta ttccagaagt 32220

ttcagaggtt cgggttatgt ctttgatcca tttagggtta ctttttgtga aaggtaatgg 32280

acacagttct gtttcattca ttattctaca tgtggacatc tacttttccc agcaccagtt 32340

ttgaagatgt tatcttttct gcaggttgtt tgtttgcttg ttttgtctct tcagaaaatc 32400

ccagatggcg gtagctgtga gtgcttaggc ttggcctacc tgtttcatta tgttggcttg 32460

catgtctgtt ttgtgcagtg ccaccatatt gtcttaattg ctatagctct gcaatctatc 32520

ttgacatctg tgttggcaat cctgcagttt cgacccttct gctcagcagt gctttggcca 32580

tctggggtct tttctgggtt cataatgaat tttaggattt ttttttctat ttctgagaaa 32640

gtattgttga tattttgatt gcgattgaat tgaatctgta aattgctttt ggtagaatgg 32700

tcattttcac aatattaatt ttactgatcc atgaacatag gatgactcca gtctctcatg 32760

tctccctata gccctgtctt aagagatttg gagtcttcat tgtagaagtc cttcacctcc 32820

ttggttaagt ttatttctag atattgtatt gtctttggta ttataaatgg tagtatgtcc 32880

atgatcttgt tctcagtgtt tttttagttt agtttttttt aatttatgtg tatgagtgtt 32940

tgttttatat atgtgtatat gtgcattcat gtcctctggg catcagatcc cctgggactg 33000

gatttacaga cagctttgag ctgcctgtag gtgctgagaa ttgaacccag gtcctctgca 33060

agaacagcca gtgctcctac tccccagccc cagaagtact aatttttaag agctgatttt 33120

ctacctttgc tgacattgtt gattgtttct agaagtttag tgatagagtt tttgagattt 33180

cttatatatc ttatgttatc tgtaaaaagg gataatttga ctccttttcc tatttatatc 33240

cttttatttc tttcatttgc catattgttc tagctagtgc ttcccgctca gtattgaaaa 33300

gagtggtgat tgtgaacagc ttttcttatt tcttatttta atgggattat tcacccattt 33360

aagataatgt tggttatggg tttgtcatac acagcccttc ttatattgag gtatgttcct 33420

tccagtcctg ttctctctag gacttttttt tttttaatca agaaagcata ttgggttttt 33480

tgttgttgtt attttgtttt gtttttctag acaaggtttc tctgtgcagc cctggctgtc 33540

ctggaattca ctctgtagac caggctggcc ttgaactcag aaatccacct gcctctgcct 33600

cccgagtgct gggattaaag gcgtgcacca ccactgcctg gcacatgttg gttattttgc 33660

aagccctttc tacatctact aagatgagca tgtggtttca tctttgtctg tttatattgt 33720

ctgttgtatt tattgactta tgtgtgttga gccaacctga agttctggga taaaacccac 33780

atgctttgga tgatttttgt gctatgtgct tatattgtgt ttgttagtgc tttattgagg 33840

acgtctgcat ccgtgttcat ctggggtact gtctgtagtt tgcttatttt gttgtcttta 33900

cctgctctgc attttagagt aatcctggat ttatagaaag catttgggag tagtccttct 33960

gtttattaaa aaaaaaaatt aagaatgatt ggttgttgtg tggtggaatt ctgctgtgaa 34020

cccatctggt tctggactct attcggaagg ctttttatta ctgtttcagt ctccttgttt 34080

gcagtgatct atttaggttg ctaatctcct tatgattcat ttggatgaat caagaaatta 34140

atccatctct ttagatttcc agcttaatgg aatatgagtg ttaaagtatt tctttatagc 34200

attctgtatt ttttggcatc tgttgtaata tttccctgtt ctttctgtta atctctttct 34260

ttcttgtggt tagttgggct aagaggctct tggttttttt tttttttttt tatcttttta 34320

aaggaccagc tcttagattc attaattctt tgtattattt tccttgtttc tttttcactg 34380

atttcatttt agattttatt atttcttgcc atctactgcg tttgggttgg ttttagttat 34440

ttttccaaga ttttcagttt catcactaag tcattcattt gggctctttt gggtttcttc 34500

acgagaaccc acttgggact gttaccttcc cttttagacc tgcttttaat gtgccccaga 34560

gatttgttac attgtctttt cgatttaact tagtttcagg aatattttga tttcttcttt 34620

gacccattca tcattcggta atgagttgtt taatctctag tgagtttata catttattag 34680

aattttgttt actgatgatt ttaaggtgtt tggctttgtt tgtttgtttg tttgtttgtt 34740

tttcgagaca gggtttctct gttgtagctc tggctttctc tatgtagaac agtctgacct 34800

caaattcaca gagatccacc tgcctctgcc actgaagttc tgggattaaa ggtgtgtgcc 34860

accactacct ggctgatttt aagttttatt acataataga caggtagggt acatagattt 34920

tctacatttg tgaaggtttg cgttgtttgt cagcatgtaa ttctgtgtgc tgctgaggga 34980

atgtatgttg ttttgacagt taggtggaaa agtctgtaga catctgttag atccatttta 35040

catttcaaga agccatttaa ttctgaagtt tctctgctta ttttttccca ggtgacttac 35100

ctattggaga aaatagggtg ctaaaatcat ttactattat tgtttttttt aagaagaaaa 35160

taattaattt aaaaaaccct ggaaagaaag ataccaaatg tgaatcatgt ttcctggata 35220

gtggggttat atttgatcat ttatttttcc tctcaaatac tgtgagtttt tacaatgaat 35280

aacaacataa atatttttat gttgctgtgg actttaactt tgctttgata atatatttgg 35340

ttttttgaga ctaatttctt tttgatattt tattttctca tactagtttt tagtaaactt 35400

tggttttgtt ttgtttgtat tttttagact ggccaccaac ttgctatgtt gtcaagggtg 35460

gccttaaaat ccacacccaa tactttgtcc tctctttctt tctttctttt ttttttttat 35520

tggaacaaaa tttctaggtg ggaatctcac tatgttaccc aggctgacct gaaacttctg 35580

ggcttaagca agatgggtgc acatgatcag agacgctgcg ctgcccgcct cagcccctgc 35640

tagttggaac tataggcaca gacagctgta cttcactcat ttcaatgatt taacatttag 35700

actatatgca aataaatatg aaatgtattc accaagttct cctatgggag aaacagagcc 35760

cttaagattt tttcctttca gcttgccagt gcaacggaca cagcaaatgc atcaaccaga 35820

gtatctgtga gaagtgtgag gacctgacca cgggcaagca ctgcgagacc tgcatatctg 35880

gcttctatgg tgacccgact aatggaggca aatgtcagcg taagtcacac aggtcaagtt 35940

agtcacaagt caggtacaat agtacagtac ctgcagttga cttaaatatc ttaaagggaa 36000

aaggcctctt ggtttgggat attgcctttc ttaattatgt taaattgtta aaagtttaac 36060

tgaggggcta gaaatgtggc tcagttggct aagaacactg actgttcttc tagaggaccg 36120

aggttcaatt cccagcaccc acatggcagc tcacaagtgc ttgtaacacc tgggatccaa 36180

caacctcata cagacataca tgcatgcaaa acactaatat acataaaata aatccattaa 36240

aaagtgtttg atgatgctgg aagaggaaaa aaggctcaac ttgtgggttt gggagcagtt 36300

agttaaagca acaaaccgac agtaaaggag ctaagctttt atttcttcag cagaggcata 36360

aacaaggggc cgaagtcact gaggcaccag ctgcctttat tccatttccc tcccatggaa 36420

gcacatcagc tcaagtcaag cagagcagcc tgggatggga ggtcatctca ttggagaagg 36480

aggcaggagg cattgtgagg ggagggagga caaggctggg aatgggaagt cctgagctca 36540

gaatcagaat gaggacaaga tcttcagttt ccttcttaat ataaagaggt atcacagagg 36600

tctctataga agtctactgg aagcctcaca caggcacaag ggtacatttg aaaaactgtg 36660

acagccaggg agagtcccct tctgaagtgt ccttcctcag agactgcagc acctgactgt 36720

gccccagtct gcaagaggtt tggggagagc aactgacctc ctgaggaccc cagatgaatc 36780

tttaagatgg cctgcttttg gttttggttg gttggttttt agacagatct aggagagttg 36840

gtgatgagct tgaattctct gtcctcctgc ctgacctcca aatgcccagc ttcacatggg 36900

ctcccattaa gttgtgagtt tcggtgtctg gctcctgctc tcacagccag tgcagtacat 36960

tgagctccat agagatagcg ccggggcaaa tgagagctgg acgggcactg ggtgactctg 37020

tgccttgtgc cggaaaatca actaaacatg ggcaaaggag atcctaagaa gccgagaggc 37080

aaaatgtcct catatgcact ctttgtgaaa acctgctggg aggagcacaa gaagaagcac 37140

ccggatgctt ctgtcaactt ctcagagttc tccaagaagt gctcagagag gtggaagacc 37200

atgtctgcta aagaaaaggg gaaatttgaa gatatggcaa aggctgacaa ggctcgttat 37260

gaaagagaaa tgaaaaccta catcccctgc ccccaaacag gagaccaaaa cgaagtacta 37320

ggaccccaat gcacccaatg ccttcttcgg ccttcttgtt ctgttctgag tacctcccca 37380

aaatcaaagg tgagcaccca gcttatccat tggtgatgtt gcaaagaaac taggagagat 37440

gtgaacaacg ctgcagcaga tgacaagcaa ccctaggaga agaaggctgc caagctgaag 37500

gaaaagtacg agaaggatat tgctgcctac agagctaaag gaaaacctga tgcagcaaaa 37560

aaaaaaaaaa ggggggtggc caaggctgaa aagagcaaga aaaagaagga agaggaagat 37620

gggaggagta tgaggaagag gaggaagaag aaagatgaag aagaatatga tgatgatgaa 37680

taagctggtt ctagtttttt tctcatctat aaagcattta acccccctgt atacaattca 37740

ctccttttaa agaaaaaaat tgaaatgtaa gcctgtgtta gatttgtttt taaactttac 37800

agtgtctttt ttttgtataa ttaacatact gccgaatatg tctttagata gccctgttct 37860

ggtggtattt tcaatagcca gtaaccttgc ctggtacagt ctgggggttg taaattggca 37920

tggaaattta aagcaggttc ttgttggtgc acagcataaa ttagttatat atggggacag 37980

tagtttggtt ttggttttat ttttgggttt ttttttttca tcttcagtcg cctctgatgc 38040

agcttatatg aatatgattg ttgttctgtt aactgaatac cactctgtaa ttgaaaaaaa 38100

aaaatcgtgg ctgtcttgac atcctgaatg tttctaagta aatacagttt tgtttttatt 38160

aatattgtcc tttcgacagg tctgaaagtt ttcttcttga gggaaagcag tcttttgctt 38220

ttgtcccttt tgggtcacat gggttactgc agtgtgtatc ttttcatata gttagctgga 38280

agaaagcttt tgtccacaca ccctgcatat tgtggtaggg gtaacacttt catccatatt 38340

caaagaatct ccaaaatcgt gatcagttgg ataagaaata ttatataacc tacttggcaa 38400

agcaaggtgt gatcaattct gtcacaccat gggatcatta gaatcaagca atctgaaaat 38460

ctgtccttaa aggactgata gaaaagtatt ttctaatcct tatacaaagg ctctccttta 38520

actgccactg ctatgtaatg acagttatgt tttgcagttt ccctactaaa gaagacctga 38580

gaatgtatcc ccaaaagcgt gagcctaaac tacacaactg cagtactatt tgttgacctt 38640

agtcccagcg aaggctatca cgagaatgct agctataata taatgcctct gcccctctat 38700

ctaaatatgg attgctcagg aaacttgact gcttaaaggt atttttttca tattgttgtt 38760

cctcctatag ggttgcagac ccctttagct ccttgggtac tctctctatc tatctatcta 38820

tctatctatc tatctatcta tctatctatc tatctcttgt cagatttctt ttttcttttc 38880

tctttctttc tttctttttt taagatttat ttattattat ttctaagtac actgtagctg 38940

tcttcagatg caccagaaga gggtgtcaga tctcattacg gatggttgtg agccaccatg 39000

tggttgctgg gctttgaact caggacctta ggaagagcag tcggtgctct taaccactga 39060

gccatctcta caacccttaa aggtattttt aagtagttga gtcagctttt aaaattatgc 39120

cagaagtgtc aaaagttcaa aagtttagga ccatcctcta ttgaagtaca gggtcatcct 39180

gggctacatg agaccctgcc ttaaaaccaa aatcaaacaa acaacaggaa aaacaagagt 39240

taagaaagag aaaaagaagc acttggaaac aaagatctgt ggagtatgta taggcttctc 39300

tacaacaggt gtatgtagga tcttgatggc ttttgagtct attaccctca aagaggtact 39360

gagaaaccta aatgtgatca ccgtggtctc tgaggggcac ctggcaggat tatgggagat 39420

aactaaagct tgctaatcac agagtttagg gagggaggac gtctctaagg caagttaact 39480

gtctggtttg agatgcttag gtgatgtctg aggaagtaat aaggcctgtc cattttcata 39540

cacactcagg ccttaagtct gggtaatggc tacttgaaca taaaatagtc ctctatgaaa 39600

ggaataatat ctctgtgtca gcagccttca cggctaatgt taattgtgca ggaaccctgc 39660

ttctcagtca gacagaagct caatcaggca ggggcaggac ttctttgcct ttcccatgtc 39720

cttgtaattt ccctggcttt tcatcttggt tcaaacatac ttacctgtta ggtaattata 39780

agaacaccaa atattactga ataaaatgtg tttatgactt tgtggtgact gccattcaag 39840

aattagatgc cttagccagc aatgatggca cacgccttta atcccagcac ttgggaggca 39900

gagataggca gatttctgag ttccaggaca gccagggcta cacagagaaa ccctgtctcg 39960

aaaaaacaaa acaaacaaac aaaaagattt cgatgtcttt atcacccaaa tcaagtaact 40020

ttccaaagtc tcacagtgag atgtagccta gttgggagcc acatctaata tatgctgatg 40080

atcttaacaa gtagcctgct tgtgtcttca ggtgaccacc ccggtgtcct cagctacctc 40140

tagaaagatc acactttcct ctgtggtctc tgcagggtcc ctgtatgatt ctggaacctt 40200

gctgtacttc tcagagtcct gattcataaa gcactgagtt tttgcttgtt tgtttgtttt 40260

gatactattg gtaagaatat atattgaacc ttgacatgcc tttttaaaat aacattattt 40320

ttacaatagt actttagcct tgattatgtt aactgcttac tgtttcagat gacattcgta 40380

catcttttaa tcctcaaacc agtcctatga gatggctagc atcattgtca catcatttag 40440

gcaaggaaac aggtcttggg ttaagcttca tgctcagagc tccttggaac acagtggact 40500

caagtgcaag cagactgacg cgactgggtt ttactaattc agtaagcctg tactctatgg 40560

aggaagagtt tctgaccact ggatgcagtc tgatgacctc tgactgttct gtttgaaagg 40620

tttctttcag tgattttatt tttctccatg tggacttttt ttccagcttt taaaatatat 40680

atatatatct tattcgcttc acatcctgct cactgtcctc cctcccctgt catcccctcc 40740

tacaatcctt catatccccc cttaccttct gagcagctgg gagcccctct gggtatcccc 40800

acactcgggc acatcaagtc tgtgaggctg gacgcatctt cccccactgt ggccagacaa 40860

ggcagcccaa ctagaacata tcccacagac aggcaacagc ttttaggata gcccctgctc 40920

cagttgttca gcacccacat gaagaccaag ctgcacatct gctacatatg tgcagggagg 40980

cctaagttca gcccatgtat gttctttggt ttgtggttca gtctctgaga accccaagga 41040

tacaagttat ctgactctct taatcttcct atagagttcc tatctcctct ggggcccacg 41100

attggtgtcc ctattgcttc actgggattc ctgcctggct acacccacta tgaccaaggc 41160

aagtcttaga aaagacaaca tttaactggg gctggcttac aggttcagag gttcagttca 41220

gtatcatcaa ggcaggaaca tggcatcatc caagcaggca tagtatagaa agagctgaga 41280

gttctacaac ttatctgaag gctgctagca gaataccgac ttccaggcag ctaggatggg 41340

ggtcttcaga cccacaccca cagttggtgt ccctattgct tcactggggt tcctgcctgg 41400

ctacaggagg tagcctcttc aggttccata tccccaatgc tgtgagccac agttaaggtc 41460

acccactatt gattctaggg tgtctccctc atcccaggtc tctttcattg tggagatgcc 41520

ccccacttcc ccaccactgt cagttgcaga tttccattct cgggaccatc tggccatgcc 41580

ttctgtttct cctcacacct gatcccgaca cccccgccca ttccttctcc tacctagttc 41640

cctccctcca tatgcttcct atgactattt tattccccct tctaagtgag attcaagcat 41700

cctcacttgg gccggccttc ttgttttgtt tctttgggac tgtggagtgt agcttgggta 41760

tcccattttt ttatggctaa tatctgctta taagtgagta cataccattc gtgtcctttt 41820

gggattgagt tacctcactc aggatggtat tcttaagttc tattcatttg cctgcaaaat 41880

tcatgatgtt tttgttttta gtaactgaat agtagtccac tgtatagatg taccacagtt 41940

tctttatcca ttcttcagtt gagtgaaatc taggttgttt ccagtttctg gctattacaa 42000

ataaagctgc tatgaacata gtggagcatg tgtccttgtg ggatggtaga gcatcttttg 42060

ggcatatgcc caggagtgat gatatagctg agtcttgaag tagaactatt cttagttttc 42120

taaaaaacca cgaaattgat ttccaaagta gttgtacaaa tttgcactcc ctctaaccaa 42180

gcaagtgaaa gatctgtatg acaagaacta caagtccctg aagaaataaa ctgaagaaga 42240

tatcaaaaga tggaaagatc tcccatgatc gtgaataggt aggattaaca aggtgaaact 42300

ggacatctta ccaaaagcaa tctagagatt cagtgcaatc cccatcaaaa ttccaacaca 42360

atttttctgt agaccttgaa agagcaattc tcagtttcat ataggaaaac ataaagccca 42420

ggagagccaa aacagttctg agccataaac gaacttgtgg aggaatcacc atccctgacc 42480

ttaaagccgc actacagagc agtcgtgatt aaaacaacaa caaaggctgc gcacttttgg 42540

tacagaaaca gacgtgctga ccaatggcat ccaatccaag atccagaaag aaacccacac 42600

actatagttt ttttttaaat ataaagttct tcagcttaat gcttctcatt attcatgaga 42660

gaagaagact caacagcaaa gaaggtgaaa caagggtgac aagtaccaca gggctctcga 42720

gtgtctcttg tgatggacta gggagcccgt cagttctgaa tgctcaggaa tgtggttcac 42780

agtgtggcca cagtacagaa gatccccgag ataaggcaga agacagtcac cacaggtcat 42840

ctccacaggg caaggactca gtatatggca tattactaat gctcttaaat atttactgaa 42900

caaaggaaca aaatgctgag tctgtcacag agatgaaaat agccgttgct tcaggggaca 42960

gcagaagata gccttttttt ctccttgaat ggtagttaat ttaatgttgc ctctatatta 43020

ttagaaataa attacaagct gaaaaataat gagtcatacg cagtgatttc tcttgcttta 43080

ggctgtcttt actacaaacc catttcaggc taaatgattt tgtcttaatc acagtctatg 43140

gtaatctgtc aagccagttg tgacctgtct tcctttcctt cttcccagca tgcaagtgca 43200

atgggcacgc atcactgtgc aacaccaaca ccggcaagtg cttctgtacc accaaaggtg 43260

tcaaggggga cgagtgccag ctgtgagtac cacacacact ctgtgtctcc agtgggggac 43320

tgggccttgc agctgcctgg gccctgtcgg ccacctgctt gcctgggcat tgttgccctt 43380

cactcccagg gtctttgagt ggactagtgt ggaggtttac cttttttcct tcagacaggt 43440

tatctcagtt actttaatat tgctctgata aaacatatga ccaaggcaac ttacaaaata 43500

aagcctttaa ttgggcttat gacttaagag cattggagtc tacattgagt tccagggcaa 43560

tagagctaca tagtaagact gtatcaatca atcaataaat aggactacat agtaagactg 43620

tatcaatcaa tcagtagatg aagagaaaga aagaaagaaa gaaagaaaga aagaaagaaa 43680

ggaaggaagg aaggaaggaa ggaaggaagg aaggaagggg aagaacaaaa caagcttaga 43740

taggaagaac aggatagaat gaatgacaaa tgcttgaaaa atgttttagc tgtactttta 43800

gaagcatact caatccacac agaagtaaaa atgttgttcc ttatgagtag tacctagcat 43860

tattacatat gtacttgcct gtgtccttgg gcaagtattt gtttatttgt tgtttttata 43920

ctgttgctgg tgtaaattac tgagcagtta gcagaaacat tcctgcaaat gggatagtct 43980

ctctgatctg aataatgata tagtttatgt aaaaggattt acttggttta aaaataaata 44040

tagagtctgt gctttaaatg tcaatagaag ataatttctt ttttccctag atgtgaggta 44100

gaaaatcgat accagggaaa ccctctcaaa ggaacatgct actgtaagtt tttgtaattg 44160

tttctagaga gtaattgaac aaaacgacat tgcttttttt ttttaccatt gtctgagaat 44220

gataaatgct tgggggatga agcaaatact catagccatg cccctgactt ggtgaacact 44280

gttctaactg aggcatggtc tctgctggtc atccagagca gttagcaggg gtgctgtcct 44340

gcctgtcctt gttcagctcc cgcggaggcg tgctcattca ccattgccca gtgtagctta 44400

tcatgtccaa tcttcagaca gccaggaagg agtttctaag atagaggtgc gttccaccat 44460

tctctctgca gctgatttgt gctcacaaac aagtaaataa aacaccaaat taataccttg 44520

gtgtgaaagt gaatctggta agcttacagc tttatcataa atatattttt tgtctatgag 44580

aatctacata gtaggttcta gactatagaa caataaaaaa aggaattaac atttggcata 44640

tgcagcataa tggtatatat aaattgtaga agaaaatgga tggttctaga cctgaaaaga 44700

caagaaaatt gcttgtgtgt aatctgggca ggtcttaagt tgtgaccttc aacatctgct 44760

tcccaagcag ctggaaccac caggcctaca gaattcttag ctatgattct aaaggtcatt 44820

catcaaatat aatgttaatg tgtattttat taaagtttca aacttctatc tttaataatc 44880

tgcaaatgta gctcagtaga ggagagctct cgctgtaagg tcctgtgttc tatccccagc 44940

acaacaaaac aagacattta agaaaaaatt aaaacaagtt ggctgtattg tctcagtatc 45000

tcatccttga gatagtgagg caggaggact tttagtttga ggcctatgtg ggttatgtag 45060

tgtgaaacct ttctcaaata atatttacac ttttttcttt aaaaaacaac ttttttctta 45120

atttatgtgt tttgcaacat gtaagtctgt gcaatgtgaa catatctgtt gcctttgaat 45180

gccagagagg gtttcagttt tcctggatct ggagttacca agggttgtga gctgccatag 45240

tgggtgctgg taatgaactg agtcctctgg aagagcagcc agtgctctta actgctgagc 45300

catctctgct gctaggtact cccccttccc cccttaaatt taagacaaag gtctcactgt 45360

gtagcctcag atggtctaga actcaatttg tagaatggtt gacctttgaa ctcacaaaac 45420

tctgcctgct tctgcctcct gagtgttgag attaaagttg tatgtcacca cacctgcccc 45480

tatgatttct atatttaata aagatcatga ctaggatata gagaacactt ttagaactga 45540

agaagaagac agttacagtt aaaagcaaaa caaaaacaaa aacaaaacaa aacccagaaa 45600

aaaaaagaat gaaaactagc actgaagaaa aaataaattt taaaaatagg caaagagtca 45660

ctattatatt gtgatggatg tgttatatgt ttaaaaccac aagtgagata caggcctgaa 45720

atgactttaa tcgaagctac accagcctgg ggtggtagtt cagttggtaa agttcttgct 45780

atgcaagcac aagaagctgg gtttgatgcc caggacccat gctgaaaccc aggagtgctg 45840

ctgagtgctt cagctctggg gtggcagggc tcactggcag gaagcctagg ctaagagaga 45900

ctctgtctcg aaaaacaagg ccgatggcac ctgatgaacg gcatctcagc atgacctttg 45960

ctcggcatat aatgtgtaca cacaaattca tagtttagta gaagacaagt atgatctgct 46020

tttcatgaag tctgttgtaa tacgccttct ttagttaacc atagttgctt aaaaaaagaa 46080

aaaaatcgac ctcactggac agaaaatgga tagagtgttc taatagccaa ttcaattcat 46140

catcattatc aaaacctata acttaggggg ctggagagat ggctcagcgg gtaagagcac 46200

tgactcctct tctgaaggtc ctgagttcaa atcccagcaa ccagatggtg gctcacaacc 46260

atccataaag agatctgatg ccctcttctg gagtgtctga agacagctac agtgtactta 46320

cataaaataa ataaataaat ctttaaaaaa aaacacctat aacttaaact tatcaataac 46380

tttaactttc ctaccccatg cttcctagtt acccattctg ctttctgttt gtatgatcct 46440

gggtatggca tcttaatgga accacagtgt ttgactttgt atctacttaa tattaggcat 46500

gatgcctctg actctcatcc ctgatatagc acagttcaaa attgcctttc tttggtgctg 46560

tacatatagc tgagcgtttg agtgcttccc tgcatgcaca ggtttctgaa ttcaatcccc 46620

agcacaaaaa atgataaaaa gaaagcaaaa aggcttattt ttacagctgg acagatcatc 46680

ctgcattgtg cctgtcatgt tttgcttgtt tcttctgtca gtggacactg tgttacttct 46740

accttttggt tgttgtcagg aatattgtaa acatgagtga atatacaccc agaagtacaa 46800

ctggatgtgg taattctatg agtgttttgt tttttgaggg atggttatta ttgtttccat 46860

acaataaatt acatttcctt acagttcatt acatttccaa aagccatgca tagcatttct 46920

gttgttctac attcttattg acaccagttt tcaatttaca tttattttgt gagtttttta 46980

attggtaacc atcataatgg acataaaaaa tagctcattg tagttttggt atttgtattt 47040

cagtaatgct tggtgtgatt atctttttat attcttatta accattagtg tgtatctttt 47100

tttggaaaaa cacctcttca agggttttac tatgtagctc tggctggcct ggaacttgtg 47160

cagaccaggc ttgcctccgg ttcccactgt cttaggtagg tttccattgc tgtgaagagg 47220

caccatgacc agagcaactc ttacgaagga catttaattg gggctggctt acagtttcag 47280

aggtttaatc cattatcatc atggcaggaa gcatggcagc atccaggcag atgtggtgct 47340

ggaggagccg agagagttct atatcttgat tcaaaaatag ccaggaaaag actgtctaca 47400

gcaggcaacc aggaggagac tgtcttccat attgggcaga acttgagcac taggagtgtt 47460

ccaaagccac ctacacagtg acacagtaca tccaaaaagg ccacacctat tccaacaagg 47520

ccacacctcc taatagttct acttctcatg ggccaagcat actcaaacca ctacatccac 47580

ctacttctgt ctcccgaatg ctgggattaa aggcatatgt tgccattacc caattttaaa 47640

ccagattatt attgtttttt tgtacaacag acttttaagg ttaaagtttg cagcaatagg 47700

cattctttga agctgtatca cactgatata tgtctgttgt tttcttcctt cctagattaa 47760

aatagtacag tatattcaag tttcaattgt ccctttccat aagaagtcct ggtttctgtt 47820

ccattattag tttatatctt agtgttctta agtaaaaata ctcagtattt atagatgagt 47880

tagattagag ccaaacccca atcagggtat tggtaatgaa ggtttgctgg ataattcaaa 47940

ggatactgca aagatctggt ttctaatgga aagaacatgt aagttggcca ttagtggacc 48000

acacatctgt atttcttatt ctttggaacc ttgggcagga tagacagatg agctaagatt 48060

ccttcatagc tattgaattt gtgagaaaaa caaattgtgt ttccagaaac ctgctttagt 48120

ttgtatcaac acttactttc tttctgtgtg tggtgtgtgt gatgtgcctg taccattttc 48180

aagtttttct tccttctttc catagatacc cttctcattg actatcagtt cacctttagc 48240

ctgtcccagg aagacgaccg ctactacaca gccatcaact ttgtggctac tcctgatgaa 48300

gtaagctttt cttttaagct gtcttatttt gtgttaaatt ttgtataggt ttttttcttg 48360

gtcatcctgg acaaaagtac tacatagaag cagacagtat cagggtggga atataaaagg 48420

caaccagttt ttaagtattt ttttatttac ttgttgacag ttttatatga ttatataatg 48480

tgcttgatga tattcaacct gtgacctttt gtctccctca tacttagttc cttctctccc 48540

caccaagtca ccttcactcc ctctcgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg 48600

