Polynucleotides encoding the fkbA gene of the FK-520 polyketide synthase gene cluster

The present application claims priority to related U.S. patent application Serial No. 60/102,748, filed Oct. 2, 1998; No. 60/139,650, filed Jun. 17, 1999; and No. 60/123,810, filed Mar. 11, 1999, each of which is incorporated herein by reference.

The contents of the following submission on compact discs are incorporated herein by reference in their entirety: a compact disc copy of the substitute Sequence Listing (COPY 1) (file name: 3006220026.txt, date recorded: Feb. 19, 2002, size: 521 KB); a duplicate compact disc copy of the substitute Sequence Listing (COPY 2) (file name: 3006220026.txt, date recorded: Feb. 19, 2002, size: 521 KB); and a computer readable form copy of the substitute Sequence Listing (CRF COPY) (file name: 3006220026.txt, date recorded: Feb. 19, 2002, size: 521 KB).

The present invention relates to polyketides and the polyketide synthase (PKS) enzymes that produce them. The invention also relates generally to genes encoding PKS enzymes and to recombinant host cells containing such genes and in which expression of such genes leads to the production of polyketides. The present invention also relates to compounds useful as medicaments having immunosuppressive and/or neurotrophic activity. Thus, the invention relates to the fields of chemistry, molecular biology, and agricultural, medical, and veterinary technology.

Polyketides are a class of compounds synthesized from 2-carbon units through a series of condensations and subsequent modifications. Polyketides occur in many types of organisms, including fungi and mycelial bacteria, in particular, the actinomycetes. Polyketides are biologically active molecules with a wide variety of structures, and the class encompasses numerous compounds with diverse activities. Tetracycline, erythromycin, epothilone, FK-506, FK-520, narbomycin, picromycin, rapamycin, spinocyn, and tylosin are examples of polyketides. Given the difficulty in producing polyketide compounds by traditional chemical methodology, and the typically low production of polyketides in wild-type cells, there has been considerable interest in finding improved or alternate means to produce polyketide compounds.

This interest has resulted in the cloning, analysis, and manipulation by recombinant DNA technology of genes that encode PKS enzymes. The resulting technology allows one to manipulate a known PKS gene cluster either to produce the polyketide synthesized by that PKS at higher levels than occur in nature or in hosts that otherwise do not produce the polyketide. The technology also allows one to produce molecules that are structurally related to, but distinct from, the polyketides produced from known PKS gene clusters. See, e.g., PCT publication Nos. WO 93/13663; 95/08548; 96/40968; 97/02358; 98/27203; and 98/49315; U.S. Pat. Nos. 4,874,748; 5,063,155; 5,098,837; 5,149,639; 5,672,491; 5,712,146; 5,830,750; and 5,843,718; and Fu et al., 1994, Biochemistry 33: 9321-9326; McDaniel et al., 1993 , Science 262: 1546-1550; and Rohr, 1995, Angew. Chem. Int. Ed. Engl. 34(8): 881-888, each of which is incorporated herein by reference.

Polyketides are synthesized in nature by PKS enzymes. These enzymes, which are complexes of multiple large proteins, are similar to the synthases that catalyze condensation of 2-carbon units in the biosynthesis of fatty acids. PKSs catalyze the biosynthesis of polyketides through repeated, decarboxylative Claisen condensations between acylthioester building blocks. The building blocks used to form complex polyketides are typically acylthioesters, such as acetyl, butyryl, propionyl, malonyl, hydroxymalonyl, methylmalonyl, and ethylmalonyl CoA. Other building blocks include amino acid like acylthioesters. PKS enzymes that incorporate such building blocks include an activity that functions as an amino acid ligase (an AMP ligase) or as a non-ribosomal peptide synthetase (NRPS). Two major types of PKS enzymes are known; these differ in their composition and mode of synthesis of the polyketide synthesized. These two major types of PKS enzymes are commonly referred to as Type I or “modular” and Type II “iterative” PKS enzymes.

In the Type I or modular PKS enzyme group, a set of separate catalytic active sites (each active site is termed a “domain”, and a set thereof is termed a “module”) exists for each cycle of carbon chain elongation and modification in the polyketide synthesis pathway. The typical modular PKS is composed of several large polypeptides, which can be segregated from amino to carboxy termini into a loading module, multiple extender modules, and a releasing (or thioesterase) domain. The PKS enzyme known as 6-deoxyerythronolide B synthase (DEBS) is a Type I PKS. In DEBS, there is a loading module, six extender modules, and a thioesterase (TE) domain. The loading module, six extender modules, and TE of DEBS are present on three separate proteins (designated DEBS-1, DEBS-2, and DEBS-3, with two extender modules per protein). Each of the DEBS polypeptides is encoded by a separate open reading frame (ORF) or gene; these genes are known as eryAI, eryAII, and eryAIII. See Caffrey et al., 1992 , FEBS Letters 304: 205, and U.S. Pat. No. 5,824,513, each of which is incorporated herein by reference.

Generally, the loading module is responsible for binding the first building block used to synthesize the polyketide and transferring it to the first extender module. The loading module of DEBS consists of an acyltransferase (AT) domain and an acyl carrier protein (ACP) domain. Another type of loading module utilizes an inactivated ketosynthase (KS) domain and AT and ACP domains. This inactivated KS is in some instances called KSQ, where the superscript letter is the abbreviation for the amino acid, glutamine, that is present instead of the active site cysteine required for ketosynthase activity. In other PKS enzymes, including the FK-506 PKS, the loading module incorporates an unusual starter unit and is composed of a CoA ligase like activity domain. In any event, the loading module recognizes a particular acyl-CoA (usually acetyl or propionyl but sometimes butyryl or other acyl-CoA) and transfers it as a thiol ester to the ACP of the loading module.

The AT on each of the extender modules recognizes a particular extender-CoA (malonyl or alpha-substituted malonyl, i.e., methylmalonyl, ethylmalonyl, and 2-hydroxymalonyl) and transfers it to the ACP of that extender module to form a thioester. Each extender module is responsible for accepting a compound from a prior module, binding a building block, attaching the building block to the compound from the prior module, optionally performing one or more additional functions, and transferring the resulting compound to the next module.

Each extender module of a modular PKS contains a KS, AT, ACP, and zero, one, two, or three domains that modify the beta-carbon of the growing polyketide chain. A typical (non-loading) minimal Type I PKS extender module is exemplified by extender module three of DEBS, which contains a KS domain, an AT domain, and an ACP domain. These three domains are sufficient to activate a 2-carbon extender unit and attach it to the growing polyketide molecule. The next extender module, in turn, is responsible for attaching the next building block and transferring the growing compound to the next extender module until synthesis is complete.

Once the PKS is primed with acyl- and malonyl-ACPs, the acyl group of the loading module is transferred to form a thiol ester (trans-esterification) at the KS of the first extender module; at this stage, extender module one possesses an acyl-KS and a malonyl (or substituted malonyl) ACP. The acyl group derived from the loading module is then covalently attached to the alpha-carbon of the malonyl group to form a carbon-carbon bond, driven by concomitant decarboxylation, and generating a new acyl-ACP that has a backbone two carbons longer than the loading building block (elongation or extension).

The polyketide chain, growing by two carbons each extender module, is sequentially passed as covalently bound thiol esters from extender module to extender module, in an assembly line-like process. The carbon chain produced by this process alone would possess a ketone at every other carbon atom, producing a polyketone, from which the name polyketide arises. Most commonly, however, additional enzymatic activities modify the beta keto group of each two carbon unit just after it has been added to the growing polyketide chain but before it is transferred to the next module.

Thus, in addition to the minimal module containing KS, AT, and ACP domains necessary to form the carbon-carbon bond, and as noted above, other domains that modify the beta-carbonyl moiety can be present. Thus, modules may contain a ketoreductase (KR) domain that reduces the keto group to an alcohol. Modules may also contain a KR domain plus a dehydratase (DH) domain that dehydrates the alcohol to a double bond. Modules may also contain a KR domain, a DH domain, and an enoylreductase (ER) domain that converts the double bond product to a saturated single bond using the beta carbon as a methylene function. An extender module can also contain other enzymatic activities, such as, for example, a methylase or dimethylase activity.

After traversing the final extender module, the polyketide encounters a releasing domain that cleaves the polyketide from the PKS and typically cyclizes the polyketide. For example, final synthesis of 6-dEB is regulated by a TE domain located at the end of extender module six. In the synthesis of 6-dEB, the TE domain catalyzes cyclization of the macrolide ring by formation of an ester linkage. In FK-506, FK-520, rapamycin, and similar polyketides, the TE activity is replaced by a RapP (for rapamycin) or RapP like activity that makes a linkage incorporating a pipecolate acid residue. The enzymatic activity that catalyzes this incorporation for the rapamycin enzyme is known as RapP, encoded by the rap? gene. The polyketide can be modified further by tailoring enzymes; these enzymes add carbohydrate groups or methyl groups, or make other modifications, i.e., oxidation or reduction, on the polyketide core molecule. For example, 6-dEB is hydroxylated at C-6 and C-12 and glycosylated at C-3 and C-5 in the synthesis of erythromycin A.

In Type I PKS polypeptides, the order of catalytic domains is conserved. When all beta-keto processing domains are present in a module, the order of domains in that module from N-to-C-terminus is always KS, AT, DH, ER, KR, and ACP. Some or all of the beta-keto processing domains may be missing in particular modules, but the order of the domains present in a module remains the same. The order of domains within modules is believed to be important for proper folding of the PKS polypetides into an active complex. Importantly, there is considerable flexibility in PKS enzymes, which allows for the genetic engineering of novel catalytic complexes. The engineering of these enzymes is achieved by modifying, adding, or deleting domains, or replacing them with those taken from other Type I PKS enzymes. It is also achieved by deleting, replacing, or adding entire modules with those taken from other sources. A genetically engineered PKS complex should of course have the ability to catalyze the synthesis of the product predicted from the genetic alterations made.

Alignments of the many available amino acid sequences for Type I PKS enzymes has approximately defined the boundaries of the various catalytic domains. Sequence alignments also have revealed linker regions between the catalytic domains and at the N- and C-termini of individual polypeptides. The sequences of these linker regions are less well conserved than are those for the catalytic domains, which is in part how linker regions are identified. Linker regions can be important for proper association between domains and between the individual polypeptides that comprise the PKS complex. One can thus view the linkers and domains together as creating a scaffold on which the domains and modules are positioned in the correct orientation to be active. This organization and positioning, if retained, permits PKS domains of different or identical substrate specificities to be substituted (usually at the DNA level) between PKS enzymes by various available methodologies. In selecting the boundaries of, for example, an AT replacement, one can thus make the replacement so as to retain the linkers of the recipient PKS or to replace them with the linkers of the donor PKS AT domain, or, preferably, make both constructs to ensure that the correct linker regions between the KS and AT domains have been included in at least one of the engineered enzymes. Thus, there is considerable flexibility in the design of new PKS enzymes with the result that known polyketides can be produced more effectively, and novel polyketides useful as pharmaceuticals or for other purposes can be made.

By appropriate application of recombinant DNA technology, a wide variety of polyketides can be prepared in a variety of different host cells provided one has access to nucleic acid compounds that encode PKS proteins and polyketide modification enzymes. The present invention helps meet the need for such nucleic acid compounds by providing recombinant vectors that encode the FK-520 PKS enzyme and various FK-520 modification enzymes. Moreover, while the FK-506 and FK-520 polyketides have many useful activities, there remains a need for compounds with similar useful activities but with better pharmacokinetic profile and metabolism and fewer side-effects. The present invention helps meet the need for such compounds as well.

In one embodiment, the present invention provides recombinant DNA vectors that encode all or part of the FK-520 PKS enzyme. Illustrative vectors of the invention include cosmid pKOS034-120, pKOS034-124, pKOS065-C31, pKOS065-C3, pKOS065-M27, and pKOS065-M21. The invention also provides nucleic acid compounds that encode the various domains of the FK-520 PKS, i.e., the KS, AT, ACP, KR, DH, and ER domains. These compounds can be readily used, alone or in combination with nucleic acids encoding other FK-520 or non-FK-520 PKS domains, as intermediates in the construction of recombinant vectors that encode all or part of PKS enzymes that make novel polyketides.

The invention also provides isolated nucleic acids that encode all or part of one or more modules of the FK-520 PKS, each module comprising a ketosynthase activity, an acyl transferase activity, and an acyl carrier protein activity. The invention provides an isolated nucleic acid that encodes one or more open reading frames of FK-520 PKS genes, said open reading frames comprising coding sequences for a CoA ligase activity, an NRPS activity, or two or more extender modules. The invention also provides recombinant expression vectors containing these nucleic acids.

In another embodiment, the invention provides isolated nucleic acids that encode all or a part of a PKS that contains at least one module in which at least one of the domains in the module is a domain from a non-FK-520 PKS and at least one domain is from the FK-520 PKS. The non-FK-520 PKS domain or module originates from the rapamycin PKS, the FK-506 PKS, DEBS, or another PKS. The invention also provides recombinant expression vectors containing these nucleic acids.

In another embodiment, the invention provides a method of preparing a polyketide, said method comprising transforming a host cell with a recombinant DNA vector that encodes at least one module of a PKS, said module comprising at least one FK-520 PKS domain, and culturing said host cell under conditions such that said PKS is produced and catalyzes synthesis of said polyketide. In one aspect, the method is practiced with a Streptomyces host cell. In another aspect, the polyketide produced is FK-520. In another aspect, the polyketide produced is a polyketide related in structure to FK-520. In another aspect, the polyketide produced is a polyketide related in structure to FK-506 or rapamycin.

In another embodiment, the invention provides a set of genes in recombinant form sufficient for the synthesis of ethylmalonyl CoA in a heterologous host cell. These genes and the methods of the invention enable one to create recombinant host cells with the ability to produce polyketides or other compounds that require ethylmalonyl CoA for biosynthesis. The invention also provides recombinant nucleic acids that encode AT domains specific for ethylmalonyl CoA. Thus, the compounds of the invention can be used to produce polyketides requiring ethylmalonyl CoA in host cells that otherwise are unable to produce such polyketides.

In another embodiment, the invention provides a set of genes in recombinant form sufficient for the synthesis of 2-hydroxymalonyl CoA and 2-methoxymalonyl CoA in a heterologous host cell. These genes and the methods of the invention enable one to create recombinant host cells with the ability to produce polyketides or other compounds that require 2-hydroxymalonyl CoA for biosynthesis. The invention also provides recombinant nucleic acids that encode AT domains specific for 2-hydroxymalonyl CoA and 2-methoxymalonyl CoA. Thus, the compounds of the invention can be used to produce polyketides requiring 2-hydroxymalonyl CoA or 2-methoxymalonyl CoA in host cells that are otherwise unable to produce such polyketides.

In another embodiment, the invention provides a compound related in structure to FK-520 or FK-506 that is useful in the treatment of a medical condition. These compounds include compounds in which the C-13 methoxy group is replaced by a moiety selected from the group consisting of hydrogen, methyl, and ethyl moieties. Such compounds are less susceptible to the main in vivo pathway of degradation for FK-520 and FK-506 and related compounds and thus exhibit an improved pharmacokinetic profile. The compounds of the invention also include compounds in which the C-15 methoxy group is replaced by a moiety selected from the group consisting of hydrogen, methyl, and ethyl moieties. The compounds of the invention also include the above compounds further modified by chemical methodology to produce derivatives such as, but not limited to, the C-18 hydroxyl derivatives, which have potent neurotrophin but not immunosuppresion activities.

Thus, the invention provides polyketides having the structure:

wherein, R 1 is hydrogen, methyl, ethyl, or allyl; R 2 is hydrogen or hydroxyl, provided that when R 2 is hydrogen, there is a double bond between C-20 and C-19; R 3 is hydrogen or hydroxyl; R 4 is methoxyl, hydrogen, methyl, or ethyl; and R 5 is methoxyl, hydrogen, methyl, or ethyl; but not including FK-506, FK-520, 18-hydroxy-FK-520, and 18-hydroxy-FK-506. The invention provides these compounds in purified form and in pharmaceutical compositions.

In another embodiment, the invention provides a method for treating a medical condition by administering a pharmaceutically efficacious dose of a compound of the invention. The compounds of the invention may be administered to achieve immunosuppression or to stimulate nerve growth and regeneration.

These and other embodiments and aspects of the invention will be more fully understood after consideration of the attached Drawings and their brief description below, together with the detailed description, examples, and claims that follow.

FIG. 1 shows a diagram of the FK-520 biosynthetic gene cluster. The top line provides a scale in kilobase pairs (kb). The second line shows a restriction map with selected restriction enzyme recognition sequences indicated. K is KpnI; X is XhoI, S is SacI; P is PstI; and E is EcoRI. The third line indicates the position of FK-520 PKS and related genes. Genes are abbreviated with a one letter designation, i.e., C fkbC. Immediately under the third line are numbered segments showing where the loading module (L) and ten different extender modules (numbered 1-10) are encoded on the various genes shown. At the bottom of the Figure, the DNA inserts of various cosmids of the invention (i.e., 34-124 is cosmid pKOS034-124) are shown in alignment with the FK-520 biosynthetic gene cluster.

FIG. 2 shows the loading module (load), the ten extender modules, and the peptide synthetase domain of the FK-520 PKS, together with, on the top line, the genes that encode the various domains and modules. Also shown are the various intermediates in FK-520 biosynthesis, as well as the structure of FK-520, with carbons 13, 15, 21, and 31 numbered. The various domains of each module and subdomains of the loading module are also shown. The darkened circles showing the DH domains in modules 2, 3, and 4 indicate that the dehydratase domain is not functional as a dehydratase; this domain may affect the stereochemistry at the corresponding position in the polyketide. The substituents on the FK-520 structure that result from the action of non-PKS enzymes are also indicated by arrows, together with the types of enzymes or the genes that code for the enzymes that mediate the action. Although the methyltransferase is shown acting at the C-13 and C-15 hydroxyl groups after release of the polyketide from the PKS, the methyltransferase may act on the 2-hydroxymalonyl substrate prior to or contemporaneously with its incorporation during polyketide synthesis.

FIG. 3 shows a close-up view of the left end of the FK-520 gene cluster, which contains at least ten additional genes. The ethyl side chain on carbon 21 of FK-520 (FIG. 2) is derived from an ethylmalonyl CoA extender unit that is incorporated by an ethylmalonyl specific AT domain in extender module 4 of the PKS. At least four of the genes in this region code for enzymes involved in ethylmalonyl biosynthesis. The polyhydroxybutyrate depolymerase is involved in maintaining hydroxybutyryl-CoA pools during FK-520 production. Polyhydroxybutyrate accumulates during vegetative growth and disappears during stationary phase in other Streptomyces (Ranade and Vining, 1993 , Can. J Microbiol. 39:377). Open reading frames with unknown function are indicated with a question mark.

FIG. 4 shows a biosynthetic pathway for the biosynthesis of ethylmalonyl CoA from acetoacetyl CoA consistent with the function assigned to four of the genes in the FK-520 gene cluster shown in FIG. 3 .

FIG. 5 shows a close-up view of the right-end of the FK-520 PKS gene cluster (and of the sequences on cosmid pKOS065-C31). The genes shown include fkbD, fkbM (a methyl transferase that methylates the hydroxyl group on C-31 of FK-520), fkbN (a homolog of a gene described as a regulator of cholesterol oxidase and that is believed to be a transcriptional activator), fkbQ (a type II thioesterase, which can increase polyketide production levels), and fkbS (a crotonyl-CoA reductase involved in the biosynthesis of ethylmalonyl CoA).

FIG. 6 shows the proposed degradative pathway for tacrolimus (FK-506) metabolism.

FIG. 7 shows a schematic process for the construction of recombinant PKS genes of the invention that encode PKS enzymes that produce 13-desmethoxy FK-506 and FK-520 polyketides of the invention, as described in Example 4, below.

FIG. 8, in Parts A and B, shows certain compounds of the invention preferred for dermal application in Part A and a synthetic route for making those compounds in Part B.

Given the valuable pharmaceutical properties of polyketides, there is a need for methods and reagents for producing large quantities of polyketides, as well as for producing related compounds not found in nature. The present invention provides such methods and reagents, with particular application to methods and reagents for producing the polyketides known as FK-520, also known as ascomycin or L-683,590 (see Holt et al., 1993 , JACS 115:9925), and FK-506, also known as tacrolimus. Tacrolimus is a macrolide immunosuppressant used to prevent or treat rejection of transplanted heart, kidney, liver, lung, pancreas, and small bowel allografts. The drug is also useful for the prevention and treatment of graft-versus-host disease in patients receiving bone marrow transplants, and for the treatment of severe, refractory uveitis. There have been additional reports of the unapproved use of tacrolimus for other conditions, including alopecia universalis, autoimmune chronic active hepatitis, inflammatory bowel disease, multiple sclerosis, primary biliary cirrhosis, and scleroderma. The invention provides methods and reagents for making novel polyketides related in structure to FK-520 and FK-506. and structurally related polyketides such as rapamycin.

The FK-506 and rapamycin polyketides are potent immunosuppressants, with chemical structures shown below.

FK-520 differs from FK-506 in that it lacks the allyl group at C-21 of FK-506, having instead an ethyl group at that position, and has similar activity to FK-506, albeit reduced immunosuppressive activity.

These compounds act through initial formation of an intermediate complex with protein “immunophilins” known as FKBPs (FK-506 binding proteins), including FKBP-12. Immunophilins are a class of cytosolic proteins that form complexes with molecules such as FK-506, FK-520, and rapamycin that in turn serve as ligands for other cellular targets involved in signal transduction. Binding of FK-506, FK-520, and rapamycin to FKBP occurs through the structurally similar segments of the polyketide molecules, known as the “FKBP-binding domain” (as generally but not precisely indicated by the stippled regions in the structures above). The FK-506-FKBP complex then binds calcineurin, while the rapamycin-FKBP complex binds to a protein known as RAFT-1. Binding of the FKBP-polyketide complex to these second proteins occurs through the dissimilar regions of the drugs known as the “effector” domains.

The three component FKBP-polyketide-effector complex is required for signal transduction and subsequent immunosuppressive activity of FK-506, FK-520, and rapamycin. Modifications in the effector domains of FK-506, FK-520, and rapamycin that destroy binding to the effector proteins (calcineurin or RAFT) lead to loss of immunosuppressive activity, even though FKBP binding is unaffected. Further, such analogs antagonize the immunosuppressive effects of the parent polyketides, because they compete for FKBP. Such non-immunosuppressive analogs also show reduced toxicity (see Dumont et al., 1992 , Journal of Experimental Medicine 176, 751-760), indicating that much of the toxicity of these drugs is not linked to FKBP binding.

In addition to immunosuppressive activity, FK-520, FK-506, and rapamycin have neurotrophic activity. In the central nervous system and in peripheral nerves, immunophilins are referred to as “neuroimmunophilins”. The neuroimmunophilin FKBP is markedly enriched in the central nervous system and in peripheral nerves. Molecules that bind to the neuroimmunophilin FKBP, such as FK-506 and FK-520, have the remarkable effect of stimulating nerve growth. In vitro, they act as neurotrophins, i.e., they promote neurite outgrowth in NGF-treated PC 12 cells and in sensory neuronal cultures, and in intact animals, they promote regrowth of damaged facial and sciatic nerves, and repair lesioned serotonin and dopamine neurons in the brain. See Gold et al., June 1999 , J. Pharm. Exp. Ther. 289(3): 1202-1210; Lyons et al., 1994 , Proc. National Academy of Science 91: 3191-3195; Gold et al., 1995 , Journal of Neuroscience 15: 7509-7516; and Steiner et al., 1997 , Proc. National Academy of Science 94: 2019-2024. Further, the restored central and peripheral neurons appear to be functional.

Compared to protein neurotrophic molecules (BNDF, NGF, etc.), the small-molecule neurotrophins such as FK-506, FK-520, and rapamycin have different, and often advantageous, properties. First, whereas protein neurotrophins are difficult to deliver to their intended site of action and may require intra-cranial injection, the small-molecule neurotrophins display excellent bioavailability; they are active when administered subcutaneously and orally. Second, whereas protein neurotrophins show quite specific effects, the small-molecule neurotrophins show rather broad effects. Finally, whereas protein neurotrophins often show effects on normal sensory nerves, the small-molecule neurotrophins do not induce aberrant sprouting of normal neuronal processes and seem to affect damaged nerves specifically. Neuroimmunophilin ligands have potential therapeutic utility in a variety of disorders involving nerve degeneration (e.g. multiple sclerosis, Parkinson's disease, Alzheimer's disease, stroke, traumatic spinal cord and brain injury, peripheral neuropathies).

Recent studies have shown that the immunosuppressive and neurite outgrowth activity of FK-506, FK-520, and rapamycin can be separated; the neuroregenerative activity in the absence of immunosuppressive activity is retained by agents which bind to FKBP but not to the effector proteins calcineurin or RAFT. See Steiner et al., 1997 , Nature Medicine 3: 421-428.

Available structure-activity data show that the important features for neurotrophic activity of rapamycin, FK-520, and FK-506 lie within the common, contiguous segments of the macrolide ring that bind to FKBP. This portion of the molecule is termed the “FKBP binding domain” (see VanDuyne et al., 1993 , Journal of Molecular Biology 229: 105-124.). Nevertheless, the effector domains of the parent macrolides contribute to conformational rigidity of the binding domain and thus indirectly contribute to FKBP binding.

There are a number of other reported analogs of FK-506, FK-520, and rapamycin that 110 bind to FKBP but not the effector protein calcineurin or RAFT. These analogs show effects on nerve regeneration without immunosuppressive effects.

Naturally occurring FK-520 and FK-506 analogs include the antascomycins, which are FK-506-like macrolides that lack the functional groups of FK-506 that bind to calcineurin (see Fehr et al., 1996 , The Journal of Antibiotics 49: 230-233). These molecules bind FKBP as effectively as does FK-506; they antagonize the effects of both FK-506 and rapamycin, yet lack immunosuppressive activity.

Other analogs can be produced by chemically modifying FK-506, FK-520, or rapamycin. One approach to obtaining neuroimmunophilin ligands is to destroy the effector binding region of FK-506, FK-520, or rapamycin by chemical modification. While the chemical modifications permitted on the parent compounds are quite limited, some useful chemically modified analogs exist. The FK-520 analog L-685,818 (ED 50 =0.7 nM for FKBP binding; see Dumont et al., 1992), and the rapamycin analog WAY-124,466 (IC 50 12.5 nM; see Ocain et al., 1993 , Biochemistry Biophysical Research Communications 192: 1340-134693) are about as effective as FK-506, FK-520, and rapamycin at promoting neurite outgrowth in sensory neurons (see Steiner et al., 1997).

One of the few positions of rapamycin that is readily amenable to chemical modification is the allylic 16-methoxy group; this reactive group is readily exchanged by acid-catalyzed nucleophilic substitution. Replacement of the 16-methoxy group of rapamycin with a variety of bulky groups has produced analogs showing selective loss of immunosuppressive activity while retaining FKBP-binding (see Luengo et al., 1995 , Chemistry & Biology 2: 471-481). One of the best compounds, 1, below, shows complete loss of activity in the splenocyte proliferation assay with only a 10-fold reduction in binding to FKBP.

There are also synthetic analogs of FKBP binding domains. These compounds reflect an approach to obtaining neuroimmunophilin ligands based on “rationally designed” molecules that retain the FKBP-binding region in an appropriate conformation for binding to FKBP, but do not possess the effector binding regions. In one example, the ends of the FKBP binding domain were tethered by hydrocarbon chains (see Holt et al., 1993 , Journal of the American Chemical Society 115: 9925-9938); the best analog, 2, below, binds to FKBP about as well as FK-506. In a similar approach, the ends of the FKBP binding domain were tethered by a tripeptide to give analog 3, below, which binds to FKBP about 20-fold poorer than FK-506. These compounds are anticipated to have neuroimmunophilin binding activity.

In a primate MPTP model of Parkinson's disease, administration of FKBP ligand GPI-1046 caused brain cells to regenerate and behavioral measures to improve. MPTP is a neurotoxin, which, when administered to animals, selectively damages nigral-striatal dopamine neurons in the brain, mimicking the damage caused by Parkinson's disease. Whereas, before treatment, animals were unable to use affected limbs, the FKBP ligand restored the ability of animals to feed themselves and gave improvements in measures of locomotor activity, neurological outcome, and fine motor control. There were also corresponding increases in regrowth of damaged nerve terminals. These results demonstrate the utility of FKBP ligands for treatment of diseases of the CNS.

From the above description, two general approaches towards the design of non-immunosuppressant, neuroimmunophilin ligands can be seen. The first involves the construction of constrained cyclic analogs of FK-506 in which the FKBP binding domain is fixed in a conformation optimal for binding to FKBP. The advantages of this approach are that the conformation of the analogs can be accurately modeled and predicted by computational methods, and the analogs closely resemble parent molecules that have proven pharmacological properties. A disadvantage is that the difficult chemistry limits the numbers and types of compounds that can be prepared. The second approach involves the trial and error construction of acyclic analogs of the FKBP binding domain by conventional medicinal chemistry. The advantages to this approach are that the chemistry is suitable for production of the numerous compounds needed for such interactive chemistry-bioassay approaches. The disadvantages are that the molecular types of compounds that have emerged have no known history of appropriate pharmacological properties, have rather labile ester functional groups, and are too conformationally mobile to allow accurate prediction of conformational properties.

The present invention provides useful methods and reagents related to the first approach, but with significant advantages. The invention provides recombinant PKS genes that produce a wide variety of polyketides that cannot otherwise be readily synthesized by chemical methodology alone. Moreover, the present invention provides polyketides that have either or both of the desired immunosuppressive and neurotrophic activities, some of which are produced only by fermentation and others of which are produced by fermentation and chemical modification. Thus, in one aspect, the invention provides compounds that optimally bind to FKBP but do not bind to the effector proteins. The methods and reagents of the invention can be used to prepare numerous constrained cyclic analogs of FK-520 in which the FKBP binding domain is fixed in a conformation optimal for binding to FKBP. Such compounds will show neuroimmunophilin binding (neurotrophic) but not immunosuppressive effects. The invention also allows direct manipulation of FK-520 and related chemical structures via genetic engineering of the enzymes involved in the biosynthesis of FK-520 (as well as related compounds, such as FK-506 and rapamycin); similar chemical modifications are simply not possible because of the complexity of the structures. The invention can also be used to introduce “chemical handles” into normally inert positions that permit subsequent chemical modifications.

Several general approaches to achieve the development of novel neuroimmunophilin ligands are facilitated by the methods and reagents of the present invention. One approach is to make “point mutations” of the functional groups of the parent FK-520 structure that bind to the effector molecules to eliminate their binding potential. These types of structural modifications are difficult to perform by chemical modification, but can be readily accomplished with the methods and reagents of the invention.

A second, more extensive approach facilitated by the present invention is to utilize molecular modeling to predict optimal structures ab initio that bind to FKBP but not effector molecules. Using the available X-ray crystal structure of FK-520 (or FK-506) bound to FKBP, molecular modeling can be used to predict polyketides that should optimally bind to FKBP but not calcineurin. Various macrolide structures can be generated by linking the ends of the FKBP-binding domain with “all possible” polyketide chains of variable length and substitution patterns that can be prepared by genetic manipulation of the FK-520 or FK-506 PKS gene cluster in accordance with the methods of the invention. The ground state conformations of the virtual library can be determined, and compounds that possess binding domains most likely to bind well to FKBP can be prepared and tested.

Once a compound is identified in accordance with the above approaches, the invention can be used to generate a focused library of analogs around the lead candidate, to “fine tune” the compound for optimal properties. Finally, the genetic engineering methods of the invention can be directed towards producing “chemical handles” that enable medicinal chemists to modify positions of the molecule previously inert to chemical modification. This opens the path to previously prohibited chemical optimization of lead compounds by time-proven approaches.

Moreover, the present invention provides polyketide compounds and the recombinant genes for the PKS enzymes that produce the compounds that have significant advantages over FK-506 and FK-520 and their analogs. The metabolism and pharmacokinetics of tacrolimus has been exstensively studied, and FK-520 is believed to be similar in these respects. Absorption of tacrolimus is rapid, variable, and incomplete from the gastrointestinal tract (Harrison's Principles of Internal Medicine, 14th edition, 1998, McGraw Hill, 14, 20, 21, 64-67). The mean bioavailability of the oral dosage form is 27%, (range 5 to 65%). The volume of distribution (VoID) based on plasma is 5 to 65 L per kg of body weight (L/kg), and is much higher than the VoID based on whole blood concentrations, the difference reflecting the binding of tacrolimus to red blood cells. Whole blood concentrations may be 12 to 67 times the plasma concentrations. Protein binding is high (75 to 99%), primarily to albumin and alpha1-acid glycoprotein. The half-life for distribution is 0.9 hour; elimination is biphasic and variable: terminal-11.3 hr (range, 3.5 to 40.5 hours). The time to peak concentration is 0.5 to 4 hours after oral administration.

Tacrolimus is metabolized primarily by cytochrome P4503A enzymes in the liver and small intestine. The drug is extensively metabolized with less than 1% excreted unchanged in urine. Because hepatic dysfunction decreases clearance of tacrolimus, doses have to be reduced substantially in primary graft non-function, especially in children. In addition, drugs that induce the cytochrome P4503A enzymes reduce tacrolimus levels, while drugs that inhibit these P450s increase tacrolimus levels. Tacrolimus bioavailability doubles with co-administration of ketoconazole, a drug that inhibits P4503A. See, Vincent et al., 1992, In vitro metabolism of FK-506 in rat, rabbit, and human liver microsomes: Identification of a major metabolite and of cytochrome P4503A as the major enzymes responsible for its metabolism, Arch. Biochem. Biophys. 294: 454-460; Iwasaki et al., 1993, Isolation, identification, and biological activities of oxidative metabolites of FK-506, a potent immunosuppressive macrolide lactone, Drug Metabolism & Disposition 21: 971-977; Shiraga et al., 1994, Metabolism of FK-506, a potent immunosuppressive agent, by cytochrome P4503A enzymes in rat, dog, and human liver microsomes, Biochem. Pharmacol 47: 727-735; and Iwasaki et al., 1995, Further metabolism of FK-506 (Tacrolimus); Identification and biological activities of the metabolites oxidized at multiple sites of FK-506 , Drug Metabolism & Disposition 23: 28-34. The cytochrome P4503A subfamily of isozymes has been implicated as important in this degradative process.

Structures of the eight isolated metabolites formed by liver microsomes are shown in FIG. 6 . Four metabolites of FK-506 involve demethylation of the oxygens on carbons 13, 15, and 31, and hydroxylation of carbon 12. The 13-demethylated (hydroxy) compounds undergo cyclizations of the 13-hydroxy at C-10 to give MI, MVI and MVII, and the 12-hydroxy metabolite at C-10 to give I. Another four metabolites formed by oxidation of the four metabolites mentioned above were isolated by liver microsomes from dexamethasone treated rats. Three of these are metabolites doubly demethylated at the methoxy groups on carbons 15 and 31 (M-V), 13 and 31 (M-VI), and 13 and 15 (M-VII). The fourth, M-VIII, was the metabolite produced after demethylation of the 31-methoxy group, followed by formation of a fused ring system by further oxidation. Among the eight metabolites, M-II has immunosuppressive activity comparable to that of FK-506, whereas the other metabolites exhibit weak or negligible activities. Importantly, the major metabolite of human, dog, and rat liver microsomes is the 13-demethylated and cyclized FK-506 (M-I).

Thus, the major metabolism of FK-506 proceeds via 13-demethylation followed by cyclization to the inactive M-I, this representing about 90% of the metabolic products after a 10 minute incubation with liver microsomes. Analogs of tacrolimus that do not possess a C-13 methoxy group would not be susceptible to the first and most important biotransformation in the destructive metabolism of tacrolimus (i.e. cyclization of 13-hydroxy to C-10). Thus, a 13-desmethoxy analog of FK-506 should have a longer half-life in the body than does FK-506. The C-13 methoxy group is believed not to be required for binding to FKBP or calcineurin. The C-13 methoxy is not present on the identical position of rapamycin, which binds to FKBP with equipotent affinity as tacrolimus. Also, analysis of the 3-dimensional structure of the FKBP-tacrolimus-calcineurin complex shows that the C-13 methoxy has no interaction with FKBP and only aminor interaction with calcineurin. The present invention provides C-13-desmethoxy analogs of FK-506 and FK-520, as well as the recombinant genes that encode the PKS enzymes that catalyze their synthesis and host cells that produce the compounds.

These compounds exhibit, relative to their naturally occurring counterparts, prolonged immunosuppressive action in vivo, thereby allowing a lower dosage and/or reduced frequency of administration. Dosing is more predictable, because the variability in FK-506 dosage is largely due to variation of metabolism rate. FK-506 levels in blood can vary widely depending on interactions with drugs that induce or inhibit cytochrome P4503A (summarized in USP Drug Information for the Health Care Professional). Of particular importance are the numerous drugs that inhibit or compete for CYP 3A, because they increase FK-506 blood levels and lead to toxicity (Prograf package insert, Fujisawa□US, Rev April 1997, Rec June. 1997). Also important are the drugs that induce P4503A (e.g. Dexamethasone), because they decrease FK-506 blood levels and reduce efficacy. Because the major site of CYP 3A action on FK-506 is removed in the analogs provided by the present invention, those analogs are not as susceptible to drug interactions as the naturally occurring compounds.

Hyperglycemia, nephrotoxicity, and neurotoxicity are the most significant adverse effects resulting from the use of FK-506 and are believed to be similar for FK-520. Because these effects appear to occur primarily by the same mechanism as the immunosuppressive action (i.e. FKBP-calcineurin interaction), the intrinsic toxicity of the desmethoxy analogs may be similar to FK-506. However, toxicity of FK-506 is dose related and correlates with high blood levels of the drug (Prograf package insert, Fujisawa□US, Rev April 1997, Rec June 1997). Because the levels of the compounds provided by the present invention should be more controllable, the incidence of toxicity should be significantly decreased with the 13-desmethoxy analogs. Some reports show that certain FK-506 metabolites are more toxic than FK-506 itself, and this provides an additional reason to expect that a CYP 3A resistant analog can have lower toxicity and a higher therapeutic index.

Thus, the present invention provides novel compounds related in structure to FK-506 and FK-520 but with improved properties. The invention also provides methods for making these compounds by fermentation of recombinant host cells, as well as the recombinant host cells, the recombinant vectors in those host cells, and the recombinant proteins encoded by those vectors. The present invention also provides other valuable materials useful in the construction of these recombinant vectors that have many other important applications as well. In particular, the present invention provides the FK-520 PKS genes, as well as certain genes involved in the biosynthesis of FK-520 in recombinant form.

FK-520 is produced at relatively low levels in the naturally occurring cells, Streptomyces hygroscopicus var. ascomyceticus , in which it was first identified. Thus, another benefit provided by the recombinant FK-520 PKS and related genes of the present invention is the ability to produce FK-520 in greater quantities in the recombinant host cells provided by the invention. The invention also provides methods for making novel FK-520 analogs, in addition to the desmethoxy analogs described above, and derivatives in recombinant host cells of any origin.

The biosynthesis of FK-520 involves the action of several enzymes. The FK-520 PKS enzyme, which is composed of the fkbA, fkbB, fkbC, and fkbP gene products, synthesizes the core structure of the molecule. There is also a hydroxylation at C-9 mediated by the P450 hydroxylase that is the fkbD gene product and that is oxidized by the fkbO gene product to result in the formation of a keto group at C-9. There is also a methylation at C-31 that is mediated by an O-methyltransferase that is the fkbM gene product. There are also methylations at the C-13 and C-15 positions by a methyltransferase believed to be encoded by the fkbG gene; this methyltransferase may act on the hydroxymalonyl CoA substrates prior to binding of the substrate to the AT domains of the PKS during polyketide synthesis. The present invention provides the genes encoding these enzymes in recombinant form. The invention also provides the genes encoding the enzymes involved in ethylmalonyl CoA and 2-hydroxymalonyl CoA biosynthesis in recombinant form. Moreover, the invention provides Streptomyces hygroscopicus var. ascomyceticus recombinant host cells lacking one or more of these genes that are useful in the production of useful compounds.

The cells are useful in production in a variety of ways. First, certain cells make a useful FK-520-related compound merely as a result of inactivation of one or more of the FK-520 biosynthesis genes. Thus, by inactivating the C-310-methyltransferase gene in Streptomyces hygroscopicus var. ascomyceticus , one creates a host cell that makes a desmethyl (at C-31) derivative of FK-520. Second, other cells of the invention are unable to make FK-520 or FK-520 related compounds due to an inactivation of one or more of the PKS genes. These cells are useful in the production of other polyketides produced by PKS enzymes that are encoded on recombinant expression vectors and introduced into the host cell.

Moreover, if only one PKS gene is inactivated, the ability to produce FK-520 or an FK-520 derivative compound is restored by introduction of a recombinant expression vector that contains the functional gene in a modified or unmodified form. The introduced gene produces a gene product that, together with the other endogenous and functional gene products, produces the desired compound. This methodology enables one to produce FK-520 derivative compounds without requiring that all of the genes for the PKS enzyme be present on one or more expression vectors. Additional applications and benefits of such cells and methodology will be readily apparent to those of skill in the art after consideration of how the recombinant genes were isolated and employed in the construction of the compounds of the invention.