tgtgtgtgag agagagagag agagagagag agagagagag agagagagaa agacagacag 48660

acagatagac agagagacag agattgattg attgattgat tgattgattg attgattgat 48720

ttacctacct agtttaccag ctgactgcag gagcatgctg ggtgggaagt tcttactgga 48780

gcatagacac attacagtga ctacaccact gaagaaagtg actccctctc aggtagtctt 48840

cactgccact aggtcctcag ggatcaagag aatgtttgga gtctacattt tatctttttt 48900

ccactcagaa ggcaaacatt actgaatgtt tttaagtagt agaataatgt tcatgatagt 48960

ctgtttaata ttaaattaag aatttgttcc taattataaa atttttagaa gatagacaag 49020

aagacaaaat ttttgagtta acagtttgaa aggtttattt ttattttatt ttatatgtat 49080

gaatatttta gcttcttgta tccctgtgca tcatgtgtgt gcagtgcctg tggaggccag 49140

aaatagatat tggatccctg gaactagagt gatagatcat tgtgagccat catatgggtg 49200

ctagaaccaa cccagggtcc tctgcaagag cagtgagtgc tcttaactgc taggccattt 49260

ctttagcccc taaatgtgaa caactcttta aataaatgta agtgatctta aatactctgg 49320

agaaaaatct gtagctatac cttacttttt aaaaattatt ttgttttata ttatgagtgt 49380

tttgcctaca tatatgtgtg tctgatgcct gcagaggtca gaagagggtg ttggatcccc 49440

tagaactggg gttacagatg gctgtgagca gctatgtggt gcctgggagt tgaaccctgg 49500

ttctctgtta gggcaacaac tgcttttaac catcaaccca tctctttggc acatgggtgc 49560

attgttggtt tggctgcttg agttgtgtgt gaggggtgtg tgtgcataca tatgtgggtc 49620

catgcttatc cagtggaggc cagaggtcag agtcatgtat ctctctgtta ctttctacct 49680

tatgttttgg aagcaagatt agatagaccc ctgggacctt cctgtcttct cctcagcact 49740

aggactacaa gtccacacct gactttttac atggggcttc agatctaact cagtcccaac 49800

acttgtttca tttccttagc accttggcta gattcttagg attttagaag gagcttatag 49860

caaaatacca caagtgaaat ttactactgc cttagtcata agcaaatatt gaaggctcag 49920

tctttaaggg tataattgat agtgttcttt ttttttttaa gtaaacaaat agcctgtcat 49980

ggtaactatc gctgtagtcc cattacttgt gagagatgtc agctcaaggc cagcctccgc 50040

tacataagta agggaagacc agcctgagct atatgggact ctatcaaaac aaataaacat 50100

tgtagaattt ttgtaatact tattagaagg tagctgatga tcatgagagt ctttagacat 50160

ttcttcattc cactgttttg tgtgtgtgtg tttcatgaca gatttcttac tagatttatc 50220

tctttgtgtg tgtgtgtgtg tgtgtgtgtt ttacaaaatg acaaagattt tagtccttct 50280

cgtggaaagt agttgctagt ggtcagcaga tacttgctag tataaataaa tgagcataga 50340

tctgcgcttg caaaggaaga caaagggaaa aaaggttttc ttgaacataa ttcctacttt 50400

gtgaaagaaa cttctcattt ggaaattaca ttttgaaaat aggtattgtg aatgtttcca 50460

ttgtggtttg tggtataact atcaaataac acttttttaa aaagaaaaat cttaattttc 50520

taagattttt aaataccctt ttaaaatgag catttccagc atggtttgat taatttgtaa 50580

aatgtaagaa tatagtatct aaggctacag aaatgactca gtggttaaga gcactggctg 50640

ctttacagag gacccaggtt ccatccccag caccctcatg acagttcaca gccatctgta 50700

tttctagttc cagggcatct gatgcccttc tctgattttc tccagtacta gtgacacaca 50760

gcatacattt gaacaaaacc actgatacac ataaaataaa ttgttttcaa gaaacaatat 50820

agcatctaat tagcttacaa aactaattat ttgtttctgt actaattacg tttctattgg 50880

catgactaag gcaacttata agagaaagca tttaatttgg ggttcacact tctagtgcct 50940

tagattctat gagcatcatg gtagggagtg tggcagtagg caggcaggca tggtgctgga 51000

gcagaggctg agagctcaca tttgattttc tactagaaga cacagagaga gctaactgga 51060

aaaggcatgg gcttttcaaa cctcaaagcc cccctctagg aacacacacc tccaccaagg 51120

ccatacctcc taatcaaaca gtcctaccaa ctgaggacta accattcaga gatagatgag 51180

tctatggagg ccattgtcat ccaaaccacc acaggcccca agaaagattt gttagtgaaa 51240

tttcagtgaa aactaaaaca gcattagaat ttacctggca tagccagcaa tgatctcttc 51300

tgttcagtgc cacagatttc tttgagttaa aactcagttg ttaaaaccaa aaatcaaaat 51360

gtaattggca ctttaaattg ctataagggg aaacaaggtt ttcaaagcca tgaaaccata 51420

ttcagaataa ttttagcgag agaaatattt tttctttttt ttgtcgtttc ttttttttct 51480

tggagagaaa tatttttatt attttatatt attttaatta catatttaat tattaaccat 51540

ttctgacaga gggcaaaagg tgaggatctt catggaacta atatctgata aagcaccaaa 51600

ttcttcccaa ctctgggatg caaatgacag ttcaacttca gtttattgct tgtattgaag 51660

aaaattgaca agaaatgtca tgtcttaaca taagcatgga tttcttttaa gatgtagaat 51720

agtctataat taatgttttt gagactagta agacctgatt attgttgtat cttaaaatct 51780

agaaggtact aacaattttc taatgtgtat tttttttttc atcagcaaaa cagggatttg 51840

gacatgttca tcaatgcctc caaaaacttc aacctcaaca tcacctgggc caccagcttc 51900

ccaggtacag acacacctag agagatggat tggcaagttt agtgtaggag ttggggaagg 51960

aggctctgaa ggctggtgag tgagttcaga gcccacctct gcctcttagt agccatggca 52020

ccttgaacaa gccatgcttg aacaagcatg tacaattccc tctctacctt aggctactca 52080

gagtgaggag tcacagctct tgcctccagc gttgctggtt caggttggtt ggatggctgc 52140

tccctgcttt gccaccacct tccagcacta tgactatctc tatgtttgtg cttcacaggg 52200

gaaaaactaa agtgactcat agttttaaga aatgaaaact ctttaaggga agggggataa 52260

ctctaatatg tagaggtatt catactttgg gataactcct aaaagtacag cttttccatt 52320

cttgtttatc ttatagtgac tataaaattc tgatggccct aatgtagcag ttactataaa 52380

taaccactcc ataacttgat agccctgaag atagacctag gtttgaattt acctgcacgg 52440

tgttgaacaa gttactgaag ctttcttttc tttgtttttt aagtttgttt tattttatgt 52500

gtgtgtttgc ctttgcctgt atgtgtataa gtgtaccatg tatgtgcagt gcttgagaag 52560

gtcagaagag gacatcagct ccccaccctc aacgagttac agacaattat gaactactat 52620

atctgtgctg gcaacagaac ccaggtcttc tgaaagagca accagtgctc ttaactgctg 52680

agccatctct ctctagcccc caagttacct aaactttctg atccagtttc cttctttata 52740

aaatgataca gtgaaaatag ctttgctatg tacagagata ttccaacttt ttaatattac 52800

aacatgacat ctacaaatat gttagccctc attcataatc ttgcctgaat tgtagagtgt 52860

tgcaaggaat aaatgaaata aaggaggtac ttattataga gtttgaggtt tgccttcatg 52920

cataaagaga agcttttttg agtctgtact actcatgttc ttagccaatg gagtatataa 52980

aatatggtag aaccatttag aaatggagtc tcactgggta caggcctgaa tgcagtggta 53040

gcaggtagca gaaagaaggc ctgagtggct gcttgagcac cttctccatc aagacttgag 53100

gacctttctg cttaggaagt gatgagcgag taagtgtccc tgaacaggag ccttgagcat 53160

attctacagt gtgaagcaga aatacaaagg agttgaggta tcatgtgcaa aatgaatgca 53220

gtgtctgttt tatatgtatg attgttttac atacatgtat gtctgtgcat cgcttatata 53280

tctggagcct ctgagacaga ttacttaatc tattgggact tgagtttttc caatctgtag 53340

atggagatag gaaggtgttg tgtgggttag agactgaagc tcataaggct atattctttt 53400

gacactgtaa gtgctcaata aacttttacc ctcattacta gtgcgcaaag attctttctg 53460

attggcatac ccgcctccca agtctttatt tttattcttg cttctttcta gccggaaccc 53520

agactggaga agaggtgcct gttgtttcaa aaaccaacat caaggaatac aaagatagct 53580

tctctaatga gaaatttgat tttcgcaacc atccaaacat cactttcttt gtttatgtca 53640

gtaatttcac ttggcccatc aaaattcagg taagaactgc tttttaactt cattcccgta 53700

aagatggtga catctcttta gtggagacta acttcactca tttggaatct gtggtgactg 53760

aaagatagtg ttgctttgcc tttgagggat ctttgccata gactgagtag caggtgagtg 53820

ctgttcttag gttggagaga tgttcagtga gtggagtgct tgctacacaa gcctgaggac 53880

atgcagttca tctgcagcct ctcatacaaa gcgggacacg cagggtgtgc ctgtcacctc 53940

agcactggac atgcagtgtg tgcctgtcac cccagcacag gacacgcagg gtgtgcctgt 54000

cacctcagca ctggacatgc agtgtgtgcc tgtcacccca gcacaggaca cgcagtgtgt 54060

gcctgtcacc ccagcactgg acacgcagtg tgtgcctgtc accccagcac tgggaagcag 54120

gggacagaaa gatcttgctt gctggccagc cactcaaagc tggatctgtg agttctagat 54180

tcagttagag accctgtctc aagtaaaata aggtagagag gaattgagga agacacctga 54240

ttacctctgg cttctgtatg catgtgcaca tatatatacc ttcacacata tacacactca 54300

gagaaaaaat tctgagagtg tcatatcact tgtgaagaaa gttttaaagc acttttaaaa 54360

gcaagatgaa agctatgcaa ggtatgcaag gtagtatact tttgtaatcc caggatgtgg 54420

aagaccaatg caggaggatc accctgagtt tgaggccata ggaagaccct gcctcaaaag 54480

gagggaagga gggagggagg gagagagaga aagagaaaga gaaagagaga gagaaagaga 54540

aagagaaaaa gaaagaaaga aagaaagaag gaaggaagga gaaagaaaat caaattgatt 54600

ggcatatagt tatgtgttta ttttttgagt aattgctatg taaaagcctt tagaaataca 54660

cagttttaat tatggaattg agtataaata aaacaagtac atgtttgtaa ccaataaagt 54720

ataaaaatga cacataagat gtcaaagtgg tatgatggct ataatgtgga gtccatagag 54780

gaagcagtag gcagtatgag gtactgtgta aaaacacata gctttactat tgcacagaca 54840

agtgtggatt cttgttctgt gtgtggttca tggaggctct ccagtttgca gattctctgt 54900

gcatgtgtcc tgaaggattg gtcttcctgc tatgacctct ggtgttatta gcctgaactg 54960

agtcctaagg agacaggtag tggaaatgtt tgtattgcaa agacagtatg ggtagttgtt 55020

tttagaaaca ggagttcaac agaattgata gaacttgtga tcaagaagct aacagctgga 55080

ctgggatgta gctcagttga aagaacgctt gtctaacatt aagaagccct gggtaccatc 55140

actaccacag cataaactga gagtagtgac agactcatgt gtcccagcac tgggaaggta 55200

gaggtaggag gatcagaggc tgcccaggga ggttgagagt gacttacgct aggagataga 55260

tctaaaaatg aaaaggaaaa agaacttggt agctgctaga gctaccatga agagagtgga 55320

gcttaaggat tcagctgaag aatgtaaact gccttctgat gacaactgag agtcgctgag 55380

ttatttaaag tcaggaagtg aacaaagatc agtgtttcag aaagacctct gtggcaacag 55440

tattgactag aagtagcccc tcctatgtca ggtactggtt tagactgtat ttggaagtgt 55500

cctctttctt gatggccctc agacaccttt catggccact cctctgcatt tgtaccccat 55560

agccacacac ttgatggttc tttattacat aaatagctcc ttataggcaa tgatagattt 55620

tatatttttg ataattttaa gataaactct atgtcattgc atagaattta gtagttgtag 55680

gtactcagta aatgtatata ggatgaatac aaaagcttta gggtaacagt attttgttct 55740

tcttcccccg catttttaac tatctcatag tagcacagac taacccataa ctgaccatga 55800

agccaaggat gaccttgaac tcctgtacct tctacctctt ccccgaaagt gctgaagtta 55860

ctggcatgtg ctgctcaccc aactaatagc aagtttttct tataaaggtg ctgatgccct 55920

ttccctgttt gtgttaattg ctgacactta aaagctcttt atcccaaccc acagtgttaa 55980

agagtttagt taaattttgt ggaaattttg tcccaaatga agtggttgat ggcaggcctg 56040

gtggctcctt cctataattc caacactcag gagacagagt caggacgatg gccaagaatt 56100

caaggccttg ggcctacaga gtagaagaga gaagaatgag gattcgaaca cctgattaaa 56160

tagataccat ttcctgctac caacctgtgc cttagctact cttctattgc cgtgacaaaa 56220

catcataccc aaggcagctt ataaaagaaa gcatttatta ggactcacag tttcaagggt 56280

tatactccaa aaccatcatg gccgggagca ggcagcaggc aggaacatct gctgtgagga 56340

agagctgaga gctcacttct ttatccacaa ataggaggca gagagaaagc taactaggaa 56400

tagaatgagc tttgcagacc tcaaagccca cctccttccc aaacatttcc accaattggg 56460

aactaagtat tctaatctgt gagcctctgg aggcccattc ttatttaaac taccacactt 56520

tataagttaa tactacatgt gatgaggaaa ctggtatggg aattctgaaa agtagttcac 56580

aggagtggga ggggctgaac gtgagtagat gctagcatgt gtgtcaggag tgaagtgttc 56640

agagcattgc ctggtttgac ttctctccag agctgaggtg aacatgcttt gtgccaatac 56700

aaacccgtat taaagcggtg gtagttactg aaaatcagtg cagggctgtg gtctcaacac 56760

aatgtttgaa aaagaaaaca gggcatccac atcaggcagt gtacagctgc ttataattcc 56820

agtcctctgg cctctgctca catgcacata cccccccata catacacaca tgattaaaca 56880

taatgaaaaa ttaaaaatta atgctataaa aatggaaaga gccgggcgtg gtggtgcatg 56940

cctttaatcc cagcacttgg gaggcagagg caggcggatt tctgagttcg aggccagcct 57000

gatctacaga gtgagttcca gtacagctag ggctacacag agaaaccctg tctcgaaaaa 57060

caaaaacaaa aacaaaaaca aaaaaaaaag tggaaagaaa ggttcactgt ttcacaggaa 57120

aactctgaga ggtgataatc caatcccagt ttaaaatata ctccatagtg cacacagcct 57180

ctcccatcct tggcaactga ggcctgtgag aagactcagt cctctcctgg cttccaacct 57240

tacagtgttc aaaactcttc tgcaagatcc acatggtcct accaagaccc tgaaggtcag 57300

gcatgctgat taggctgtct ctgggcctga agtgaaaggt aaacacttcc gagatctcca 57360

aagccttggg aagattctga aatgtatggg tgttggttca ggtagactct cagccttggt 57420

gaagctgccc ccggagctgt agggttatct gcagaaagtc agccaggtgc acttaccctg 57480

gaatcctctc ccattcacag acacctccct gaggctttgt ggcttcacct cactgtgcag 57540

ctagctcctg ttttacatgc ttatataatg aatggtcttg gtaaagaaga tgataaaggc 57600

aagctagagg cctttttttt cccctcttca aattttgatt ggcctttccc tactgttaca 57660

ctgtctactc aaggttttga gcatttactt tgtgtacata gtaaaagcaa agtacatatt 57720

tttaagtaga aaagaaagca tctgtggtct ttgatatagg tgcttttctt tattttaata 57780

gtaatactta ttccatgctt gttaagaaat tcattcacag cgtgttttca tagagacttt 57840

ctctatagag atatatagaa atctagacat gaggacagcc cactaaccca ctcttcagac 57900

actagctgct tctcttagag ccctgggctc tcaccctttg gaggacagcc atcctcactc 57960

atatgtgaca agcttagaca cagaataatc acagagactc cagcctcccc cacaaaccca 58020

caatgccaat atcccatatt cccaggaact tttaataagc catccactct aatactccat 58080

ctcttatctc aggcataggc cctggttttg gtttgcttca gagtactgcc ttttctctac 58140

cacgcccttc ccactctttg ctgaccctcc agagatgtca tttccaaatg aagggggttt 58200

ttggttctgt gggtgttttg tttttcagtg cagttcctta actgctattc aggggacgga 58260

gcaggcaaac cagatctcta acttctgagg cctgtgaaga gaagcatcag aacctcccag 58320

gggagctgta ggagcaggag tcaggcctag atatgactgt gagagagtgg ggaccattac 58380

cagtgtctta caaatgaggg gaaggactac cgtgctgggc cctgaaagat aaggaggacc 58440

aggcttcagg aaggtaggac acattctgct gactgtctgg gattgaggac agtaacacaa 58500

ctacttagac atactttgaa tgaaggacag acttagtgct tcagaactgt aaatccatta 58560

tatctttccc aagtcttagg ctagccaagt ttctcaacat ttatctacct catcccaaag 58620

ggttcccagg acaaatattt cttactcaaa catttgatgg gagttggaat caggttgagg 58680

aaatgcaggg gtgtagattt tagatttctg ggaatatgta tagatagcta ccttctgttg 58740

gatagaaaat gagattgtaa gtttttcagt gtttttttac acgagtttgt gtgcccatgt 58800

atgcacatgt ggaggccacg ggtctacctt aggtgtcttc ttcaggaacc agccatctta 58860

tttttaagat gatctctctc cagacctcag ggctatcaac acacctcagg gatccatcct 58920

cctgactgta tgtccctagc atttgggtta ctgtaccacc atgctcaggt ctttgtgtag 58980

gtcctgggga tcacagttag gttctcatac tgcagggcaa gcactttgta aacaactatc 59040

tcccctgcat atggaagtat taccactaaa ttacaacaag attttcttct attaaaatta 59100

tattttagaa gctggatata gtaatgcgtt ggggcaaaag gagggaggga aatgaagagg 59160

ataggaagag ggggagggag aagggaaaga gtggaggcgg gatcagaagt ccaatgttat 59220

tcaagggcag cctgacctag ataaatccct attaaaaagt tttcagtata gaaacttctc 59280

atcaccttca ttatcagaaa agcccctaaa ttcagaacac tttttaatct taattagttg 59340

acaatttcat aaatgtatta tttatatata tgaataacat tttcctccta cctttttttc 59400

ccttcccctc tgatgattcc catcctccca accaagcccc ccttctgcat ttgtttgttg 59460

ctttaatgac ccactgagtt ccattgggct cacttccatg agtgtgacta gaagagctat 59520

ttatcagaat gtgggcaact taccagtagt gacactgatg aagaaagtgt ttccctctta 59580

cccagtaacc attaatggcc aggagctcct gggaggggtg ggcgccttat gagccccttc 59640

tccaaaatgc tttcaaactg tgaccagcta tatttaatgt ttttattatg cctgtgtatc 59700

catgtgggac aagaaagctt gagagtatca tagcatgcat gtggaggtca aagaacaact 59760

gtgtaaagtc agatctcact tcccaccttc acatgggctc tggcactgaa ctcatgtcag 59820

tgacctgaga ggcactttat cctctaacac gcaccctgtg cccagcctaa aatttgacct 59880

ttgcaaggtt tagtgtgtgt tatctgactg tctgagtaag gatgacaaaa tgaaaccaaa 59940

cttatgggat aaagcttggt ggttgtatca gtacattttt attgttgtga taaaacatta 60000

tgaccaagac agcttataga agagtttatt tgggtgtata gttccagaga ggtaagagtc 60060

tgtcctgaca aggaagctgt ggcagcaagt ggcaggtatg gctacaggag caggaagcag 60120

aaagagcaaa ctagaaacag ttgaggtttt ttaataggaa agcccactcc cctaatgatg 60180

tccttcccct agcagaccac aagtcctaac cctccctaca cagcaccacc agctggggag 60240

ttcaaatgtc tgggactgca ggggacatct cattcagacc acctcagtgg gagaatgctt 60300

gccttcatag tatgtgcaag gccctaggtt caattctagc caagaaaaga gaacatgagg 60360

aaagaaaaga aggtgggaga gagtagagaa agaagagaag aagaggaaaa aggaagggaa 60420

gggggagaca gaggaaagca gggaagcaga ggagaggaga agagaaagaa aagattaacc 60480

agcctggttt ttaatagcac ccctcccact ctcagtagtt cccaatttga gcattaagtt 60540

caagactgat agatatttct gggtgggtga ccagtgtggt cataaacatg gtgacttttg 60600

ctctccgtac aacttgtgat tatgaacttg ttagatgatc agcttcaaca ggagagggcc 60660

tcctttagtc tcaggtgccc cctccagcca ccctgggact cgcagcctct ctgtgatgag 60720

acacaggaca ttaactggta tggttctgct ttgccaaaac gtcagtccat ggttgaactc 60780

tccacaatga gaaagaagct ttgagaatca ttacatggca tcaggcaagc caggactgat 60840

ggagcctgag aaagggccag gagcatccgc aggttttggc acccagtact aactagtaaa 60900

agcacctcat aggtttcttt aaaatgcaaa cactaaggaa aatctaactt ttttttattt 60960

attaaggcca ttcattttat tttataagta ttttgcctgt atacatatgt accacatgca 61020

tacaaggtca aaagatagta ttgggtcttc gaactggagg tacagatgat tgtgagctgc 61080

catgtggatc ctcgaaattg aacctaggtc gtctacaaga gcaggaagtg ctcttaactt 61140

ctgagccatc tctccagctc cagaaaagct actcataaaa gtcaaatcta agccatgtgt 61200

ctggtgatgt acacctttaa ttgtagcaca tggaaggcgg aagtaggcgg attgttattc 61260

atccaaggcc agtcttctct taacagtgac aaaaacaaaa ccaaacccga aacctgttac 61320

tttgcacttt agagtataag tgatagagaa aagacacaga aattttagaa tctatacctt 61380

aaaatacctt atggcttata tgatactgtt gggaccatat ttacttatgg aatgcaaaaa 61440

aaaaaaaaaa aaaaaaagat ggggggggag ctgaaggtct cctttctatt ctgttgtaaa 61500

tctagctata aaaagagtaa gaggcatgag tgtgtctcag tggtagagca cctgcttagc 61560

ttgtgtggga ttgaatgatc ctcagcacca cagaagaagg gtggggcaat aaatttagga 61620

aaataagatg ctaatcattg actttcttga tttttttaaa aaaagttatt attttatgtt 61680

tattgtatat gtttatattt tctatgtgtg tttttgatgt gtgctggagg gatgggggcc 61740

acttgctgaa cttcccaatt gttatcataa ctaccatctt tagtgaaaca gttaccatct 61800

acttagtaat tgtttcattc gaatagatac tgaacactct taatctgaaa ctaatgctca 61860

gaaagttcca ctttgccaag caagcaggat aatgtaagcc tataatttta gcactgggag 61920

ggtgaggcag aattgtgagc tcaagggcac cctgagcttt tgagatcctg tctcaaataa 61980

aattaaatta tatagatatc agattttcag aataggtgtg ttcagctgct gaataaatct 62040

aagcaaatat ccccaaagaa ccctgaaatc tgaaacgtat tagttctaag ccctatgttg 62100

tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg taagatagat 62160

tctcactatg taaccctagc ttgcctggat cttgctatat agagagacca ggcatatgct 62220

atcgtgcctg ggagtcccaa acgttttaga tgaaagattt cagttgtacc attatcttcc 62280

taatgagggc tctggtctag tgaggcaggt gacattaggc cagtagtaag tattaggaat 62340

tggtgatgac ggtcaattct gagacacaca gtagatacat ctaatctacc aatacaacca 62400

atgatttaga aagaattagg ccatagttaa atttgcagtg tttttcttct ccacaaaata 62460

atgttacttc tttcagttct tagttcaaat acagtaggaa ttttttatat tcttggtgct 62520

aaacactatt attttatagt aaagttagta agatagaaat gacgccctgt gggttgtctg 62580

gtcgtagtct gtagctgagg ccattttgct gagaagcagc gtaggctgtc actggctttg 62640

tcacccatat tttctgtatt tttgctgcag attgccttct cccagcacag caacttcatg 62700

gacctggtac agttcttcgt gactttcttc aggtaatttc tctatgctaa ttgtacacat 62760

tccatcgaga cagtccctta actgcagctt gctttgtata tccctacaaa gctgcttttc 62820

actcacagtg atgtaaattt agtctgatgt gataaaactc tccgtttgta tgattcggct 62880

ctttgcatgg ggagaggttt gggctcaagc agttattaat aatatagcta ctgctgtgag 62940

ctacatgtct taatctgtct taatcaagat atgactgtga ttttccatag ggaaaggtaa 63000

ggtttacttg caaactcctg gggttctcct ttttttatag ttttcttatt agtagggttt 63060

tttttttttt tgagaatact atgcagaaat gattgaaaag aacaaattag tcattgcata 63120

ttggtaagag aagcagcaag agccacctca cctccctctg ctctccccaa atagaaactg 63180