The FK-520 biosynthetic genes were isolated by the following procedure. Genomic DNA was isolated from Streptomyces hygroscopicus var. ascomyceticus (ATCC 14891) using the lysozyme/proteinase K protocol described in Genetic Manipulation of Streptomyces —A Laboratory Manual (Hopwood et al., 1986). The average size of the DNA was estimated to be between 80-120 kb by electrophoresis on 0.3% agarose gels. A library was constructed in the SuperCos™ vector according to the manufacturer's instructions and with the reagents provided in the commercially available kit (Stratagene). Briefly, 100 μg of genomic DNA was partially digested with 4 units of Sau3A I for 20 min. in a reaction volume of 1 mL, and the fragments were dephosphorylated and ligated to SuperCos vector arms. The ligated DNA was packaged and used to infect log-stage XL1-BlueMR cells. A library of about 10,000 independent cosmid clones was obtained.

Based on recently published sequence from the FK-506 cluster (Motamedi and Shafiee, 1998 , Eur. J. Biochem. 256: 528), a probe for the fkbO gene was isolated from ATCC 14891 using PCR with degenerate primers. With this probe, a cosmid designated pKOS034-124 was isolated from the library. With probes made from the ends of cosmid pKOS034-124, an additional cosmid designated pKOS034-120 was isolated. These cosmids (pKOS034-124 and pKOS034-120) were shown to contain DNA inserts that overlap with one another. Initial sequence data from these two cosmids generated sequences similar to sequences from the FK-506 and rapamycin clusters, indicating that the inserts were from the FK-520 PKS gene cluster. Two EcoRI fragments were subcloned from cosmids pKOS034-124 and pKOS034-120. These subclones were used to prepare shotgun libraries by partial digestion with Sau3AI, gel purification of fragments between 1.5 kb and 3 kb in size, and ligation into the pLitmus28 vector (New England Biolabs). These libraries were sequenced using dye terminators on a Beckmann CEQ2000 capillary electrophoresis sequencer, according to the manufacturer's protocols.

To obtain cosmids containing sequence on the left and right sides of the sequenced region described above, a new cosmid library of ATCC 14891 DNA was prepared essentially as described above. This new library was screened with a new fkbM probe isolated using DNA from ATCC 14891. A probe representing the fkbP gene at the end of cosmid pKOS034-124 was also used. Several additional cosmids to the right of the previously sequenced region were identified. Cosmids pKOS065-C31 and pKOS065-C3 were identified and then mapped with restriction enzymes. Initial sequences from these cosmids were consistent with the expected organization of the cluster in this region. More extensive sequencing showed that both cosmids contained in addition to the desired sequences, other sequences not contiguous to the desired sequences on the host cell chromosomal DNA. Probing of additional cosmid libraries identified two additional cosmids, pKOS065-M27 and pKOS065-M21, that contained the desired sequences in a contiguous segment of chromosomal DNA. Cosmids pKOS034-124, pKOS034-120, pKOS065-M27, and pKOS065-M21 have been deposited with the American Type Culture Collection, Manassas, Va., USA. The complete nucleotide sequence of the coding sequences of the genes that encode the proteins of the FK-520 PKS are shown below but can also be determined from the cosmids of the invention deposited with the ATCC using standard methodology.

Referring to FIGS. 1 and 3, the FK-520 PKS gene cluster is composed of four open reading frames designated fkbB, fkbC, fkbA, and fkbP. The fkbB open reading frame encodes the loading module and the first four extender modules of the PKS. The fkbC open reading frame encodes extender modules five and six of the PKS. The fkbA open reading frame encodes extender modules seven, eight, nine, and ten of the PKS. The fkbP open reading frame encodes the NRPS of the PKS. Each of these genes can be isolated from the cosmids of the invention described above. The DNA sequences of these genes are provided below (SEQ ID NO:1) preceded by the following table identifying the start and stop codons of the open reading frames of each gene and the modules and domains contained therein.

Those of skill in the art will recognize that, due to the degenerate nature of the genetic code, a variety of DNA compounds differing in their nucleotide sequences can be used to encode a given amino acid sequence of the invention. The native DNA sequence encoding the FK-520 PKS of Streptomyces hygroscopicus is shown herein merely to illustrate a preferred embodiment of the invention, and the present invention includes DNA compounds of any sequence that encode the amino acid sequences of the polypeptides and proteins of the invention. In similar fashion, a polypeptide can typically tolerate one or more amino acid substitutions, deletions, and insertions in its amino acid sequence without loss or significant loss of a desired activity. The present invention includes such polypeptides with alternate amino acid sequences, and the amino acid sequences shown merely illustrate preferred embodiments of the invention.

The recombinant nucleic acids, proteins, and peptides of the invention are many and diverse. To facilitate an understanding of the invention and the diverse compounds and methods provided thereby, the following general description of the FK-520 PKS genes and modules of the PKS proteins encoded thereby is provided. This general description is followed by a more detailed description of the various domains and modules of the FK-520 PKS contained in and encoded by the compounds of the invention. In this description, reference to a heterologous PKS refers to any PKS other than the FK-520 PKS. Unless otherwise indicated, reference to a PKS includes reference to a portion of a PKS. Moreover, reference to a domain, module, or PKS includes reference to the nucleic acids encoding the same and vice-versa, because the methods and reagents of the invention provide or enable one to prepare proteins and the nucleic acids that encode them.

The FK-520 PKS is composed of three proteins encoded by three genes designated fkbA, fkbB, and fkbC. The fkbA ORF encodes extender modules 7-10 of the PKS. The fkbB ORF encodes the loading module (the CoA ligase) and extender modules 1-4 of the PKS. The fkbC ORF encodes extender modules 5-6 of the PKS. The fkbP ORF encodes the NRPS that attaches the pipecolic acid and cyclizes the FK-520 polyketide.

The loading module of the FK-520 PKS includes a CoA ligase, an ER domain, and an ACP domain. The starter building block or unit for FK-520 is believed to be a dihydroxycyclohexene carboxylic acid, which is derived from shikimate. The recombinant DNA compounds of the invention that encode the loading module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of methods and in a variety of compounds. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 loading module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for the loading module of the heterologous PKS is replaced by the coding sequence for the FK-520 loading module, provides a novel PKS coding sequence. Examples of heterologous PKS coding sequences include the rapamycin, FK-506, rifamycin, and avermectin PKS coding sequences. In another embodiment, a DNA compound comprising a sequence that encodes the FK-520 loading module is inserted into a DNA compound that comprises the coding sequence for the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion of the loading module coding sequence is utilized in conjunction with a heterologous coding sequence. In this embodiment, the invention provides, for example, either replacing the CoA ligase with a different CoA ligase, deleting the ER, or replacing the ER with a different ER. In addition, or alternatively, the ACP can be replaced by another ACP. In similar fashion, the corresponding domains in another loading or extender module can be replaced by one or more domains of the FK-520 PKS. The resulting heterologous loading module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide.

The first extender module of the FK-520 PKS includes a KS domain, an AT domain specific for methylmalonyl CoA, a DH domain, a KR domain, and an ACP domain. The recombinant DNA compounds of the invention that encode the first extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 first extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the first extender module of the FK-520 PKS or the latter is merely added to coding sequences for modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the first extender module of the FK-520 PKS is inserted into a DNA compound that comprises the remainder of the coding sequence for the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, all or only a portion of the first extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the methylmalonyl CoA specific AT with a malonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; deleting either the DH or KR or both; replacing the DH or KR or both with another DH or KR; and/or inserting an ER. In replacing or inserting KR, DH, and ER domains, it is often beneficial to replace the existing KR, DH, and ER domains with the complete set of domains desired from another module. Thus, if one desires to insert an ER domain, one may simply replace the existing KR and DH domains with a KR, DH, and ER set of domains from a module containing such domains. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a gene for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous first extender module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the first extender module of the FK-520 PKS.

In an illustrative embodiment of this aspect of the invention, the invention provides recombinant PKSs and recombinant DNA compounds and vectors that encode such PKSs in which the KS domain of the first extender module has been inactivated. Such constructs are especially useful when placed in translational reading frame with the remaining modules and domains of an FK-520 or FK-520 derivative PKS. The utility of these constructs is that host cells expressing, or cell free extracts containing, the PKS encoded thereby can be fed or supplied with N-acylcysteamine thioesters of novel precursor molecules to prepare FK-520 derivatives. See U.S. patent application Ser. No. 60/117,384, filed Jan. 27, 1999, and PCT patent publication Nos. US97/02358 and US99/03986, each of which is incorporated herein by reference.

The second extender module of the FK-520 PKS includes a KS, an AT specific for methylmalonyl CoA, a KR, an inactive DH, and an ACP. The recombinant DNA compounds of the invention that encode the second extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 second extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the second extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the second extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, all or a portion of the second extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the methylmalonyl CoA specific AT with a malonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; deleting the KR and/or the inactive DH; replacing the KR with another KR; and/or inserting an active DH or an active DH and an ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous second extender module coding sequence can be utilized in conjunction with a coding sequence from a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the second extender module of the FK-520 PKS.

The third extender module of the FK-520 PKS includes a KS, an AT specific for malonyl CoA, a KR, an inactive DH, and an ACP. The recombinant DNA compounds of the invention that encode the third extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 third extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the third extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the third extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, all or a portion of the third extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the malonyl CoA specific AT with a methylmalonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; deleting the KR and/or the inactive DH; replacing the KR with another KR; and/or inserting an active DH or an active DH and an ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous third extender module coding sequence can be utilized in conjunction with a coding sequence from a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the third extender module of the FK-520 PKS.

The fourth extender module of the FK-520 PKS includes a KS, an AT that binds ethylmalonyl CoA, an inactive DH, and an ACP. The recombinant DNA compounds of the invention that encode the fourth extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 fourth extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the fourth extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the fourth extender module of the FK-520 PKS is inserted into a DNA compound that comprises the remainder of the coding sequence for the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion of the fourth extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the ethylmalonyl CoA specific AT with a malonyl CoA, methylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; and/or deleting the inactive DH, inserting a KR, a KR and an active DH, or a KR, an active DH, and an ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, a PKS for a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous fourth extender module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the fourth extender module of the FK-520 PKS.

As illustrative examples, the present invention provides recombinant genes, vectors, and host cells that result from the conversion of the FK-506 PKS to an FK-520 PKS and vice-versa. In one embodiment, the invention provides a recombinant set of FK-506 PKS genes but in which the coding sequences for the fourth extender module or at least those for the AT domain in the fourth extender module have been replaced by those for the AT domain of the fourth extender module of the FK-520 PKS. This recombinant PKS can be used to produce FK-520 in recombinant host cells. In another embodiment, the invention provides a recombinant set of FK-520 PKS genes but in which the coding sequences for the fourth extender module or at least those for the AT domain in the fourth extender module have been replaced by those for the AT domain of the fourth extender module of the FK-506 PKS. This recombinant PKS can be used to produce FK-506 in recombinant host cells.

Other examples of hybrid PKS enzymes of the invention include those in which the AT domain of module 4 has been replaced with a malonyl specific AT domain to provide a PKS that produces 21-desethyl-FK520 or with a methylmalonyl specific AT domain to provide a PKS that produces 21-desethyl-21-methyl-FK520. Another hybrid PKS of the invention is prepared by replacing the AT and inactive KR domain of FK-520 extender module 4 with a methylmalonyl specific AT and an active KR domain, such as, for example, from module 2 of the DEBS or oleandolide PKS enzymes, to produce 21-desethyl-21-methyl-22-desoxo-22-hydroxy-FK520. The compounds produced by these hybrid PKS enzymes are neurotrophins.

The fifth extender module of the FK-520 PKS includes a KS, an AT that binds methylmalonyl CoA, a DH, a KR, and an ACP. The recombinant DNA compounds of the invention that encode the fifth extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 fifth extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the fifth extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS. In another embodiment, a DNA compound comprising a sequence that encodes the fifth extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion of the fifth extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the methylmalonyl CoA specific AT with a malonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; deleting any one or both of the DH and KR; replacing any one or both of the DH and KR with either a KR and/or DH; and/or inserting an ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous fifth extender module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the fifth extender module of the FK-520 PKS.

In an illustrative embodiment, the present invention provides a set of recombinant FK-520 PKS genes in which the coding sequences for the DH domain of the fifth extender module have been deleted or mutated to render the DH non-functional. In one such mutated gene, the KR and DH coding sequences are replaced with those encoding only a KR domain from another PKS gene. The resulting PKS genes code for the expression of an FK-520 PKS that produces an FK-520 analog that lacks the C-19 to C-20 double bond of FK-520 and has a C-20 hydroxyl group. Such analogs are preferred neurotrophins, because they have little or no immunosuppressant activity. This recombinant fifth extender module coding sequence can be combined with other coding sequences to make additional compounds of the invention. In an illustrative embodiment, the present invention provides a recombinant FK-520 PKS that contains both this fifth extender module and the recombinant fourth extender module described above that comprises the coding sequence for the fourth extender module AT domain of the FK-506 PKS. The invention also provides recombinant host cells derived from FK-506 producing host cells that have been mutated to prevent production of FK-506 but that express this recombinant PKS and so synthesize the corresponding (lacking the C-19 to C-20 double bond of FK-506 and having a C-20 hydroxyl group) FK-506 derivative. In another embodiment, the present invention provides a recombinant FK-506 PKS in which the DH domain of module 5 has been deleted or otherwise rendered inactive and thus produces this novel polyketide.

The sixth extender module of the FK-520 PKS includes a KS, an AT specific for methylmalonyl CoA, a KR, a DH, an ER, and an ACP. The recombinant DNA compounds of the invention that encode the sixth extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 sixth extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the sixth extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the sixth extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion of the sixth extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the methylmalonyl CoA specific AT with a malonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; deleting any one, two, or all three of the KR, DH, and ER; and/or replacing any one, two, or all three of the KR, DH, and ER with another KR, DH, and ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous sixth extender module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the sixth extender module of the FK-520 PKS.

In an illustrative embodiment, the present invention provides a set of recombinant FK-520 PKS genes in which the coding sequences for the DH and ER domains of the sixth extender module have been deleted or mutated to render them non-functional. In one such mutated gene, the KR, ER, and DH coding sequences are replaced with those encoding only a KR domain from another PKS gene. This can also be accomplished by simply replacing the coding sequences for extender module six with those for an extender module having a methylmalonyl specific AT and only a KR domain from a heterologous PKS gene, such as, for example, the coding sequences for extender module two encoded by the eryAI gene. The resulting PKS genes code for the expression of an FK-520 PKS that produces an FK-520 analog that has a C-18 hydroxyl group. Such analogs are preferred neurotrophins, because they have little or no immunosuppressant activity. This recombinant sixth extender module coding sequence can be combined with other coding sequences to make additional compounds of the invention. In an illustrative embodiment, the present invention provides a recombinant FK-520 PKS that contains both this sixth extender module and the recombinant fourth extender module described above that comprises the coding sequence for the fourth extender module AT domain of the FK-506 PKS. The invention also provides recombinant host cells derived from FK-506 producing host cells that have been mutated to prevent production of FK-506 but that express this recombinant PKS and so synthesize the corresponding (having a C-18 hydroxyl group) FK-506 derivative. In another embodiment, the present invention provides a recombinant FK-506 PKS in which the DH and ER domains of module 6 have been deleted or otherwise rendered inactive and thus produces this novel polyketide.

The seventh extender module of the FK-520 PKS includes a KS, an AT specific for 2-hydroxymalonyl CoA, a KR, a DH, an ER, and an ACP. The recombinant DNA compounds of the invention that encode the seventh extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 seventh extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the seventh extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the seventh extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion or all of the seventh extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the 2-hydroxymalonyl CoA specific AT with a methylmalonyl CoA, ethylmalonyl CoA, or malonyl CoA specific AT; deleting the KR, the DH, and/or the ER; and/or replacing the KR, DH, and/or ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous seventh extender module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the seventh extender module of the FK-520 PKS.

In an illustrative embodiment, the present invention provides a set of recombinant FK-520 PKS genes in which the coding sequences for the AT domain of the seventh extender module has been replaced with those encoding an AT domain for malonyl, methylmalonyl, or ethylmalonyl CoA from another PKS gene. The resulting PKS genes code for the expression of an FK-520 PKS that produces an FK-520 analog that lacks the C-15 methoxy group, having instead a hydrogen, methyl, or ethyl group at that position, respectively. Such analogs are preferred, because they are more slowly metabolized than FK-520. This recombinant seventh extender module coding sequence can be combined with other coding sequences to make additional compounds of the invention. In an illustrative embodiment, the present invention provides a recombinant FK-520 PKS that contains both this seventh extender module and the recombinant fourth extender module described above that comprises the coding sequence for the fourth extender module AT domain of the FK-506 PKS. The invention also provides recombinant host cells derived from FK-506 producing host cells that have been mutated to prevent production of FK-506 but that express this recombinant PKS and so synthesize the corresponding (C-15-desmethoxy) FK-506 derivative. In another embodiment, the present invention provides a recombinant FK-506 PKS in which the AT domain of module 7 has been replaced and thus produces this novel polyketide.

In another illustrative embodiment, the present invention provides a hybrid PKS in which the AT and KR domains of module 7 of the FK-520 PKS are replaced by a methylmalonyl specific AT domain and an inactive KR domain, such as, for example, the AT and KR domains of extender module 6 of the rapamycin PKS. The resulting hybrid PKS produces 15-desmethoxy-15-methyl-16-oxo-FK-520, a neurotrophin compound.

The eighth extender module of the FK-520 PKS includes a KS, an AT specific for 2-hydroxymalonyl CoA, a KR, and an ACP. The recombinant DNA compounds of the invention that encode the eighth extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 eighth extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the eighth extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the eighth extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion of the eighth extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the 2-hydroxymalonyl CoA specific AT with a methylmalonyl CoA, ethylmalonyl CoA, or malonyl CoA specific AT; deleting or replacing the KR; and/or inserting a DH or a DH and an ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous eighth extender module coding sequence can be utilized in conjunction with a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the eighth extender module of the FK-520 PKS.

In an illustrative embodiment, the present invention provides a set of recombinant FK-520 PKS genes in which the coding sequences for the AT domain of the eighth extender module has been replaced with those encoding an AT domain for malonyl, methylmalonyl, or ethylmalonyl CoA from another PKS gene. The resulting PKS genes code for the expression of an FK-520 PKS that produces an FK-520 analog that lacks the C-13 methoxy group, having instead a hydrogen, methyl, or ethyl group at that position, respectively. Such analogs are preferred, because they are more slowly metabolized than FK-520. This recombinant eighth extender module coding sequence can be combined with other coding sequences to make additional compounds of the invention. In an illustrative embodiment, the present invention provides a recombinant FK-520 PKS that contains both this eighth extender module and the recombinant fourth extender module described above that comprises the coding sequence for the fourth extender module AT domain of the FK-506 PKS. The invention also provides recombinant host cells derived from FK-506 producing host cells that have been mutated to prevent production of FK-506 but that express this recombinant PKS and so synthesize the corresponding (C-13-desmethoxy) FK-506 derivative. In another embodiment, the present invention provides a recombinant FK-506 PKS in which the AT domain of module 8 has been replaced and thus produces this novel polyketide.

The ninth extender module of the FK-520 PKS includes a KS, an AT specific for methylmalonyl CoA, a KR, a DH, an ER, and an ACP. The recombinant DNA compounds of the invention that encode the ninth extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 ninth extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the ninth extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the ninth extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion of the ninth extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the methylmalonyl CoA specific AT with a malonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; deleting any one, two, or all three of the KR, DH, and ER; and/or replacing any one, two, or all three of the KR, DH, and ER with another KR, DH, and/or ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous ninth extender module coding sequence can be utilized in conjunction with a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the ninth extender module of the FK-520 PKS.

The tenth extender module of the FK-520 PKS includes a KS, an AT specific for malonyl CoA, and an ACP. The recombinant DNA compounds of the invention that encode the tenth extender module of the FK-520 PKS and the corresponding polypeptides encoded thereby are useful for a variety of applications. In one embodiment, a DNA compound comprising a sequence that encodes the FK-520 tenth extender module is inserted into a DNA compound that comprises the coding sequence for a heterologous PKS. The resulting construct, in which the coding sequence for a module of the heterologous PKS is either replaced by that for the tenth extender module of the FK-520 PKS or the latter is merely added to coding sequences for the modules of the heterologous PKS, provides a novel PKS coding sequence. In another embodiment, a DNA compound comprising a sequence that encodes the tenth extender module of the FK-520 PKS is inserted into a DNA compound that comprises the coding sequence for the remainder of the FK-520 PKS or a recombinant FK-520 PKS that produces an FK-520 derivative.

In another embodiment, a portion or all of the tenth extender module coding sequence is utilized in conjunction with other PKS coding sequences to create a hybrid module. In this embodiment, the invention provides, for example, either replacing the malonyl CoA specific AT with a methylmalonyl CoA, ethylmalonyl CoA, or 2-hydroxymalonyl CoA specific AT; and/or inserting a KR, a KR and DH, or a KR, DH, and an ER. In addition, the KS and/or ACP can be replaced with another KS and/or ACP. In each of these replacements or insertions, the heterologous KS, AT, DH, KR, ER, or ACP coding sequence can originate from a coding sequence for another module of the FK-520 PKS, from a coding sequence for a PKS that produces a polyketide other than FK-520, or from chemical synthesis. The resulting heterologous tenth extender module coding sequence can be utilized in conjunction with a coding sequence for a PKS that synthesizes FK-520, an FK-520 derivative, or another polyketide. In similar fashion, the corresponding domains in a module of a heterologous PKS can be replaced by one or more domains of the tenth extender module of the FK-520 PKS.

The FK-520 polyketide precursor produced by the action of the tenth extender module of the PKS is then attached to pipecolic acid and cyclized to form FK-520. The enzyme FkbP is the NRPS like enzyme that catalyzes these reactions. FkbP also includes a thioesterase activity that cleaves the nascent FK-520 polyketide from the NRPS. The present invention provides recombinant DNA compounds that encode the fkbP gene and so provides recombinant methods for expressing the fkbP gene product in recombinant host cells. The recombinant fkbP genes of the invention include those in which the coding sequence for the adenylation domain has been mutated or replaced with coding sequences from other NRPS like enzymes so that the resulting recombinant FkbP incorporates a moiety other than pipecolic acid. For the construction of host cells that do not naturally produce pipecolic acid, the present invention provides recombinant DNA compounds that express the enzymes that catalyze at least some of the biosynthesis of pipecolic acid (see Nielsen et al., 1991 , Biochem. 30: 5789-96). The fkbL gene encodes a homolog of RapL, a lysine cyclodeaminase responsible in part for producing the pipecolate unit added to the end of the polyketide chain. The fkbB and fkbL recombinant genes of the invention can be used in heterologous hosts to produce compounds such as FK-520 or, in conjunction with other PKS or NRPS genes, to produce known or novel polyketides and non-ribosmal peptides.

The present invention also provides recombinant DNA compounds that encode the P450 oxidase and methyltransferase genes involved in the biosynthesis of FK-520. FIG. 2 shows the various sites on the FK-520 polyketide core structure at which these enzymes act. By providing these genes in recombinant form, the present invention provides recombinant host cells that can produce FK-520. This is accomplished by introducing the recombinant PKS, P450 oxidase, and methyltransferase genes into a heterologous host cell. In a preferred embodiment, the heterologous host cell is Streptomyces coelicolor CH999 or Streptomyces lividans K4-114, as described in U.S. Pat. No. 5,830,750 and U.S. patent application Ser. No. 08/828,898, filed Mar. 31, 1997, and Ser. No. 09/181,833, filed Oct. 28, 1998, each of which is incorporated herein by reference. In addition, by providing recombinant host cells that express only a subset of these genes, the present invention provides methods for making FK-520 precursor compounds not readily obtainable by other means.

In a related aspect, the present invention provides recombinant DNA compounds and vectors that are useful in generating, by homologous recombination, recombinant host cells that produce FK-520 precursor compounds. In this aspect of the invention, a native host cell that produces FK-520 is transformed with a vector (such as an SCP2* derived vector for Streptomyces host cells) that encodes one or more disrupted genes (i.e., a hydroxylase, a methyltransferase, or both) or merely flanking regions from those genes. When the vector integrates by homologous recombination, the native, functional gene is deleted or replaced by the non-functional recombinant gene, and the resulting host cell thus produces an FK-520 precursor. Such host cells can also be complemented by introduction of a modified form of the deleted or mutated non-functional gene to produce a novel compound.

In one important embodiment, the present invention provides a hybrid PKS and the corresponding recombinant DNA compounds that encode those hybrid PKS enzymes. For purposes of the present invention a hybrid PKS is a recombinant PKS that comprises all or part of one or more modules and thioesterase/cyclase domain of a first PKS and all or part of one or more modules, loading module, and thioesterase/cyclase domain of a second PKS. In one preferred embodiment, the first PKS is all or part of the FK-520 PKS, and the second PKS is only a portion or all of a non-FK-520 PKS.

One example of the preferred embodiment is an FK-520 PKS in which the AT domain of module 8, which specifies a hydroxymalonyl CoA and from which the C-13 methoxy group of FK-520 is derived, is replaced by an AT domain that specifies a malonyl, methylmalonyl, or ethylmalonyl CoA. Examples of such replacement AT domains include the AT domains from modules 3, 12, and 13 of the rapaymycin PKS and from modules 1 and 2 of the erythromycin PKS. Such replacements, conducted at the level of the gene for the PKS, are illustrated in the examples below. Another illustrative example of such a hybrid PKS includes an FK-520 PKS in which the natural loading module has been replaced with a loading module of another PKS. Another example of such a hybrid PKS is an FK-520 PKS in which the AT domain of module three is replaced with an AT domain that binds methylmalonyl CoA.

In another preferred embodiment, the first PKS is most but not all of a non-FK-520 PKS, and the second PKS is only a portion or all of the FK-520 PKS. An illustrative example of such a hybrid PKS includes an erythromycin PKS in which an AT specific for methylmalonyl CoA is replaced with an AT from the FK-520 PKS specfic for malonyl CoA.

Those of skill in the art will recognize that all or part of either the first or second PKS in a hybrid PKS of the invention need not be isolated from a naturally occurring source. For example, only a small portion of an AT domain determines its specificity. See U.S. provisional patent application Serial No. 60/091,526, incorporated herein by reference. The state of the art in DNA synthesis allows the artisan to construct de novo DNA compounds of size sufficient to construct a useful portion of a PKS module or domain. For purposes of the present invention, such synthetic DNA compounds are deemed to be a portion of a PKS.

Thus, the hybrid modules of the invention are incorporated into a PKS to provide a hybrid PKS of the invention. A hybrid PKS of the invention can result not only:

(i) from fusions of heterologous domain (where heterologous means the domains in that module are from at least two different naturally occurring modules) coding sequences to produce a hybrid module coding sequence contained in a PKS gene whose product is incorporated into a PKS, but also:

(ii) from fusions of heterologous module (where heterologous module means two modules are adjacent to one another that are not adjacent to one another in naturally occurring PKS enzymes) coding sequences to produce a hybrid coding sequence contained in a PKS gene whose product is incorporated into a PKS,

(iii) from expression of one or more FK-520 PKS genes with one or more non-FK-520 PKS genes, including both naturally occurring and recombinant non-FK-520 PKS genes, and

(iv) from combinations of the foregoing.

Various hybrid PKSs of the invention illustrating these various alternatives are described herein.

Examples of the production of a hybrid PKS by co-expression of PKS genes from the FK-520 PKS and another non-FK-520 PKS include hybrid PKS enzymes produced by coexpression of FK-520 and rapamycin PKS genes. Preferably, such hybrid PKS enzymes are produced in recombinant Streptomyces host cells that produce FK-520 or FK-506 but have been mutated to inactivate the gene whose function is to be replaced by the rapamycin PKS gene introduced to produce the hybrid PKS. Particular examples include (i) replacement of the fkbC gene with the rapB gene; and (ii) replacement of the fkbA gene with the rapC gene. The latter hybrid PKS produces 13,15-didesmethoxy-FK-520, if the host cell is an FK-520 producing host cell, and 13,15-didesmethoxy-FK-506, if the host cell is an FK-506 producing host cell. The compounds produced by these hybrid PKS enzymes are immunosuppressants and neurotrophins but can be readily modified to act only as neurotrophins, as described in Example 6, below.

Other illustrative hybrid PKS enzymes of the invention are prepared by replacing the fkbA gene of an FK-520 or FK-506 producing host cell with a hybrid fkbA gene in which: (a) the extender module 8 through 10, inclusive, coding sequences have been replaced by the coding sequnces for extender modules 12 to 14, inclusive, of the rapamycin PKS; and (b) the module 8 coding sequences have been replaced by the module 8 coding sequence of the rifamycin PKS. When expressed with the other, naturally occurring FK-520 or FK-506 PKS genes and the genes of the modification enzymes, the resulting hybrid PKS enzymes produce, respectively, (a) 13-desmethoxy-FK-520 or 13-desmethoxy-FK-506; and (b) 13-desmethoxy-13-methyl-FK-520 or 13-desmethoxy-13-methyl-FK-506. In a preferred embodiment, these recombinant PKS genes of the invention are introduced into the producing host cell by a vector such as pHU204, which is a plamsid pRM5 derivative that has the well-characterized SCP2* replicon, the colE1 replicon, the tsr and bla resistance genes, and a cos site. This vector can be used to introduce the recombinant fkbA replacement gene in an FK-520 or FK-506 producing host cell (or a host cell derived therefrom in which the endogenous fkbA gene has either been rendered inactive by mutation, deletion or homologous recombination with the gene that replaces it) to produce the desired hybrid PKS.

In constructing hybrid PKSs of the invention, certain general methods may be helpful. For example, it is often beneficial to retain the framework of the module to be altered to make the hybrid PKS. Thus, if one desires to add DH and ER functionalities to a module, it is often preferred to replace the KR domain of the original module with a KR, DH, and ER domain-containing segment from another module, instead of merely inserting DH and ER domains. One can alter the stereochemical specificity of a module by replacement of the KS domain with a KS domain from a module that specifies a different stereochemistry. See Lau et al., 1999, “Dissecting the role of acyltransferase domains of modular polyketide synthases in the choice and stereochemical fate of extender units,” Biochemistry 38(5):1643-1651, incorporated herein by reference. Stereochemistry can also be changed by changing the KR domain. Also, one can alter the specificity of an AT domain by changing only a small segment of the domain. See Lau et al., supra. One can also take advantage of known linker regions in PKS proteins to link modules from two different PKSs to create a hybrid PKS. See Gokhale et al., Apr. 16, 1999, “Dissecting and Exploiting Intermodular Communication in Polyketide Synthases,” Science 284: 482-485, incorporated herein by reference.

The following Table lists references describing illustrative PKS genes and corresponding enzymes that can be utilized in the construction of the recombinant PKSs and the corresponding DNA compounds that encode them of the invention. Also presented are various references describing tailoring enzymes and corresponding genes that can be employed in accordance with the methods of the present invention.

Avermectin

U.S. Pat. No. 5,252,474 to Merck.

MacNeil et al., 1993 , Industrial Microorganisms: Basic and Applied Molecular Genetics , Baltz, Hegeman, & Skatrud, eds. (ASM), pp. 245-256, A Comparison of the Genes Encoding the Polyketide Synthases for Avermectin, Erythromycin, and Nemadectin.

MacNeil et al., 1992 , Gene 115: 119-125, Complex Organization of the Streptomyces avermitilis genes encoding the avermectin polyketide synthase.

Ikeda et al., August 1999, Organization of the biosynthetic gene cluster for the polyketide anthelmintic macrolide avermectin in Streptomyces avermitilis, Proc. Natl. Acad. Sci. USA 96: 9509-9514.

Candicidin (FR008)

Hu et al., 1994 , Mol. Microbiol. 14: 163-172.

Epothilone

U.S. Pat. App. Ser. No. 60/130,560, filed Apr. 22, 1999.

Erythromycin

PCT Pub. No. 93/13663 to Abbott.

U.S. Pat. No. 5,824,513 to Abbott.

Donadio et al., 1991 , Science 252:675-9.

Cortes et al., Nov. 8, 1990 , Nature 348:176-8, An unusually large multifunctional polypeptide in the erythromycin producing polyketide synthase of Saccharopolyspora erythraea.

Glycosylation Enzymes

PCT Pat. App. Pub. No. 97/23630 to Abbott. FK-506

Motamedi et al., 1998, The biosynthetic gene cluster for the macrolactone ring of the immunosuppressant FK-506 , Eur. J. biochem. 256: 528-534.

Motamedi et al., 1997, Structural organization of a multifunctional polyketide synthase involved in the biosynthesis of the macrolide immunosuppressant FK-506 , Eur. J. Biochem. 244: 74-80.

Methyltransferase

U.S. Pat. No. 5,264,355, issued Nov. 23, 1993, Methylating enzyme from Streptomyces MA6858. 31-O-desmethyl-FK-506 methyltransferase.

Motamedi et al., 1996, Characterization of methyltransferase and hydroxylase genes involved in the biosynthesis of the immunosuppressants FK-506 and FK-520 , J. Bacteriol. 178: 5243-5248.

Streptomyces hygroscopicus

U.S. patent application Ser. No. 09/154,083, filed Sep. 16, 1998.

Lovastatin

U.S. Pat. No. 5,744,350 to Merck.

Narbomycin

U.S. patent application Ser. No. 60/107,093, filed Nov. 5, 1998, and Serial No. 60/120,254, filed Feb. 16, 1999.

Nemadectin

MacNeil et al., 1993, supra.

Niddamycin

Kakavas et al., 1997, Identification and characterization of the niddamycin polyketide synthase genes from Streptomyces caelestis, J. Bacteriol. 179: 7515-7522.

Oleandomycin

Swan et al., 1994, Characterisation of a Streptomyces antibioticus gene encoding a type I polyketide synthase which has an unusual coding sequence, Mol. Gen. Genet. 242: 358-362.

U.S. patent application Ser. No. 60/120,254, filed Feb. 16, 1999.

Olano et al., 1998, Analysis of a Streptomyces antibioticus chromosomal region involved in oleandomycin biosynthesis, which encodes two glycosyltransferases responsible for glycosylation of the macrolactone ring, Mol. Gen. Genet. 259(3): 299-308.

Picromycin

PCT patent application US99/15047, filed Jul. 2, 1999.

Xue et al., 1998, Hydroxylation of macrolactones YC-17 and narbomycin is mediated by the pikC-encoded cytochrome P450 in Streptomyces venezuelae, Chemistry & Biology 5(11): 661-667.

Xue et al., October 1998, A gene cluster for macrolide antibiotic biosynthesis in Streptomyces venezuelae : Architecture of metabolic diversity, Proc. Natl. Acad. Sci. USA 95: 12111 12116.

Platenolide

EP Pat. App. Pub. No. 791,656 to Lilly.

Rapamycin

Schwecke et al., August 1995, The biosynthetic gene cluster for the polyketide rapamycin, Proc. Natl. Acad. Sci. USA 92:7839-7843.

Aparicio et al., 1996, Organization of the biosynthetic gene cluster for rapamycin in Streptomyces hygroscopicus : analysis of the enzymatic domains in the modular polyketide synthase, Gene 169: 9-16.

Rifamycin

August et al., Feb. 13, 1998, Biosynthesis of the ansamycin antibiotic rifamycin: deductions from the molecular analysis of the rif biosynthetic gene cluster of Amycolatopsis mediterranei S 669 , Chemistry & Biology, 5(2): 69-79.

Sorangium PKS

U.S. patent application Ser. No. 09/144,085, filed Aug. 31, 1998.

Soraphen

U.S. Pat. No. 5,716,849 to Novartis.

Schupp et al., 1995 , J. Bacteriology 177: 3673-3679. A Sorangium cellulosum (Myxobacterium) Gene Cluster for the Biosynthesis of the Macrolide Antibiotic Soraphen A: Cloning, Characterization, and Homology to Polyketide Synthase Genes from Actinomycetes.

Spiramycin

U.S. Pat. No. 5,098,837 to Lilly.

Activator Gene

U.S. Pat. No. 5,514,544 to Lilly.

Tylosin

EP Pub. No. 791,655 to Lilly.

U.S. Pat. No. 5,876,991 to Lilly.

Kuhstoss et al., 1996 , Gene 183:231-6., Production of a novel polyketide through the construction of a hybrid polyketide synthase.

Tailoring Enzymes

Merson-Davies and Cundliffe, 1994 , Mol. Microbiol. 13: 349-355. Analysis of five tylosin biosynthetic genes from the tylBA region of the Streptomyces fradiae genome.

As the above Table illustrates, there are a wide variety of polyketide synthase genes that serve as readily available sources of DNA and sequence information for use in constructing the hybrid PKS-encoding DNA compounds of the invention. Methods for constructing hybrid PKS-encoding DNA compounds are described without reference to the FK-520 PKS in PCT patent publication No. 98/51695; U.S. Pat. Nos. 5,672,491 and 5,712,146 and U.S. patent application Ser. No. 09/073,538, filed May 6, 1998, and Ser. No. 09/141,908, filed Aug. 28, 1998, each of which is incorporated herein by reference.

The hybrid PKS-encoding DNA compounds of the invention can be and often are hybrids of more than two PKS genes. Moreover, there are often two or more modules in the hybrid PKS in which all or part of the module is derived from a second (or third) PKS. Thus, as one illustrative example, the present invention provides a hybrid FK-520 PKS that contains the naturally occurring loading module and FkbP as well as modules one, two, four, six, seven, and eight, nine, and ten of the FK-520 PKS and further contains hybrid or heterologous modules three and five. Hybrid or heterologous module three contains an AT domain that is specific of methylmalonyl CoA and can be derived for example, from the erythromycin or rapamycin PKS genes. Hybrid or heterologous module five contains an AT domain that is specific for malonyl CoA and can be derived for example, from the picromycin or rapamycin PKS genes.

While an important embodiment of the present invention relates to hybrid PKS enzymes and corresponding genes, the present invention also provides recombinant FK-520 PKS genes in which there is no second PKS gene sequence present but which differ from the FK-520 PKS gene by one or more deletions. The deletions can encompass one or more modules and/or can be limited to a partial deletion within one or more modules. When a deletion encompasses an entire module, the resulting FK-520 derivative is at least two carbons shorter than the gene from which it was derived. When a deletion is within a module, the deletion typically encompasses a KR, DH, or ER domain, or both DH and ER domains, or both KR and DH domains, or all three KR, DH, and ER domains.

To construct a hybrid PKS or FK-520 derivative PKS gene of the invention, one can employ a technique, described in PCT Pub. No. 98/27203 and U.S. patent application Ser. No. 08/989,332, filed Dec. 11, 1997, now U.S. Pat. No. 6,033,883, each of which is incorporated herein by reference, in which the large PKS gene is divided into two or more, typically three, segments, and each segment is placed on a separate expression vector. In this manner, each of the segments of the gene can be altered, and various altered segments can be combined in a single host cell to provide a recombinant PKS gene of the invention. This technique makes more efficient the construction of large libraries of recombinant PKS genes, vectors for expressing those genes, and host cells comprising those vectors.

Thus, in one important embodiment, the recombinant DNA compounds of the invention are expression vectors. As used herein, the term expression vector refers to any nucleic acid that can be introduced into a host cell or cell-free transcription and translation medium. An expression vector can be maintained stably or transiently in a cell, whether as part of the chromosomal or other DNA in the cell or in any cellular compartment, such as a replicating vector in the cytoplasm. An expression vector also comprises a gene that serves to produce RNA that is translated into a polypeptide in the cell or cell extract. Furthermore, expression vectors typically contain additional functional elements, such as resistance-conferring genes to act as selectable markers.

The various components of an expression vector can vary widely, depending on the intended use of the vector. In particular, the components depend on the host cell(s) in which the vector will be used or is intended to function. Vector components for expression and maintenance of vectors in E. coli are widely known and commercially available, as are vector components for other commonly used organisms, such as yeast cells and Streptomyces cells.

In a preferred embodiment, the expression vectors of the invention are used to construct recombinant Streptomyces host cells that express a recombinant PKS of the invention. Preferred Streptomyces host cell/vector combinations of the invention include S. coelicolor CH999 and S. lividans K4-114 host cells, which do not produce actinorhodin, and expression vectors derived from the pRM1 and pRM5 vectors, as described in U.S. Pat. No. 5,830,750 and U.S. patent application Ser. No. 08/828,898, filed Mar. 31, 1997, and Ser. No. 09/181,833, filed Oct. 28, 1998, each of which is incorporated herein by reference.

The present invention provides a wide variety of expression vectors for use in Streptomyces. For replicating vectors, the origin of replication can be, for example and without limitation, a low copy number vector, such as SCP2* (see Hopwood et al., Genetic Manipulation of Streptomyces: A Laboratory manual (The John Innes Foundation, Norwich, U.K., 1985); Lydiate et al., 1985 , Gene 35: 223-235; and Kieser and Melton, 1988 , Gene 65: 83-91, each of which is incorporated herein by reference), SLP1.2 (Thompson et al., 1982 , Gene 20: 51-62, incorporated herein by reference), and SG5(ts) (Muth et al., 1989 , Mol. Gen. Genet. 219: 341-348, and Bierman et al., 1992 , Gene 116: 43-49, each of which is incorporated herein by reference), or a high copy number vector, such as pIJI01 and pJV1 (see Katz et al., 1983 , J. Gen. Microbiol. 129: 2703-2714; Vara et al., 1989 , J. Bacteriol. 171: 5782-5781; and Servin-Gonzalez, 1993 , Plasmid 30: 131-140, each of which is incorporated herein by reference). Generally, however, high copy number vectors are not preferred for expression of genes contained on large segments of DNA. For non-replicating and integrating vectors, it is useful to include at least an E. coli origin of replication, such as from pUC, p1P, p1I, and pBR. For phage based vectors, the phages phiC31 and KC515 can be employed (see Hopwood et al., supra).