ctctgctgtg ctgcttctct accttcacac caatgctcgg cctgccaact cagttatctt 63240

tcctttcctt ttaagatagg gtctctcctt atagtagtta tgactgtcct ggaattctaa 63300

atagaagagg ttggctttca aatcacagat cctcctgcct ctgccttctg agtactggaa 63360

gtatggtgta tgccaccgtg ccacagctaa ctcagttatt ttttggtgtt ctataactgc 63420

cttacataca tacagaccag gtacacacaa aattcctttc cattaattta atagttatat 63480

cacaatgcat tgaccaacta aaaaatccta aattgactta tgattctact tgctcatgtt 63540

ttaaaggaaa ggttactctt tgcttatctt aaatgtaata tttttccttt gcagttgctg 63600

tttaaatttt ccctataagt cgaccccaaa tttacatcta taatctggca aaacaaaaag 63660

acctctagtg atggttgtct cttagcttta gtctctcttg gactccattc cctccaccca 63720

taatgttcca tcctctgtcc ttaagtgtac tagtctccaa ggcctgctat gtggttgtca 63780

ttgttgtagt tacttttcta tgttgtgaca aagcaccctg acagtggcaa tttagaaagc 63840

atataatttg aggatcacag ttcctggtta gaatccatga ccatcttagc aaaggcagac 63900

aggcaggcct ggcactgaac aagtagctga gatcgtccat ctggtccaca agcataaggc 63960

agagaagcta attgggaatg gcatgggctt tggaaacctc agagtccact cttagtgata 64020

cctccttatc cttccaaaca gtattacaca ttcaaacttc aaatgtgtga gcctctgggg 64080

accactctca tttaaaccac cacagtgatc ttggcaactt cttttgtgtt cgtcccatgc 64140

cacagtcttt ccatgtattt ctccttttgc tggaactttt tccctcgaag gttcctgagg 64200

aaagaaacat agataacttt tgtatgtact tctacaactg aaagtatctt aatttttgcc 64260

ctaacaaatt tttgtttgct tacttgcttg cttacttgat tctgcgtgca tgcatttatt 64320

tgtttgtttg tttgtttgtt tgtttgagac aagatctctc tttgtagttc tggctgcctc 64380

aaactcagag agattcatct gcctctgcct ccagaatgct gggataaagg catgctccac 64440

catacctaat ccaacctcac aattttttaa gtgtgtattt atatgtgtgt gtggtatatg 64500

taaaggtgtg tgtgttcatg cacacatgtg cagagatcag aggagtcagg ttttctcatc 64560

tatcactctc tgccttatta ttttgagaca gggtctcttg ttcgatatta catatactag 64620

gtgagatagc ccaggagctt gtaggaattc tctcccattt ctaccttcca aatgtgtgct 64680

actgcatctg gctttaagca agttctggga atctgaggtc aggtccttac acctatgtag 64740

caactctgcc tactgagtca tcttactagt attcacaagg tcaaaggttg ggaccaacag 64800

ccaaggttgt cctcagatct ccacacagat gtacccacaa ttatacaaac actcaacata 64860

aacctattta cacacccaca tcacacgcac acacatacat gcacatacaa aaaaatgctt 64920

tttgaaagaa gtagagaatg ctagatatgg tattacacgt atataatcca agccactctg 64980

gaagctgagg caggaggatt tcaagtttga aaccagcttg accacataat tataccatgc 65040

ctcaaaaatt gtatagagaa taagaatgaa tatgaatgag actaaagtca tatctcagtt 65100

acttttctat tgctgtggca aaacaccatg acaaaggtaa tttacagaag agattattgg 65160

ggcatatagt ttcagagggt gagtccatga caattatgat atggcactga agtaatagct 65220

gagagcttaa atctggtcca caacattagg cagacagaga gctaactgga aatagccatg 65280

agattttgaa acctcaagcc ccactcctag tgatgtccca cacctcctaa tccttcccaa 65340

acagttccat cagctgggaa caagatattc aacatataag cctatggggg tcattctcat 65400

tcaaaccacc agtagtaatt attagagccc agcaaagaag gaagggatag aaagaaatga 65460

ttgatgggaa ctggggtgaa gtctgataca gagagatctt tatgtactgc agcgtagctc 65520

aggaagataa ctatggttaa ggacaattag ctaagtgatt agtagagagg attttaatat 65580

ttccaataca aagaaatgct gcaggcctga aatagggtac gtttcagtga cccagatctg 65640

attattacaa ctcatacact tgtaccaacc acataaatat gtacaataat tgtgtcagtt 65700

ttatattaaa taaaaatgtg gagcaagtta aaaaatgcct gttttaaact gatcacagtt 65760

atatgccagc ttttctttgc tgtgacaaaa taccataggg agtagtttat aaggaaagag 65820

atttcctcca gctcataatt ccagaatttt cagtctagag tcagttagtt ctatcatatt 65880

gggcccacag ctagaccaaa tacaatgatg gggagaatgt ggtaaagaaa agtatttacc 65940

tcagagtggt caggaggaac acaagacaaa atatacattt cagtcccata cctccagtga 66000

cttgcttcat ccaaacagac gccaccatcc aatagccatt aaaatacaag tcaaccagtt 66060

gattgacatc cattgatctt agtcatatcc ctaaattcaa cctctaagct ctgatgctct 66120

gggggccaag cctctattgc ataaatctct ggagcatatt tcataatatg aaatattaaa 66180

caggtctctc aggagctgtt tggtagactt agttgttttt tttttttttt tgtttaaggt 66240

ttttttggtt gggttttgtt ttgttttgtt ttgttttgtt tgtttgtttg tttgtttttt 66300

ttcgagaccg ggtttctctg tatagccctg gcggtcctgg aactcacttg tagaccaggc 66360

tggcctcgga ctcagaaatt tacctgcctc ctcctcccaa gtgctgggat taaaggtgtg 66420

cgacaccact gcctggccta gacttatttt tttaatcaga tttgagtctt tgcctctgga 66480

atcacagtag cttttcccat tcaacaccta gtttacagaa gaaagaaaac ccaatttttt 66540

tttttataat cattagacaa ctagaagttt tccctcctat taagaaaaca tattaacggg 66600

ctggcgagat ggctcagtgg gtaagagcac ccgactgctc ttcccaaggt ccagagttca 66660

aatcccagca accacatggt ggctcacaac catccgtaac gagatctgac tccctcttct 66720

ggagtgtctg aagacagcta cagtgtactt acatataatc aataaataaa tctttttaaa 66780

aaaaaaaaaa aagaaaaaga aaaagaaaac atattaacag tattgagaaa actgttggct 66840

taaatttgat gatttgaatt ttattttact aataaatgca tgtattgctg ggcatggcag 66900

cacatcccag cactcaggat tccgagataa gagatcataa gtccacgcta gctggaatag 66960

caaaataaaa tcttttttta aaaaatatac atacatacat acatacatac atacatacat 67020

acatacacac acacacacac ttttctcagt agtacggcca attagttgac ttgtctaacg 67080

gagggaggaa gaggaggcag agagcatgct gttcagatca cgttctcttt tgcattcagt 67140

ctgggacccc agcccatagt gtggtgctgc ccacattgat tattggtatt cagttaaccc 67200

agtgtagaaa ctctctcaga gacatgccca gatgcttgcc tcataaccac tgtgtatgta 67260

tatatgctta cagaaaatat actcatcatt acacataaat ttcatccact tacctcttat 67320

gaaaagttga ttatttactg agatttttct cattctgaaa atccataaag tctaccacat 67380

tgattaaatt acttgttttt tacctgttat tgctcatgtt agaattgctt tccttatttg 67440

gggtaagctg tcgttggcca ctgtgagggg cttatcaaga agtcagaaat gggaacacct 67500

tctaggaagt caggactgga agcttagctg agccagcaag tgtttctcac actgcacttc 67560

ctgtgagcct acctgtgcgg catcaggaac tggagttggg accttgagga ttgttccctg 67620

gaggcagggg tggagtcagg caggggtgaa gctgactcac aagatggtct tgcctttcag 67680

ttgtttcctc tcgctgcttc tggtggctgc agtggtctgg aagatcaagc agagctgttg 67740

ggcatccagg cggagagagg taagcccaag tagacaaact ccacataaaa ctcatttttt 67800

tccttctttc taggcagatc acttttacct gttgagtgat gactaatatt catatgagaa 67860

gcatgctgtt taacctgcat tctgtggttc cactatgtgc catcagtaga ttttaattat 67920

tcttgcataa agtgtcatta gttttgccac tgcttgattc aagtcttcct aagagtcttt 67980

cctaagaata tgagtgtaga gacaagttca gctcagtgac agagcacttg cctggcataa 68040

actgagtccc tggattctag tctcagcacc ctctaatagc acaacactag agacaaagct 68100

tctaacctgt gggtcttggg cagcaggtag gggaggggga tttaaaaaac aaaaacaaac 68160

ctctagctgt agcctgtgtc atttgttatg actaagcact agagtgggta ctagtagaca 68220

tgccatgtgg acattgagca cctctccatc ccaggcactg atccaggtgg ttctgcttta 68280

tcttcatctc caccctagga tataagggag gctacgtaac tacccatcac cacacagatg 68340

ctgaggtaca gaactgaggg gtaactagtg cctctgcctt cacagcacag gttcctaaac 68400

acgttttcta caaacacttc atttgttcta gtctgttcat ttaagaatct catgttctga 68460

ctgaatgagc tagacaactc accctagact atacattcta aagaagggca acaaggcagt 68520

tttgttactg ttgagaagaa aacaaagtta tttccgtatg agttattgag atagaatagt 68580

agagatttgt ctgaatacaa aatagaaagt atataaaagt atataagtgg atcataaaga 68640

aagcaacaat caactggaaa atatttgcag tatcatgaga gagagaaaac tagaagatga 68700

accccctcaa aaaaggattt ttaaaatatg cttagactgt attcagtcag ctaataaact 68760

ttttttacct ttatttggaa tttacgaata gcactgaacc tgaccattgt aaatgcacga 68820

ggtcaggcat gacttgttcc cagtaggaag ttgtttttag ttcttgctgt ggcctgggtc 68880

ctgatggaag ttctttaccc accttatctc ctgtcctctt ggcagaggtt ctagaatagt 68940

gctgtgatgg ggtagcaact gtcttcctgt gaccctgcac ctagattatt acagaaccca 69000

gactgggttt gctgagttaa tggaaattct ttctaggttc agtagagaga tgtgctgaca 69060

catactaggc catctagttt ttcagtaatg ctcagagacc gcaataggat atgtaacagc 69120

aacaaaattt ttaacataaa atttcccttc taaaacagag tgatgattta tgtagcttca 69180

ggatcctgcc tcctagaaga tggtttgaag caaggccagt ttgtcttccc tagcataacc 69240

tcagaagacc tctcatatta ttgatggtat aggaatgaat gcccacattc tgtatttgag 69300

atgtgtgcta tagtatctca tctgacccaa catgaaaaca tttcaagcca tgtgtgcttg 69360

ggtaaggtag gagttcaaag tcatccagtg agttcaaggc cagcctgggc tgcatgagac 69420

actgtctcat aaacagacac ttgaatctca tttaaagaag acattgaaga cttgatactt 69480

tgaacaccta tcctaacgta tccaccccca aatccagagt ccttcatgtt cttgtcctct 69540

gcagttccac tttcattgtg ttctcagcag cagctctctc cgaggagagt tgtctcccat 69600

cctatcagcc atctttttta ttgttgttgc tctgacaatg tctggttcag gttttaacac 69660

aaagcaagct agagtgattt taatctagca acaaaaatat aaaaaggtaa gtttttgccc 69720

ttttatatat tcaatcaaca gatatcatag cattatatcc tccactttaa cttttatttc 69780

ttactggtaa gggcttttta taaaaatata atagtgttac cacatgtaac aaaatttgat 69840

accttgtgct acctagcacc ttgtcatgtc cagttttcct cagctgtcac agaagcgaca 69900

ctgcatctga tcagtttgaa tcagagagag tgtagcatgt ctaatatcta gtattcacta 69960

ataaaatctc agtactaagc atattaataa tactatatta ttcattagca acttcttcgg 70020

gagatgcaac agatggccag ccgccccttt gcttctgtaa acgttgcctt ggaaacagat 70080

gaagaacctc ctgatctcat tgggggaagt ataaaggtga gaagtggctc aaaggtccat 70140

atagcttttc agaactcagg cctcagtttg ctaggctaca gacagcaagc gctctgtgtg 70200

tcactcctgt ctcctctcta acagttagtc agcagaagca accccgagcg accgtaaggg 70260

gctctgtgtg tggctttact tttcgagttg ttgcatgtca gattttaaca tgcaaattaa 70320

gcttgttatt cttactttgt ggcataatac tttatagttt ttatttggaa atatctaatc 70380

tgggctaggt gtgatggtgc acatctttaa tctcagttca gaggaggcag agacagaggc 70440

aggcaggatc tccttgagtt ctagaacagc tggtctacat aatgagaccc tatatgttag 70500

aaaaaaagaa agagggggtg ggggaaggca gctaacttta accattaatt gaaccaacac 70560

acacacattt tgttcagagc cccagtactc aattaaaagc caggcaggca tggtaacagt 70620

acttagggag tcagaaacag gattcccaga gtaagcagtc tgactaggct agcaggaaat 70680

ggtgagtttc aggttcagca agaggccctg cctcagtaag taaattgaag aacaactgag 70740

ggagacttgc atgtgcactt gtgcatgcac ccacacatgc acttgcacac ataccatatg 70800

tcaccatgct tagactataa aatgtagtca ctactggcag cacatgccta caatacagat 70860

gcaggagaat cactgcaaat ttgagatcag cctgggctac tggacaagat tttgtctcaa 70920

gaaaactaaa acaatacaaa agtgtactgg gggggttatt ctaatgccag tgtttatgac 70980

agcacattca gaactgacag taaaggcaat caaggactgt cagtggtggg tatatacata 71040

ggcagaggag caactgctac tagaaactgt ttatccttta aaagactaat gtatgctgca 71100

gcatagacaa acgttaagtt gtgttaagta aaagatgctg tatcattcca cttacccatc 71160

gagaataatc aaatacaaga cagagtaaaa tagtgactgc tagaggctta aaagaaaaga 71220

ccagggggtg gggaaaggga gggaaggaag tgggagaggg aggaagggag agagggaggg 71280

agggagccag actttgtggc ttacagcatc aagaggctga ggcagaaggg ttacaaattc 71340

aaggccctac tgggctacat agtgagaagt aggatttcct tgagctgtct ttctaggtca 71400

taatctctca ttgggggaag tcagggcagg gacttgaggc agaaaccatg gggaatgcta 71460

tttgctggct ccttcccagg ctcctctcta gctttgtttt ctcattttgt ttttactgtc 71520

tatgggtgtt ttacctgctt gtttttctgt gtaccatata catgcctgct acccacagag 71580

gcactgatgc ctggaactgg agttacagat ggttgcaggc tgccctgtga gtgctgggaa 71640

ctaaactcgg gtcctctaca tgagcaagtg ttcttaacca ttgagccatc tctccagcct 71700

ataaaattct tttttaaaaa taaagtctgc aacagaaaat gaatattttc tagagctgaa 71760

gcattcaatg agtggataaa gaatccattt gatgagctat ctacctttca caagctctta 71820

acccctacag actcaggact tagtggctgg aagatgaatg taaaacaggt agctctctcc 71880

ataatatctg gtctgtttgt gccaggtgtg cagaactgtg caacaggtca ccatacaaac 71940

cggcgtgggc ctttcctgac actcacacag ctctcgggac agtgcccgtg gggacctctt 72000

attgacctta taagcacctg actgtgcagt gtagcaggga gttaaggtgc ttctgttttc 72060

ttcctccaga ccgttcctaa gcccattgcc ctggagccct gctttggtaa caaagccgca 72120

gtcctctctg tattcgtgag gctccctcga ggactgggag gaatccctcc tcctggtcag 72180

tcaggtgagt agacaggaga caatgacaga tattggtctg tgaaggactg agtcttagac 72240

acttcttctg gtatagaacc tgggtctggg cacagtgctt agtggtacag agctttggtg 72300

gaacaattct atagtcccca aactgtgttc tgagcactga cattcctgtc ctggggtgga 72360

agttcaggac cttcctcacg gtgcacagcg tcctcagaca ttcatgctct ggtccccttg 72420

actctattga tccctgcttt cttttttttt ttaacccctt gttcttatct caaatttagg 72480

ctttttcttc cttgatacaa gctcctattc atctccatgc ctctggcttc cagccatgtc 72540

ctcaaagctt gtgttgccaa gtacagagtt ctagtcatgc tccacatctt cttaaggtct 72600

tgctatgcag ccttagctgg acgagtgctc gttataggcc aggcagtggt ggtacacgcc 72660

ttatgtccta gcactgagga ggcagaggca ggcagatctc tgagttcaag accaccctgg 72720

tctacagagt aaattccagg acaaccagag ctacataggg aaaccctgtc tcaaaaaaat 72780

aaaaacaaca acaggaacaa ccccaaaaac tcattatatt gcccaggctg gcttcaaact 72840

catagttatc ctcctacttc agcctccaaa gtgctgggat tatgggtgtg acccttcatg 72900

cccagattgt cttaaatatg aggcatgaag aagtattatg aaaacataaa ggatattttg 72960

aaaattataa ttctactggg ttaatgcaga tccattttca tttcattgaa ataatgatac 73020

agcctttgga ggttagggga gcctctcctg ttttcaaact gactttgaac ttctgatcat 73080

cccgccacca ccgccacctc ctcctcctcc tcctcctccc cagtgctgag atacatcact 73140

actcctggtt tatgtggcac agaggctcaa acccagggcc tcatgcatgc taggcagaca 73200

ctctaccagc caacctaccc acagctccta gatgtgcacc gtattacaaa catttattct 73260

tcagcatgtt tttttttttt tttcctaaaa atcatctcta caggaaacaa gtaccagtgg 73320

tgttttaggg caggaatagg aagaaaatat ttttactata tactcttttt ttttaatcat 73380

tttttagatt ttatttattt taaaatttat ttactattat taataagtac actgtagctg 73440

tcttcagaca acccagcaga gggcatcaga tctcattacg gatggttgtg agccaccacg 73500

tagttgctgg gatttgaact caggaccttt ggaagagcag tcagtgctct taactggtga 73560

gccatctctc cagcccctac tatatactct tttaaatgac ttatttgctt ttatttttat 73620

gtgcattggt aatctgcctg catgtatgtc tctgagagag gatcagattc cttggaattt 73680

gagttacctt gtgggtgctg ggaattgaac ccaggtcctc tggaagaaca gccagtgctc 73740

ataactgctg agccgtctct gcagccccta ctatatactt tttttatagt tttgaatttt 73800

tttttctttt tgggtattgc taaggatcaa atatagatct actatttatt ttttataaca 73860

tccattagta tttttataac ttactacata gtttgccaat tcttttatac atgtccatca 73920

aacatgtaag tcataattta tataaacctt gtgttaaagc tggaggcaca gaaggaagat 73980

tgctacagag tgaagtctag actagccagg gctatatagt gggaccctgt tgcaaagaaa 74040

aagttctctc tttaaacaca aaggcagtat gaaaagacat accttgattc tgaagctgtg 74100

cataggaatg cctcacacag tgttctgctc aggactatac tcagatgcag tggtctgagg 74160

gacttggtgg tgtctcagcc aaaataacct ggagtttagt aggaaagtct cctttatccg 74220

tgtccagtcc tgaagggaag ccttatttat gtatgatgag tcaggaccca ttgtcttcat 74280

cttacttggc atccccccag cactgagtct ctgagttagc cttacttgga cagagtgact 74340

ctctgggcac tctggacagc atctcctgct tcaaaagggc aagatcttta gaagacacag 74400

agatggagca ggtcttacat ggagatatag cagcttttcc ttcctgaccc ttgacccaat 74460

gcttctttgg aaatcctcat gaaaccctgc tcctttctgg agacccaccc cacagcaggg 74520

ttatccatgc caagcttcct gtactttctc tttttgagga agcacataca cacaaagttt 74580

tagtagctcg cacatctcac tgtgaagtag tgatactttc attgctatct tctggaaaca 74640

ggcaggagta ggcacacgct cagagcatag ctgcactctc attcacttgc caccctgagg 74700

cagagcacac gactttgtga tctgctatgg aggagagaga aatgagtagt taggtgtgta 74760

taaataagct aacaccatca cccctttatc tttcactagg gaaatgtaaa aagaaatctg 74820

aaattatttt gtaaaaaagt aagctgcttc atgacacatg tcccctcttg tgggttcttc 74880

caaggtctcg ctgtggccag tgccctggtg gacatttctc agcagatgcc aatagtgtac 74940

aaggagaagt caggagctgt aagaaaccgg aagcagcagc cgcctgcaca gcctggaacc 75000

tgcatttgat actggggcag gaattcgccc tcacagaggg cgtgtggtcc acgaagctgt 75060

ctacagggga ggctgcaggc aggaagcagg cgtggggcag aagactgggg acccttgaag 75120

cgtccaactc atgtgcatga tcatgcaagc tgttttcatg gctcacccct ctgtgtccag 75180

catctaacct tttacttctg tgtaggaaat aatttaatta caagtccagg aatggtctgc 75240

tctactcatg ggtggaggag accagtgccg accccgtgag agctgaaggt gatgctgagg 75300

tcccttgtgg aagcctctct tgggaatctc aactgcagag gagctgccct ctgtcagcag 75360

ctctccagca tggtcctctg acactcctca gatgaactgt tctcatcgga agcttgctgt 75420

ctttttacaa gatgagcttt tactctcttc caggaagtag ctttttttct agctgagaat 75480

taataatggt ctttctcttt ggaagtcata tcaaagtata attgatgggg gccttgtttt 75540

gttttgtttt ggtttttgga gacagggtct cactgtgtag tcctagctgg cctggaactc 75600

actatgtaga tcaggctgga ctgaactcac aaagatccac ctgcctctgc ctcacaaatg 75660

ctgggataaa aagcatgaac caccaggccc agcaaagagg gctattctaa atgtcaaggt 75720

caatggagtt agaatatata taaaaaaatg caattgataa ttctctatag aaacttgatt 75780

aattttaatc cattctttcc ttctctttct ctcactctgt cttacacaca tgcacacata 75840

cacacacact aagtgcctag actttgaata gatctagcaa ttggacatta gtaagcctaa 75900

gtttttacat gattgcattc ctacattctt gtaaacttta agtaactacc attgcagttt 75960

gttctttttt taaagtctaa tttgcagcca agaacgagta attctcaccc caagcaacat 76020

ctaataggga ctgagtgacc ccagcccagc ctagtgtcac tttaggcctg acgtttgagc 76080

aaccctcggc tcttgccaag gcaccacaga atgcacttgc tcatgccctg tgcctcttga 76140

gcagaaaaga gcactgacaa ctgggacacc tggctctgtc ttcctacagc tgctcgcact 76200

gacctgtggg aacctgtggg tcatccccag gctgaatgga gtacacacta gaagagggat 76260

gatgcctagc attggggcag catctgctca gcacatggaa agggacctgg ttccatctcc 76320

cctgggcagg agttggtcca gcctcctccc agacccagct ggtggctgtg aggaggtggg 76380

gaatgctaat gagaatgaaa agcacatggg ttgatgggaa gggacaagat taccacgtta 76440

ggagggtgag cagccctctg ctatgtgccc aggaccctgc ctggacattg catttcccca 76500

tttatggtgc tccgtattct ggcattatgc agcagcctca cacacctgtc ctctccttct 76560

tcatgtccta cagttctgct atcacctgac tagaatagcc ctctaggcaa cagtgctcaa 76620

atgtatgagt ttggagaagt taacaatcag aagaacaaaa actgtagtgt ttcaccttta 76680

aatgcagtgt tgaagaggga gcctttctct aagccctgca ctaacccact cctcccaaga 76740

ctcttgtgga gtgacagttc caagctgaac cataaatcac tgatgcacaa aacactgcta 76800

gaaggctcac ctctcaaaac acgactcttt gcatcactat taaagagcag aaagttctag 76860

aaatgatccc agcctcatcc cctatacagt taggagctcc ccacatctct accaaaaccc 76920

agcacataag tatctgcgtg gtctagcctt tcatctccgt aacaagccag gggactcttg 76980

gccaaaagaa agaaagggaa gttgcactag ggcttgtccg tccataagga attcccctct 77040

gctttgctca aaggaccaaa tttctttggc caaagaagtt gcttctatgt tagtcccata 77100

ccctgaagta atatgtacca tggctcccac ctacctgttt atgctctccc tgcccccagg 77160

gaaactgttt attctttcaa aagaagcaaa cagcgttcat ttctgctcct gtaatggaga 77220

aacagccagc tcccctgcat cccttacagc caacagctcc cttcaggctt agagcagggg 77280

gaatggcagg gattaagagc tcagctcaga gccagttacc aagatggaat ggagttgtga 77340

cccagtaact gtgtcacgag agaccatgta tataaaatag tcatgacgac actgacctct 77400

tgcacttgta cataactata ctgtagtgtc cagaatgttc agacattcag ggtgtacata 77460

aacagaagag tatcataatg tatttttatt aaacactaac atctgagttt cacctaatct 77520

gtttctgtgc catatactgg gtatccaagc tctgggaagt tatcctacca ggccctgatc 77580

tgttgataag gcactataca ccatgctggt gtgttctgta gccttgtgcc cattaggtaa 77640

ctgaacaatg attcagctct tagaatacct aggaagacag caagcagggt gacacacggc 77700