Typically, the expression vector will comprise one or more marker genes by which host cells containing the vector can be identified and/or selected. Useful antibiotic resistance conferring genes for use in Streptomyces host cells include the ermE (confers resistance to erythromycin and other macrolides and lincomycin), tsr (confers resistance to thiostrepton), aadA (confers resistance to spectinomycin and streptomycin), aacC4 (confers resistance to apramycin, kanamycin, gentamicin, geneticin (G418), and neomycin), hyg (confers resistance to hygromycin), and vph (confers resistance to viomycin) resistance conferring genes.

The recombinant PKS gene on the vector will be under the control of a promoter, typically with an attendant ribosome binding site sequence. The present invention provides the endogenous promoters of the FK-520 PKS and related biosynthetic genes in recombinant form, and these promoters are preferred for use in the native hosts and in heterologous hosts in which the promoters function. A preferred promoter of the invention is the fkbO gene promoter, comprised in a sequence of about 270 bp between the start of the open reading frames of the fkbO and fkbB genes. The fkbO promoter is believed to be bi-directional in that it promotes transcription of the genes fkbO, fkbP, and fkbA in one direction and fkbB, fkbC, and fkbL in the other. Thus, in one aspect, the present invention provides a recombinant expression vector comprising the promoter of the fkbO gene of an FK-520 producing organism positioned to transcribe a gene other than fkbO. In a preferred embodiment the transcribed gene is an FK-520 PKS gene. In another preferred embodiment, the transcribed gene is a gene that encodes a protein comprised in a hybrid PKS.

Heterologous promoters can also be employed and are preferred for use in host cells in which the endogenous FK-520 PKS gene promoters do not function or function poorly. A preferred heterologous promoter is the actI promoter and its attendant activator gene actI1-ORF4, which is provided in the pRM1 and pRM5 expression vectors, supra. This promoter is activated in the stationary phase of growth when secondary metabolites are normally synthesized. Other useful Streptomyces promoters include without limitation those from the ermE gene and the melC1 gene, which act constitutively, and the tipA gene and the merA gene, which can be induced at any growth stage. In addition, the T7 RNA polymerase system has been transferred to Streptomyces and can be employed in the vectors and host cells of the invention. In this system, the coding sequence for the T7 RNA polymerase is inserted into a neutral site of the chromosome or in a vector under the control of the inducible merA promoter, and the gene of interest is placed under the control of the T7 promoter. As noted above, one or more activator genes can also be employed to enhance the activity of a promoter. Activator genes in addition to the acti-ORF4 gene discussed above include dnrI, redD, and ptpA genes (see U.S. patent application Ser. No. 09/181,833, supra) to activate promoters under their control.

In addition to providing recombinant DNA compounds that encode the FK-520 PKS, the present invention also provides DNA compounds that encode the ethylmalonyl CoA and 2-hydroxymalonyl CoA utilized in the synthesis of FK-520. Thus, the present invention also provides recombinant host cells that express the genes required for the biosynthesis of ethylmalonyl CoA and 2-hydroxymalonyl CoA. FIGS. 3 and 4 show the location of these genes on the cosmids of the invention and the biosynthetic pathway that produces ethylmalonyl CoA.

For 2-hydroxymalonyl CoA biosynthesis, the fkbH, fkbI, fkbJ, and fkbK genes are sufficient to confer this ability on Streptomcyces host cells. For conversion of 2-hydroxymalonyl to 2-methoxymalonyl, the fkbG gene is also employed. While the complete coding sequence for fkbH is provided on the cosmids of the invention, the sequence for this gene provided herein may be missing a T residue, based on a comparison made with a similar gene cloned from the ansamitocin gene cluster by Dr. H. Floss. Where the sequence herein shows one T, there may be two, resulting in an extension of the fkbH reading frame to encode the amino acid sequence (SEQ ID NO:2):

For ethylmalonyl CoA biosynthesis, one requires only a crotonyl CoA reductase, which can be supplied by the host cell but can also be supplied by recombinant expression of the frbS gene of the present invention. To increase yield of ethylmalonyl CoA, one can also express the fkbE and fkbU genes as well. While such production can be achieved using only the recombinant genes above, one can also achieve such production by placing into the recombinant host cell a large segment of the DNA provided by the cosmids of the invention. Thus, for 2-hydroxymalonyl and 2-methoxymalonyl CoA biosynthesis, one can simply provide the cells with the segment of DNA located on the left side of the FK-520 PKS genes shown in FIG. 1 . For ethylmalonyl CoA biosynthesis, one can simply provide the cells with the segment of DNA located on the right side of the FK-520 PKS genes shown in FIG. 1 or, alternatively, both the right and left segments of DNA.

The recombinant DNA expression vectors that encode these genes can be used to construct recombinant host cells that can make these important polyketide building blocks from cells that otherwise are unable to produce them. For example, Streptomyces coelicolor and Streptomyces lividans do not synthesisze ethylmalonyl CoA or 2-hydroxymalonyl CoA. The invention provides methods and vectors for constructing recombinant Streptomyces coelicolor and Streptomyces lividans that are able to synthesize either or both ethylmalonyl CoA and 2-hydroxymalonyl CoA. These host cells are thus able to make polyketides, those requiring these substrates, that cannot otherwise be made in such cells.

In a preferred embodiment, the present invention provides recombinant Streptomyces host cells, such as S. coelicolor and S. lividans , that have been transformed with a recombinant vector of the invention that codes for the expression of the ethylmalonyl CoA biosynthetic genes. The resulting host cells produce ethylmalonyl CoA and so are preferred host cells for the production of polyketides produced by PKS enzymes that comprise one or more AT domains specific for ethylmalonyl CoA. Illustrative PKS enzymes of this type include the FK-520 PKS and a recombinant PKS in which one or more AT domains is specific for ethylmalonyl CoA.

In a related embodiment, the present invention provides Streptomyces host cells in which one or more of the ethylmalonyl or 2-hydroxymalonyl biosynthetic genes have been deleted by homologous recombination or rendered inactive by mutation. For example, deletion or inactivation of the fkbG gene can prevent formation of the methoxyl groups at C-13 and C-15 of FK-520 (or, in the corresponding FK-506 producing cell, FK-506), leading to the production of 13,15-didesmethoxy-13,15-dihydroxy-FK-520 (or, in the corresponding FK-506 producing cell, 13,15-didesmethoxy-13,15-dihydroxy-FK-506). If the fkbG gene product acts on 2-hydroxymalonyl and the resulting 2-methoxymalonyl substrate is required for incorporation by the PKS, the AT domains of modules 7 and 8 may bind malonyl CoA and methylmalonyl CoA. Such incorporation results in the production of a mixture of polyketides in which the methoxy groups at C-13 and C-15 of FK-520 (or FK-506) are replaced by either hydrogen or methyl.

This possibility of non-specific binding results from the construction of a hybrid PKS of the invention in which the AT domain of module 8 of the FK-520 PKS replaced the AT domain of module 6 of DEBS. The resulting PKS produced, in Streptomyces lividans, 6-dEB and 2-desmethyl-6-dEB, indicating that the AT domain of module 8 of the FK-520 PKS could bind malonyl CoA and methylmalonyl CoA substrates. Thus, one could possibly also prepare the 13,15-didesmethoxy-FK-520 and corresponding FK-506 compounds of the invention by deleting or otherwise inactivating one or more or all of the genes required for 2-hydroxymalonyl CoA biosynthesis, i.e., the fkbH, fkbI, fkbJ, and fkbK genes. In any event, the deletion or inactivation of one or more biosynthetic genes required for ethylmalonyl and/or 2-hydroxymalonyl production prevents the formation of polyketides requiring ethylmalonyl and/or 2-hydroxymalonyl for biosynthesis, and the resulting host cells are thus preferred for production of polyketides that do not require the same.

The host cells of the invention can be grown and fermented under conditions known in the art for other purposes to produce the compounds of the invention. See, e.g., U.S. Pat. Nos. 5,194,378; 5,116,756; and 5,494,820, incorporated herein by reference, for suitable fermentation processes. The compounds of the invention can be isolated from the fermentation broths of these cultured cells and purified by standard procedures. Preferred compounds of the invention include the following compounds: 13-desmethoxy-FK-506; 13-desmethoxy-FK-520; 13,15-didesmethoxy-FK-506; 13,15-didesmethoxy-FK-520; 13-desmethoxy-18-hydroxy-FK-506; 13-desmethoxy-18-hydroxy-FK-520; 13,15-didesmethoxy-18-hydroxy-FK-506; and 13,15-didesmethoxy-18-hydroxy-FK-520. These compounds can be further modified as described for tacrolimus and FK-520 in U.S. Pat. Nos. 5,225,403; 5,189,042; 5,164,495; 5,068,323; 4,980,466; and 4,920,218, incorporated herein by reference.

Other compounds of the invention are shown in FIG. 8, Parts A and B. In FIG. 8, Part A, illustrative C-32-substituted compounds of the invention are shown in two columns under the heading R. The substituted compounds are preferred for topical administration and are applied to the dermis for treatment of conditions such as psoriasis. In FIG. 8, Part B, illustrative reaction schemes for making the compounds shown in FIG. 8, Part A, are provided. In the upper scheme in FIG. 8, Part B, the C-32 substitution is a tetrazole moiety, illustrative of the groups shown in the left column under R in FIG. 8, Part A. In the lower scheme in FIG. 8, Part B, the C-32 substitution is a disubstituted amino group, where R 3 and R 4 can be any group similar to the illustrative groups shown attached to the amine in the right column under R in FIG. 8, Part A. While FIG. 8 shows the C-32-substituted compounds in which the C-15-methoxy is present, the invention includes these C-32-substituted compounds in which C-15 is ethyl, methyl, or hydrogen. Also, while C-21 is shown as substituted with ethyl or allyl, the compounds of the invention includes the C-32-substituted compounds in which C-21 is substituted with hydrogen or methyl.

To make these C-32-substituted compounds, FIG. 8, Part B, provides illustrative reaction schemes. Thus, a selective reaction of the starting compound (see FIG. 8, Part B, for an illustrative starting compound) with trifluoromethanesulfonic anhydride in the presence of a base yields the C-32 O-triflate derivative, as shown in the upper scheme of FIG. 8, Part B. Displacement of the triflate with 1H-tetrazole or triazole derivatives provides the C-32 tetrazole or teiazole derivative. As shown in the lower scheme of FIG. 8, Part B, reacting the starting compound with p-nitrophenylchloroformate yields the corresponding carbonate, which, upon displacement with an amino compound, provides the corresponding carbamate derivative.

The compounds can be readily formulated to provide the pharmaceutical compositions of the invention. The pharmaceutical compositions of the invention can be used in the form of a pharmaceutical preparation, for example, in solid, semisolid, or liquid form. This preparation contains one or more of the compounds of the invention as an active ingredient in admixture with an organic or inorganic carrier or excipient suitable for external, enteral, or parenteral application. The active ingredient may be compounded, for example, with the usual non-toxic, pharmaceutically acceptable carriers for tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, and any other form suitable for use. Suitable formulation processes and compositions for the compounds of the present invention are described with respect to tacrolimus in U.S. Pat. Nos. 5,939,427; 5,922,729; 5,385,907; 5,338,684; and 5,260,301, incorporated herein by reference. Many of the compounds of the invention contain one or more chiral centers, and all of the stereoisomers are included within the scope of the invention, as pure compounds as well as mixtures of stereoisomers. Thus the compounds of the invention may be supplied as a mixture of stereoisomers in any proportion.

The carriers which can be used include water, glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea, and other carriers suitable for use in manufacturing preparations, in solid, semi-solid, or liquified form. In addition, auxiliary stabilizing, thickening, and coloring agents and perfumes may be used. For example, the compounds of the invention may be utilized with hydroxypropyl methylcellulose essentially as described in U.S. Pat. No. 4,916,138, incorporated herein by reference, or with a surfactant essentially as described in EPO patent publication No. 428,169, incorporated herein by reference.

Oral dosage forms may be prepared essentially as described by Hondo et al., 1987 , Transplantation Proceedings XIX , Supp. 6: 17-22, incorporated herein by reference. Dosage forms for external application may be prepared essentially as described in EPO patent publication No. 423,714, incorporated herein by reference. The active compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect upon the disease process or condition.

For the treatment of conditions and diseases relating to immunosuppression or neuronal damage, a compound of the invention may be administered orally, topically, parenterally, by inhalation spray, or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvant, and vehicles. The term parenteral, as used herein, includes subcutaneous injections, and intravenous, intramuscular, and intrasternal injection or infusion techniques.

Dosage levels of the compounds of the present invention are of the order from about 0.01 mg to about 50 mg per kilogram of body weight per day, preferably from about 0.1 mg to about 10 mg per kilogram of body weight per day. The dosage levels are useful in the treatment of the above-indicated conditions (from about 0.7 mg to about 3.5 mg per patient per day, assuming a 70 kg patient). In addition, the compounds of the present invention may be administered on an intermittent basis, i.e., at semi-weekly, weekly, semi-monthly, or monthly intervals.

The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. For example, a formulation intended for oral administration to humans may contain from 0.5 mg to 5 g of active agent compounded with an appropriate and convenient amount of carrier material, which may vary from about 5 percent to about 95 percent of the total composition. Dosage unit forms will generally contain from about 0.5 mg to about 500 mg of active ingredient. For external administration, the compounds of the invention can be formulated within the range of, for example, 0.00001% to 60% by weight, preferably from 0.001% to 10% by weight, and most preferably from about 0.005% to 0.8% by weight. The compounds and compositions of the invention are useful in treating disease conditions using doses and administration schedules as described for tacrolimus in U.S. Pat. Nos. 5,542,436; 5,365,948; 5,348,966; and 5,196,437, incorporated herein by reference. The compounds of the invention can be used as single therapeutic agents or in combination with other therapeutic agents. Drugs that can be usefully combined with compounds of the invention include one or more immunosuppressant agents such as rapamycin, cyclosporin A, FK-506, or one or more neurotrophic agents.

It will be understood, however, that the specific dosage level for any particular patient will depend on a variety of factors. These factors include the activity of the specific compound employed; the age, body weight, general health, sex, and diet of the subject; the time and route of administration and the rate of excretion of the drug; whether a drug combination is employed in the treatment; and the severity of the particular disease or condition for which therapy is sought.

A detailed description of the invention having been provided above, the following examples are given for the purpose of illustrating the present invention and shall not be construed as being a limitation on the scope of the invention or claims.

A  G  A  V  E  L  L  T  S  A  R  P  W  P  E  T  D

R  P  R  R  A  A  V  S  S  F  G  V  S  G  T  N  A

H  V  I  L  E  A  G  P  V  T  E  T  P  A  A  S

P  S  G  D  L  P  L  L  V  S  A  R  S  R  E  A  L

D  E  Q  I  R  R  L  R  A  Y  L  D  T  T  P  D  V

D  R  V  A  V  A  Q  T  L  A  R  R  T  H  F  A

H  R  A  V  L  L  G  D  T  V  I  T  T  P  P  A  D

R  P  D  E  L  V  F  V  Y  S  G  Q  G  T  Q  H  P

A  M  G  E  Q  L  A  A  A  H  P  V  F  A  D  A

The sequences shown below provide the location of the AT/DH boundary chosen in the FK-520 module 8 coding sequences. The region where an XhoI site was engineered is indicated by lower case and underlining (SEQ ID NOS:66-67).

I  L  G  A  G  S  R  H  D  A  D  V  P  A  Y  A  F

Q  R  R  H  Y  W  I  E  S  A  R  P  A  A  S  D  A

The sequences shown below provide the location of the KS/AT boundaries chosen in the FK-506 module 8 coding sequences. Regions where AvrII and NheI sites were engineered are indicated by lower case and underlining (SEQ ID NOS:68-69).

S  A  R  P  W  P  R  T  G  R  P  R  R  A  A  V  S

S  F  C  V  S  G  T  N  A  H  I  I  L  E  A  G

P  D  Q  E  E  P  S  A  E  P  A  G  D  L  P  L  L

V  S  A  R  S  P  E  A  L  D E  Q  I  G  R  L  R

D  Y  L  D  A  A  P  G  V  D  L  A  A   V  A  R

T  L  A  T  R  T  H  F  S  H  R  A  V  L  L  G  D

T  V  I  T  A  P  P  V  E  Q  P  G  E  L  V  F  V

Y  S  G  Q  G  T  Q  H  P  A  M  G  E  R  L  A

A  A  F  P  V  F  A  D  P  D  V  P  A  Y  A  F  Q

The sequences shown below provide the location of the AT/DH boundary chosen in the FK-506 module 8 coding sequences. The region where an XhoI site was engineered is indicated by lower case and underlining (SEQ ID NOS:70-71).

D  P  D  V  P  A  Y  A  F  Q  R  R  P  Y  W  T  E

<160> NUMBER OF SEQ ID NOS: 72

<210> SEQ ID NO 1

<211> LENGTH: 77536

<212> TYPE: DNA

<213> ORGANISM: Streptomyces hygroscopicus

<220> FEATURE:

<221> NAME/KEY: CDS

<222> LOCATION: (52275)...(71465)

<400> SEQUENCE: 1

gatctcaggc atgaagtcct ccaggcgagg cgccgaggtg gtgaacacct cg

#ccgctgct     60

tgtacggacc acttcagtca gcggcgattg cggaaccaag tcatccggaa ta

#aagggcgg    120

ttacaagatc ctcacattgc gcgaccgcca gcatacgctg agttgcctca ga

#ggcaaacc    180

gaaagggcgc gggcggtccg caccagggcg gagtacgcga cgagagtggc gc

#acccgcgc    240

accgtcacct ctctcccccg ccggcgggat gcccggcgtg acacggttgg gc

#tctcctcg    300

acgctgaaca cccgcgcggt gtggcgtcgg ggacaccgcc tggcatcggc cg

#ggtgacgg    360

tacggggagg gcgtacggcg gccgtggctc gtgctcacgg ccgccgggcg gt

#catccgtc    420

gagacggcac tcggcgagca gggacgcctg gtcggcacct gcgggccgga cg

#accgtgtg    480

gttcgcgggc gggcggtggc cggtggtgag ccagctctcc agggcggtga ag

#gctgagcg    540

gtgacacggc agcaaaggcc ggagtcggtc ggggaaggtg tcgacgaggg cg

#tcggtgtg    600

cgtgccgtcc tcgatgcggt agtagcggta ccggccgcca ggccgctgcc gg

#acatacgc    660

gcgtacacgt cggagcccgg gcggcaggca gcagcacgtc gagagtgcct gg

#atggtgat    720

cagcggcttg ccgatacgac cggtcaacgc gatgcgttcc acggccgcgt gg

#acgccgga    780

ggagcgggtg gcgtagtcgt agtcggcatc gcagcccggg accgtccccg gg

#gcgcaata    840

cggtgtgccg gcttccttct ccccatcgaa gccggggtcg aactcctcgc gg

#tagacgcg    900

ctgcgtcaga tcccagtaga cctcgtggtg gtacggccac aagaactcgg ag

#tcggccgg    960

gaacccggcg cggagcagcg cctcgcgcgc ctggccggct gcggggccgc ct

#gccgcgta   1020

ggtggggtag tcgcgcaggg cggccggcag gaaggtgaag aggttgggac cc

#tccgcgcg   1080

ccacagggtg ccttcccagt cgactcctcc gtcgtacagc tcgggatggt tc

#tccagctg   1140

ccagcgcacg aggtagccgc cgttggacat cccggtgacc agggtgcgct cg

#agcggccg   1200

gtggtagcgc tgggcgaccg acgcgcgggc ggcccgggtc agctgggtga gg

#cgggtgtt   1260

ccactcggcg acggcgtcgc ccggccggga gccatcacgg tagaacgcgg gg

#ccggtgtt   1320

gcccttgtcg gtggcggcgt aggcgtaacc gcgggcgagc acccagtcgg cg

#atggcccg   1380

gtcgttggcg tactgctcgc ggttaccggg ggtgccggcc acgaccaggc ca

#ccgttcca   1440

gcggtcgggc agccggatga cgaactgggc gtcgtggttc cacccgtggt tg

#gtgttggt   1500

ggtggaggtg tcggggaagt agccgtcgat ctggatcccg ggcactccgg tg

#ggagtggc   1560

caggttcttg ggcgtcagcc ctgcccagtc cgccgggtcg gtgtggccgg tg

#gccgccgt   1620

tcccgccgtg gtcagctcgt ccaggcagtc ggcctgctga cgtgccgccg cc

#gggacacg   1680

cagctgggac agacgggcgc agtgaccgtc cggggcatcg ggagcaggcc gg

#gccgtggc   1740

cggtgagggg agcaggacgg cgactgcggc cagggtgaga gcgccgaggc cg

#gtgcgtct   1800

tctcggggcc cgtccgacac cgaggggcag aaccatggag agcctccaga cg

#tgcggatg   1860

gatgacggac tggaggctag gtcgcgcacg gtggagacga acatgggtgc gc

#ccgccatg   1920

actgaggccc ctcagaggtg ggccgccgcc atgacgggcg cgggaccgcg gg

#cgctccgg   1980

ggcggtgccc gcggccgcca ccggttccgg gtccccgggt cagggacagg tg

#tcgttcgc   2040

gacggtgaag tagccggtcg gcgactcttt caaggtggtc gtgacgaagg tg

#ttgtacag   2100

gcccatgttc tggccggagc ccttggcgta ggtgtaaccg gcgctcgtcg tg

#gcgcggcc   2160

cgcctggacg tgagcgtagt tgccggcggt ccagcagacg gccgtggcac cg

#gtcgtctg   2220

cgcggtgacc gcgcccgaga gcggtccggc cttgccgtcc gcgtcccggg cg

#gcgaccgc   2280

gtaggtgtgc gatgtgcccg ccctcaggcc ggtgtccgtg tacgacgtcg tg

#gcggacgt   2340

ggtgatctgg gcaccgtcgc ggtggacggc gtagtcggtg gcgccgtcga cg

#ggtttcca   2400

ggtcaggctg atggtggtgt cggtggcgcc ggtggcggcc aggccggacg ga

#gcgggcag   2460

cgaaccgggg tcggaggcgg atccgctcag gccgaagaac tgcgtgatcc ag

#tagctgga   2520

acagatcgag tccaggaagt aggcggcgcc ggtgctgccg cactgctgtg ct

#ccggtgcc   2580

gggatcgacc ggggtgccgt gcccgatgcc cggcacccgg ttcacctcca cg

#gccaccga   2640

tccgtccgcg gccaggtact cctcgtgccg ggtggagttc gggccgatca cc

#gaggtacg   2700

gtccggcgtc tgggacacgc cgtgcacagc ggtccactgg tcgcgcaact cg

#tcggcgtt   2760

gcgcggcgcg acggtggtgt ccttgtcgcc gtgccagatg gccacgcgcg gc

#cacgggcc   2820

cgaccacgag gggtagccgt cacggacccg ccgcgcccac tggtccgcgg tc

#aggtcggt   2880

cccggggttc atgcacaggt acgcgctgct gacgtcggtg gcacagccga ag

#ggcaggcc   2940

ggcgacgacc gcgccggcct ggaagacgtc cggataggtg gcgagcatca cc

#gacgtcat   3000

ggcaccgccg gcggacagcc cggtgatgta ggtgcgctgg gggtccgcgc cg

#taggcgga   3060

gacggtgtga gcggccatct gccggatcga cgcggcttcg ccctggcccc tg

#cggttgtc   3120

gctgctctgg aaccagttga agcacctgtt cgcgttgttc gacgacgtgg tc

#tcggcgaa   3180

cacgagcagg aagccatagc ggtccgcgaa tgagagcagg ccggagttgt cg

#gcgtagcc   3240

ctgggcgtcc tgggtgcaac cgtgcagggc gaacaccacc gccggctccg cg

#ggcaggga   3300

cgcgggccgg tagacgtaca tgttcagccg gcccgggttc gtgccgaagt cc

#gcgacctc   3360

ggtcaggtcc gccttggtca gaccgggctt ggccaggccc gccgcggcgt gg

#gccgtcgg   3420

cgccgggccg agcagggccg ctccgagtac gagggccacg acggccacga ga

#cgggtgag   3480

caccccccgc cgtcccggac gcgacaacga cccgaccggc ggcgaggagg ag

#agggggaa   3540

cagcggggtg aggattcccc ggaacggcgg cggctgcatg gcggctccct cg

#atgtcgtg   3600

ggggggacac ggagggctcc ctgacgtcga tcagtgggag cgccccggtg cc

#cggcaccg   3660

taggggtggt tcaacccgca acggtatggc ccggagcacc acaccccgca cc

#gcgcgatg   3720

tgcgcccgga cggattgtgt cgccttgcgg aatctgatac ccggacgcga cg

#aacgcccc   3780

acccgacacg ggtagggcgt catggtgtcc gactcggccg gtcggccttg cc

#tgccctgg   3840

acggaccggg cgtcggcgga ccgggcgtcg gcgggctggg cggtatggcg gc

#cgaggacg   3900

ccagccgcgt ggggcggccg cgcccaagtg cagtacgccg accgtggccg gc

#gggagggc   3960

cggaccggtc agtgcagtcc cgcggccctg cgggaccgct cgtcccagac gg

#gttccacc   4020

gcggcgaacc ggggtccgtg tccgcggcgg tagaccatca gtgtccgctc ga

#aggtgatg   4080

acgatgacac cgtcctggtt gtagccgatg gtgcgcacgc tgatgatgcc ta

#cgtcaggt   4140

cggctggcgg actcccgggt gttcaggacc tcggactgcg agtagatggt gt

#cgccctcg   4200

aagaccgggt tcggcagcct gacccggtcc cagccgaggt tggccatcac at

#gctgggag   4260

atgtcggtga cgctctgccc ggtgaccagg gcgagggtga aggtggagtc ca

#ccagcggc   4320

ttgccccagg tggtgcccgc cgagtagtgg cggtcgaagt gcagcggcgc gg

#tgttctgc   4380

gtcaggagcg tgagccagga gttgtcggtc tccaggaccg tgcggcccag gg

#ggtggcgg   4440

tacacgtcgc cggtggtgaa gtcctcgaag tagcggccct gccagccctc ga

#ccacagcg   4500

gtgcgggtgg cgtcctggtc cgggttctca gtcgtcatgg cgctcattct gg

#gaagtccc   4560

cggtccgctg tgaaatgccg aaccttcacc gggctcatac gtgcggcgca tg

#agccctgg   4620

accgtacgta gtcgtagaac ctcgccacca ctggcgcgcg tggtcctccg gc

#gagtgtga   4680

ccacgccgac cgtgcgccgc gcctgcgggt cgtcgagcgg cacggcgacg gc

#gtggtcac   4740

cgggcccgga cgggctgccg gtgagggggg cgacggccac accgaggccg gc

#ggcgacca   4800

gggcccgcag cgtgctcagc tcggtgctct ccaggacgac ccgcggcacg aa

#tccggccg   4860

cggcgcacag ccggtcggtg atctggcgca gtccgaagac cggctccagt gc

#cacgaacg   4920

cctcatcggc cagctccgcg gtccgcaccc ggcggcgtct ggccagccgg tg

#tccgggtg   4980

ggacgagcag gcacagtgcc tcgtcccgca gtggtgtcca ctccacatcg tc

#cccggcgg   5040

gtcgtgggct ggtcagcccc aggtccagcc tgctgttgcg gacgtcgtcg ac

#cacggcgt   5100

cggcggcgtc gccgcgcagt tcgaaggtgg tgccgggagc cagccggcgg ta

#cccggcga   5160

ggaggtcggg caccagccag gtgccgtagg agtgcaggaa acccagtgcc ac

#ggtgccgg   5220

tgtcggggtc gatcagggcg gtgatgcgct gctcggcgcc ggagacctca ct

#gatcgcgc   5280

gcagggcgtg ggcgcggaag acctcgccgt acttgttgag ccggagccgg tt

#ctggtgcc   5340

ggtcgaacag cggcacgccc actcgtcgct ccagccgccg gatggccctg ga

#cagggtcg   5400

gctgggagat gttgagccgt tccgcggtga tcgtcacgtg ctcgtgctcg gc

#caaggccg   5460

tgaaccactg caactcccgt atctccatgc agggactata cgtaccgggc at

#ggtcctgg   5520

cgaggtttcg tcatttcaca gcggccgggc ggcggcccac agtgagtcct ca

#ccaaccag   5580

gaccccatgg gagggacccc atgtccgagc cgcatcctcg ccctgaacag ga

#acgccccg   5640

ccgggcccct gtccggtctg ctcgtggttt ctttggagca ggccgtcgcc gc

#tccgttcg   5700

ccacccgcca cctggcggac ctgggcgccc gtgtcatcaa gatcgaacgc cc

#cggcagcg   5760

gcgacctcgc ccgcggctac gaccgcacgg tgcgtggcat gtccagccac tt

#cgtctggc   5820

tgaaccgggg gaaggagagc gtccagctcg atgtgcgctc gccggagggc aa

#ccggcacc   5880

tgcacgcctt ggtggaccgg gccgatgtcc tggtgcagaa tctggcaccc gg

#cgccgcgg   5940

gccgcctggc atcggccacc aggtcctcgc gcggagccac cgaggctgat ca

#cctgcgga   6000

catatccggc tacggcagta ccggctgcta ccgcggaccg caaggcgtac ga

#cctcctgg   6060

tccagtgcga agcggggctg gtctccatca ccggcacccc cgagaccccg tc

#caaggtgg   6120

gcctgtccat cgcggacatc tgtgcgggga tgtacgcgta ctccggcatc ct

#cacggccc   6180

tgctgaagcg ggcccgcacc ggccggggct cgcagttgga ggtctcgatg ct

#cgaagccc   6240

tcggtgaatg gatgggatac gccgagtact acacgcgcta cggcggcacc gc

#tccggccc   6300

gcgccggcgc cagccacgcg acgatcgccc cctacggccc gttcaccacg cg

#cgacgggc   6360

agacgatcaa tctcgggctc cagaacgagc gggagtgggc ttccttctgc gg

#tgtcgtgc   6420

tacaacgccc cggtctctgc gacgacccgc gcttttccgg caacgccgac cg

#ggtggcgc   6480

accgcaccga gctcgacgcc ctggtgagcg aggtgacggg cacgctcacc gg

#cgaggaac   6540

tggtggcgcg gctggaggag gcgtcgatcg cctacgcacg ccagcgcacc gt

#gcgggagt   6600

tcagcgaaca cccccaactg cgtgaccgtg gacgctgggc tccgttcgac ag

#cccggtcg   6660

gtgcgctgga gggcctgatc cccccggtca ccttccacgg cgagcacccg cg

#gcggctgg   6720

gccgggtccc ggagctgggc gagcataccg agtccgtcct ggcgtggctg gc

#cgcgcccc   6780

acagcgccga ccgcgaagag gccggccatg ccgaatgaac tcaccggagt cc

#tgatcctg   6840

gccgccgtgt tcctgctcgc cggcgtacgg gggctgaaca tgggcctgct cg

#cgctggtc   6900

gccacctttc tgctcggggt ggtcgcactc gaccgaacgc cggacgaggt gc

#tggcgggt   6960

ttccccgcga gcatgttcct ggtgctggtc gccgtcacgt tcctcttcgg ga

#tcgcccgc   7020

gtcaacggca cggtggactg gctggtacgt gtcgcggtgc gggcggtggg gg

#cccgggtg   7080

ggagccgtcc cctgggtgct cttcggcctg gcggcactgc tctgcgcgac ag

#gcgcggcc   7140

tcgcccgcgg cggtggcgat cgtggcgccg atcagcgtcg cgttcgccgt ca

#ggcaccgc   7200

atcgatccgc tgtacgccgg actgatggcg gtgaacgggg ccgcagccgg ca

#gtttcgcc   7260

ccctccggga tcctgggcgg catcgtccac tcggcgctgg agaagaacca tc

#tgcccgtc   7320

agcggcgggc tgctcttcgc aggcaccttc gccttcaacc tggcggtcgc cg

#cggtgtca   7380

tggctcgtcc tcgggcgcag gcgcctcgaa ccacatgacc tggacgagga ca

#ccgatccc   7440

acggaagggg acccggcttc ccgccccggc gcggaacacg tgatgacgct ga

#ccgcgatg   7500

gccgcgctgg tgctgggaac cacggtcctc tccctggaca ccggcttcct gg

#ccctcacc   7560

ttggcggcgt tgctggcgct gctcttcccg cgcacctccc agcaggccac ca

#aggagatc   7620

gcctggcccg tggtgctgct ggtatgcggg atcgtgacct acgtcgccct gc

#tccaggag   7680

ctgggcatcg tggactccct ggggaagatg atcgcggcga tcggcacccc gc

#tgctggcc   7740

gccctggtga tctgctacgt gggcggtgtc gtctcggcct tcgcctcgac ca

#ccgggatc   7800

ctcggtgccc tgatgccgct gtccgagccg ttcctgaagt ccggtgccat cg

#ggacgacc   7860

ggcatggtga tggccctggc ggccgcggcg accgtggtgg acgcgagtcc ct

#tctccacc   7920

aatggtgctc tggtggtggc caacgctccc gagcggctgc ggcccggcgt gt

#accagggg   7980

ttgctgtggt ggggcgccgg ggtgtgcgca ctggctcccg cggccgcctg gg

#cggccttc   8040

gtggtggcgt gagcgcagcg gagcgggaat cccctggagc ccgtttcccg tg

#ctgtgtcg   8100

ctgacgtagc gtcaagtcca cgtgccgggc gggcagtacg cctagcatgt cg

#ggcatggc   8160

taatcagata accctgtccg acacgctgct cgcttacgta cggaaggtgt cc

#ctgcgcga   8220

tgacgaggtg ctgagccggc tgcgcgcgca gacggccgag ctgccgggcg gt

#ggcgtact   8280

gccggtgcag gccgaggagg gacagttcct cgagttcctg gtgcggttga cc

#ggcgcgcg   8340

tcaggtgctg gagatcggga cgtacaccgg ctacagcacg ctctgcctgg cc

#cgcggatt   8400

ggcgcccggg ggccgtgtgg tgacgtgcga tgtcatgccg aagtggcccg ag

#gtgggcga   8460

gcggtactgg gaggaggccg gggttgccga ccggatcgac gtccggatcg gc

#gacgcccg   8520

gaccgtcctc accgggctgc tcgacgaggc gggcgcgggg ccggagtcgt tc

#gacatggt   8580

gttcatcgac gccgacaagg ccggctaccc cgcctactac gaggcggcgc tg

#ccgctggt   8640

acgccgcggc gggctgatcg tcgtcgacaa cacgctgttc ttcggccggg tg

#gccgacga   8700

agcggtgcag gacccggaca cggtcgcggt acgcgaactc aacgcggcac tg

#cgcgacga   8760

cgaccgggtg gacctggcga tgctgacgac ggccgacggc gtcaccctgc tg

#cggaaacg   8820

gtgaccgggg cgatgtcggc ggcggtcagc gtcagcgtcg tcggcgcggg cc

#tcgcggag   8880

ggctccagat gcaggcgttc gacgccggcg gcggaagcgc ccgccacctc gg

#acacgcag   8940

gggcagtcgg agtccgcgaa gcccgcgaac cggtaggcga tctccatcat gc

#ggttgcgg   9000

tccgtacgcc ggaagtccgc caccaggtgc gcccccgcgc gggcgccctg gt

#ccgtgagc   9060

cagttcagga tcgtcgcacc ggcaccgaac gacacgaccc ggcaggacgt gg

#cgagcagt   9120

ttcaggtgcc acgtcgacgg cttcttctcc agcaggatga tgccgacggc gc

#cgtgcggg   9180

ccgaagcggt cgcccatggt gacgacgagg acctcatggg cgggatcggt ga

#gcacgcgc   9240

gcaggtcggc gtcggagtag tgcacgccgg tcgcgttcat ctggctggtc cg

#cagcgtca   9300

gttcctcgac gcggctgagt tcctcctccc ccgcgggtgc gatcgtcatg ga

#gaggtcga   9360

gcgagcgcag gaagtcctcg tcgggaccgg agtacgcctc ccgggcctgg tc

#gcgcgcga   9420

aacccgcctg gtacatcagg cggcgccgac gcgagtcgac cgtggacacc gg

#cgggctga   9480

actccggcag cgacaggagc gtggccgcct gctcggccgg gtagcaccgc ac

#ctcgggca   9540

ggtggaacgc cacctcggca cgctcggcgg gctggtcgtc gatgaacgcg at

#cgtggtcg   9600

gtgcgaagtt cagctccgtg gcgatctcgc ggacggactg cgacttcggc cc

#ccatccga   9660

tgcgggccag cacgaagtac tccgccacac cgaggcgttc cagacgctcc ca

#cgcgaggt   9720

cgtggtcgtt cttgctcgcc accgcctgga ggatgccgcg gtcgtcgagc gt

#ggtgatca   9780

cctcgcggat ctcgtcggtg aggaccacct cgtcgtcctc cagcacggtg cc

#ccgccaca   9840

aggtgttgtc caggtcccag accagacact tgacaatggt catggctgtc ct

#ctcaagcc   9900

gggagcgcca gcgcgtgctg ggccagcatc acccggcaca tctcgctgct gc

#cctcgatg   9960

atctccatga gcttggcgtc gcggtacgcc cgttcgacga cgtgtccctc tc

#tcgcgcct  10020

gccgacgcga gcacctgtgc ggcggtcgcg gccccggcgg cggctcgttc gg

#cggcgacg  10080

tgcttggcca ggatcgtcgc gggcaccatc tcgggcgagc cctcgtccca gt

#ggtcgctg  10140

gcgtactcgc acacgcgggc cgcgatctgc tccgcggtcc acaggtcggc ga

#tgtgcccg  10200

gcgacgagtt ggtggtcgcc gagcggccgg ccgaactgct cccgggtccg gg

#cgtgggcc  10260

accgcggcgg tgcggcaggc ccgcaggatc ccgacgcagc cccaggcgac cg

#acttgcgc  10320

ccgtaggcga gtgacgccgc gaccagcatc ggcagtgacg cgccggagcc gg

#ccaggacc  10380

gcgccggccg gcacacgcac ctggtccagg tgcagatcgg cgtggccggc gg

#cgcggcag  10440

ccggacggct tcgggacgcg ctcgacgcgt acgccggggg tgtcggcggg ca

#cgaccacc  10500

accgcaccgg aaccatcctc ctggagaccg aagacgacca ggtggtccgc gt

#aggcggcg  10560

gcagtcgtcc agaccttgtg gccgtcgacg acagcggtgt ccccgtcgag cc