tgtgatctaa gcattcagaa gacagaggca ggaagaaaat tcaaaaatgg ggctggagag 77760

atggctcagt ggttaaaagc actggctgct cttggtcagg acactagttc agttcccagt 77820

acccacatgg tggctcacaa ccttctgtga ctacagttcc agataacctg acaccctcct 77880

ctggcctcct cgggtgcctg tggtggtcca cctggtgcac agacaaacac ccaatacaca 77940

caaaacaaaa gtaactcaag aatagcctgg gctacatagc aagagcctgt ctcaaaacaa 78000

acgaacctat gaagagccag gcagtctatc tatttacatg gcagtatact agagaaactc 78060

aggaagcaag agtgttcatc actgttgtaa tttcaaatgc tccttgtgat ttctggcatc 78120

tctgtggggt gaggtgttct gttactcttc acattcaaag actgtcaccc atgaacgtca 78180

gactttgcaa aggggctctc taagctgcac tgttgtggct ttgtctaaaa ttttaatgac 78240

gtttctgaga accatgttct ttttatacta aaatctgggg atgggagggc tcatttgttg 78300

ataaatagca ctattttccc acacctcagc ctcctgtccc cgtcctggtc ttccctacac 78360

agtctggaga gggctctgaa aggtccacag agtttgacag acacgaaagc aacccattgc 78420

cccgttgacc tgacctggaa gaagactgtc agcaaaagga aaataccaga atatctggaa 78480

agcttgaagt gtaagatggg atctcgttgg ggaattggat gaagaaaagc agagcgcctc 78540

tggtaggtga ctctgcagcc tgccagcgcc cgccctcttt ctacacagca gagtgtgcat 78600

ggcaaggaaa tgagtcacct ccttggggga tggtgctgtt tttatgaaaa cctctgatcc 78660

ttggtgtcct ttaattgatc tgttcaacaa atatttacta aacacttcta agctaacatt 78720

agggcagtga ctgaggtgga aacccagctc tttagacagc tgtcatccta ggatagcttc 78780

ctggaagcag aaccaagaag ccagaaggtt cttcctaggg tggccttggc tccctgaagg 78840

aatctgaaat gctgaccctg tcacaacctc ccagcacagc tttggaatga gacatcagcc 78900

tggcctccag cagagcagag gctctggagc tccacatcct gcctgcaggg agccctcagg 78960

gtgccctcca gagtacaggg agaaactaaa ggcaataaca gaagctgctc tcagagcctg 79020

actgtgcaca aaacactagt gaagcctgct gaactaattc tgcctctgga aatcttttct 79080

ggttctttac agtttgttgt tttgttttga tccaagctta gtttgttact atgtgtgatt 79140

tagcatctgt cgcacttgtg taaatatgga gtaagtattg taaactattt aattgctgcg 79200

attgttgggt tatacataca tttaggactg caattttttg gtattttttg tattgtaaaa 79260

taacagctaa tttcatcagg aacaagagaa ttaagggggt ctgcatttta aatgcagatg 79320

tgaagcactt gtatataaat aaaagtaaat actataatac aaagttcctt ctgaaataaa 79380

agtagatctg gtaaaaatgt gcgtgcgttt cgttctgaat gttcaatgct aattttgttt 79440

tattttatat ttacatttta gtccttattt tagcagtgag gagacaggca cagcagtgca 79500

ttctcacctt ggcagctgag gaatccccta gagtagactg caactcaaga ctcttggctt 79560

ccacactgaa aagagtttca gtttatgaag cagagtttag gaagtttagt gaggaattta 79620

aggacttctt ttaatgtttg tgtctacata tgtgggtaca tatatgacac agcatgcatg 79680

tggaaggcaa acaacacctt aatggaagtg gcctgaagaa caaactcagg acttcaatct 79740

tggcagcata aacctttacc taatgagtca tctccagtct atacggggtg tgtgtgtgaa 79800

cacatgtgca acagcacaca gtggaggtca gcacaactct tgcgagtcaa ttctccctta 79860

ccttgtaaga cctagaattc cacattgccc aggctctgaa agttaggttg ggtccacact 79920

gggccatggc tgatgaaatg ttggaaaagt gataacacca aacttttgca cagaaaatat 79980

tttcatctgg ggccttccct ggagttcaca ggctaaagtg ttggaaggaa catgggtccc 80040

tgagccacca ctttcacaaa aactacctga tcaagaagaa ctattctggg tttctgttgc 80100

taaaattcct tcccagagag aaatgtaagc aatgtctgcc ccttcaaggg tcccagcaag 80160

aaaccaaggc acaattccac caaagttcac tagaaaacca gtgagtttat tgggcttccg 80220

tgcagaacat acatgagggg ttacttagag aagtgtggat actcctcccc ctaacaatcc 80280

acaccctgaa aaagccttac ccagcaggga tgagggcttc cccagaccca cattgatggt 80340

gctcccattc catttttccc tggcatgcaa agagatagac agaaaaatag attatatata 80400

atatacacat aaattagaaa aatagattat ataatataca cataaattat atattatata 80460

tataatatat aatacacaga tagattatat atgatatata aaacacacag aaatagggta 80520

tatataatat ataatacaca aactactcag ctattaaaaa cagtggattc atgaaattct 80580

taggcaaatg gatggaacta gaaaatattc tgagtgaggt aacccaatca caaaagaaca 80640

cacatggtat gcactcactg ataagtggat attagcccag aagcttggaa tacccaagat 80700

accattcaca gaccacatga agctcaagaa aggaagatca acgtgtgggt gcttctgttc 80760

ttcttagagg aacaccctca taaagtagtg gtggggggtg gggggagaca gaataggtgg 80820

tttccaggag aggaggaaaa caggaaaggg aataaataac atttgaaatg taaataaaga 80880

aaatacccaa taataaaaga aaaagaattt tgaaacagag ggtaaaaaat aatacacaaa 80940

ccaggtagat agattatata taatatatat aacacagaga tagatagata gatagataga 81000

tagatagata gatagataga tagatagata gatagatagg tcaactgctc gcccctccac 81060

taggtaacat gcagttaagg cagagctgca tacaacagat gttagggata ctcaggtgag 81120

aatctcaggc tttgctccat ccatctatgc tggggtgtaa gctgtcaaca agtttagctg 81180

ggatgatgct ttgcaagagg gcacagctga atgccctaag atggtagatg cttggctcaa 81240

aggagacact acagctctgc atcaaggcaa actaactgag atgagggcct ttattttcca 81300

gatctgtatc ctggagcatc attcacctgt tactacactg aaaacatttg gtgttggttt 81360

catggcagat gacaggcagt gagagaagta cagcagcgga ctgctagagg tgggggttct 81420

gtcaggacgt gggaggctgt ttggttagta acttggaaag caacaagttt ttagctagag 81480

ggagaaaagc tggagataac tgtacttgct tgatttctta aatatcaaat tttattttat 81540

gcatatgggt attttgcttg catgtatggc tatacactac atgcttgtgg tgcccacaga 81600

gaccagagga agtagtgtga gcctctgaaa ctgaagttac agacattacg acttgagtgc 81660

ctgaaactga accttggtcc tctggaagaa cagccagggc tcctaaccac tgagctatct 81720

ctccagccct gacagaacat catgtactcc aggctggtct caaatttgct ttatagccaa 81780

gaacggtctt aaattctgat cctcctgttc tctcaagtag tggggttaca ggtctacact 81840

gccgttttct tgagcaaatc attacaaatt gagttctaag ccaggtgtaa tagttcatgt 81900

agtaacaatc tggaattttg gtctcttaaa aaaacaaata ttataagaat gtattttcat 81960

tttaatccca ggtgtatggc atatatcgaa ctgctttgga ctgactacag cagctatgat 82020

tttttcttgt tctagcagag gtatggtttt gccagctaca gatagtttct gtgattgtgt 82080

gacatttgga attctggaaa cttttcagat ggtatataaa tattagagcc ccaataggca 82140

gagttgatga ttgttggtca ttcaggggta ttggttgtgg ttagtagtct tgcttgaaga 82200

agaaacaaga acaaattaga ttcagagatc tctatatctc tctctatctt cctttctgtc 82260

ctatctagta atagggggta aaaccaggat gataaagggt tgggggaacc cacaaagtaa 82320

caaagactgg ctacaagtgg cacccaactt ggaactcaaa attgccatag aggaagcagc 82380

aggggatgaa ggaatggatt gtggctgttg ttgctgggat attcctcact ttgctcccag 82440

aggggatttt tctgaggttt tgttgttttg ctttgctttg ctttggtttt cttctacata 82500

ttctgttttt taagtaagtt gaaataatag ccgagaagct ggaaaagttt ggtgtggaaa 82560

tggagcagcc tgagaaacaa acaatgtatg aaatgggaaa actaaagggg ccactcttct 82620

ctcttttctg aaaggcttgc agacttggtg gtgcacctgg agagtttatg gatggagatg 82680

gaagctctta ggagacaaga agcatggaaa aagagaacaa aggctcagtc ccagtgactg 82740

aagagagcag gagttttcca aagaaggtgc atgggagggc cactggtcag aaaaaaaagg 82800

ctgaaaaata ccaaaggaca atgtgctgaa atagcccatt tcaagagaaa gggttcatct 82860

caaaccagca ttctgacaga gtggaaggag gggtggctca gggttatgag atcaccatca 82920

gcttttccag ttttcccata tagcatatgc ctgctaatgg tatggaacaa gagtaaggca 82980

aaataggatg gtgtcctata gaaatgatag ctctaaggtg tttttaaaag gccttgattt 83040

catatggaat gcactctcct tatgtggaac ggatattaaa taaccggggt acacaaacta 83100

gaatcccttc ccaagattgg aagggattgg taacagctgt actagagact gtcagccgtt 83160

gcaatggtta acatgctgga ggaaagaagc tgtgaacatt gaacagtaaa acagagcaag 83220

gggtattaat atagtgaacg aacagctgct aggtgaaggg cggtactcta gtgtacaagc 83280

acagactcgg tgtcatgaaa ctactataga acaaggttgc ctcagtggct ataacacctt 83340

gggacaaagg aggagccagg aaaaagtcca gttcatttac aaagattata taaggctctg 83400

gagaagcctt cactgatttt tttttttaca aagattagtc tcagctatga acaaagccat 83460

atcagaccct gacacaaggc aggtgttgat agagaccttg gtgtatgaca atgcaaatac 83520

caaatataaa aatgtcatta gacttttaaa ggcacaagta atgcctatgg atgagtggat 83580

aagggataag accaatatta gttctaatgt gtactgtgct aatatcattg atcaagctat 83640

agctagagat ctctgatgtc aaaatgcctt gtgcttcagt tgcagcaaat acagtaattt 83700

gcaaggagtc attgtggcca aaactaaagg tctcagatct caaaatgcct gatgctttgt 83760

gggaaatagg gtcatttgca acaaaaatgt gaacaagaca tctttaaggg caatggtttt 83820

tctaaatata aaccagaaag acggcctagg cttccaaggt tgtgctggcg atgtggccag 83880

ggttgccact ggaccaatga gtgtaggtcc aaaagagata ttcaaggtaa cgtattacca 83940

tcatgaaatg gtcttggggg cctatcttga ggccctgcag caaagagtat gagccattcc 84000

aaccagagag tggcatggag actcaaaacc ttcactgggc actggagatt taatgcacac 84060

tagctattgc aggcagcatg gctctagact tggccacaga taaacatctt gctctatccc 84120

ccaaaattca aagttataac atagctactg gagtgtatgg tctttttccc tcagggacag 84180

taaggataat cttgggaagg agtggattga cttcctaaga attcactgtg catcaggaag 84240

tatagatgaa tatttcaaag gagaaattaa aattgtggca tatgtaaagg tagagctgca 84300

acttaacaca ggcgataggg ttgctcagct gctgctgttt ccctatatca aaggcaaagc 84360

aactgcagca gaaagaggag aggcctgaaa accttgggca ctgacacaaa aattgcttat 84420

ttcattgaaa atgtctgttt ataacttccc actatacagc acaacaggag ggggcttaaa 84480

acataatggg gaaaatgtca caattctgca atttttgttt ccttaaaaaa aacacacaca 84540

cagaatttta ataatgtgtt ctcatcttaa tcccgggtgt gggaattagg gctgctttgg 84600

accattccca gcagctgact atgatttgcc tcatgctcta gcagaagtat gatttttgcc 84660

acctgcagat agtttctggg attgtgtgac atttggaatt ttgggaactt ttctgaaggt 84720

atataaatgc taaggccctg gtggggaggg ttggtggttg gtggtcattc aggggggtgg 84780

ttgtggttag tggtcttgct caaagaacaa acaagaaagt catttgattc agatgtatct 84840

ttcttccttc ccccactctt tctctcctcc ccccggcacc ctgccccctg ccccgacctc 84900

tacccttctt tttctatcta gtgacaagga tgaaaccagg gggataaagg gtgggaaaaa 84960

gaagagccca caaagtaact caggttggct acaagttcat gccaagaatc ctaggacctt 85020

gttgtttaaa ggcttgtttt attttgtgaa catgaatgtt aaatgtacat acatgttaag 85080

tgtatgtatg tacaccatat gcatgcatac agaatccaga agaaagtaca ttataccctg 85140

gaatggaact tagagttgtg agacagcatg aggatgctgg gaactgaacc cagtttctcc 85200

acaagaggag tagttgctct tcactgctta acctttcctc cagccccaat cctagcattt 85260

tggaggctga tgtaggaaga ttatcccaag tgtgaggtca tcttgggctc cataataagt 85320

ttaagaccaa tctcagctcc agagtaggac cctgcctcaa aaacacacag gtggaaagat 85380

gggtcggcaa tgaagagcac acactgtgcc tccaggggac ccaagcttgg gtccaagcac 85440

ccttgttggg cagctcacaa ctgcctgtaa ctccacctcc agaggatcct aagccacctt 85500

ctggcttggc ttcatggagg gaacaggtat gtgggtatct gagtgtgacg aatgagcagc 85560

aagtgagtct cgctgtggct agcacaaagt atgggctgaa gagcaggagg acagctgaaa 85620

agtggccttt cctggtgact aagttggtct gagcagctga gtcagtttct tcctggctgc 85680

ttggctggtc tcagtgctta taagctgctc acttgtaagt cttttcctag gagcccagct 85740

tgtctagggg ttgtctttgc aactggcctt gtctgacagt gactttcagc agtcttagct 85800

gcttatatac acagtcttag gaaagaaggc tggtgaatct gatccatttc aggaactttc 85860

tgaagctatt ctgaatttac tttacaagct tacctgcagg atagaggatc tcagctcttt 85920

ataaacatcc tgtcctaaaa caccctgttg ttcctcttct cttttacatc ctgtgtcttg 85980

agaagtttgc ctccaggatg gaagttgttc aattcagagg acactgttgc acaagctccc 86040

agcacccaca tgtgagctca gtgctctcct tggctctagc tctgccctat gaggtttttt 86100

attttgtcat cataatcttt tcctatatcc ttccttgttc tgggaactca tctggttcat 86160

ttttttggca ttttgagaaa agctctcact atacaaatca ggctgcctcc aaatcatctt 86220

tttgccttaa cctcctcagt accaagatca cgagtggatc ttaacacttg actgactcgt 86280

ttaagtgtga ggaaatgtgg accaataaga gagcccagga aagcccagga gaatctgtag 86340

ccccatggct gttgtgtcag aacccagagt tttgtcaaca gaatttggtt cctaatttct 86400

ccactttata aaaacgagtg agagaaacag gaacctattc agatctggcg tctgagcaat 86460

cagtgggtga acatctagag atctgttctg catctcctcg ccagctggca gagcatgcgt 86520

aaggcgggag ggaacaaggg caatcactca ctctggggct caggcttgcc ccttgggtca 86580

ggtgtttctg agagacgtga tgtctgcttc tcttgttacc atccctcatc ctctcccctc 86640

cttctgtccc ctacttacca atttcactgg ccagtgtcca tatttcctgc aaaagcgatt 86700

tggtttaatg agcttgacta tgcccgactc ctttagggag ggtggggaaa gggcaacgag 86760

ggcagtaagt ggtttccaca accactttgc acccggctgc tgggccccaa gccagaggaa 86820

cgtgcatgag ccatgaagtt tccactgata aatccacaga tgcttctagc acctgccttt 86880

ctgactcagc ctcaccgtgc cgcctgccag ctgtgaaatc agtgccaaca acaggtaacc 86940

gagacccagg cgcagggcca ggacagctgt ctgacacttc cagacaggat gtggaggctg 87000

acagttgtga tggagaggag atggggagga cagagacggg ctcagcttta agacaccgag 87060

ccacagagca ccaaacaaaa gccagggcct tctgaggtag aagtaacaga aaccaaacag 87120

gcaattctac tagtttcctg ggactgtttg ctgcatttgc caatcttggt agttttaaaa 87180

aacaaaaaca gtttgttctc agcactggca gagctttcct cctctggagg ctccaggggt 87240

ccagactctc ctctgtggta cactggcttc agacatatct cttgcctatg gctgcctcac 87300

tctaaactct gcctgtcctt gaattacctc tctctgcact ggctttataa aggaaacatg 87360

agattgtgtt tagggcctgt ttgggtgacc tcctcaggat ctataacata atcacatctc 87420

taccgtatga agtgacgctt ccgtcccagt gtgtaataca tttgccggcg cctgtcctta 87480

ggacagtgac caccaccaac tgtggaactt gactatgtcc acgtcatctt cctactagct 87540

ttagaaggct tatacccaca ctttctatcc agaattgtat ttttatttag aatcattcct 87600

acttttaaaa aagtctctgt ggttaaaagc attgcagagg gcttgggttt tggtccccag 87660

gacccacatc aagtggctca cagtgtcctg gaactcttgt tccaataccc tcttctggtc 87720

tccataggca ctacatacat atggcacata tatgtatact caggcacacg tgtaaatttt 87780

aatgtctact ttttatgcta aatatcaaag tcactcgagc agtggagttg agcacactca 87840

cataaggaaa tcatcagaca gacacttcat cctgtgttgg agccactttg tggctggagt 87900

aagcagggca gagtgatgtt ttcattactc tctggcccca gcaccccctg cctctcccca 87960

cccattcgtc catgcaggtg gggaagagaa ttctctttgt gaaattggaa gtttggaccc 88020

agcttcactc ttactctgcc cagtacctcc tgtgagaaac cctcctatcc caggtgacct 88080

gctggctgtg actctcctca gcaaaaggcc cgtgacccac actgcgccac taatgtatca 88140

tccccaaatg ctgaaaagga agcgtgtctt cctctctctc tctttttctt ttggtctttt 88200

tgagacagag tttctctgta tagccctggc tgtcctggaa ctcactttgt agaccaggct 88260

ggcctcgaac tcagaactcc gcctgcctct gcctcccgag tgctgggatt aaaggcgtgc 88320

atcaccactg cccggctgcg tgtctttctc ttagcggtct ctgtggagat gctgagtatg 88380

aagctcatcc tacccaccct tcagtggggc cttttctagc tactgagcag ctgtgtgagg 88440

actcgtgatc acaaggtcct ttgaaccctt gagacagatg tgcctgagcc cagtttgacc 88500

tgacaaaagc ctagagctca ctgataatgc cagcaaacac catctttgag tttgcaaagg 88560

aatcgcaaca catgcattca gtttccgttg ctggctgctg ctccagagat ggctatattc 88620

attctcaggt actcagactc aagagtagtt ctggccacac aggtctccac atttcgaggt 88680

caaatgacag aaaaccaggt tggtctcagt gcacatgggt ttattgagcc actgcaggtg 88740

ctggggaaac catggcaggg agatcctggg aagccagtgg ggtgctgagc aggagggacc 88800

tcagtctctc cttaatgtct acacactgtg tcataggtga caagccacgt cagtgctgtg 88860

acacgggtaa gcttaatggt gagtaatggc taactgggag ggtatttagg cagccttgtc 88920

tgtcagcctg ttcatatgat ctccttagtg ccttgtcatc ttggaaaagg acagttccaa 88980

attctaggag cgggggctag tctctgtcct gctctgtaag cccaggggac ccaatgaggc 89040

ctcatctatg ggtgctcagc tctaggatgg ggaagaaaat ggacaagatg cctactgacg 89100

ggaacacagg cttttcagtc agaccctagc ctccagcccc caatccagag gacagccaca 89160

caggggtcca ggcctgcaaa gggcagcaga cctgagggca agggagtttc agctcagtga 89220

gcagtcatcg ggagacatgg cagtcagctg tgtcgtccac ggttcatgtt cctaatcaga 89280

gcagggcctg gagagccagg gcagtgagtg catacagcca ggacaccttg ggcgttagga 89340

caaaacaagg actgtttctg cctccagctc ttctcaggcc actcgtgcct tgcctaggaa 89400

gggtaagaga gcacagatgg gaaggattcg gaaactgtca actccctgtc ctctccccat 89460

acctacccgc gggaaacagc acccagcagt ctggtcctgc agaactgatg gctgcaagct 89520

gtcaaaggct tgtatggcac catctgcgga gtgcagagat ccagagaagg cttggccagg 89580

aaaccctaga aactacccca ctcccttggg acaaaaaata agacaccctg gaacctgcaa 89640

ggcatggcct gagatggaag gtcactgtgc taagaatgac ccacaaactg ctagtgaggt 89700

tgacaagggc tgccccctct ccctttacag gtgaacacaa tcgggattaa taagagttta 89760

actctcagct actaagtggc agagacaggc ttcaaacaga cccccagaaa tctggaactg 89820

agccattcca cccagaggca agaacagcag aggtaagttg ggcacacatg gaagaaaggg 89880

ccaccccatt agtgtcaaaa gggaggccaa cttcaggcca ttggacacgt tttaacgctg 89940

acttccaccc atgtaccatg gcatgtgcac actgtccatc gcccacacca aacatgatgc 90000

gacgtaaata agacccacgg gccaggcagc ttggattggg ccacagacat 90050

6

226

Mus musculus

6

agcgctattc agctgtgcct cctttgctgt cttggctcct cctggagcac tatatgcacc 60

catgtcctta ccaggccttt cacagacgct gccattgaga gggttgatgc aggttgcagc 120

ctttaatccc cgagtactag gctctgacaa gatcccacag aagccagcat cactgggctc 180

agatggcatc cactgcagca aactatttgt gaatggagac atatcc 226

7

221

Mus musculus

7

agcgctattc agctgtgcct cctttgctgt cttggctcct cctggagcac tatatgcacc 60

catgtcctta ccaggccttt cacagaccat tgagagggtt gatgcaggtt gcagccttta 120

atccccgagt actaggctct gacaagatcc cacagaagcc agcatcactg ggctcagatg 180

gcatccactg cagcaaacta tttgtgaatg gagacatatc c 221

8

2419

Mus musculus

8

gaattccggg cgaaggggag ccggcgtgcg gggtgtgtat gtgttcgctg ggcgccggct 60

cagccccagg aagatggtgg cggtggcggc ggcggcggcg actgaggcgc ggctgagggg 120

gagcacgagg acgacagcag cgcctgcggg caggaagggc aggcagcacc gaccctgcac 180

cgcgacaggg gcctggaggc cgggaccgcg cgcccggctg tgtctcccgc gggtgctgtc 240

gcgggcgctg cccccgccgc cgctgctgcc gctgctcttt tcgctgctgc tgctgccgct 300

gccccgggag gccgaggccg ctgcggtggc ggcggcggtg tccggctcgg ccgcagccga 360

ggccaaggaa tgtgaccggc cgtgtgtcaa cggcggccgc tgcaaccctg gcaccggcca 420

gtgcgtctgc cccacgggct gggtgggcga gcaatgccag cactgcgggg gccgcttcag 480

gacatctgtc tcacgcctat aatcacagct gttcggaagg tgaggctgga ggaacagttc 540

gaggcaagct tcggctacag aataagttca agagtaacct ggggcaactt gggcttgtct 600

ccaaaaccaa aatgagcgaa aaggagcaag ctagagtctt ttgggaaaat tttagctgac 660

taatttttca ccgagaacta actggctctt ctggatttgt aacagatgga cctgggaatt 720

ataaatataa gacgaagtgc acatggctca ttgaaggaca gccaaataga ataatgagac 780

ttcgcttcaa ccattttgct acagaatgta gctgggacca tttatatgtt tatgatgggg 840

actcaatcta cgcacctctg attgctgcct ttagtggcct cattgttcct gaaagagatg 900

gcaatgagac ggctcctgag gtcactgtca cttcaggtta tgcactgctg cattttttca 960

gtgatgctgc ttataatctg actggattta atatcactta caattttgac atgtgtccga 1020

ataattgctc aggccgagga gagtgtaaga gcagtaacag cagcagcgct gttgagtgtg 1080

aatgttctga aaactggaaa ggggagtcgt gtgacattcc tcactgtaca gacaactgtg 1140

gctttcctca ccgaggcatc tgtaatgcaa gcgataccag agggtgctcc tgctttcctc 1200

actggcaggg tcctggatgt tcaattcctg tgccagctaa ccagtctttt tggactcgag 1260

aagaatattc tgatttaaag cttcccagag cctctcataa agctgtggtc aatggaaata 1320

taatgtgggt tgttggcgga tatatgttca accattcaga ttacagcatg gttttagcgt 1380

atgacctgac ttctagggaa tggcttccac taaaccattc tgtgaacagt gtggttgtaa 1440

gatatggtca ttctttggca ttacataagg ataaaatcta catgtatgga ggaaaaattg 1500

attcaacagg gaacgtgacc aatgagctga gagtatttca tattcataat gaatcatggg 1560

tattgttaac tccgaaagct aaggatcagt atgcagtggt tggacactca gcacacattg 1620

ttacactggc atctggccgt gtggtcatgt tggtcatctt cggtcattgc ccactctatg 1680

gatatataag cgttgtgcag gaatatgact tggaaaagaa cacatggagt atattacata 1740