#gaacccgc  10620

gtccgcatcg ccgacagatc gctgcccgcc tgccgctcac tgaagccgac gg

#ccgcgagt  10680

ttcccgctgg tcagctcctt caggaaggtc gcccgctgac cggcgtcgcc ga

#gccgctgc  10740

acggtccacg cggccatgcc ctgcgacgtc atgacactgc gcagcgaact gc

#agaggctg  10800

ccgacgtgtg cggtgaactc gccgttctcc cggctgccga gtcccagacc gc

#cgtgctcg  10860

gccgccactt ccgcgcagag caggccgtcg gcgccgagcc ggacgagcag gt

#cgcgcggc  10920

agttcgccgg acgtgtccca ctcggcggcc cggtcaccga caaggtcggt ca

#gcagcgcg  10980

tcacgctcag gcatcgacgg cccgcagccg gtggacgagt gcgaccatgg ac

#tcgacggt  11040

acggaagttc gcgagctgga ggtccgggcc ggcgatcgtg acgtcgaacg tc

#ttctccag  11100

gtacacgacc agttccatcg cgaacagcga cgtgaggccg ccctccgcga ac

#aggtcgcg  11160

gtccacgggc cagtccgacc tggtcttcgt cttgaggaac gcgaccaacg cg

#tgcgcgac  11220

ggggtcgtcc ttgacgggtg cggtcatgag aacaccttct cgtattcgta ga

#agccccgg  11280

ccggtcttcc ggccgtggtg tccctcgcgg accttgccca gcagcaggtc ac

#aggggcgg  11340

ctgcgctcgt cgccggtgcg tttgtgcagc acccacagcg cgtcgacgag gt

#tgtcgatg  11400

ccgatcaggt ccgcggtgcg cagcggcccg gtcggatggc cgaggcaccc cg

#tcatgagc  11460

gcgtcgacgt cctcgacgga cgcggtgccc tcctgcacga tccgcgccgc gt

#cgttgatc  11520

atcgggtgga gcagccggct cgtgacgaag ccgggcgcgt cccggacgac ga

#tcggcttg  11580

cgccgcagcg ccgcgagcag gtccccggcg gcggccatgg ccttctcacc gg

#tccggggt  11640

ccgcggatca cctcgaccgt cgggatcagg tacgacgggt tcatgaagtg cg

#tgccgagc  11700

aggtcctcgg gccgggccac ggagtcggcc agttcgtcaa ccgggatcga cg

#acgtgttc  11760

gtgatgaccg ggataccggg cgccgctgcc gagaccgtgg cgagtacctc cg

#ccttgacc  11820

tcggcgtcct cgacgacggc ctcgatcacc gcggtggccg taccgatcgc gg

#gcagcgcg  11880

gacgtggccg tccgcagcac accggggtcg gcctcggcgg gcccggccac ga

#gttgtgcc  11940

gtccgcagtt cggtggcgat ccgcgcccgc gccgccgtaa ggatctcctc gg

#acgtgtcg  12000

acgagtgtca ccgggacgcc gtggcgcagc gcgagcgtgg tgatgccggt gc

#ccatcact  12060

cccgcgccga gcacgatcag ctggtggtcc acgctgtttc ctccctccgg gg

#tcaccatg  12120

gcagcgagta cgggtcgagg acgtcttccg gggtcgaccc gatcgcgtcc tt

#gcggccga  12180

ggccgagttc gtcggcgaag ccgagcagca cgtcgaacgc gatgtggtcg gc

#gaacgcgc  12240

tgcccgtcga gtcgaggacg ctcaggctgt cccggtggtc cgccgcggtg tc

#cggtgccg  12300

cgcacagggc cgccagcgac gggccgagct cgcggtccgg cagttgctgg ta

#ctcgccct  12360

cggcgcgggc ctgccccgga tggtcgacgc agatgaacgc gtcgtcgagc ag

#ggtcttcg  12420

gcagttcggt cttgcccggc tcgtcggcgc cgatggcgtt cacatgcagg tg

#cggcagcc  12480

gcggctcggc gggcagcacc ggccctttgc ccgagggcac cgaggtgacg gt

#ggacagga  12540

catccgcggc ggcggcggcc tccgccggat cggtcacctt gaccggcagt cc

#gaggaacg  12600

cgatgcggtc cgcgaacgac gccgcgtggc cggggtcggt gtcgctgacc ag

#gatccgct  12660

cgatgggcag gaccctgctg agcgcgtgcg cctgggtcac cgcctgtgcg cc

#cgcgccga  12720

tcagcgtgag cgtggcgctg tcggaccggg ccagcagccg gctcgcgacg gc

#ggcgaccg  12780

cgccggtccg catcgcggtg atcacgcctg cgtcggcgag ggcggtcaga ct

#gccgctgt  12840

cgtcgtcgag gcgcgacatc gtgccgacga tcgtcggcag ccggaagcgc gg

#atagttgt  12900

gcggactgta cgaaaccgtc ttcatggtca cgccgacacc ggggacccgg ta

#cggcatga  12960

actcgatgac gccgggaatg tcgccgccgc ggacgaatcc ggtacgcggc gg

#cgcctcgg  13020

cgaactcgcc gcggccgagc gcggcgaacc cgtcgtgcag ctcgctgatc ag

#ccggtcca  13080

tcatcacgtc gcggccgatc acggagagaa tccgcttgat gtcacgttgg cg

#caggaccc  13140

tggtctgcat gtgtcacctc cctttcgtgg ccggagctgt cttggtggtg cc

#gctcgggg  13200

cggcttccgt tctcatcgca gctccctgtc gatgaggtcg aaaatctcgt cc

#gcggtcgc  13260

gtccgcggac agcacgccgg ccggcgtggt cgggcgggtc tcccgccgcc ag

#cggttgag  13320

cagggcgtcc agccgggttc cgatcgcgtc cgcctggcgg gcgcccgggt cg

#acaccggc  13380

aacgagtgct tccagccggt cgagctgcgc gagcaccacg gtcaccgggt cg

#tccgggga  13440

cagcagttca ccgatgcggt cggcgagtgc gcgcggcgac gggtagtcga ag

#acgagcgt  13500

ggcggacagt cgcagaccgg tcgcctcgtt gaggccgttg cgcagctgca cc

#gcgatgag  13560

cgagtccaca ccgagttccc ggaacgccgc gtcctccggg atgtcctccg gg

#tcggcgtg  13620

gcccaggacg gccgctgcct tctgccggac gagggcgagc aggtcggtgg gg

#cgttcctg  13680

ctcgttgcgg gcgctccggc gggccgacgg cttgggccgg ccacgcagca gc

#gggaggtc  13740

cggcggcagg tcgcccgcca cggcgacgac actgcccgtt ccggtgtgga cg

#gcggcgtc  13800

gtacatgcgc atgccctgtt cggcggtgag cgcgctcgcc ccacccttgc gc

#atacggcg  13860

ccggtcggcg tcggtcaggt ccgcggtcag gccactcgcc tggtcccaca gc

#ccccacgc  13920

gatcgacagc cctggcagcc cttgtgcacg ccggtgttcg gcgagcgcgt cg

#aggaacgc  13980

gttcgccgcc gcgtagttgc cctgaccggg ggtgcccagc acaccggccg cc

#gacgagta  14040

gacgacgaat gcggcgaggt cggtgtcgcg ggtgagccgg tgcaggtgcc ag

#gcggcgtc  14100

ggccttgggt ttgaggacgg tgtcgatgcg gtcgggggtg aggttgtcga gc

#agggcgtc  14160

gtcgagggtt ccggcggtgt ggaagacggc ggtgaggggt tgagggatgt gg

#gcgagggt  14220

ggtggcgagt tggtgggggt cgccgacgtc gcaggggagg tgggtgccgg gg

#gtggtgtc  14280

ggggggtggg gtgcgggaga ggaggtaggt gtgggggtgg ttcaggtggc gg

#gcgaggat  14340

gccggcgagg gtgccggagc cgccggtgat gacgacggcc ccctcggggt cc

#agcggccg  14400

cgggaccgtg aggacgatct tgccggtgtg ctcgccgcgg ctcatggtcg cc

#agcgcctc  14460

gcggacctgc cgcatgtcgt gcaccgtcac cggcagcggg tgcagcacac cg

#cgcgcgaa  14520

caggccgagc agctccgcga tgatctcctt gagccggtcg ggccccgcgt cc

#atcaggtc  14580

gaacggtcgc tggacggcgt gccggatgtc cgtcttcccc atctcgatga ac

#cggccacc  14640

cggcgcgagc aggccgacgg acgcgtcgag gagttcaccg gtgagcgagt tg

#agcacgac  14700

gtcgaccggc gggaacgcgt cggcgaacgc ggtgctgcgg gaatcggcca ga

#tgcgctcc  14760

gtccaggtcc accagatggc gcttcgcggc gctggtggtc gcgtacacct cc

#gcgcccag  14820

gtgccgcgcg atctgccggg cggcggaacc gacaccgccg gtggccgcgt gg

#atcaggac  14880

cttctcgccg gggcgcagcc cggcgaggtc gaccaggccg taccacgcgg tc

#gcgaacgc  14940

ggtcatcacg gacgccgcct gcgggaacgt ccagccgtcc ggcatccggc cg

#agcatccg  15000

gtggtcggcg atgaccgtgg ggccgaagcc ggtgccgacg aggccgaaga cg

#cggtcgcc  15060

cggtgccaga ccggagacgt cggcgccggt ctccaggacg atgcccgcgg cc

#tcgccgcc  15120

gagcacgccc tgaccggggt aggtgccgag cgcgatcagc acatcgcgga ag

#ttgaggcc  15180

cgccgcacgc acaccgatcc ggacctcggc cggggcgagg gggcgccggg gc

#tccgccga  15240

gtcggccgcg gtgaggccgt cgagggtgcc cgtccgcgcc ggccggatca gc

#cacgtgtc  15300

gctgtccggc acggtgagcg gctccggcac ccgggtgagg cgggccgcct cg

#aaccggcc  15360

gccgcgcagc cgcagacgcg gctcgccgag tgcgacggcg atgcgctgct gc

#tcgggggc  15420

gagcgtgacg ccggactcgg tctcgacgtg gacgaaccgg ccgggctgct cg

#gcctgggc  15480

ggcgcgcagc agtccggccg ccgcgccggt ggcgaggccc gcggtggtgt gc

#acgagcag  15540

atccccgccg gagccggtca gggcggtcag cagccgggtg gtgagcgcac gc

#gtctcggc  15600

caccgggtcg tcgccatcag cggcaggcaa cgtgatgacg tccacgtcgg tc

#gcggggac  15660

atccgtgggt gcggcgacct cgatccaggt gagacgcatc aggccggtgc cg

#acgggtgg  15720

ggacagcggg cgggtgcgga ccgtccggat ctcggcgacg agttggccgg cg

#gagtcggc  15780

gacgcgcaga ctcagctcgt cgccgtcacg agtgatcacg gctcggagca tg

#gccgagcc  15840

cgtggcgacg aaccgggccc ccttccaggc gaacggcaga cccgcagcgc tg

#tcgtccgg  15900

cgtggtgagg gcgacggcgt gcagggccgc gtcgagcagc gccggatgca ca

#ccgaaacc  15960

gtccgcctcg gcggcctgct cgtcgggcag cgccacctcg gcatacacgg tg

#tcaccatc  16020

acgccaggca gcccgcaacc cctggaacgc cgacccgtac tcataaccgg ca

#tcccgcag  16080

ttcgtcatag aaccccgaga cgtcgacggc cacggccgtg accggcggcc ac

#tgcgagaa  16140

cggctccaca ccgacaacac cgggggtgtc gggggtgtcg ggggtcaggg tg

#ccgctggc  16200

gtgccgggtc cagctgcccg tgccctcggt acgcgcgtgg acggtcaccg gc

#cgccgtcc  16260

ggcctcatca gccccttcca cggtcaccga cacatccacc gctgcggtca cc

#ggcaccac  16320

aaggggggat tcgatgacca gctcgtccac tatcccgcaa ccggtctcgt ca

#ccggcccg  16380

gatgaccagc tccacaaacg ccgtacccgg cagcaggacc gtgccccgca cc

#gcgtgatc  16440

agccagccag gggtgagtgc gcaatgagat ccggccagtg agaacaacac ca

#ccatcgtc  16500

ggcgggcagc gctgtgacag cggccagcat cggatgcgcc gcacccgtca ac

#cccgccgc  16560

cgacagatcg gtggcaccgg ccgcctccag ccagtaccgc ctgtgctcga ac

#gcgtacgt  16620

gggcagatcc agcagccgtc ccggcaccgg ttcgaccacc gtgtcccagt cc

#actgccgt  16680

gcccagggtc cacgcctgcg ccaacgccgt cagccaccgc tcccagccgc cg

#tcaccggt  16740

ccgcaacgac gccaccgtgt gagcctgctc catcgccggc agcagcaccg ga

#tgggcact  16800

gcactccacg aacaccgacc catccagctc cgccaccgcc gcgtccaacg cc

#accggacg  16860

acgcagattc cggtaccagt acccctcatc caccggctcc gtcacccagg cg

#ctgtccac  16920

ggtcgaccac cacgccaccg acgcggcctt ccctgccacc ccctccagta cc

#ttggccag  16980

ttcatcctcg atggcttcca cgtggggcgt gtgggaggcg tagtcgaccg cg

#atacgacg  17040

cacccgcacg ccttcggcct cataccgcgc caccacctcc tccaccgccg ac

#gggtcccc  17100

cgccaccacc gtcgaagccg ggccgttacg cgccgcgatc cacacaccct cg

#accagacc  17160

gacctcaccg gccggcaacg ccaccgaagc catcgctccc cgcccggcca gt

#cgcgccgc  17220

gatgacctga ctgcgcaatg ccaccacgcg ggcggcgtcc tcgaggctga gg

#gctccggc  17280

cacgcacgcc gccgcgatct cgccctggga gtgtccgatc accgcgtccg gc

#acgacccc  17340

atgcgcctgc cacagcgcgg ccaggctcac cgcgaccgcc cagctggccg gc

#tggaccac  17400

ctccacccgc tccgccacat ccggccgcgc caacatctcc cgcacatccc ag

#cccgtgtg  17460

cggcagcaac gcctgagcgc actcctccat acgcgcggcg aacaccgcgg ag

#tgggccat  17520

gagttccacg cccatgccga cccactgggc gccctggccg gggaagacga ac

#accgtacg  17580

cggctggtcc accgccacac ccgtcacccg ggcatcgccc agcagcaccg ca

#cggtgacc  17640

gaagacagca cgctcccgca ccaacccctg cgcgaccgcg gccacatcca ca

#ccaccccc  17700

gcgcagatac ccctccagcc gctccacctg cccccgcaga ctcacctcac ca

#cgagccga  17760

caccggcaac ggcaccaacc cgtcaacaac cgactcccca cgcgacggcc ca

#ggaacacc  17820

ctcaaggatc acgtgcgcgt tcgtaccgct caccccgaac gacgacacac cc

#gcatgcgg  17880

tgcccgatcc gactcgggcc acggcctcgc ctcggtgagc agctccaccg ca

#ccggccga  17940

ccagtccaca tgcgacgacg gctcgtccac atgcagcgtc ttcggcgcga tc

#ccgtaccg  18000

catcgccatg accatcttga tcacaccggc gacacccgcc gccgcctgcg ca

#tgaccgat  18060

gttcgacttc aacgaaccca gcagcagcgg aacctcacgc tcctgcccgt ac

#gtcgccag  18120

aatggcctgc gcctcgatgg gatcgcccag cgtcgtcccc gtcccgtgcg cc

#tccaccac  18180

gtccacatcg gcggcgcgca gtccggcgtt caccaacgcc tgctggatga ca

#cgctgctg  18240

ggacgggccg ttgggggcgg acagcccgtt ggaggcaccg tcctggttca cc

#gccgaccc  18300

gcggacgacc gcgagaacgg tgtgtccgtt gcgctcggcg tcggagagcc gc

#tccagcac  18360

aagaacgccg gcgccctccg cccagccggt gccgttggcg gcgtccgcga ac

#gcgcggca  18420

gcggccgtcg ggggagagtc cgccctgctg ctggaattcc acgaacccgg tc

#ggggtcgc  18480

catgacggtg acaccgccga ccagcgccag cgagcactcc ccgtggcgca gt

#gcgtgccc  18540

ggcctggtgc agcgcgacca gcgacgacga gcacgccgtg tccaccgtga ac

#gccggtcc  18600

ctggagccca tagaagtacg agatccggcc ggtgagcacg ctgggctgca tg

#ccgatcga  18660

gccgaacccg tccaggtccg cgccgacgcc gtacccgtac gagaaggcgc cc

#atgaacac  18720

gccggtgtcg ctgccgcgca gtgtgcccgg cacgatgccc gcgctctcga ac

#gcctccca  18780

tgtcgtttcc agcaggatcc gctgctgggg gtccatggcc cgtgcctcac gg

#gggctgat  18840

gccgaagaac gcggcatcga agccggcggc gtcggagagg aagccgccgc gg

#tccgtgtc  18900

cgatccgccg gtgaggccgg acgggtccca gccacggtcg gccgggaagc cg

#gtgaccgc  18960

gtcgccgcca ctgtccacca tgcgccacag gtcgtcgggc gaggtgacgc cg

#cccggcag  19020

tcggcaggcc atgcccacga tggccagcgg ttcgtcacgg gtcgcggcgg ct

#gtgggaac  19080

agcgaccggt gcggcaccac cgaccagagc ctcgtccaac cgcgacgcga tg

#gcccgcgg  19140

cgtcgggtag tcgaagacaa gcgtggcggg cagtcggaca ccggtcgccg cg

#gcgagtcg  19200

gttccgcagt tcgacggcgg tcagcgagtc gatacccagt tccttgaagg cc

#gcgtccgc  19260

ggacacgtcc gcggcgtccg cgtggccgag caccgccgcc gcgttgtcgc gg

#accagtgc  19320

cagcagcgcg gtgtcccgct cagcgccgga catggtgccg agccggtcgg cg

#agcggaac  19380

ggcggtggcc gccgccgggc gcgatacggc gcggcgcaga tcggcgaaaa gc

#ggcgatgt  19440

gtgcgcggtg aggtccatcg tggccgccac ggcgaacgcg gtgccggttc cg

#gccgcggc  19500

ttccagcagg cgcatgccca caccggccga catggggcgg aaaccgccgc gg

#cggacacg  19560

ggtgcggttg gtgccgctca tgctgccggt gagtccgctg tcatcggccc ag

#aggcccca  19620

ggccagcgac agcgcgggca gtccttcggc atggcgcagc gtcgcgagtc cg

#tcgaggaa  19680

cccgttcgcc gccgagtagt tgccctggcc gcggccgccc atgatgcccg cg

#acggacga  19740

gtagaggacg aacgagcgca ggtccgcgtc ccgggtcagc tcgtgcaggt gc

#caggcgcc  19800

gtcggctttg gggcgcagtg tggtggcgag ccgctccggg gtgagtgccg tg

#gtcacgcc  19860

gtcgtcgagc acggctgccg tgtggaagac cgccgtgagc ggcctgccgg cg

#gcggcgag  19920

cgcggcggcg agctggtccc ggtcggcgac gtcacagcgg atgtggacac cg

#ggagtgtc  19980

cgccggcggt tcgctgcgcg acagcaacag gaggtggcgg gcgccatgct cg

#gcgacgag  20040

atgccgggcg aggagacctg ccagcacacc cgagccgccg gtgatgacca cc

#gtgccgtc  20100

cgggtcgagc agcggttcgg gcgtttccgc ggcggccgtg cgggtgaacc gc

#ggcgcttc  20160

gtaccggccg tcggtgacgc ggacgtacgg ctcggccagt gtcgtggcgg cg

#gccagcgc  20220

ctcgatgggg gtgtcggtgc cggtctccac cagcacgaac cggcccgggt gc

#tcggcctg  20280

ggcggaccgg acgaggccgg cgaccgctcc tccgaccggt cccgcgtcga tc

#cggacgac  20340

gagggtggtc tccgcagggc cgtcctcggc gatcacccgg tgcagctcgc cg

#agcacgaa  20400

ctcggtgagc cggtacgtct cgtcgaggac atccgcgccc ggttccggga gc

#gcggagac  20460

gatgtggacc gcgtccgcag gaccgggccc gggagtgggc agctcggtcc ag

#gagaggcc  20520

gtacaaggag ttccgtacga cggcggcgtc gccgtcgacg ttcaccggtc gc

#gcggtcag  20580

cgcggcgacg gtcaccaccg gttggccgac cgggtccgtc gcatgcacgg ca

#gcgccgtc  20640

cgggccctga gtgatcgtga cgcgcagcgt ggtggccccg gtcgtgtgga ac

#cgcacgcc  20700

gctccacgag aacggcagcc gcacctccgc ttcctgttcc gcgagcagcg gc

#aggcaggt  20760

gacgtgcaag gccgcgtcga acagcgccgg gtggacgcca tagtgcggcg tg

#tcgtccgc  20820

ctgttccccg gcgatctcca cctcggcgta cagggtttcg ccgtcgcgcc ag

#gcggtgcg  20880

cagtccctgg aacgctgggc cgtagctgta gccggtctcg gccagccgct cg

#tagaacgc  20940

gctcacgtcg acgcgtcgcg cgcccggcgg cggccacgcg ggcggcggga cc

#gccgcgac  21000

gcttccggcc cggccgaggg tgccgctggc gtgccgggtc cagctgtccg tg

#ccctcggt  21060

acgcgcgtgg acggtcactc gccgccgtcc ggcctcatcg gccccttcga cg

#gtcaccga  21120

cacatccacc gcgccggtca ccggcaccac gagcggggtc tcgatgacca gt

#tcatccac  21180

caccccgcaa ccggtctcgt caccggcccg gatgaccagc tccacaaacg cc

#gtacccgg  21240