ctcagggtgc tcttgtgcaa gggggttatg gccacagtag tgtttatgat gacaggacca 1800

aggctctgta cgttcatggt ggctacaagg ctttcagcgc caacaaatac cggcttgcag 1860

atgacctcta cagatacgat gtggatactc agatgtggac cattcttaag gacagccgat 1920

ttttccgtta cttgcataca gctgtgatag tgagtggaac catgctggtg tttggaggga 1980

acacacacaa tgacacttcc atgagccacg gtgccaaatg cttctcctcg gacttcatgg 2040

cttatgacat tgcttgtgac cgatggtcag tgcttcccag acctgagctc catcatgatg 2100

tcaacagatt tggccattca gcagtcttgt acaacagcac catgtatgtg ttcggcggct 2160

tcaacagcct cctcctcagt gacgtcttgg tctttacctc ggagcagtgc gatgcacacc 2220

gcagtgaagc tgcttgtgtg gcagcaggac ctggtatccg gtgtctgtgg gacacacagt 2280

cgtctcgatg tacctcctgg gagttggcaa ctgaagaaca agcagaaaag ttaaaatcag 2340

agtgtttttc taaaagaacc cttgaccatg acagatgtga ccagcacaca gattgttaca 2400

gctgcacagc caataccaa 2419

9

Met Arg Leu Arg Phe Asn His Phe Ala Thr Glu Cys Ser Trp Asp His

1 5 10 15

Leu Tyr Val Tyr Asp Gly Asp Ser Ile Tyr Ala Pro Leu Ile Ala Ala

20 25 30

Phe Ser Gly Leu Ile Val Pro Glu Arg Asp Gly Asn Glu Thr Ala Pro

35 40 45

Glu Val Thr Val Thr Ser Gly Tyr Ala Leu Leu His Phe Phe Ser Asp

50 55 60

Ala Ala Tyr Asn Leu Thr Gly Phe Asn Ile Thr Tyr Asn Phe Asp Met

65 70 75 80

Cys Pro Asn Asn Cys Ser Gly Arg Gly Glu Cys Lys Ser Ser Asn Ser

85 90 95

Ser Ser Ala Val Glu Cys Glu Cys Ser Glu Asn Trp Lys Gly Glu Ser

100 105 110

Cys Asp Ile Pro His Cys Thr Asp Asn Cys Gly Phe Pro His Arg Gly

115 120 125

Ile Cys Asn Ala Ser Asp Thr Arg Gly Cys Ser Cys Phe Pro His Trp

130 135 140

Gln Gly Pro Gly Cys Ser Ile Pro Val Pro Ala Asn Gln Ser Phe Trp

145 150 155 160

Thr Arg Glu Glu Tyr Ser Asp Leu Lys Leu Pro Arg Ala Ser His Lys

165 170 175

Ala Val Val Asn Gly Asn Ile Met Trp Val Val Gly Gly Tyr Met Phe

180 185 190

Asn His Ser Asp Tyr Ser Met Val Leu Ala Tyr Asp Leu Thr Ser Arg

195 200 205

Glu Trp Leu Pro Leu Asn His Ser Val Asn Ser Val Val Val Arg Tyr

210 215 220

Gly His Ser Leu Ala Leu His Lys Asp Lys Ile Tyr Met Tyr Gly Gly

225 230 235 240

Lys Ile Asp Ser Thr Gly Asn Val Thr Asn Glu Leu Arg Val Phe His

245 250 255

Ile His Asn Glu Ser Trp Val Leu Leu Thr Pro Lys Ala Lys Asp Gln

260 265 270

Tyr Ala Val Val Gly His Ser Ala His Ile Val Thr Leu Ala Ser Gly

275 280 285

Arg Val Val Met Leu Val Ile Phe Gly His Cys Pro Leu Tyr Gly Tyr

290 295 300

Ile Ser Val Val Gln Glu Tyr Asp Leu Glu Lys Asn Thr Trp Ser Ile

305 310 315 320

Leu His Thr Gln Gly Ala Leu Val Gln Gly Gly Tyr Gly His Ser Ser

325 330 335

Val Tyr Asp Asp Arg Thr Lys Ala Leu Tyr Val His Gly Gly Tyr Lys

340 345 350

Ala Phe Ser Ala Asn Lys Tyr Arg Leu Ala Asp Asp Leu Tyr Arg Tyr

355 360 365

Asp Val Asp Thr Gln Met Trp Thr Ile Leu Lys Asp Ser Arg Phe Phe

370 375 380

Arg Tyr Leu His Thr Ala Val Ile Val Ser Gly Thr Met Leu Val Phe

385 390 395 400

Gly Gly Asn Thr His Asn Asp Thr Ser Met Ser His Gly Ala Lys Cys

405 410 415

Phe Ser Ser Asp Phe Met Ala Tyr Asp Ile Ala Cys Asp Arg Trp Ser

420 425 430

Val Leu Pro Arg Pro Glu Leu His His Asp Val Asn Arg Phe Gly His

435 440 445

Ser Ala Val Leu Tyr Asn Ser Thr Met Tyr Val Phe Gly Gly Phe Asn

450 455 460

Ser Leu Leu Leu Ser Asp Val Leu Val Phe Thr Ser Glu Gln Cys Asp

465 470 475 480

Ala His Arg Ser Glu Ala Ala Cys Val Ala Ala Gly Pro Gly Ile Arg

485 490 495

Cys Leu Trp Asp Thr Gln Ser Ser Arg Cys Thr Ser Trp Glu Leu Ala

500 505 510

Thr Glu Glu Gln Ala Glu Lys Leu Lys Ser Glu Cys Phe Ser Lys Arg

515 520 525

Thr Leu Asp His Asp Arg Cys Asp Gln His Thr Asp Cys Tyr Ser Cys

530 535 540

Thr Ala Asn Thr Xaa

545

10

1051

Mus musculus

10

gaattccgga aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 120

aaaaaaaaaa aaaaaaaagg agggcgaagg ggagccggcg tgcggggtgt gtatgtgttc 180

gctgggcgcc ggctcagccc caggaagatg gtggcggtgg cggcggcggc ggcgactgag 240

gcgcggctga gggggagcac gacgacgaca gcagcgcctg cgggcaggaa gggcaggcag 300

caccgaccct gcaccgcgac aggggcctgg aggccgggac cgcgcgcccg gctgtgtctc 360

ccgcgggtgc tgtcgcgggc gctgcccccg ccgccgctgc tgccgctgct cttttcgctg 420

ctgctgctgc cgctgccccg ggaggccgag gccgctgcgg tggcggcggc ggtgtccggc 480

tcggccgcag ccgaggccaa ggaatgtgac cggccgtgtg tcaacggcgg ccgctgcaac 540

cctggcaccg gccagtgcgt ctgccccacg ggctgggtgg gcgagcaatg ccagcactgc 600

gggggccgct tcagactaac tggctcttct ggatttgtaa cagatggacc tgggaattat 660

aaatataaga cgaagtgcac atggctcatt gaaggacagc caaatagaat aatgagactt 720

cgcttcaacc attttgctac agaatgtagc tgggaccatt tatatgttta tgatggggac 780

tcaatctacg cacctctgat tgctgccttt agtggcctca ttgttcctga aagagatggc 840

aatgagacgg ctcctgaggt cactgtcact tcaggttatg cactgctgca ttttttcagt 900

gatgctgctt ataatctgac tggatttaat atcacttaca attttgacat gtgtccgaat 960

aattgctcag gccgaggaga gtgtaagagc agtaacagca gcagcgctgt tgagtgtgaa 1020

tgttctgaaa actggaaagg ggccggaatt c 1051

11

Glu Phe Arg Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys

1 5 10 15

Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys

20 25 30

Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Gly Gly

35 40 45

Arg Arg Gly Ala Gly Val Arg Gly Val Tyr Val Phe Ala Gly Arg Arg

50 55 60

Leu Ser Pro Arg Lys Met Val Ala Val Ala Ala Ala Ala Ala Thr Glu

65 70 75 80

Ala Arg Leu Arg Gly Ser Thr Thr Thr Thr Ala Ala Pro Ala Gly Arg

85 90 95

Lys Gly Arg Gln His Arg Pro Cys Thr Ala Thr Gly Ala Trp Arg Pro

100 105 110

Gly Pro Arg Ala Arg Leu Cys Leu Pro Arg Val Leu Ser Arg Ala Leu

115 120 125

Pro Pro Pro Pro Leu Leu Pro Leu Leu Phe Ser Leu Leu Leu Leu Pro

130 135 140

Leu Pro Arg Glu Ala Glu Ala Ala Ala Val Ala Ala Ala Val Ser Gly

145 150 155 160

Ser Ala Ala Ala Glu Ala Lys Glu Cys Asp Arg Pro Cys Val Asn Gly

165 170 175

Gly Arg Cys Asn Pro Gly Thr Gly Gln Cys Val Cys Pro Thr Gly Trp

180 185 190

Val Gly Glu Gln Cys Gln His Cys Gly Gly Arg Phe Arg Leu Thr Gly

195 200 205

Ser Ser Gly Phe Val Thr Asp Gly Pro Gly Asn Tyr Lys Tyr Lys Thr

210 215 220

Lys Cys Thr Trp Leu Ile Glu Gly Gln Pro Asn Arg Ile Met Arg Leu

225 230 235 240

Arg Phe Asn His Phe Ala Thr Glu Cys Ser Trp Asp His Leu Tyr Val

245 250 255

Tyr Asp Gly Asp Ser Ile Tyr Ala Pro Leu Ile Ala Ala Phe Ser Gly

260 265 270

Leu Ile Val Pro Glu Arg Asp Gly Asn Glu Thr Ala Pro Glu Val Thr

275 280 285

Val Thr Ser Gly Tyr Ala Leu Leu His Phe Phe Ser Asp Ala Ala Tyr

290 295 300

Asn Leu Thr Gly Phe Asn Ile Thr Tyr Asn Phe Asp Met Cys Pro Asn

305 310 315 320

Asn Cys Ser Gly Arg Gly Glu Cys Lys Ser Ser Asn Ser Ser Ser Ala

325 330 335

Val Glu Cys Glu Cys Ser Glu Asn Trp Lys Gly Ala Gly Ile Xaa

340 345 350

12

6370

Homo sapiens

12

atgtactact gtaacaagaa gaccagctgc aggagctgtg ccctggacca gaactgccag 60

tgggagcccc ggaatcagga gtgcattgcc ctgcccgaaa atatctgtgg cattggctgg 120

catttggttg gaaactcatg tttgaaaatt actactgcca aggagaatta tgacaatgct 180

aaattgttct gtaggaacca caatgccctt ttggcttctc ttacaaccca gaagaaggta 240

gaatttgtcc ttaagcagct gcgaataatg cagtcatctc agagcatgtc caagctcacc 300

ttaaccccat gggtcgggcc ttcgggaagg tycaatgtgt cctactkggt gctggggaag 360

gatatgktcc ccattttaca aatagtttta ctacagtggg atgsccgtct tgaggcccag 420

tgttgcttgg rattctgtgg gaattttatt caggaaccca gttacttcgg ggactgaagg 480

ctgcaacctg cattcaaccc actymaatgg tagtgtctgt gaaaggcctg caaaccacag 540

tgctaaggca gtgccggaca ccatgtgcct tgaggacagc atgtggagat tgcaccagcg 600

gcagctctga gtgcatgtgg tgcagcaaca tgaagcagtg tgtggactcc aatgcctatg 660

tggcctcctt cccttttggc cagtgtatgg aatggtatac gatgagcacc tgcccccctg 720

aaaattgttc aggctactgt acctgtagtc attgcttgga gcaaccaggc tgtggctggt 780

gtactgatcc cagcaatact ggcaaaggga aatgcataga gggttcctat aaaggaccag 840

tgaagatgcc ttcgcaagcc cctacaggaa atttctatcc acagcccctg ctcaattcca 900

gcatgtgtct agaggacagc agatacaact ggtctttcat tcactgtcca gcttgccaat 960

gcaacggcca cagtaaatgc atcaatcaga gcatctgtga gaagtgtgag aacctgacca 1020

caggcaagca ctgcgagacc tgcatatctg gcttctacgg tgatcccacc aatggaggga 1080

aatgtcagcc atgcaagtgc aatgggcacg cgtctctgtg caacaccaac acgggcaagt 1140

gcttctgcac caccaagggc gtcaaggggg acgagtgcca gctatgtgag gtagaaaatc 1200

gataccaagg aaaccctctc agaggaacat gttattatac tcttcttatt gactatcagt 1260

tcacctttag tctatcccag gaagatgatc gctattacac agctatcaat tttgtggcta 1320

ctcctgacga acaaaacagg gatttggaca tgttcatcaa tgcctccaag aatttcaacc 1380

tcaacatcac ctgggctgcc agtttctcag ctggaaccca ggctggagaa gagatgcctg 1440

ttgtttcaaa aaccaacatt aaggagtaca aagatagttt ctctaatgag aagtttgatt 1500

ttcgcaacca cccaaatatc actttctttg tttatgtcag taatttcacc tggcccatca 1560

aaattcagat tgccttctct cagcacagca attttatgga cctggtacag ttcttcgtga 1620

ctttcttcag ttgtttcctc tctttgctcc tggtggctgc tgtggtttgg aagatcaaac 1680

aaagttgttg ggcctccaga cgtagagagc aacttcttcg agagatgcaa cagatggcca 1740

gccgtccctt tgcctctgta aatgtcgcct tggaaacaga tgaggagcct cctgatctta 1800

ttggggggag tataaagact gttcccaaac ccattgcact ggagccgtgt tttggcaaca 1860

aagccgctgt cctctctgtg tttgtgaggc tccctcgagg cctgggtggc atccctcctc 1920

ctgggcagtc aggtcttgct gtggccagcg ccctggtgga catttctcag cagatgccga 1980

tagtgtacaa ggagaagtca ggagccgtga gaaaccggaa gcagcagccc cctgcacagc 2040

ctgggacctg catctgatgc tggggccagg gactctccca cgcacgagct agtgagtggc 2100

acaccagagc catctgcagg gaagggcgtg gcggggaaat ggctgtgcgg tgcgggacgg 2160

aagactggaa accctcaaag catctgactc acctgcatga tcacaagctt tctttgacgg 2220

tttctcccat ccgtgttcca gcatctaacc ttttactttt gcataggaaa tacttgattt 2280

aattacaggt ccagggatga gctgatggtt gctggaggag gccagtgtag agccagtgag 2340

agaactagga atgacactca ggttcactgt ggaaaactgt tcttgggact gtctcaactg 2400

tgcaaaaaac aaaagatgga gtgtttacaa gtagacattc gtcatcagtt gttcttgaac 2460

atggtctttt aaaaactagt cagatgaatt aacttgtttt catctgaagc ctgctatctt 2520

ttttaaaaga tgtgctattt attcttgcac gatttaggca attatctctc ttccagggag 2580

tacctttttt tctagttgag aattaataat ggtccatctc ttttgatcat atcaagctag 2640

gatagaaggg gggctatttt aaatgtcaag gtcagcagtg ttactttgaa tgtaaactgg 2700

tataataggt agttttctat agtaacttga ttaatttagt cttaatccat ttgaaactct 2760

ctcttccttt ctctctgcct gtccctctcc ttctccatct caccctccct ctctcacaca 2820

tacacacaca aacacataca cacaacacta agtgcctaga ctttaaatag atctagcaat 2880

tggaaagtta gtaagcctaa gtttttacat aattgcattc ctacattctt gtaaaattta 2940

aatagctacc attggcaatc tgcttttttt ctaaaatctg atttgcagcc aggaaagaat 3000

tttctcaccc aaggaacatt tgatctagca gcagggatga gaggaaagca gaaatgaatg 3060

aactgtgaaa gctcctgttt ttattatcaa aaaggacact gtcaagaagg cgccccctgc 3120

ccccaccccc gtgtcaccct aggcctgata agcgatcaga ggaaaggact cattcatgtc 3180

acgcttcctt gagcagaaaa gagcactgag agcacttggg acccctggat cagagagcat 3240

ctgtgtgtcc tgcagcctcc tctgaacttg tggttcattc tcaggctggg gtggactcag 3300

atgccaggaa agggacagcc tcccattgtc aggcagaagc tgcccaaagc ctggagaagg 3360

acttgtttgc cctctttccc ccaggagggg ctcgacccac ccaccctccc tctcagacca 3420

aggtggtggc tgtgaggagg gcagcaaatg ctgacaagga tgaaaagcac atggaaaaaa 3480

atggacgagg agggaaaact ctgccaaatg gaaaatgacc aaatttaaga gggtgggaca 3540

gtcccctgct cctctcccag agggcactgc ttggaaattg tgttttcccc atttatggtg 3600

ctctgtattc tggcattatg cagcagcctc ccagaagctc tcttctgctt caaaacctgg 3660

gatctctggc attaccctat tgggatggac cgctggacag caatgctcga gtttgtgaat 3720

ttggagagat actcaaaaga gctaaaactg cagcatttta cctttaaatg cagtgcctag 3780

agagagagta ttgtctcttc cccaacacta accccactcc catgaagaat tgcctggaaa 3840

gatgttttca aggaatttga accataaaac actatctgat gcacagaaca cctctacttt 3900

gagactcacc tctcataaag cttctttttc acattactgt taaagaccag acgttctaga 3960

aaagacccct cctctcatga gctcccccat ccctgctaca gaacacagca cccatggcgc 4020

ctgcagtgga ctggcccctt aattcccaca ggccccccca gcaaggccaa agggaggccc 4080

ctgggtattg tcctcctaca aggaagatcc tctttgtttg ttcaaaggac cagttttcct 4140

aggccaaaga agtctcttcc ccatgttagt cctatgcctt gaaatatcat gcaccatgac 4200

ccacagccat ctggttatgt cttatttttt tcctaaaaga taatgtttat ttttaaaaag 4260

gaaggaagaa gcaagtgaag tttcattctg ctccagcggt ggggaagccg ctgaatccac 4320

ctgcttctcc tttgcaaccg acagcaaaca gctttctccg gcctcagggc agaaaaaggg 4380

aatggcaggg agtaagaggc gctgggctcg gagcctgttt ccaagaagga attggttgtc 4440

atctggcagt gttgcgcgtc acaagagagc ctgtatataa attaaaatag tcaagacaac 4500

actgaccttg cacttgtaca taactataca gtagtgtcca gaatgttcag acattcggag 4560

tgtacataaa acagaaaaaa tcttcatgta tttttattaa atataacaat gtctgagttt 4620

cacctaagat gtttttgtgc catatgctgg atatccaggt tctcgccagg ccccgataca 4680

tgaataacaa acccaagaaa cgcatcccca ttgtgtgatg tgttcagatg catctggcac 4740

caattaggta tttcttaaaa caggactcat ctgtcagagt gcacatgaaa aatcaggcag 4800

ggaatcgaaa cgacagcgct ggaggagact caggaagcag aggcgtccct gccgctgccc 4860

ttggccctgc aagcacatca tgaccctttc tggcagcctc ttggtgctct gggtagtgag 4920

ggatgaccag tcttgtcctg agaaatgttt ctcttagtct ttaagttcaa agactaacct 4980

gtagcaatca gactttccaa aagggggttc tccatttttt gtagttttgt ctaaattttt 5040

aatgaccatt tcctggaatc agtttattat actgaaaact gggggtggga gtagggagct 5100

agtttgttga taaatagttc ccatttcccc gtggagaatt tgacataccc tggactcctg 5160

tgtgcctcct gccatccctg cacacagcct ggggagaagc ctgtgcctcc ccgtgtggag 5220

agaaggcaac cccagatccc ctgagctaac ccggaggaaa ggcagtcctg gacagaagac 5280

tgtcagcaga aggaaagtac tggactaccc gtgggtaagt cctgccattc aagactggag 5340

acacctggga aataaaaaga gcagggcact gctggtggga agaggcattt taccttccag 5400

tgcaaatcct gctcctttga tttaatgggg tgtactgggg ccaggggctg attcacttcc 5460

ttgggagatg gtggtgtttt catgaacatc tttgatcctt ccatttcatt tattcatcca 5520

tccattcaac aagtatttgc taaacactaa cttaagctaa tgctagggta gtgactgaga 5580

tgtaaaaata gattttagaa ttaaaacaaa atccaagtcc tcacacccct gtcatcccag 5640

gagatctttc cttgtggtgg tttctgtgag aattggccat cctgaggaca cagccaggac 5700

ggcagaggcc tcctggcctc agggcatgcc ctgcctacct tctgaaatgt ttaccccatt 5760

gaccaaactt ggctccagcc attgcggtgg tttctagata gccaggccca ccaagagata 5820

ttgccccttg atgagagtca aacaccctgc ctacaaggag atgttttgaa atggagagga 5880

aaattggcac ctcatctttt aaaggcagta atggaattga ttttcagtaa ctgaatttgt 5940

gcacaaaaca ttctaaacac tagtgaagcc tgtttcgttg aactaattct ggctctggaa 6000

atgtttttgt tttatagtta tttacgattt cgtttgtttg gattcaagct tagtttgtta 6060

atatgtataa tttagcatct attacactca tgtaaatatg gagtaagtat tgtaaactat 6120

ttcattgcgg ggattgtggg tgttatacat acatttagga ctgcaatttt ttggtatttt 6180

ttgtattgta aaataacagc taatttaagc aggaacaaga gaactaaggg aggtctgtgc 6240

attttaaaca caaatgtgaa gaacttgtat ataaacaaaa gtaaatacta taatacaaac 6300

ttccttctga aataaaagta gatctggtaa aaaaaaaaaa agaaaaaaaa aaaaaaaaaa 6360

gggcggccgc 6370

13

Met Tyr Tyr Cys Asn Lys Lys Thr Ser Cys Arg Ser Cys Ala Leu Asp

1 5 10 15

Gln Asn Cys Gln Trp Glu Pro Arg Asn Gln Glu Cys Ile Ala Leu Pro

20 25 30

Glu Asn Ile Cys Gly Ile Gly Trp His Leu Val Gly Asn Ser Cys Leu

35 40 45

Lys Ile Thr Thr Ala Lys Glu Asn Tyr Asp Asn Ala Lys Leu Phe Cys

50 55 60

Arg Asn His Asn Ala Leu Leu Ala Ser Leu Thr Thr Gln Lys Lys Val

65 70 75 80

Glu Phe Val Leu Lys Gln Leu Arg Ile Met Gln Ser Ser Gln Ser Met

85 90 95

Ser Lys Leu Thr Leu Thr Pro Trp Val Gly Pro Ser Gly Arg Xaa Asn

100 105 110

Val Ser Tyr Xaa Val Leu Gly Lys Asp Met Xaa Pro Ile Leu Gln Ile

115 120 125

Val Leu Leu Gln Trp Asp Xaa Arg Leu Glu Ala Gln Cys Cys Leu Xaa

130 135 140

Phe Cys Gly Asn Phe Xaa Ser Gly Thr Gln Leu Leu Arg Gly Leu Lys

145 150 155 160

Ala Ala Thr Cys Ile Gln Pro Thr Xaa Met Val Val Ser Val Lys Gly

165 170 175

Leu Gln Thr Thr Val Leu Arg Gln Cys Arg Thr Pro Cys Ala Leu Arg

180 185 190

Thr Ala Cys Gly Asp Cys Thr Ser Gly Ser Ser Glu Xaa His Val Val

195 200 205

Gln Gln His Glu Xaa Ser Val Trp Thr Pro Met Pro Met Trp Pro Pro

210 215 220

Ser Leu Leu Xaa Gln Cys Met Glu Trp Tyr Thr Met Ser Thr Cys Pro

225 230 235 240

Pro Glu Asn Cys Ser Gly Tyr Cys Thr Cys Ser His Cys Leu Glu Gln

245 250 255

Pro Gly Cys Gly Trp Cys Thr Asp Pro Ser Asn Thr Gly Lys Gly Lys

260 265 270

Cys Ile Glu Gly Ser Tyr Lys Gly Pro Val Lys Met Pro Ser Gln Ala

275 280 285

Pro Thr Gly Asn Phe Tyr Pro Gln Pro Leu Leu Asn Ser Ser Met Cys

290 295 300

Leu Glu Asp Ser Arg Tyr Asn Trp Ser Phe Ile His Cys Pro Ala Cys

305 310 315 320

Gln Cys Asn Gly His Ser Lys Cys Ile Asn Gln Ser Ile Cys Glu Lys

325 330 335

Cys Glu Asn Leu Thr Thr Gly Lys His Cys Glu Thr Cys Ile Ser Gly

340 345 350

Phe Tyr Gly Asp Pro Thr Asn Gly Gly Lys Cys Gln Pro Cys Lys Cys

355 360 365

Asn Gly His Ala Ser Leu Cys Asn Thr Asn Thr Gly Lys Cys Phe Cys

370 375 380

Thr Thr Lys Gly Val Lys Gly Asp Glu Cys Gln Leu Cys Glu Val Glu

385 390 395 400

Asn Arg Tyr Gln Gly Asn Pro Leu Arg Gly Thr Cys Tyr Tyr Thr Leu

405 410 415

Leu Ile Asp Tyr Gln Phe Thr Phe Ser Leu Ser Gln Glu Asp Asp Arg

420 425 430

Tyr Tyr Thr Ala Ile Asn Phe Val Ala Thr Pro Asp Glu Gln Asn Arg

435 440 445

Asp Leu Asp Met Phe Ile Asn Ala Ser Lys Asn Phe Asn Leu Asn Ile

450 455 460

Thr Trp Ala Ala Ser Phe Ser Ala Gly Thr Gln Ala Gly Glu Glu Met

465 470 475 480

Pro Val Val Ser Lys Thr Asn Ile Lys Glu Tyr Lys Asp Ser Phe Ser

485 490 495

Asn Glu Lys Phe Asp Phe Arg Asn His Pro Asn Ile Thr Phe Phe Val

500 505 510

Tyr Val Ser Asn Phe Thr Trp Pro Ile Lys Ile Gln Ile Ala Phe Ser

515 520 525

Gln His Ser Asn Phe Met Asp Leu Val Gln Phe Phe Val Thr Phe Phe

530 535 540

Ser Cys Phe Leu Ser Leu Leu Leu Val Ala Ala Val Val Trp Lys Ile

545 550 555 560

Lys Gln Ser Cys Trp Ala Ser Arg Arg Arg Glu Gln Leu Leu Arg Glu

565 570 575

Met Gln Gln Met Ala Ser Arg Pro Phe Ala Ser Val Asn Val Ala Leu

580 585 590

Glu Thr Asp Glu Glu Pro Pro Asp Leu Ile Gly Gly Ser Ile Lys Thr

595 600 605

Val Pro Lys Pro Ile Ala Leu Glu Pro Cys Phe Gly Asn Lys Ala Ala

610 615 620

Val Leu Ser Val Phe Val Arg Leu Pro Arg Gly Leu Gly Gly Ile Pro

625 630 635 640

Pro Pro Gly Gln Ser Gly Leu Ala Val Ala Ser Ala Leu Val Asp Ile

645 650 655

Ser Gln Gln Met Pro Ile Val Tyr Lys Glu Lys Ser Gly Ala Val Arg

660 665 670

Asn Arg Lys Gln Gln Pro Pro Ala Gln Pro Gly Thr Cys Ile

675 680 685

14

8589

Homo sapiens

14

atggtggcgg tggccgcagc ggcggcaact gaggcaaggc tgaggaggag gacggcggcg 60

acggcagcgc tcgcgggcag gagcggcggg ccgcaccgac cctgcaccgc gacaggggcc 120

tggaggccgg gaccgcgcgc ccggctgtgt ctcccgcggg tgctgtcgcg ggcgctgccc 180

ccgccgccgc tgctgccgct gctcttttcg ctgctgctgc tgccgctgcc ccgggaggcc 240

gaggccgctg cggtggcggc ggcggtgtcc ggctcggccg cagccgaggc caaggaatgt 300

gaccggccgt gtgtcaacgg cggtcgctgc aaccctggca ccggccagtg cgtctgcccc 360

gccggctggg tgggcgagca atgccagcac tgcgggggcc gcttcagact aactggatct 420

tctgggtttg tgacagatgg acctggaaat tataaataca aaacgaagtg cacgtggctc 480

attgaaggac agccaaatag aataatgaga cttcgtttca atcattttgc tacagagtgt 540

agttgggacc atttatatgt ttatgatggg gactcaattt atgcaccgct agttgctgca 600

tttagtggcc tcattgttcc tgagagagat ggcaatgaga ctgtccctga ggttgttgcc 660

acatcaggtt atgccttgct gcattttttt agtgatgctg cttataattt gactggattt 720

aatattactt acagttttga tatgtgtcca aataactgct caggccgagg agagtgtaag 780

atcagtaata gcagcgatac tgttgaatgt gaatgttctg aaaactggaa aggtgaagca 840

tgtgacattc ctcactgtac agacaactgt ggttttcctc atcgaggcat ctgcaattca 900

agtgatgtca gaggatgctc ctgcttctca gactggcagg gtcctggatg ttcagttcct 960

gtaccagcta accagtcatt ttggactcga gaggaatatt ctaacttaaa gctccccaga 1020

gcatctcata aagctgtggt caatggaaac attatgtggg ttgttggagg atatatgttc 1080

aaccactcag attataacat ggttctagcg tatgaccttg cttctaggga gtggcttcca 1140

ctaaaccgtt ctgtgaacaa tgtggttgtt agatatggtc attctttggc attatacaag 1200

gataaaattt acatgtatgg aggaaaaatt gattcaactg ggaatgtgac caatgagttg 1260

agagtttttc acattcataa tgagtcatgg gtgttgttga cccctaaggc aaaggagcag 1320

tatgcagtgg ttgggcactc tgcacacatt gttacactga agaatggccg agtggtcatg 1380

ctggtcatct ttggtcactg ccctctctat ggatatataa gcaatgtgca ggaatatgat 1440

ttggataaga acacatggag tatattacac acccagggtg cccttgtgca agggggttac 1500

ggccatagca gtgtttacga ccataggacc agggccctat acgttcatgg tggctacaag 1560

gctttcagtg ccaataagta ccggcttgca gatgatctct accgatatga tgtggatacc 1620

cagatgtgga ccattcttaa ggacagccga tttttccgtt acttgcacac agctgtgata 1680

gtgagtggaa ccatgctggt gtttggggga aacacacaca atgacacatc tatgagccat 1740

ggcgccaaat gcttctcttc agatttcatg gcctatgaca ttgcctgtga ccgctggtca 1800

gtgcttccca gacctgatct ccaccatgat gtcaacagat ttggccattc agcagtctta 1860

cacaacagca ccatgtatgt gttcggtggt ttcaatagtc tcctcctcag cgacatcctg 1920

gtattcacct cggaacagtg tgatgcgcat cggagtgaag ccgcttgttt agcagcagga 1980

cctggtattc ggtgtgtgtg gaacacaggg tcgtctcagt gtatctcgtg ggcgctggca 2040

actgatgaac aagaagaaaa gttaaaatca gaatgttttt ccaaaagaac tcttgaccat 2100

gacagatgtg accagcacac agattgttac agctgcacag ccaacaccaa tgactgccac 2160

tggtgcaatg accattgtgt ccccaggaac cacagctgct cagaaggcca gatctccatt 2220

tttaggtatg agaattgccc caaggataac cctatgtact actgtaacaa gaagaccagc 2280

tgcaggagct gtgccctgga ccagaactgc cagtgggagc cccggaatca ggagtgcatt 2340

gccctgcccg aaaatatctg tggcattggc tggcatttgg ttggaaactc atgtttgaaa 2400

attactactg ccaaggagaa ttatgacaat gctaaattgt tctgtaggaa ccacaatgcc 2460

cttttggctt ctcttacaac ccagaagaag gtagaatttg tccttaagca gctgcgaata 2520

atgcagtcat ctcagagcat gtccaagctc accttaaccc catgggtcgg ccttcggaag 2580

atcaatgtgt cctactggtg ctgggaagat atgtccccat ttacaaatag tttactacag 2640

tggatgccgt ctgagcccag tgatgctgga ttctgtggaa ttttatcaga acccagtact 2700

cggggactga aggctgcaac ctgcatcaac ccactcaatg gtagtgtctg tgaaaggcct 2760

gcaaaccaca gtgctaagca gtgccggaca ccatgtgcct tgaggacagc atgtggagat 2820

tgcaccagcg gcagctctga gtgcatgtgg tgcagcaaca tgaagcagtg tgtggactcc 2880

aatgcctatg tggcctcctt cccttttggc cagtgtatgg aatggtatac gatgagcacc 2940

tgcccccctg aaaattgttc aggctactgt acctgtagtc attgcttgga gcaaccaggc 3000

tgtggctggt gtactgatcc cagcaatact ggcaaaggga aatgcataga gggttcctat 3060

aaaggaccag tgaagatgcc ttcgcaagcc cctacaggaa atttctatcc acagcccctg 3120

ctcaattcca gcatgtgtct agaggacagc agatacaact ggtctttcat tcactgtcca 3180

gcttgccaat gcaacggcca cagtaaatgc atcaatcaga gcatctgtga gaagtgtgag 3240

aacctgacca caggcaagca ctgcgagacc tgcatatctg gcttctacgg tgatcccacc 3300

aatggaggga aatgtcagcc atgcaagtgc aatgggcacg cgtctctgtg caacaccaac 3360

acgggcaagt gcttctgcac caccaagggc gtcaaggggg acgagtgcca gctatgtgag 3420

gtagaaaatc gataccaagg aaaccctctc agaggaacat gttattatac tcttcttatt 3480

gactatcagt tcacctttag tctatcccag gaagatgatc gctattacac agctatcaat 3540

tttgtggcta ctcctgacga acaaaacagg gatttggaca tgttcatcaa tgcctccaag 3600

aatttcaacc tcaacatcac ctgggctgcc agtttctcag ctggaaccca ggctggagaa 3660

gagatgcctg ttgtttcaaa aaccaacatt aaggagtaca aagatagttt ctctaatgag 3720

aagtttgatt ttcgcaacca cccaaatatc actttctttg tttatgtcag taatttcacc 3780

tggcccatca aaattcagat tgccttctct cagcacagca attttatgga cctggtacag 3840

ttcttcgtga ctttcttcag ttgtttcctc tctttgctcc tggtggctgc tgtggtttgg 3900

aagatcaaac aaagttgttg ggcctccaga cgtagagagc aacttcttcg agagatgcaa 3960

cagatggcca gccgtccctt tgcctctgta aatgtcgcct tggaaacaga tgaggagcct 4020

cctgatctta ttggggggag tataaagact gttcccaaac ccattgcact ggagccgtgt 4080

tttggcaaca aagccgctgt cctctctgtg tttgtgaggc tccctcgagg cctgggtggc 4140

atccctcctc ctgggcagtc aggtcttgct gtggccagcg ccctggtgga catttctcag 4200

cagatgccga tagtgtacaa ggagaagtca ggagccgtga gaaaccggaa gcagcagccc 4260

cctgcacagc ctgggacctg catctgatgc tggggccagg gactctccca cgcacgagct 4320

agtgagtggc acaccagagc catctgcagg gaagggcgtg gcggggaaat ggctgtgcgg 4380

tgcgggacgg aagactggaa accctcaaag catctgactc acctgcatga tcacaagctt 4440

tctttgacgg tttctcccat ccgtgttcca gcatctaacc ttttactttt gcataggaaa 4500

tacttgattt aattacaggt ccagggatga gctgatggtt gctggaggag gccagtgtag 4560

agccagtgag agaactagga atgacactca ggttcactgt ggaaaactgt tcttgggact 4620

gtctcaactg tgcaaaaaac aaaagatgga gtgtttacaa gtagacattc gtcatcagtt 4680

gttcttgaac atggtctttt aaaaactagt cagatgaatt aacttgtttt catctgaagc 4740

ctgctatctt ttttaaaaga tgtgctattt attcttgcac gatttaggca attatctctc 4800

ttccagggag tacctttttt tctagttgag aattaataat ggtccatctc ttttgatcat 4860

atcaagctag gatagaaggg gggctatttt aaatgtcaag gtcagcagtg ttactttgaa 4920

tgtaaactgg tataataggt agttttctat agtaacttga ttaatttagt cttaatccat 4980

ttgaaactct ctcttccttt ctctctgcct gtccctctcc ttctccatct caccctccct 5040

ctctcacaca tacacacaca aacacataca cacaacacta agtgcctaga ctttaaatag 5100

atctagcaat tggaaagtta gtaagcctaa gtttttacat aattgcattc ctacattctt 5160

gtaaaattta aatagctacc attggcaatc tgcttttttt ctaaaatctg atttgcagcc 5220

aggaaagaat tttctcaccc aaggaacatt tgatctagca gcagggatga gaggaaagca 5280

gaaatgaatg aactgtgaaa gctcctgttt ttattatcaa aaaggacact gtcaagaagg 5340

cgccccctgc ccccaccccc gtgtcaccct aggcctgata agcgatcaga ggaaaggact 5400

cattcatgtc acgcttcctt gagcagaaaa gagcactgag agcacttggg acccctggat 5460

cagagagcat ctgtgtgtcc tgcagcctcc tctgaacttg tggttcattc tcaggctggg 5520

gtggactcag atgccaggaa agggacagcc tcccattgtc aggcagaagc tgcccaaagc 5580

ctggagaagg acttgtttgc cctctttccc ccaggagggg ctcgacccac ccaccctccc 5640

tctcagacca aggtggtggc tgtgaggagg gcagcaaatg ctgacaagga tgaaaagcac 5700

atggaaaaaa atggacgagg agggaaaact ctgccaaatg gaaaatgacc aaatttaaga 5760

gggtgggaca gtcccctgct cctctcccag agggcactgc ttggaaattg tgttttcccc 5820

atttatggtg ctctgtattc tggcattatg cagcagcctc ccagaagctc tcttctgctt 5880

caaaacctgg gatctctggc attaccctat tgggatggac cgctggacag caatgctcga 5940

gtttgtgaat ttggagagat actcaaaaga gctaaaactg cagcatttta cctttaaatg 6000

cagtgcctag agagagagta ttgtctcttc cccaacacta accccactcc catgaagaat 6060

tgcctggaaa gatgttttca aggaatttga accataaaac actatctgat gcacagaaca 6120

cctctacttt gagactcacc tctcataaag cttctttttc acattactgt taaagaccag 6180

acgttctaga aaagacccct cctctcatga gctcccccat ccctgctaca gaacacagca 6240

cccatggcgc ctgcagtgga ctggcccctt aattcccaca ggccccccca gcaaggccaa 6300

agggaggccc ctgggtattg tcctcctaca aggaagatcc tctttgtttg ttcaaaggac 6360

cagttttcct aggccaaaga agtctcttcc ccatgttagt cctatgcctt gaaatatcat 6420

gcaccatgac ccacagccat ctggttatgt cttatttttt tcctaaaaga taatgtttat 6480

ttttaaaaag gaaggaagaa gcaagtgaag tttcattctg ctccagcggt ggggaagccg 6540

ctgaatccac ctgcttctcc tttgcaaccg acagcaaaca gctttctccg gcctcagggc 6600

agaaaaaggg aatggcaggg agtaagaggc gctgggctcg gagcctgttt ccaagaagga 6660

attggttgtc atctggcagt gttgcgcgtc acaagagagc ctgtatataa attaaaatag 6720

tcaagacaac actgaccttg cacttgtaca taactataca gtagtgtcca gaatgttcag 6780

acattcggag tgtacataaa acagaaaaaa tcttcatgta tttttattaa atataacaat 6840

gtctgagttt cacctaagat gtttttgtgc catatgctgg atatccaggt tctcgccagg 6900

ccccgataca tgaataacaa acccaagaaa cgcatcccca ttgtgtgatg tgttcagatg 6960

catctggcac caattaggta tttcttaaaa caggactcat ctgtcagagt gcacatgaaa 7020

aatcaggcag ggaatcgaaa cgacagcgct ggaggagact caggaagcag aggcgtccct 7080

gccgctgccc ttggccctgc aagcacatca tgaccctttc tggcagcctc ttggtgctct 7140

gggtagtgag ggatgaccag tcttgtcctg agaaatgttt ctcttagtct ttaagttcaa 7200

agactaacct gtagcaatca gactttccaa aagggggttc tccatttttt gtagttttgt 7260

ctaaattttt aatgaccatt tcctggaatc agtttattat actgaaaact gggggtggga 7320

gtagggagct agtttgttga taaatagttc ccatttcccc gtggagaatt tgacataccc 7380

tggactcctg tgtgcctcct gccatccctg cacacagcct ggggagaagc ctgtgcctcc 7440

ccgtgtggag agaaggcaac cccagatccc ctgagctaac ccggaggaaa ggcagtcctg 7500

gacagaagac tgtcagcaga aggaaagtac tggactaccc gtgggtaagt cctgccattc 7560

aagactggag acacctggga aataaaaaga gcagggcact gctggtggga agaggcattt 7620

taccttccag tgcaaatcct gctcctttga tttaatgggg tgtactgggg ccaggggctg 7680

attcacttcc ttgggagatg gtggtgtttt catgaacatc tttgatcctt ccatttcatt 7740

tattcatcca tccattcaac aagtatttgc taaacactaa cttaagctaa tgctagggta 7800

gtgactgaga tgtaaaaata gattttagaa ttaaaacaaa atccaagtcc tcacacccct 7860

gtcatcccag gagatctttc cttgtggtgg tttctgtgag aattggccat cctgaggaca 7920

cagccaggac ggcagaggcc tcctggcctc agggcatgcc ctgcctacct tctgaaatgt 7980

ttaccccatt gaccaaactt ggctccagcc attgcggtgg tttctagata gccaggccca 8040

ccaagagata ttgccccttg atgagagtca aacaccctgc ctacaaggag atgttttgaa 8100

atggagagga aaattggcac ctcatctttt aaaggcagta atggaattga ttttcagtaa 8160

ctgaatttgt gcacaaaaca ttctaaacac tagtgaagcc tgtttcgttg aactaattct 8220

ggctctggaa atgtttttgt tttatagtta tttacgattt cgtttgtttg gattcaagct 8280

tagtttgtta atatgtataa tttagcatct attacactca tgtaaatatg gagtaagtat 8340

tgtaaactat ttcattgcgg ggattgtggg tgttatacat acatttagga ctgcaatttt 8400

ttggtatttt ttgtattgta aaataacagc taatttaagc aggaacaaga gaactaaggg 8460

aggtctgtgc attttaaaca caaatgtgaa gaacttgtat ataaacaaaa gtaaatacta 8520

taatacaaac ttccttctga aataaaagta gatctggtaa aaaaaaaaaa agaaaaaaaa 8580

aaaaaaaaa 8589

15

2787

Homo sapiens

15

Met Val Ala Val Ala Ala Ala Ala Ala Thr Glu Ala Arg Leu Arg Arg

1 5 10 15

Arg Thr Ala Ala Thr Ala Ala Leu Ala Gly Arg Ser Gly Gly Pro His

20 25 30

Arg Pro Cys Thr Ala Thr Gly Ala Trp Arg Pro Gly Pro Arg Ala Arg

35 40 45

Leu Cys Leu Pro Arg Val Leu Ser Arg Ala Leu Pro Pro Pro Pro Leu

50 55 60

Leu Pro Leu Leu Phe Ser Leu Leu Leu Leu Pro Leu Pro Arg Glu Ala

65 70 75 80

Glu Ala Ala Ala Val Ala Ala Ala Val Ser Gly Ser Ala Ala Ala Glu

85 90 95

Ala Lys Glu Cys Asp Arg Pro Cys Val Asn Gly Gly Arg Cys Asn Pro

100 105 110

Gly Thr Gly Gln Cys Val Cys Pro Ala Gly Trp Val Gly Glu Gln Cys

115 120 125

Gln His Cys Gly Gly Arg Phe Arg Leu Thr Gly Ser Ser Gly Phe Val

130 135 140

Thr Asp Gly Pro Gly Asn Tyr Lys Tyr Lys Thr Lys Cys Thr Trp Leu

145 150 155 160

Ile Glu Gly Gln Pro Asn Arg Ile Met Arg Leu Arg Phe Asn His Phe

165 170 175

Ala Thr Glu Cys Ser Trp Asp His Leu Tyr Val Tyr Asp Gly Asp Ser

180 185 190

Ile Tyr Ala Pro Leu Val Ala Ala Phe Ser Gly Leu Ile Val Pro Glu

195 200 205

Arg Asp Gly Asn Glu Thr Val Pro Glu Val Val Ala Thr Ser Gly Tyr

210 215 220

Ala Leu Leu His Phe Phe Ser Asp Ala Ala Tyr Asn Leu Thr Gly Phe

225 230 235 240

Asn Ile Thr Tyr Ser Phe Asp Met Cys Pro Asn Asn Cys Ser Gly Arg

245 250 255

Gly Glu Cys Lys Ile Ser Asn Ser Ser Asp Thr Val Glu Cys Glu Cys

260 265 270

Ser Glu Asn Trp Lys Gly Glu Ala Cys Asp Ile Pro His Cys Thr Asp

275 280 285

Asn Cys Gly Phe Pro His Arg Gly Ile Cys Asn Ser Ser Asp Val Arg

290 295 300

Gly Cys Ser Cys Phe Ser Asp Trp Gln Gly Pro Gly Cys Ser Val Pro

305 310 315 320

Val Pro Ala Asn Gln Ser Phe Trp Thr Arg Glu Glu Tyr Ser Asn Leu

325 330 335

Lys Leu Pro Arg Ala Ser His Lys Ala Val Val Asn Gly Asn Ile Met

340 345 350

Trp Val Val Gly Gly Tyr Met Phe Asn His Ser Asp Tyr Asn Met Val

355 360 365

Leu Ala Tyr Asp Leu Ala Ser Arg Glu Trp Leu Pro Leu Asn Arg Ser

370 375 380

Val Asn Asn Val Val Val Arg Tyr Gly His Ser Leu Ala Leu Tyr Lys

385 390 395 400

Asp Lys Ile Tyr Met Tyr Gly Gly Lys Ile Asp Ser Thr Gly Asn Val

405 410 415

Thr Asn Glu Leu Arg Val Phe His Ile His Asn Glu Ser Trp Val Leu

420 425 430

Leu Thr Pro Lys Ala Lys Glu Gln Tyr Ala Val Val Gly His Ser Ala

435 440 445

His Ile Val Thr Leu Lys Asn Gly Arg Val Val Met Leu Val Ile Phe

450 455 460

Gly His Cys Pro Leu Tyr Gly Tyr Ile Ser Asn Val Gln Glu Tyr Asp

465 470 475 480

Leu Asp Lys Asn Thr Trp Ser Ile Leu His Thr Gln Gly Ala Leu Val

485 490 495

Gln Gly Gly Tyr Gly His Ser Ser Val Tyr Asp His Arg Thr Arg Ala

500 505 510

Leu Tyr Val His Gly Gly Tyr Lys Ala Phe Ser Ala Asn Lys Tyr Arg

515 520 525

Leu Ala Asp Asp Leu Tyr Arg Tyr Asp Val Asp Thr Gln Met Trp Thr

530 535 540

Ile Leu Lys Asp Ser Arg Phe Phe Arg Tyr Leu His Thr Ala Val Ile

545 550 555 560

Val Ser Gly Thr Met Leu Val Phe Gly Gly Asn Thr His Asn Asp Thr

565 570 575

Ser Met Ser His Gly Ala Lys Cys Phe Ser Ser Asp Phe Met Ala Tyr

580 585 590

Asp Ile Ala Cys Asp Arg Trp Ser Val Leu Pro Arg Pro Asp Leu His

595 600 605

His Asp Val Asn Arg Phe Gly His Ser Ala Val Leu His Asn Ser Thr

610 615 620

Met Tyr Val Phe Gly Gly Phe Asn Ser Leu Leu Leu Ser Asp Ile Leu

625 630 635 640

Val Phe Thr Ser Glu Gln Cys Asp Ala His Arg Ser Glu Ala Ala Cys

645 650 655

Leu Ala Ala Gly Pro Gly Ile Arg Cys Val Trp Asn Thr Gly Ser Ser

660 665 670

Gln Cys Ile Ser Trp Ala Leu Ala Thr Asp Glu Gln Glu Glu Lys Leu

675 680 685

Lys Ser Glu Cys Phe Ser Lys Arg Thr Leu Asp His Asp Arg Cys Asp

690 695 700

Gln His Thr Asp Cys Tyr Ser Cys Thr Ala Asn Thr Asn Asp Cys His

705 710 715 720

Trp Cys Asn Asp His Cys Val Pro Arg Asn His Ser Cys Ser Glu Gly

725 730 735

Gln Ile Ser Ile Phe Arg Tyr Glu Asn Cys Pro Lys Asp Asn Pro Met

740 745 750

Tyr Tyr Cys Asn Lys Lys Thr Ser Cys Arg Ser Cys Ala Leu Asp Gln

755 760 765

Asn Cys Gln Trp Glu Pro Arg Asn Gln Glu Cys Ile Ala Leu Pro Glu

770 775 780

Asn Ile Cys Gly Ile Gly Trp His Leu Val Gly Asn Ser Cys Leu Lys

785 790 795 800

Ile Thr Thr Ala Lys Glu Asn Tyr Asp Asn Ala Lys Leu Phe Cys Arg

805 810 815

Asn His Asn Ala Leu Leu Ala Ser Leu Thr Thr Gln Lys Lys Val Glu

820 825 830

Phe Val Leu Lys Gln Leu Arg Ile Met Gln Ser Ser Gln Ser Met Ser

835 840 845

Lys Leu Thr Leu Thr Pro Trp Val Gly Leu Arg Lys Ile Asn Val Ser

850 855 860

Tyr Trp Cys Trp Glu Asp Met Ser Pro Phe Thr Asn Ser Leu Leu Gln

865 870 875 880

Trp Met Pro Ser Glu Pro Ser Asp Ala Gly Phe Cys Gly Ile Leu Ser

885 890 895

Glu Pro Ser Thr Arg Gly Leu Lys Ala Ala Thr Cys Ile Asn Pro Leu

900 905 910

Asn Gly Ser Val Cys Glu Arg Pro Ala Asn His Ser Ala Lys Gln Cys

915 920 925

Arg Thr Pro Cys Ala Leu Arg Thr Ala Cys Gly Asp Cys Thr Ser Gly

930 935 940

Ser Ser Glu Cys Met Trp Cys Ser Asn Met Lys Gln Cys Val Asp Ser

945 950 955 960

Asn Ala Tyr Val Ala Ser Phe Pro Phe Gly Gln Cys Met Glu Trp Tyr

965 970 975

Thr Met Ser Thr Cys Pro Pro Glu Asn Cys Ser Gly Tyr Cys Thr Cys

980 985 990

Ser His Cys Leu Glu Gln Pro Gly Cys Gly Trp Cys Thr Asp Pro Ser

995 1000 1005

Asn Thr Gly Lys Gly Lys Cys Ile Glu Gly Ser Tyr Lys Gly Pro Val

1010 1015 1020

Lys Met Pro Ser Gln Ala Pro Thr Gly Asn Phe Tyr Pro Gln Pro Leu

1025 1030 1035 1040

Leu Asn Ser Ser Met Cys Leu Glu Asp Ser Arg Tyr Asn Trp Ser Phe

1045 1050 1055

Ile His Cys Pro Ala Cys Gln Cys Asn Gly His Ser Lys Cys Ile Asn

1060 1065 1070

Gln Ser Ile Cys Glu Lys Cys Glu Asn Leu Thr Thr Gly Lys His Cys

1075 1080 1085

Glu Thr Cys Ile Ser Gly Phe Tyr Gly Asp Pro Thr Asn Gly Gly Lys

1090 1095 1100

Cys Gln Pro Cys Lys Cys Asn Gly His Ala Ser Leu Cys Asn Thr Asn

1105 1110 1115 1120

Thr Gly Lys Cys Phe Cys Thr Thr Lys Gly Val Lys Gly Asp Glu Cys

1125 1130 1135

Gln Leu Cys Glu Val Glu Asn Arg Tyr Gln Gly Asn Pro Leu Arg Gly

1140 1145 1150

Thr Cys Tyr Tyr Thr Leu Leu Ile Asp Tyr Gln Phe Thr Phe Ser Leu

1155 1160 1165

Ser Gln Glu Asp Asp Arg Tyr Tyr Thr Ala Ile Asn Phe Val Ala Thr

1170 1175 1180

Pro Asp Glu Gln Asn Arg Asp Leu Asp Met Phe Ile Asn Ala Ser Lys

1185 1190 1195 1200

Asn Phe Asn Leu Asn Ile Thr Trp Ala Ala Ser Phe Ser Ala Gly Thr

1205 1210 1215

Gln Ala Gly Glu Glu Met Pro Val Val Ser Lys Thr Asn Ile Lys Glu

1220 1225 1230

Tyr Lys Asp Ser Phe Ser Asn Glu Lys Phe Asp Phe Arg Asn His Pro

1235 1240 1245

Asn Ile Thr Phe Phe Val Tyr Val Ser Asn Phe Thr Trp Pro Ile Lys

1250 1255 1260

Ile Gln Ile Ala Phe Ser Gln His Ser Asn Phe Met Asp Leu Val Gln

1265 1270 1275 1280

Phe Phe Val Thr Phe Phe Ser Cys Phe Leu Ser Leu Leu Leu Val Ala

1285 1290 1295

Ala Val Val Trp Lys Ile Lys Gln Ser Cys Trp Ala Ser Arg Arg Arg

1300 1305 1310

Glu Gln Leu Leu Arg Glu Met Gln Gln Met Ala Ser Arg Pro Phe Ala

1315 1320 1325

Ser Val Asn Val Ala Leu Glu Thr Asp Glu Glu Pro Pro Asp Leu Ile

1330 1335 1340

Gly Gly Ser Ile Lys Thr Val Pro Lys Pro Ile Ala Leu Glu Pro Cys

1345 1350 1355 1360

Phe Gly Asn Lys Ala Ala Val Leu Ser Val Phe Val Arg Leu Pro Arg

1365 1370 1375

Gly Leu Gly Gly Ile Pro Pro Pro Gly Gln Ser Gly Leu Ala Val Ala

1380 1385 1390

Ser Ala Leu Val Asp Ile Ser Gln Gln Met Pro Ile Val Tyr Lys Glu

1395 1400 1405

Lys Ser Gly Ala Val Arg Asn Arg Lys Gln Gln Pro Pro Ala Gln Pro

1410 1415 1420

Gly Thr Cys Ile Cys Trp Gly Gln Gly Leu Ser His Ala Arg Ala Ser

1425 1430 1435 1440

Glu Trp His Thr Arg Ala Ile Cys Arg Glu Gly Arg Gly Gly Glu Met

1445 1450 1455

Ala Val Arg Cys Gly Thr Glu Asp Trp Lys Pro Ser Lys His Leu Thr

1460 1465 1470

His Leu His Asp His Lys Leu Ser Leu Thr Val Ser Pro Ile Arg Val

1475 1480 1485

Pro Ala Ser Asn Leu Leu Leu Leu His Arg Lys Tyr Leu Ile Leu Gln

1490 1495 1500

Val Gln Gly Ala Asp Gly Cys Trp Arg Arg Pro Val Ser Gln Glu Asn

1505 1510 1515 1520

Glu His Ser Gly Ser Leu Trp Lys Thr Val Leu Gly Thr Val Ser Thr

1525 1530 1535

Val Gln Lys Thr Lys Asp Gly Val Phe Thr Ser Arg His Ser Ser Ser

1540 1545 1550

Val Val Leu Glu His Gly Leu Leu Lys Thr Ser Gln Met Asn Leu Val

1555 1560 1565

Phe Ile Ser Leu Leu Ser Phe Leu Lys Asp Val Leu Phe Ile Leu Ala

1570 1575 1580

Arg Phe Arg Gln Leu Ser Leu Phe Gln Gly Val Pro Phe Phe Leu Val

1585 1590 1595 1600

Glu Asn Trp Ser Ile Ser Phe Asp His Ile Lys Leu Gly Lys Gly Gly

1605 1610 1615

Tyr Phe Lys Cys Gln Gly Gln Gln Cys Tyr Phe Glu Cys Lys Leu Val

1620 1625 1630

Val Val Phe Tyr Ser Asn Leu Ile Asn Leu Val Leu Ile His Leu Lys

1635 1640 1645

Leu Ser Leu Pro Phe Ser Leu Pro Val Pro Leu Leu Leu His Leu Thr

1650 1655 1660

Leu Pro Leu Ser His Ile His Thr Gln Thr His Thr His Asn Thr Lys

1665 1670 1675 1680

Cys Leu Asp Phe Lys Ile Gln Leu Glu Ser Ala Val Phe Thr Leu His

1685 1690 1695

Ser Tyr Ile Leu Val Lys Phe Lys Leu Pro Leu Ala Ile Cys Phe Phe

1700 1705 1710

Ser Lys Ile Phe Ala Ala Arg Lys Glu Phe Ser His Pro Arg Asn Ile

1715 1720 1725

Ser Ser Ser Arg Asp Glu Arg Lys Ala Glu Met Asn Glu Leu Lys Leu

1730 1735 1740

Leu Phe Leu Leu Ser Lys Arg Thr Leu Ser Arg Arg Arg Pro Leu Pro

1745 1750 1755 1760

Pro Pro Pro Cys His Pro Arg Pro Asp Lys Arg Ser Glu Glu Arg Thr

1765 1770 1775

His Ser Cys His Ala Ser Leu Ser Arg Lys Glu His Glu His Leu Gly

1780 1785 1790

Pro Leu Asp Gln Arg Ala Ser Val Cys Pro Ala Ala Ser Ser Glu Leu

1795 1800 1805

Val Val His Ser Gln Ala Gly Val Asp Ser Asp Ala Arg Lys Gly Thr

1810 1815 1820

Ala Ser His Cys Gln Ala Glu Ala Ala Gln Ser Leu Glu Lys Asp Leu

1825 1830 1835 1840

Phe Ala Leu Phe Pro Pro Gly Gly Ala Arg Pro Thr His Pro Pro Ser

1845 1850 1855

Gln Thr Lys Val Val Ala Val Arg Arg Ala Ala Asn Ala Asp Lys Asp

1860 1865 1870

Glu Lys His Met Glu Lys Asn Gly Arg Gly Gly Lys Thr Leu Pro Asn

1875 1880 1885

Gly Lys Pro Asn Leu Arg Gly Trp Asp Ser Pro Leu Leu Leu Ser Gln

1890 1895 1900

Arg Ala Leu Leu Gly Asn Cys Val Phe Pro Ile Tyr Gly Ala Leu Tyr

1905 1910 1915 1920

Ser Gly Ile Met Gln Gln Pro Pro Arg Ser Ser Leu Leu Leu Gln Asn

1925 1930 1935

Leu Gly Ser Leu Ala Leu Pro Tyr Trp Asp Gly Pro Leu Asp Ser Asn

1940 1945 1950

Ala Arg Val Cys Glu Phe Gly Glu Ile Leu Lys Arg Ala Lys Thr Ala

1955 1960 1965

Ala Phe Tyr Leu Met Gln Cys Leu Glu Arg Glu Tyr Cys Leu Phe Pro

1970 1975 1980

Asn Thr Asn Pro Thr Pro Met Lys Asn Cys Leu Glu Arg Cys Phe Gln

1985 1990 1995 2000

Gly Ile Thr Ile Lys His Tyr Leu Met His Arg Thr Pro Leu Leu Asp

2005 2010 2015

Ser Pro Leu Ile Lys Leu Leu Phe His Ile Thr Val Lys Asp Gln Thr

2020 2025 2030

Phe Lys Arg Pro Leu Leu Ser Ala Pro Pro Ser Leu Leu Gln Asn Thr

2035 2040 2045

Ala Pro Met Ala Pro Ala Val Asp Trp Pro Leu Asn Ser His Arg Pro

2050 2055 2060

Pro Gln Gln Gly Gln Arg Glu Ala Pro Gly Tyr Cys Pro Pro Thr Arg

2065 2070 2075 2080

Lys Ile Leu Phe Val Cys Ser Lys Asp Gln Phe Ser Ala Lys Glu Val

2085 2090 2095

Ser Ser Pro Cys Ser Tyr Ala Leu Lys Tyr His Ala Pro Pro Thr Ala

2100 2105 2110

Ile Trp Leu Cys Leu Ile Phe Phe Leu Lys Asp Asn Val Tyr Phe Lys

2115 2120 2125

Gly Arg Lys Lys Gln Val Lys Phe His Ser Ala Pro Ala Val Gly Lys

2130 2135 2140

Pro Leu Asn Pro Pro Ala Ser Pro Leu Gln Pro Thr Ala Asn Ser Phe

2145 2150 2155 2160

Leu Arg Pro Gln Gly Arg Lys Arg Glu Trp Gln Gly Val Arg Gly Ala

2165 2170 2175

Gly Leu Gly Ala Cys Phe Gln Glu Gly Ile Gly Cys His Leu Ala Val

2180 2185 2190

Leu Arg Val Thr Arg Glu Pro Val Tyr Lys Leu Lys Ser Arg Gln His

2195 2200 2205

Pro Cys Thr Cys Thr Leu Tyr Ser Ser Val Gln Asn Val Gln Thr Phe

2210 2215 2220

Gly Val Tyr Ile Lys Gln Lys Lys Ser Ser Cys Ile Phe Ile Lys Tyr

2225 2230 2235 2240

Asn Asn Val Val Ser Pro Lys Met Phe Leu Cys His Met Leu Asp Ile

2245 2250 2255

Gln Val Leu Ala Arg Pro Arg Tyr Met Asn Asn Lys Pro Lys Lys Arg

2260 2265 2270

Ile Pro Ile Val Cys Val Gln Met His Leu Ala Pro Ile Arg Tyr Phe

2275 2280 2285

Leu Lys Gln Asp Ser Ser Val Arg Val His Met Lys Asn Gln Ala Gly

2290 2295 2300

Asn Arg Asn Asp Ser Ala Gly Gly Asp Ser Gly Ser Arg Gly Val Pro

2305 2310 2315 2320

Ala Ala Ala Leu Gly Pro Ala Ser Thr Ser Pro Phe Leu Ala Ala Ser

2325 2330 2335

Trp Cys Ser Gly Gly Met Thr Ser Leu Val Leu Arg Asn Val Ser Leu

2340 2345 2350

Ser Leu Val Gln Arg Leu Thr Cys Ser Asn Gln Thr Phe Gln Lys Gly

2355 2360 2365

Val Leu His Phe Leu Phe Cys Leu Asn Phe Pro Phe Pro Gly Ile Ser

2370 2375 2380

Leu Leu Tyr Lys Leu Gly Val Gly Val Gly Ser Phe Val Asp Lys Phe

2385 2390 2395 2400

Pro Phe Pro Arg Gly Glu Phe Asp Ile Pro Trp Thr Pro Val Cys Leu

2405 2410 2415

Leu Pro Ser Leu His Thr Ala Trp Gly Glu Ala Cys Ala Ser Pro Cys

2420 2425 2430

Gly Glu Lys Ala Thr Pro Asp Pro Leu Ser Pro Gly Gly Lys Ala Val

2435 2440 2445

Leu Asp Arg Arg Leu Ser Ala Glu Gly Lys Tyr Trp Thr Thr Arg Gly

2450 2455 2460

Val Leu Pro Phe Lys Thr Gly Asp Thr Trp Glu Ile Lys Arg Ala Gly

2465 2470 2475 2480

His Cys Trp Trp Glu Glu Ala Phe Tyr Leu Pro Val Gln Ile Leu Leu

2485 2490 2495

Leu Phe Asn Gly Val Tyr Trp Gly Gln Gly Leu Ile His Phe Leu Gly

2500 2505 2510

Arg Trp Trp Cys Phe His Glu His Leu Ser Phe His Phe Ile Tyr Ser

2515 2520 2525

Ser Ile His Ser Thr Ser Ile Cys Thr Leu Thr Ala Asn Ala Arg Val

2530 2535 2540

Val Thr Glu Met Lys Ile Leu Glu Leu Lys Gln Asn Pro Ser Pro His

2545 2550 2555 2560

Thr Pro Val Ile Pro Gly Asp Leu Ser Leu Trp Trp Phe Leu Glu Leu

2565 2570 2575

Ala Ile Leu Arg Thr Gln Pro Gly Arg Gln Arg Pro Pro Gly Leu Arg

2580 2585 2590

Ala Cys Pro Ala Tyr Leu Leu Lys Cys Leu Pro His Pro Asn Leu Ala

2595 2600 2605

Pro Ala Ile Ala Val Val Ser Arg Pro Gly Pro Pro Arg Asp Ile Ala

2610 2615 2620

Pro Glu Ser Asn Thr Leu Pro Thr Arg Arg Cys Phe Glu Met Glu Arg

2625 2630 2635 2640

Lys Ile Gly Thr Ser Ser Phe Lys Gly Ser Asn Gly Ile Asp Phe Gln

2645 2650 2655

Leu Asn Leu Cys Thr Lys His Ser Lys His Ser Leu Phe Arg Thr Asn

2660 2665 2670

Ser Gly Ser Gly Asn Val Phe Val Leu Leu Phe Thr Ile Ser Phe Val

2675 2680 2685

Trp Ile Gln Ala Phe Val Asn Met Tyr Asn Leu Ala Ser Ile Thr Leu

2690 2695 2700

Met Ile Trp Ser Lys Tyr Cys Lys Leu Phe His Cys Gly Asp Cys Gly

2705 2710 2715 2720

Cys Tyr Thr Tyr Ile Asp Cys Asn Phe Leu Val Phe Phe Val Leu Asn

2725 2730 2735

Asn Ser Phe Lys Gln Glu Gln Glu Asn Gly Arg Ser Val His Phe Lys

2740 2745 2750

His Lys Cys Glu Glu Leu Val Tyr Lys Gln Lys Ile Leu Tyr Lys Leu

2755 2760 2765

Pro Ser Glu Ile Lys Val Asp Leu Val Lys Lys Lys Lys Glu Lys Lys

2770 2775 2780

Lys Lys Lys

2785

16

4072

Homo sapiens

16

atggtggcgg tggccgcagc ggcggcaact gaggcaaggc tgaggaggag gacggcggcg 60

acggcagcgc tcgcgggcag gagcggcggg ccgcaccgac cctgcaccgc gacaggggcc 120

tggaggccgg gaccgcgcgc ccggctgtgt ctcccgcggg tgctgtcgcg ggcgctgccc 180

ccgccgccgc tgctgccgct gctcttttcg ctgctgctgc tgccgctgcc ccgggaggcc 240

gaggccgctg cggtggcggc ggcggtgtcc ggctcggccg cagccgaggc caaggaatgt 300

gaccggccgt gtgtcaacgg cggtcgctgc aaccctggca ccggccagtg cgtctgcccc 360

gccggctggg tgggcgagca atgccagcac tgcgggggcc gcttcagact aactggatct 420

tctgggtttg tgacagatgg acctggaaat tataaataca aaacgaagtg cacgtggctc 480

attgaaggac agccaaatag aataatgaga cttcgtttca atcattttgc tacagagtgt 540

agttgggacc atttatatgt ttatgatggg gactcaattt atgcaccgct agttgctgca 600

tttagtggcc tcattgttcc tgagagagat ggcaatgaga ctgtccctga ggttgttgcc 660

acatcaggtt atgccttgct gcattttttt agtgatgctg cttataattt gactggattt 720

aatattactt acagttttga tatgtgtcca aataactgct caggccgagg agagtgtaag 780

atcagtaata gcagcgatac tgttgaatgt gaatgttctg aaaactggaa aggtgaagca 840

tgtgacattc ctcactgtac agacaactgt ggttttcctc atcgaggcat ctgcaattca 900

agtgatgtca gaggatgctc ctgcttctca gactggcagg gtcctggatg ttcagttcct 960

gtaccagcta accagtcatt ttggactcga gaggaatatt ctaacttaaa gctccccaga 1020

gcatctcata aagctgtggt caatggaaac attatgtggg ttgttggagg atatatgttc 1080

aaccactcag attataacat ggttctagcg tatgaccttg cttctaggga gtggcttcca 1140

ctaaaccgtt ctgtgaacaa tgtggttgtt agatatggtc attctttggc attatacaag 1200

gataaaattt acatgtatgg aggaaaaatt gattcaactg ggaatgtgac caatgagttg 1260

agagtttttc acattcataa tgagtcatgg gtgttgttga cccctaaggc aaaggagcag 1320

tatgcagtgg ttgggcactc tgcacacatt gttacactga agaatggccg agtggtcatg 1380

ctggtcatct ttggtcactg ccctctctat ggatatataa gcaatgtgca ggaatatgat 1440

ttggataaga acacatggag tatattacac acccagggtg cccttgtgca agggggttac 1500

ggccatagca gtgtttacga ccataggacc agggccctat acgttcatgg tggctacaag 1560

gctttcagtg ccaataagta ccggcttgca gatgatctct accgatatga tgtggatacc 1620

cagatgtgga ccattcttaa ggacagccga tttttccgtt acttgcacac agctgtgata 1680

gtgagtggaa ccatgctggt gtttggggga aacacacaca atgacacatc tatgagccat 1740

ggcgccaaat gcttctcttc agatttcatg gcctatgaca ttgcctgtga ccgctggtca 1800

gtgcttccca gacctgatct ccaccatgat gtcaacagat ttggccattc agcagtctta 1860

cacaacagca ccatgtatgt gttcggtggt ttcaatagtc tcctcctcag cgacatcctg 1920

gtattcacct cggaacagtg tgatgcgcat cggagtgaag ccgcttgttt agcagcagga 1980

cctggtattc ggtgtgtgtg gaacacaggg tcgtctcagt gtatctcgtg ggcgctggca 2040

actgatgaac aagaagaaaa gttaaaatca gaatgttttt ccaaaagaac tcttgaccat 2100

gacagatgtg accagcacac agattgttac agctgcacag ccaacaccaa tgactgccac 2160

tggtgcaatg accattgtgt ccccaggaac cacagctgct cagaaggcca gatctccatt 2220

tttaggtatg agaattgccc caaggataac cctatgtact actgtaacaa gaagaccagc 2280

tgcaggagct gtgccctgga ccagaactgc cagtgggagc cccggaatca ggagtgcatt 2340

gccctgcccg aaaatatctg tggcattggc tggcatttgg ttggaaactc atgtttgaaa 2400

attactactg ccaaggagaa ttatgacaat gctaaattgt tctgtaggaa ccacaatgcc 2460

cttttggctt ctcttacaac ccagaagaag gtagaatttg tccttaagca gctgcgaata 2520

atgcagtcat ctcagagcat gtccaagctc accttaaccc catgggtcgg ccttcggaag 2580

atcaatgtgt cctactggtg ctgggaagat atgtccccat ttacaaatag tttactacag 2640

tggatgccgt ctgagcccag tgatgctgga ttctgtggaa ttttatcaga acccagtact 2700

cggggactga aggctgcaac ctgcatcaac ccactcaatg gtagtgtctg tgaaaggcct 2760

gcaaaccaca gtgctaagca gtgccggaca ccatgtgcct tgaggacagc atgtggagat 2820

tgcaccagcg gcagctctga gtgcatgtgg tgcagcaaca tgaagcagtg tgtggactcc 2880

aatgcctatg tggcctcctt cccttttggc cagtgtatgg aatggtatac gatgagcacc 2940

tgcccccctg aaaattgttc aggctactgt acctgtagtc attgcttgga gcaaccaggc 3000

tgtggctggt gtactgatcc cagcaatact ggcaaaggga aatgcataga gggttcctat 3060

aaaggaccag tgaagatgcc ttcgcaagcc cctacaggaa atttctatcc acagcccctg 3120

ctcaattcca gcatgtgtct agaggacagc agatacaact ggtctttcat tcactgtcca 3180

gcttgccaat gcaacggcca cagtaaatgc atcaatcaga gcatctgtga gaagtgtgag 3240

aacctgacca caggcaagca ctgcgagacc tgcatatctg gcttctacgg tgatcccacc 3300

aatggaggga aatgtcagcc atgcaagtgc aatgggcacg cgtctctgtg caacaccaac 3360

acgggcaagt gcttctgcac caccaagggc gtcaaggggg acgagtgcca gctatgtgag 3420

gtagaaaatc gataccaagg aaaccctctc agaggaacat gttattatac tcttcttatt 3480

gactatcagt tcacctttag tctatcccag gaagatgatc gctattacac agctatcaat 3540

tttgtggcta ctcctgacga acaaaacagg gatttggaca tgttcatcaa tgcctccaag 3600

aatttcaacc tcaacatcac ctgggctgcc agtttctcag ctggaaccca ggctggagaa 3660

gagatgcctg ttgtttcaaa aaccaacatt aaggagtaca aagatagttt ctctaatgag 3720

aagtttgatt ttcgcaacca cccaaatatc actttctttg tttatgtcag taatttcacc 3780

tggcccatca aaattcaggt gcaaactgaa caatgaggac gcatggacac aggaagggga 3840

acatcacaca ccagggcctg ttgtggggtg gggggaaggg gaagggatag cattagggga 3900

tatacctaat gttaaatgac gagttaatgg gtgcagcaca ccaacatggc atatgtatac 3960

atatgtaaca aacctgcatg ttgtgcacat gtaccctaaa acttaaagta taattaaaaa 4020

aaaaaaaaga aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa 4072

17

1350

Homo sapiens

17

Met Val Ala Ala Ala Ala Ala Thr Glu Ala Arg Leu Arg Arg Arg Thr

1 5 10 15

Ala Ala Thr Ala Ala Leu Ala Gly Arg Ser Gly Gly Pro His Trp Asp

20 25 30

Trp Asp Val Thr Arg Ala Gly Arg Pro Gly Leu Gly Ala Gly Leu Arg

35 40 45

Leu Pro Arg Leu Leu Ser Pro Pro Leu Arg Pro Arg Leu Leu Leu Leu

50 55 60

Leu Leu Leu Leu Pro Pro Pro Leu Leu Leu Leu Leu Leu Pro Cys Glu

65 70 75 80

Ala Glu Ala Ala Ala Ala Ala Ala Ala Val Ser Gly Ser Ala Ala Ala

85 90 95

Glu Ala Lys Glu Cys Asp Arg Pro Cys Val Asn Gly Gly Arg Cys Asn

100 105 110

Pro Gly Thr Gly Gln Cys Val Cys Pro Ala Gly Trp Val Gly Glu Gln

115 120 125

Cys Gln His Cys Gly Gly Arg Phe Arg Leu Thr Gly Ser Ser Gly Phe

130 135 140

Val Thr Asp Gly Pro Gly Asn Tyr Lys Tyr Lys Thr Lys Cys Thr Trp

145 150 155 160

Leu Ile Glu Gly Gln Pro Asn Arg Ile Met Arg Leu Arg Phe Asn His

165 170 175

Phe Ala Thr Glu Cys Ser Trp Asp His Leu Tyr Val Tyr Asp Gly Asp

180 185 190

Ser Ile Tyr Ala Pro Leu Val Ala Ala Phe Ser Gly Leu Ile Val Pro

195 200 205

Glu Arg Asp Gly Asn Glu Thr Val Pro Glu Val Val Ala Thr Ser Gly

210 215 220

Tyr Ala Leu Leu His Phe Phe Ser Asp Ala Ala Tyr Asn Leu Thr Gly

225 230 235 240

Phe Asn Ile Thr Tyr Ser Phe Asp Met Cys Pro Asn Asn Cys Ser Gly

245 250 255

Arg Gly Glu Cys Lys Ile Ser Asn Ser Ser Thr Val Glu Cys Glu Cys

260 265 270

Ser Glu Asn Trp Lys Gly Glu Ala Cys Asp Ile Pro His Cys Thr Asp

275 280 285

Asn Cys Gly Phe Pro His Arg Gly Ile Cys Asn Ser Ser Asp Val Arg

290 295 300

Gly Cys Ser Cys Phe Ser Asp Trp Gln Gly Pro Gly Cys Ser Val Pro

305 310 315 320

Val Pro Ala Asn Gln Ser Phe Trp Thr Arg Glu Glu Tyr Ser Asn Leu

325 330 335

Lys Leu Pro Arg Ala Ser His Lys Ala Val Val Asn Gly Asn Ile Met

340 345 350

Trp Val Val Gly Gly Tyr Met Phe Asn His Ser Asp Tyr Asn Met Val

355 360 365

Leu Ala Tyr Asp Leu Ala Ser Arg Glu Trp Leu Pro Leu Asn Arg Ser

370 375 380

Val Asn Asn Val Val Val Arg Tyr Gly His Ser Leu Ala Leu Tyr Lys

385 390 395 400

Asp Lys Ile Tyr Met Tyr Gly Gly Lys Ile Asp Thr Gly Asn Val Thr

405 410 415

Asn Glu Leu Arg Val Phe His Ile His Asn Glu Ser Trp Val Leu Leu

420 425 430

Thr Pro Lys Ala Lys Glu Gln Tyr Ala Val Val Gly His Ser Ala His

435 440 445

Ile Val Thr Leu Lys Asn Gly Arg Val Val Met Leu Val Ile Phe Gly

450 455 460

His Cys Pro Leu Tyr Gly Tyr Ile Ser Asn Val Gln Glu Tyr Asp Leu

465 470 475 480

Asp Lys Asn Thr Trp Ser Ile Leu His Thr Gln Gly Ala Leu Val Gln

485 490 495

Gly Gly Tyr Gly His Ser Ser Val Tyr Asp His Arg Thr Arg Ala Leu

500 505 510

Tyr Val His Gly Gly Tyr Lys Ala Phe Ser Ala Asn Lys Tyr Arg Leu

515 520 525

Ala Asp Asp Leu Tyr Arg Tyr Asp Val Asp Thr Gln Met Trp Thr Ile

530 535 540

Leu Lys Asp Ser Arg Phe Phe Arg Tyr Leu His Thr Ala Val Ile Val

545 550 555 560

Ser Gly Thr Met Leu Val Phe Gly Gly Asn Thr His Asn Asp Thr Ser

565 570 575

Met Ser His Gly Ala Lys Cys Phe Ser Ser Asp Phe Met Ala Tyr Asp

580 585 590

Ile Ala Cys Asp Arg Trp Ser Val Leu Pro Arg Pro Asp Leu His His

595 600 605

Asp Val Asn Arg Phe Gly His Ser Ala Val Leu His Asn Ser Thr Met

610 615 620

Tyr Val Phe Gly Gly Phe Asn Ser Leu Leu Leu Ser Asp Ile Leu Val

625 630 635 640

Phe Thr Ser Glu Gln Cys Asp Ala His Arg Ser Glu Ala Ala Cys Leu

645 650 655

Ala Ala Gly Pro Gly Ile Arg Cys Val Trp Asn Thr Gly Ser Ser Gln

660 665 670

Cys Ile Ser Trp Ala Leu Ala Thr Asp Glu Gln Glu Glu Lys Leu Lys

675 680 685

Ser Glu Cys Phe Ser Lys Arg Thr Leu Asp His Asp Arg Cys Asp Gln

690 695 700

His Thr Asp Cys Tyr Ser Cys Thr Ala Asn Thr Asn Asp Cys His Trp

705 710 715 720

Cys Asn Asp His Cys Val Pro Arg Asn His Ser Cys Ser Glu Gly Gln

725 730 735

Ile Ser Ile Phe Arg Tyr Glu Asn Cys Pro Lys Asp Asn Pro Met Tyr

740 745 750

Tyr Cys Asn Lys Lys Thr Ser Cys Arg Ser Cys Ala Leu Asp Gln Asn

755 760 765

Cys Gln Trp Glu Pro Arg Asn Gln Glu Cys Ile Ala Leu Pro Glu Asn

770 775 780

Ile Cys Gly Ile Gly Trp His Leu Val Gly Asn Ser Cys Leu Lys Ile

785 790 795 800

Thr Thr Ala Lys Glu Asn Tyr Asp Asn Ala Lys Leu Phe Cys Arg Asn

805 810 815

His Asn Ala Leu Leu Ala Ser Leu Thr Thr Gln Lys Lys Val Glu Phe

820 825 830

Val Leu Lys Gln Leu Arg Ile Met Gln Ser Ser Gln Ser Met Ser Lys

835 840 845

Leu Thr Leu Thr Pro Trp Val Gly Leu Arg Lys Ile Asn Val Ser Tyr

850 855 860

Trp Cys Trp Glu Asp Met Ser Pro Phe Thr Asn Ser Leu Leu Gln Trp

865 870 875 880

Met Pro Ser Glu Pro Ser Asp Ala Gly Phe Cys Gly Ile Leu Ser Glu

885 890 895

Pro Ser Thr Arg Gly Leu Lys Ala Ala Thr Cys Ile Asn Pro Leu Asn

900 905 910

Gly Ser Val Cys Glu Arg Pro Ala Asn His Ser Ala Lys Gln Cys Arg

915 920 925

Thr Pro Cys Ala Leu Arg Thr Ala Cys Gly Asp Cys Thr Ser Gly Ser

930 935 940

Ser Glu Cys Met Trp Cys Ser Asn Met Lys Gln Cys Val Asp Ser Asn

945 950 955 960

Ala Tyr Val Ala Ser Phe Pro Phe Gly Gln Cys Met Glu Trp Tyr Thr

965 970 975

Met Ser Thr Cys Pro Pro Glu Asn Cys Ser Gly Tyr Cys Thr Cys Ser

980 985 990

His Cys Leu Glu Gln Pro Gly Cys Gly Trp Cys Thr Asp Pro Ser Asn

995 1000 1005

Thr Gly Lys Gly Lys Cys Ile Glu Gly Ser Tyr Lys Gly Pro Val Lys

1010 1015 1020

Met Pro Ser Gln Ala Pro Thr Gly Asn Phe Tyr Pro Gln Pro Leu Leu

1025 1030 1035 1040

Asn Ser Ser Met Cys Leu Glu Asp Ser Arg Tyr Asn Trp Ser Phe Ile

1045 1050 1055

His Cys Pro Ala Cys Gln Cys Asn Gly His Ser Lys Cys Ile Asn Gln

1060 1065 1070

Ser Ile Cys Glu Lys Cys Glu Asn Leu Thr Thr Gly Lys His Cys Glu

1075 1080 1085

Thr Cys Ile Ser Gly Phe Tyr Gly Asp Pro Thr Asn Gly Gly Lys Cys

1090 1095 1100

Gln Pro Cys Lys Cys Asn Gly His Ala Ser Leu Cys Asn Thr Asn Thr

1105 1110 1115 1120

Gly Lys Cys Phe Cys Thr Thr Lys Gly Val Lys Gly Asp Glu Cys Gln

1125 1130 1135

Leu Cys Glu Val Glu Asn Arg Tyr Gln Gly Asn Pro Leu Arg Gly Thr

1140 1145 1150

Cys Tyr Tyr Thr Leu Leu Ile Asp Tyr Gln Phe Thr Phe Ser Leu Ser

1155 1160 1165

Gln Glu Asp Asp Arg Tyr Tyr Thr Ala Ile Asn Phe Val Ala Thr Pro

1170 1175 1180

Asp Glu Gln Asn Arg Asp Leu Asp Met Phe Ile Asn Ala Ser Lys Asn

1185 1190 1195 1200

Phe Asn Leu Asn Ile Thr Trp Ala Ala Ser Phe Ser Ala Gly Thr Gln

1205 1210 1215

Ala Gly Glu Glu Met Pro Val Val Ser Lys Thr Asn Ile Lys Glu Tyr

1220 1225 1230

Lys Asp Ser Phe Ser Asn Glu Lys Phe Asp Phe Arg Asn His Pro Asn

1235 1240 1245

Ile Thr Phe Phe Val Tyr Val Ser Asn Phe Thr Trp Pro Ile Lys Ile

1250 1255 1260

Gln Val Gln Thr Glu Gln Gly Arg Met Asp Thr Gly Arg Gly Thr Ser

1265 1270 1275 1280

His Thr Arg Ala Cys Cys Gly Val Gly Gly Arg Gly Arg Asp Ser Ile

1285 1290 1295

Arg Gly Tyr Thr Cys Met Thr Ser Trp Val Gln His Thr Asn Met Ala

1300 1305 1310

Tyr Val Tyr Ile Cys Asn Lys Pro Ala Cys Cys Ala His Val Pro Asn

1315 1320 1325

Leu Lys Tyr Asn Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys

1330 1335 1340

Lys Lys Lys Lys Lys Lys

1345 1350

18

2625

Homo sapiens

18

atggtggcgg tggccgcagc ggcggcaact gaggcaaggc tgaggaggag gacggcggcg 60

acggcagcgc tcgcgggcag gagcggcggg ccgcaccgac cctgcaccgc gacaggggcc 120

tggaggccgg gaccgcgcgc ccggctgtgt ctcccgcggg tgctgtcgcg ggcgctgccc 180

ccgccgccgc tgctgccgct gctcttttcg ctgctgctgc tgccgctgcc ccgggaggcc 240

gaggccgctg cggtggcggc ggcggtgtcc ggctcggccg cagccgaggc caaggaatgt 300

gaccggccgt gtgtcaacgg cggtcgctgc aaccctggca ccggccagtg cgtctgcccc 360

gccggctggg tgggcgagca atgccagcac tgcgggggcc gcttcagact aactggatct 420

tctgggtttg tgacagatgg acctggaaat tataaataca aaacgaagtg cacgtggctc 480

attgaaggac agccaaatag aataatgaga cttcgtttca atcattttgc tacagagtgt 540

agttgggacc atttatatgt ttatgatggg gactcaattt atgcaccgct agttgctgca 600

tttagtggcc tcattgttcc tgagagagat ggcaatgaga ctgtccctga ggttgttgcc 660

acatcaggtt atgccttgct gcattttttt agtgatgctg cttataattt gactggattt 720

aatattactt acagttttga tatgtgtcca aataactgct caggccgagg agagtgtaag 780

atcagtaata gcagcgatac tgttgaatgt gaatgttctg aaaactggaa aggtgaagca 840

tgtgacattc ctcactgtac agacaactgt ggttttcctc atcgaggcat ctgcaattca 900

agtgatgtca gaggatgctc ctgcttctca gactggcagg gtcctggatg ttcagttcct 960

gtaccagcta accagtcatt ttggactcga gaggaatatt ctaacttaaa gctccccaga 1020

gcatctcata aagctgtggt caatggaaac attatgtggg ttgttggagg atatatgttc 1080

aaccactcag attataacat ggttctagcg tatgaccttg cttctaggga gtggcttcca 1140

ctaaaccgtt ctgtgaacaa tgtggttgtt agatatggtc attctttggc attatacaag 1200

gataaaattt acatgtatgg aggaaaaatt gattcaactg ggaatgtgac caatgagttg 1260

agagtttttc acattcataa tgagtcatgg gtgttgttga cccctaaggc aaaggagcag 1320

tatgcagtgg ttgggcactc tgcacacatt gttacactga agaatggccg agtggtcatg 1380

ctggtcatct ttggtcactg ccctctctat ggatatataa gcaatgtgca ggaatatgat 1440

ttggataaga acacatggag tatattacac acccagggtg cccttgtgca agggggttac 1500

ggccatagca gtgtttacga ccataggacc agggccctat acgttcatgg tggctacaag 1560

gctttcagtg ccaataagta ccggcttgca gatgatctct accgatatga tgtggatacc 1620

cagatgtgga ccattcttaa ggacagccga tttttccgtt acttgcacac agctgtgata 1680

gtgagtggaa ccatgctggt gtttggggga aacacacaca atgacacatc tatgagccat 1740

ggcgccaaat gcttctcttc agatttcatg gcctatgaca ttgcctgtga ccgctggtca 1800

gtgcttccca gacctgatct ccaccatgat gtcaacagat ttggccattc agcagtctta 1860

cacaacagca ccatgtatgt gttcggtggt ttcaatagtc tcctcctcag cgacatcctg 1920

gtattcacct cggaacagtg tgatgcgcat cggagtgaag ccgcttgttt agcagcagga 1980

cctggtattc ggtgtgtgtg gaacacaggg tcgtctcagt gtatctcgtg ggcgctggca 2040

actgatgaac aagaagaaaa gttaaaatca gaatgttttt ccaaaagaac tcttgaccat 2100

gacagatgtg accagcacac agattgttac agctgcacag ccaacaccaa tgactgccac 2160

tggtgcaatg accattgtgt ccccaggaac cacagctgct cagaaggcca gatctccatt 2220

tttaggtatg agaattgccc caaggataac cctatgtact actgtaacaa gaagaccagc 2280

tgcaggagct gtgccctgga ccagaactgc cagtgggagc cccggaatca ggagtgcatt 2340

gccctgcccg gtaggccttg cagggtcatc ttggtgtgtg tgggtccatt acttcagcct 2400

gcttccccca acactgtgca gcctaagttg aacctagcag aggggaagag ctaattctgt 2460

ccattcatcc cccacacgag tattatgggc ttttttgttt ttaactaaaa tacagttctt 2520

aagtatttgt tcctactgtc ctttgaaata aagtgaaaca tcctttgctg ctctgtaaaa 2580

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa 2625

19

871

Homo sapiens

19

Met Val Ala Ala Ala Ala Ala Thr Glu Ala Arg Leu Arg Arg Arg Thr

1 5 10 15

Ala Ala Thr Ala Ala Leu Ala Gly Arg Ser Gly Gly Pro His Trp Asp

20 25 30

Trp Asp Val Thr Arg Ala Gly Arg Pro Gly Leu Gly Ala Gly Leu Arg

35 40 45

Leu Pro Arg Leu Leu Ser Pro Pro Leu Arg Pro Arg Leu Leu Leu Leu

50 55 60

Leu Leu Leu Leu Pro Pro Pro Leu Leu Leu Leu Leu Leu Pro Cys Glu

65 70 75 80

Ala Glu Ala Ala Ala Ala Ala Ala Ala Val Ser Gly Ser Ala Ala Ala

85 90 95

Glu Ala Lys Glu Cys Asp Arg Pro Cys Val Asn Gly Gly Arg Cys Asn

100 105 110

Pro Gly Thr Gly Gln Cys Val Cys Pro Ala Gly Trp Val Gly Glu Gln

115 120 125

Cys Gln His Cys Gly Gly Arg Phe Arg Leu Thr Gly Ser Ser Gly Phe

130 135 140

Val Thr Asp Gly Pro Gly Asn Tyr Lys Tyr Lys Thr Lys Cys Thr Trp

145 150 155 160

Leu Ile Glu Gly Gln Pro Asn Arg Ile Met Arg Leu Arg Phe Asn His

165 170 175

Phe Ala Thr Glu Cys Ser Trp Asp His Leu Tyr Val Tyr Asp Gly Asp

180 185 190

Ser Ile Tyr Ala Pro Leu Val Ala Ala Phe Ser Gly Leu Ile Val Pro

195 200 205

Glu Arg Asp Gly Asn Glu Thr Val Pro Glu Val Val Ala Thr Ser Gly

210 215 220

Tyr Ala Leu Leu His Phe Phe Ser Asp Ala Ala Tyr Asn Leu Thr Gly

225 230 235 240

Phe Asn Ile Thr Tyr Ser Phe Asp Met Cys Pro Asn Asn Cys Ser Gly

245 250 255

Arg Gly Glu Cys Lys Ile Ser Asn Ser Ser Thr Val Glu Cys Glu Cys

260 265 270

Ser Glu Asn Trp Lys Gly Glu Ala Cys Asp Ile Pro His Cys Thr Asp

275 280 285

Asn Cys Gly Phe Pro His Arg Gly Ile Cys Asn Ser Ser Asp Val Arg

290 295 300

Gly Cys Ser Cys Phe Ser Asp Trp Gln Gly Pro Gly Cys Ser Val Pro

305 310 315 320

Val Pro Ala Asn Gln Ser Phe Trp Thr Arg Glu Glu Tyr Ser Asn Leu

325 330 335

Lys Leu Pro Arg Ala Ser His Lys Ala Val Val Asn Gly Asn Ile Met

340 345 350

Trp Val Val Gly Gly Tyr Met Phe Asn His Ser Asp Tyr Asn Met Val

355 360 365

Leu Ala Tyr Asp Leu Ala Ser Arg Glu Trp Leu Pro Leu Asn Arg Ser

370 375 380

Val Asn Asn Val Val Val Arg Tyr Gly His Ser Leu Ala Leu Tyr Lys

385 390 395 400

Asp Lys Ile Tyr Met Tyr Gly Gly Lys Ile Asp Thr Gly Asn Val Thr

405 410 415

Asn Glu Leu Arg Val Phe His Ile His Asn Glu Ser Trp Val Leu Leu

420 425 430

Thr Pro Lys Ala Lys Glu Gln Tyr Ala Val Val Gly His Ser Ala His

435 440 445

Ile Val Thr Leu Lys Asn Gly Arg Val Val Met Leu Val Ile Phe Gly

450 455 460

His Cys Pro Leu Tyr Gly Tyr Ile Ser Asn Val Gln Glu Tyr Asp Leu

465 470 475 480

Asp Lys Asn Thr Trp Ser Ile Leu His Thr Gln Gly Ala Leu Val Gln

485 490 495

Gly Gly Tyr Gly His Ser Ser Val Tyr Asp His Arg Thr Arg Ala Leu

500 505 510

Tyr Val His Gly Gly Tyr Lys Ala Phe Ser Ala Asn Lys Tyr Arg Leu

515 520 525

Ala Asp Asp Leu Tyr Arg Tyr Asp Val Asp Thr Gln Met Trp Thr Ile

530 535 540

Leu Lys Asp Ser Arg Phe Phe Arg Tyr Leu His Thr Ala Val Ile Val

545 550 555 560

Ser Gly Thr Met Leu Val Phe Gly Gly Asn Thr His Asn Asp Thr Ser

565 570 575

Met Ser His Gly Ala Lys Cys Phe Ser Ser Asp Phe Met Ala Tyr Asp

580 585 590

Ile Ala Cys Asp Arg Trp Ser Val Leu Pro Arg Pro Asp Leu His His

595 600 605

Asp Val Asn Arg Phe Gly His Ser Ala Val Leu His Asn Ser Thr Met

610 615 620

Tyr Val Phe Gly Gly Phe Asn Ser Leu Leu Leu Ser Asp Ile Leu Val

625 630 635 640

Phe Thr Ser Glu Gln Cys Asp Ala His Arg Ser Glu Ala Ala Cys Leu

645 650 655

Ala Ala Gly Pro Gly Ile Arg Cys Val Trp Asn Thr Gly Ser Ser Gln

660 665 670

Cys Ile Ser Trp Ala Leu Ala Thr Asp Glu Gln Glu Glu Lys Leu Lys

675 680 685

Ser Glu Cys Phe Ser Lys Arg Thr Leu Asp His Asp Arg Cys Asp Gln

690 695 700

His Thr Asp Cys Tyr Ser Cys Thr Ala Asn Thr Asn Asp Cys His Trp

705 710 715 720

Cys Asn Asp His Cys Val Pro Arg Asn His Ser Cys Ser Glu Gly Gln

725 730 735

Ile Ser Ile Phe Arg Tyr Glu Asn Cys Pro Lys Asp Asn Pro Met Tyr

740 745 750

Tyr Cys Asn Lys Lys Thr Ser Cys Arg Ser Cys Ala Leu Asp Gln Asn

755 760 765

Cys Gln Trp Glu Pro Arg Asn Gln Glu Cys Ile Ala Leu Pro Gly Arg

770 775 780

Pro Cys Arg Val Ile Leu Val Cys Val Gly Pro Leu Leu Gln Pro Ala

785 790 795 800

Ser Pro Asn Thr Val Gln Pro Lys Leu Asn Leu Ala Glu Gly Lys Ser

805 810 815

Phe Cys Pro Phe Ile Pro His Thr Ser Ile Met Gly Phe Phe Val Phe

820 825 830

Asn Asn Thr Val Leu Lys Tyr Leu Phe Leu Leu Ser Phe Glu Ile Lys

835 840 845

Asn Ile Leu Cys Cys Ser Val Lys Lys Lys Lys Lys Lys Lys Lys Lys

850 855 860

Lys Lys Lys Lys Lys Lys Lys

865 870

20

26

Artificial Sequence

Primer

20

tctcctagtt gtagtacatg ctgttg 26

21

28

Artificial Sequence

Primer

21

aggtcctgtc tcaagaaata gcaataac 28

22

23

Artificial Sequence

Primer

22

tttgaaggcc cctgaagtca gag 23

23

29

Artificial Sequence

Primer

23

ttgagtcccc atcataaaca tataaatgg 29

24

29

Artificial Sequence

Primer

24

ttctaggcca aatagaataa tgagacttc 29

25

26

Artificial Sequence

Primer

25

agaactaatt ccatgagatg agtgtg 26

26

25

Artificial Sequence

Primer

26

tgaagttgct gtaatctggt ctgtg 25

27

22

Artificial Sequence

Primer

27

aaggagcctg actagaagcc tc 22

28

27

Artificial Sequence

Primer

28

taaactccct acagttcact aactcag 27

29

25

Artificial Sequence

Primer

29

agcgctgttg agtgtgaatg ttctg 25

30

26

Artificial Sequence

Primer

30

aaagccacag ttgtctgtac agtgag 26

31

25

Artificial Sequence

Primer

31

aggtctgcat tagttgcaat gttgc 25

32

25

Artificial Sequence

Primer

32

tatacacccc cttatataca ctcag 25

33

24

Artificial Sequence

Primer

33

agagcctctc ataaagctgt ggtc 24

34

25

Artificial Sequence

Primer

34

ttgaacatat atccgccaac aaccc 25

35

28

Artificial Sequence

Primer

35

cttggaatac tataaacttt caggctgc 28

36

29

Artificial Sequence

Primer

36

taaagcaaca ggaagagttg aacttcttg 29

37

20

Artificial Sequence

Primer

37

tgcaccctgt gtgcacatgg 20

38

26

Artificial Sequence

Primer

38

ttacggtgtc ctaataataa gggcag 26

39

28

Artificial Sequence

Primer

39

aatcatgggt attgttaact ccgaaagc 28

40

24

Artificial Sequence

Primer

40

tgtaacaatg tgtgccgagt gtcc 24

41

22

Artificial Sequence

Primer

41

tctctctcca gccctagagt tg 22

42

24

Artificial Sequence

Primer

42

agaagaggag cctgcaacat tgac 24

43

22

Artificial Sequence

Primer

43

tttgttggcg ctgaaagcct tg 22

44

25

Artificial Sequence

Primer

44

tggccacagt agtgtttatg atgac 25

45

27

Artificial Sequence

Primer

45

ttaatcaatt gcctctgcag attctag 27

46

26

Artificial Sequence

Primer

46

tggcttacgt atagggggaa atcaag 26

47

24

Artificial Sequence

Primer

47

ttgtgtgtgt tccctccaaa cacc 24

48

24

Artificial Sequence

Primer

48

ggaccattct taaggacagc cgat 24

49

26

Artificial Sequence

Primer

49

acatagtgat ctttccatca gcaaag 26

50

22

Artificial Sequence

Primer

50

tgaatgcaca gagaccctcc tg 22

51

26

Artificial Sequence

Primer

51

cctcttacca ttcagatact gttagg 26

52

24

Artificial Sequence

Primer

52

agcaacaact caaaccagcc ctac 24

53

23

Artificial Sequence

Primer

53

ttcttcagtt gccaactccc agg 23

54

21

Artificial Sequence

Primer

54

aagctgcttg tgtggcagca g 21

55

26

Artificial Sequence

Primer

55

agtaaggtga acaggaaagt acagag 26

56

24

Artificial Sequence

Primer

56

tacataagag aggctgccgc atag 24

57

23

Artificial Sequence

Primer

57

ccctacactc acactcatct agc 23

58

23

Artificial Sequence

Primer

58

ccctgtgttc cagatctcca ttg 23

59

24

Artificial Sequence

Primer

59

ttcctaggtc caccttgatc tgag 24

60

26

Artificial Sequence

Primer

60

agcacctgaa ttcaaatcag gatgag 26

61

29

Artificial Sequence

Primer

61

aaaccaaagt tctgaacaca ttaactcac 29

62

25

Artificial Sequence

Primer

62

ctggttgcat tcatagctgt gtttc 25

63

21

Artificial Sequence

Primer

63

acagaagcca gcatcactgg g 21

64

24

Artificial Sequence

Primer

64

ttactggtgc tgggaggata tgtc 24

65

26

Artificial Sequence

Primer

65

ataagtactt catcacctca gcgctc 26

66

24

Artificial Sequence

Primer

66

ttgatcttag ctgaccagtg tctc 24

67

25

Artificial Sequence

Primer

67

tctgcatgga cttgagcaga aagtc 25

68

30

Artificial Sequence

Primer

68

caaatcttgt gatagtgaat tacaagttgg 30

69

27

Artificial Sequence

Primer

69

tttatagctg ccctcaatac attttcc 27

70

22

Artificial Sequence

Primer

70

tgtacctgca gccattgctt gg 22

71

25

Artificial Sequence

Primer

71

ggatctgggc tctagtttat gtacg 25

72

24

Artificial Sequence

Primer

72

ttgaactata ggcacagaca gctg 24

73

23

Artificial Sequence

Primer

73

aacttgacct gtgtgactta cgc 23

74

26

Artificial Sequence

Primer

74

tcacagtcta tggtaatctg tcaagc 26

75

22

Artificial Sequence

Primer

75

aagggcaaca atgccctggc aa 22

76

26

Artificial Sequence

Primer

76

ttcctgcaaa tgggatagtc tctctg 26

77

26

Artificial Sequence

Primer

77

atcccccaag catttatcat tctcag 26

78

28

Artificial Sequence

Primer

78

tgtgtttcca gaaacctgct ttagtttg 28

79

26

Artificial Sequence

Primer

79

tagtactttt gtccaggatg accaag 26

80

30

Artificial Sequence

Primer

80

tgacaagaaa tgtcatgtct taacataagc 30

81

22

Artificial Sequence

Primer

81

ttcagagcct ccttccccaa ct 22

82

25

Artificial Sequence

Primer

82

tagtctgtag ctgaggccat tttgc 25

83

25

Artificial Sequence

Primer

83

aagcaagctg cagttaaggg actgt 25

84

23

Artificial Sequence

Primer

84

ttgggacctt gaggattgtt ccc 23

85

26

Artificial Sequence

Primer

85

cactcaacag gtaaaagtga tctgcc 26

86

27

Artificial Sequence

Primer

86

tgcatctgat cagtttgaat cagagag 27

87

26

Artificial Sequence

Primer

87

aaactgaggc ctgagttctg aaaagc 26

88

24

Artificial Sequence

Primer

88

caccaaagct ctgtaccact aagc 24

89

21

Artificial Sequence

Primer

89

tgactgtgca gtgatgcagg g 21

90

27

Artificial Sequence

Primer

90

ttgaccttga catttagaat agccctc 27

91

30

Artificial Sequence

Primer

91

gctgagaatt aataatggtc tttctctttg 30

92

22

Artificial Sequence

Primer

92

tacacagtga gaccctgtct cc 22

93

22

Artificial Sequence

Primer

93

tagctgaggt cccttgtgga ag 22

94

21

Artificial Sequence

Primer

94

agtgtcagag gaccatgctg g 21

95

23

Artificial Sequence

Primer

95

cttgaagcgt ccaactcatg tgc 23

96

26

Artificial Sequence

Primer

96

aactcataca tttgagcact gttgcc 26

97

23

Artificial Sequence

Primer

97

tgaggaggtg gggaatgcta atg 23

98

21

Artificial Sequence

Primer

98

acatagcaga gggctgctca c 21

99

21

Artificial Sequence

Primer

99

actgacctgt gggaacctgt g 21

100

26

Artificial Sequence

Primer

100

aatgctaggc atcatccctc ttctag 26

101

26

Artificial Sequence

Primer

101

aacatctaat agggactgag tgaccc 26

102

22

Artificial Sequence

Primer

102

ttctgtggtg ccttggcaag ag 22

103

25

Artificial Sequence

Primer

103

cacacataca cacacactaa gtgcc 25

104

32

Artificial Sequence

Primer

104

tggtagttac ttaaagttta caagaatgta gg 32

105

28

Artificial Sequence

Primer

105

aaatgctggg ataaaaagca tgaaccac 28

106

25

Artificial Sequence

Primer

106

ttcagttacc taatgggcac aaggc 25

107

23

Artificial Sequence

Primer

107

acgacactga cctcttgcac ttg 23

108

26

Artificial Sequence

Primer

108

tgtacaccct gaatgtctga acattc 26

109

25

Artificial Sequence

Primer

109

gcgttcattt ctgctcctgt aatgg 25

110

24

Artificial Sequence

Primer

110

tgagctctta atccctgcca ttcc 24

111

22

Artificial Sequence

Primer

111

tagggcttgt ccgtccataa gg 22

112

25

Artificial Sequence

Primer

112

tgttacggag atgaaaggct agacc 25

113

22

Artificial Sequence

Primer

113

taagccctgc actaacccac tc 22

114

25

Artificial Sequence

Primer

114

tgttttgaga ggtgagcctt ctagc 25

115

25

Artificial Sequence

Primer

115

catgtcctac agttctgcta tcacc 25

116

26

Artificial Sequence

Primer

116

cttttcttca tccaattccc cacgag 26

117

24

Artificial Sequence

Primer

117

tctctaagct gcactgttgt ggct 24

118

24

Artificial Sequence

Primer

118

tggaagccaa gagtcttgag ttgc 24

119

28

Artificial Sequence

Primer

119

gtctgcattt taaatgcaga tgtgaagc 28

120

25

Artificial Sequence

Primer

120

cgaaacgcac gcacattttt accag 25

121

26

Artificial Sequence

Primer

121

gtgtgattta gcatctgtcg cacttg 26

122

26

Artificial Sequence

Primer

122

tgtatgtata acccaacaat cgctgc 26

123

26

Artificial Sequence

Primer

123

tccagagtac agggagaaac taaagg 26

124

94

Homo sapiens

124

Phe Arg Asn His Pro Asn Ile Thr Phe Phe Val Tyr Val Ser Asn Phe

1 5 10 15

Thr Trp Pro Ile Lys Ile Gln Ile Ala Phe Ser Gln His Ser Asn Phe

20 25 30

Met Asp Leu Val Gln Phe Phe Val Thr Phe Phe Ser Cys Phe Leu Ser

35 40 45

Leu Leu Leu Val Ala Ala Val Val Trp Lys Ile Lys Gln Ser Cys Trp

50 55 60

Ala Ser Arg Arg Arg Glu Gln Leu Leu Arg Glu Met Gln Gln Met Ala

65 70 75 80

Ser Arg Pro Phe Ala Ser Val Glu Thr Leu Pro Trp Asn Arg

85 90

125

113

Homo sapiens

125

Phe Arg Ser Asn Pro Asn Ile Thr Phe Tyr Val Tyr Val Ser Asn Phe

1 5 10 15

Ser Trp Pro Ile Lys Ile Gln Ile Ala Phe Ser Gln His Asn Thr Ile

20 25 30

Met Asp Leu Val Gln Phe Phe Val Thr Phe Phe Ser Cys Phe Leu Ser

35 40 45

Leu Leu Leu Val Ala Ala Val Val Trp Lys Ile Lys Gln Thr Cys Trp

50 55 60

Ala Ser Arg Arg Arg Glu Gln Leu Leu Arg Glu Arg Gln Gln Met Ala

65 70 75 80

Ser Arg Pro Phe Ala Ser Val Asp Val Ala Leu Glu Val Gly Ala Glu

85 90 95

Gln Thr Glu Phe Leu Arg Gly Pro Leu Glu Gly Ala Pro Lys Pro Ile

100 105 110

Ala

126

100

Homo sapiens

126

Phe Gly Pro Asp Ser Asn Thr Thr Phe Phe Val Arg Val Tyr Asn Phe

1 5 10 15

Asn Thr Pro Val Gln Ile Val Val Ser Phe Ala Gln Ser Pro Pro Ile

20 25 30

Asn Trp Val Leu Phe Phe Val Ile Phe Ala Ala Cys Phe Ile Val Leu

35 40 45

Leu Val Val Ala Gly Leu Leu Trp Met Ile Lys Val Arg Ile Glu Ala

50 55 60

Tyr Arg Arg Asn Gln Arg Arg Ile Asp Glu Ile Glu His Met Ala Ser

65 70 75 80

Arg Pro Phe Ala Ser Thr Lys Met Glu Leu Ser Met Leu Ser Gln Phe

85 90 95

Ser Ser Ala Gly

100

127

118

Homo sapiens

127

Leu Lys Ser Ser Arg Phe Tyr Leu Leu Leu Leu Gly Val Gly Asp Pro

1 5 10 15

Ser Gly Pro Gly Ala Asn Gly Ser Ala Asp Ser Gln Gly Leu Leu Phe

20 25 30

Phe Arg Gln Asp Gln Ala His Ile Asp Leu Phe Val Phe Phe Ser Val

35 40 45

Phe Phe Ser Cys Phe Phe Leu Phe Leu Ser Leu Cys Val Leu Leu Trp

50 55 60

Lys Ala Lys Gln Ala Leu Asp Gln Arg Gln Glu Gln Arg Arg His Leu

65 70 75 80

Gln Glu Met Thr Lys Met Ala Ser Arg Pro Phe Ala Lys Val Thr Val

85 90 95

Cys Phe Pro Pro Asp Pro Thr Ala Pro Ala Ser Ala Trp Lys Pro Ala

100 105 110

Gly Leu Pro Pro Pro Ala

115

128

11

Artificial Sequence

Primer

128

ttcctcactg g 11

129

10

Artificial Sequence

Primer

129

ggacacacag 10