cagcagaacc gtgccccgca ccgcgtgatc agccagccag ggatgcgtac gc

#aacgagat  21300

ccggccagtg agaacaacac caccaccgtc gtcggcgggc agtgctgtga cg

#gcggccag  21360

catcggatgc gccgccccgg tcagcccggc cgcggacaga tcggtggcac cg

#gccgcctc  21420

cagccagtac cgcctgtgct cgaacgcgta ggtgggcaga tcgagcagcc gt

#cccggcac  21480

cggttcgacc accgtgtccc agtccactgc cgtgcccagg gtccacgcct gc

#gccaacgc  21540

cgtcagccac cgctcccagc cgccgtcacc ggtccgcaac gacgccaccg tg

#tgagcctg  21600

ttccatcgcc ggcagcagca ccggatgggc gctgcactcc acgaacacgg ac

#ccgtccag  21660

ctccgccacc gccgcgtcca gcgcgacggg gcgacgcagg ttccggtacc ag

#tagccctc  21720

atccaccggc tcggtcaccc aggcgctgtc caccgtggac caccaggcca cc

#gacccggt  21780

cccgccggaa atcccctcca gtacctcggc caactcgtcc tcgatggctt cc

#acgtgggg  21840

cgtgtgggag gcgtagtcga ccgcgatacg gcgcactcgc acgccttcgg cc

#tcgtaccg  21900

cgtcaccact tcttccaccg cggacgggtc ccccgccacc acagtcgaag ac

#gggccgtt  21960

acgcgccgcg atccacacgc cctcgaccag gtccacctca ccggccggca ac

#gccaccga  22020

agccatcgcc ccccgcccgg ccagccgccc ggcgatcacc tggctgcgca ag

#gccaccac  22080

gcgggcggcg tcctcaaggc tgagggctcc ggccacacac gccgccgcga tc

#tcgccctg  22140

ggagtgtccg accaccgcgt ccggcacgac cccatgcgcc tgccacagcg cg

#gccaggct  22200

caccgcgacc gcccagctgg ccggctggac cacctccacc cgctccgcca ca

#tccggccg  22260

cgccaacatc tcccgcacat cccagcccgt gtgcggcaac aacgcccgcg ca

#cactcctc  22320

catacgagcc gcgaacaccg cagaacacgc catcaactcc acacccatgc cc

#acccactg  22380

agcaccctgc ccgggaaaga cgaacaccgt acgcggctga tccaccgcca ca

#cccatcac  22440

ccgggcatcg cccaacaaca ccgcacggtg accgaagaca gcacgctcac gc

#accaaccc  22500

ctgcgcgacc gcggccacat ccacaccacc cccgcgcaga tacccctcca gc

#cgctccac  22560

ctgcccccgc agactcacct cactccgagc cgacaccggc aacggcacca ac

#ccatcgac  22620

agccgactcc ccacgcgacg gcccgggaac accctcaagg atcacgtgcg cg

#ttcgtacc  22680

gctcaccccg aaagcggaga caccggcccg gcgcggacgt cccgcgtcgg gc

#cacgcccg  22740

cgcctcggtg agcagttcca ccgcgccctc ggtccagtcc acatgcgacg ac

#ggctcgtc  22800

cacatgcagc gtcttcggcg cgatgccata ccgcatcgcc atgaccatct tg

#atgacacc  22860

ggcgacaccc gcagccgcct gcgcatgacc gatgttcgac ttcaacgaac cc

#agcagcag  22920

cggaacctca cgctcctgcc cgtacgtcgc cagaatcgcg tgcgcctcga tg

#ggatcgcc  22980

cagcgtcgtc cccgtcccgt gcgcctccac cacgtccacg tcggcggggg cg

#agccccgc  23040

cttgtggagg gcctggcgga tgacgcgctg ctgggagggg ccgttgggtg cg

#gagatgcc  23100

gttggaggcg ccgtcctggt tgacggcgga ggagcggacg accgcgagga cg

#gtgtgtcc  23160

gttgcgctcg gcgtcggaga gcttttcgac gacgaggacg ccggccccct cg

#gcgaaacc  23220

ggtgccgtcc gccgcgtcag cgaacgcctt gcaccgtccg tccggcgcga cg

#ccgccctg  23280

ccgggagaac tccacgaagg tctgtggtga tgccatcact gtgacaccac cg

#accagcgc  23340

cagcgagcac tccccggtcc gcagcgcctg cccggcctgg tgcagcgcga cc

#agcgacga  23400

cgaacacgcc gtgtcgaccg tgaccgccgg accctccatg ccgaagaagt ac

#gacagccg  23460

tccggcgagc accgcgggct gtgtgctgta ggcgccgaat ccgcccaggt cc

#gcgcccgt  23520

gccgtagccg tagtagaagc cgccgacgaa gacgccggtg tcgctgccgc gc

#agggtgtc  23580

cggcacgatg ccggcgtgtt cgagcgcctc ccaggcgatt tcgaggagga tc

#cgctgctg  23640

cgggtcgagt gcggtggcct cgcgcggact gatgccgaag aacgcggcat cg

#aagtcggc  23700

ggcgcccgcg agtgcgccgg cccgcccggt ggcggactcg gcggcggcgt gc

#agcgcggc  23760

cacgtcccag ccgcggtcgg tggggaagtc gccgatcgcg tcgcggccgt cc

#gcgacgag  23820

ctgccacagc tcttccggtg aggtgacgcc gcccggcagt cggcaggcca tg

#ccgacgac  23880

ggcgagcggc tcgttcgccg cggcgcgcag cgcggtgttc tcccggcgga gc

#tgcgcgtt  23940

gtccttgacc gacgtccgca gcgcctcgat caggtcgttc tcggccatcg cc

#tcatccct  24000

tcagcacgtg cgcgatgagc gcgtctgcgt ccatgtcgtc gaacagttcg tc

#gtccggct  24060

ccgcggtcgt ggtgctcgcg ggtgcctgtg ccggtggttc accgccgtcc gg

#ggtcccgt  24120

tgtcgtccgg ggtcccgttg acgtccgggg ccaggagggt cagcagatga cg

#ggtgagcg  24180

cgccggcggc gggatagtcg aagacgagcg tggccggcag cggaatgccg ag

#ggcctcgg  24240

agagccggtt gcgcaggccg agcgcggtga gcgagtcgac cccgaggtcc tt

#gaacgccg  24300

tggtggccgt gaccgccgcc gcgtcggtgt ggcccagcag ggtggcggcg gt

#gtcgcgga  24360

cgacgccgag cagcacctgt tcccgttcct tgtggggcag gtccggcagg cg

#ttccagca  24420

gggagccgcc gtcggtcgcg gagcgccggg tggggcgctg gatcggtcgc ca

#cagcggtg  24480

acgggtcgcc gggcccgggt ggggcggtcg ccacgaccac ggcttccccg gt

#ggcgcacg  24540

cggcgtcgag gaggtcggtc agccggtccg ccgcggcggt gaacgccacg gc

#cggcaggc  24600

cttgtgcccg gcgcaggtcg gccagggcct ggagcggtcc ggccgcctcg cc

#ggacggaa  24660

cggcgagaac gaacgcggtc aggtcgaggt cgcgggtcag gcggtgcagt tc

#ccaggccg  24720

actcggcggt gccgtccgcg tggacgaccg cggtcaccgg ggtttccggc ac

#tgtgcccg  24780

gctcgtaccg gatcacttcg gcgccgtgtc cgccgaggtg tccggcgagt tc

#ctccgaac  24840

cgcccgcgag gaggacggtg tcgccgtacg aggccgcggc cgtggtgggc gc

#ggcgggga  24900

cgaggcgggg cgcttcgagg cgcccgtcgg ccaggcgcag gtgcggttcg tc

#gaggcggg  24960

agagggcggc ggcgcggcgg ggggtgaccg tgtcggtggt ctccacgagc ac

#gagccggc  25020

ccggttccgc ggtgtcgagc agtgcggcga cggcaccggc gacgggcccg gc

#ctcggcgg  25080

acaccaccag cgtggcgccg gcggtcctcg ggtcgtccag tgcggtacgg ac

#ctcgtcgg  25140

gaccggatac cgggacgacg atgacgtcgg gcgtggcgtc gtcgccgagg tc

#ggtgtacc  25200

ggcgggccgt ggtgccgggt gccgccgggg cccggacgcc ggtccaggtg cg

#ccggaaca  25260

gccgcacgtc cccgtccggg cccgtcgtgg cggggggccg ggtgatgagc ga

#gccgatct  25320

gagccaccgg ccgtcccagt tcgtcggcga ggtgcacgcg ggcgccgccc tc

#gccctcgc  25380

cgtggacgaa ggtgacgcgc agtttcgtgg cgccgctggt gtggacacgg ac

#gccggtga  25440

acgcgaacgg caaccgtacc cccgcgttct cggcggccgc gccgatgctg cc

#cgcttgca  25500

gcgcggtgac gagcagcgcc gggtgcagtg tgtagcgggc ggcgtccctg gc

#gagggcgc  25560

cgtcgagggc gacttcggcg cagacggtgt ctccgtggct ccacgcggcg ga

#catgccgc  25620

ggaactcggg gccgaactcg tatcccgcgt cgtcgagtcg ctggtagaag gc

#cgcgacgt  25680

cgaccggttc cgcgtgctcg ggcggccagg gccccggcgt ggtggccggt tc

#ggtggtgg  25740

cgatgccggc gaagccggag gcgtggcggg tccatgtccg gtcgccgtcc gt

#ccgggcgt  25800

ggacgcgcac ggcacggcgt ccggtgtcgt cgggcgcggc gacggtcacg cg

#cacctgga  25860

cggcgccggt ggcgggcagg accagcggtg tctcgacgac cagttcgtcg ag

#caggtcgc  25920

agcctgcctc gtcggcgccg cgtccggcca attccaggaa ggcgggtccg gg

#cagcagta  25980

cggcgccgtc gacggagtga ccggccagcc atgggtgggt ggccagcgag aa

#ccggccgg  26040

tgagcagcac ctcgtcggag tcggggagcg ccaccgacgc ggcgagcagc gg

#gtggtcga  26100

cggcgtcgag tccgaggccg gaagcgtccg tgccggccgc ggtctcgatc ca

#gtagcgct  26160

catggtggaa ggcgtatgtg ggcaggtcgt gtgccgtcgc cgtcgcgggg ac

#gaccgccg  26220

cccagtcgac gggcacgccg gttgtgtgcg cctcggccag cgcggtgagc ag

#ccggtgga  26280

ctcccccgcc gcggcggagc gtggcgacgg tcgcgccgtc gatcgcgggc ag

#cagcacgg  26340

ggtgcgcgct gacctcgacg aacacggtgt cacccggctc gcgggcagcg gt

#cacggccg  26400

tggcgaagcc tacggggtgg cgcatgttgc ggaaccagta ctcgtcgtcg ag

#cggcgcgt  26460

cgatccagcg ttcgtcggcg gtggagaacc acgggatctc gggcgtgcgc ga

#ggtggtgt  26520

ccgcgacgat ccgctggagt tcgtcgtaca gcgggtcgac gaacggggtg tg

#ggtcgggc  26580

agtcgacggc gatgcggcgc acccagacgc cgcgggcctc gtagtcggcg at

#cagcgttt  26640

cgacggcgtc cgggcgcccg gcgacggtcg tggtggtggc gccgttgcgg cc

#cgcgaccc  26700

agacgccgtc gatccgggcg gcatccgcct cgacgtcggc ggccgggagc gc

#gaccgagc  26760

ccatcgcgcc gcgtccggcg agttcgcgca ggagcaggag aacgctgcgc ag

#cgcgacga  26820

ggcgggcacc gtcctccagg gtgagcgctc cggcgacaca ggccgcggcg at

#ctcgccct  26880

gggagtgtcc gatgacggcg tccgggcgta cgcccgcggc ctcccacacg gc

#ggccagcg  26940

acaccatgac ggcccagcag acggggtgca cgacgtcgac gcggcgggtc ac

#ctccgggt  27000

cgtcgagcat ggcgatgggg tcccagcccg tgtgcgggat cagcgcgtcg gc

#gcattggc  27060

gcatcctggc ggcgaacacc ggggaggccg ccatcagttc gacgcccatg cc

#gcgccact  27120

gcggtccttg tccggggaag acgaagacgg tgcgcggctc ggtgagcgcc gt

#gccggtga  27180

cgacgtcgtc gtcgagcagc acggcgcggt gcgggaacgt cgtacgcctg gc

#gagcaggc  27240

ccgcggcgat ggcgcgcggg tcgtggccgg gacgggcggc gaggtgctcg cg

#gagtcggc  27300

ggacctggcc gtcgagggcc gtggcggtcc gcgccgagac gggcagtggt gt

#gagcggcg  27360

tggcgatcag cggctcaccg ggcttcgagg ccgacggctc ctcggccggc gg

#ctccccgg  27420

ccgggtgggc ttccagcagg acgtgggcgt tggtgccgct gacgccgaag ga

#ggacacac  27480

cggcgcgccg cgggcggtcg gtctcgggcc agggccgggc atcggtgagg ag

#ttcgacgg  27540

cgccggccgt ccagtcgacg tgcgaggacg gcgtgtccac gtgcagggtg cg

#cggcaggg  27600

tgccgtgccg catggcgagg accatcttga tgacaccggc gacacccgcg gc

#ggcctgag  27660

tgtggccgat gttggacttc agcgagccca gcagcaccgg ggtgtcgcgc cc

#ctgcccgt  27720

aggtggccag caccgcctgt gcctcgatgg gatcgcccag cctggtgccg gt

#gccgtgcg  27780

cctccacggc gtccacgtcc gccggggtga gcccggcgtt ggccagggcc tg

#ccggatca  27840

cccgctcctg cgagggcccg ttcggcgccg acaacccgtt ggaagcaccg tc

#ctggttga  27900

ccgccgaacc ccggacaacc gccagcacac ggtggccgtt gcgctcggca tc

#ggagagcc  27960

tctcgacgat cagcacaccg gacccctcgg cgaaaccggt gccgtcagcc gc

#atccgcga  28020

acgccttgca gcgcgcgtcg ggcgcgagac cccgctgctg ggagaactcg ac

#gaagccgg  28080

acggcgaggc catcaccgtg acgccgccga ccagggcgag cgagcattcg cc

#ggagcgca  28140

gtgactgccc ggcctggtgc agcgccacca gcgacgacga acacgccgtg tc

#gaccgtga  28200

ccgccggacc ctccagaccg tagaagtacg acagccgacc ggacagcaca ct

#ggtctggg  28260

tgccggtcgc gccgaaaccg cccaggtcgg tgccgagtcc gtacccgtcg ga

#gaaggcgc  28320

ccatgaacac gccggtgtcg cttccgcgca gcgactccgg gaggatcccg gc

#gtgttcca  28380

gcgcctccca cgaggtctcc aggaccagac gctgctgcgg gtccatcgcc ag

#cgcctcac  28440

gcggactgat cccgaagaac gccgcgtcga agtccgccac cccggcgagg aa

#gccaccat  28500

gacgcacggt cgacgtgccc ggatgatccg gatcgggatc gtacagcccg tc

#cacgtccc  28560

aaccacggtc cgtcggaaac gccgtgatcc cgtcaccacc cgactccagc ag

#ccgccaca  28620

agtcctccgg cgacgcgacc ccacccggca gccggcaggc catccccacg at

#cgccaacg  28680

gctcgtcctg ccggacggcc gcggtcgtgg tgcgggtcgg cgatgccgtc cg

#gccggaca  28740

gcgccgcggt gagcttcgcc gcgacggcgc gcggcgtcgg gaagtcgaag ac

#cgcggtgg  28800

cgggcagccg tacgcccgtc gcctcggtga aggcgttgcg cagccggatc gc

#catgagcg  28860

agtcgacgcc gagttccttg aacgtggcgg tcgcctcgac ccgtgcggca cc

#gtcgtggc  28920

cgagtacggc cgcggtgcac tgccggacga cggcgagcac gtccttttcg gc

#gtccgcgg  28980

cggagagccg cgcgatccgg tcggcgaggg tggtggcgcc ggccgcccgg cg

#ccgcggct  29040

cccggcgcgg tgcgcgcagc aggggcgagc tgccgaggcc ggccgggtcg gc

#ggcgacca  29100

gcgccgggtc cgaggaccgc aacgccgcgt cgaacagcgt cagtccgcct tc

#ggcggtca  29160

gcgccgtcac gccgtcgcgg cgcatgcggg cgccggtgcc gaccgtcagc cc

#gctctccg  29220

gttcccacag gccccaggcc acggacaacg cgggcagtcc ggctgcccgg cg

#ctgttcgg  29280

ccagcgcgtc gaggaacgcg ttcgcggccg cgtagttgcc ctgtccgggg ct

#gccgagca  29340

caccggcggc cgacgagtag aggacgaacg cggccagttc cgtgtcctgg gt

#gagttcgt  29400

gcaggtgcca cgcggcgtcc accttcgggc gcagcaccgt ctcgagccgg tc

#gggggtga  29460

gcgcggtgag gacgccgtcg tcgaggacgg ccgcggtgtg cacgacggcc gt

#gagcgggt  29520

gcgccgggtc gatccccgcc agtacggagg cgagttcgtc ccggtcggcg ac

#gtcgcagg  29580

cgatcgccgt gacctcggcg ccgggcacgt cgctcgccgt gccgctgcgc ga

#cagcatca  29640

gcagccggcg cacgccgtgg cgttcgacga ggtggcggct gatgatgccg gc

#cagcgtcc  29700

cggagccacc ggtgacgagc acggtgccgt ccgggtcgag cgccggagcg tc

#acccgccg  29760

ggaccgccgg ggccagacgg cgggcgtaca cctggccgtc acgcagcacc ac

#ctggggct  29820

catcgagcgc ggtggccgct gcgagcagcg gctcggcggt gtccggggcg gc

#gtcgacga  29880

ggacgatccg gccggggtgt tcggcctgcg cggtccgcac cagtccggcg gc

#cgcggccg  29940

acgcgagacc gggcccggtg tggacggcca ggaccgcgtc ggcgtaccgg tc

#gtcggtga  30000

ggaagcgctg cacggcggtc aggacgccgg cgcccagttc gcgggtgtcg tc

#gagcgggg  30060

caccgccgcc gccgtgcgcg gggaggatca ccacgtccgg gaccgtcggg tc

#gtcgaggc  30120

ggccggtcgt cgcggtcgtg ggcggcagct ccgggagctc ggccagcacc gg

#gcgcagca  30180

ggcccggaac ggctcccgtg atcgtcaggg ggcgcctgcg cacggcgccg at

#ggtggcga  30240

cgggcccgcc ggtctcgtcc gcgaggtgta cgccgtcagc ggtgacggcg ac

#gcgtaccg  30300

ccgtggcgcc ggtggcgtgg acgcggacgt cgtcgaacgc gtacggaagg tg

#gtcccctt  30360

ccgcggcgag gcggagtgcg gcgccgagca gcgccgggtg caggccgtac cg

#tccggcgt  30420

cggcgagctg tccgtcggcg agggccactt ccgcccagac ggcgtcgtcg tc

#ggcccaga  30480

cggcgcgcgg gcggggcagc gcgggcccgt ccgtgtaccc ggctcgggcc ag

#acggtcgg  30540

cgatgtcgtc ggggtccacc ggccgggccg tggcgggcgg ccacgtcgac gg

#catctccc  30600

gcacggccgg ggccgtccgc gggtcggggg cgaggattcc gtgcgcgtgc tc

#ggtccact  30660

cccccgccgc gtgccgcgtg tgcacggtga ccgcgcggcg gccgtccgcc cc

#gggcgcgc  30720

tcaccgtgac ggagagcgcg agcgcaccgg accgcggcag cgtgaggggg gt

#gtccacgg  30780

tgaacgtgtc gagggcgccg cagccggctt cgtcgcccgc ccggatcgcc ag

#atccagga  30840

gggccgcggc gggcagcacc gcgaggccgt gcagggagtg cgccagcgga tc

#ggcggcgt  30900

cgacccggcc ggtgagcacc aggtcgccgg tgccgggcag ggtgaccgcc gc

#ggtcagcg  30960

ccgggtgcgc gaccggcgtc tgtccggccg gggccgcgtc gcccgcggtc tg

#ggtgccga  31020

gccagtagcg gacccgctcg aacgggtacg tcggcgggtg cgaggcgcgt gc

#cggcgcgg  31080

ggtcgatgac cttcggccag tcgaccgtga cgccgtcggt gtgcagccgg gc

#gagcgcgg  31140

tcagggcgga tcgcggttcg tcgtcggcgt gcagcatcgg gatgccgtcg ac

#gagtcggg  31200

tcaggctccg gtccgggccg atctccagga gcaccgcccc gtcgtgcgcg gc

#gacctgtt  31260

ccccgaaccg gacggtgtcg cggacctgtc gtacccagta ctccggcgtg gt

#gcaggcgg  31320

cgcccgcggc catcgggatc ctcggctcgt ggtacgtcag gctctccgcg ac

#cttgcgga  31380

actcctcgag catcggctcc atccgcgccg agtggaacgc gtggctggtc cg

#caggcggg  31440

tgaagcggcc gagccgggcc gcgacgtcga gcaccgcctc ctcgtcaccg ga

#gagcacga  31500

tcgacgcggg cccgttgacc gcggcgatct ccacgccgtc ccgcagcagc gg

#cagcgcgt  31560

cccgttccga cgcgatcacg gcggccatcg ccccgccgga cggcagcgcc tg

#catcaggc  31620

gggcccgtgc ggacaccagc ctgcacgcgt cctccaggga ccagacgccg gc

#gacgtacg  31680

cggcggccag ctcgccgatc gaatggccca cgaaggcgtc cgggcgtacg cc

#ccacgcct  31740

cgagctgtgc gccgagtgcg acctggagcg cgaacaccgc gggctgggcg ta

#cccggtgt  31800

cgtggaggtc gagcccggcg ggcacgtcga gggcgtccag cacctcgcgg cg

#agtgcggg  31860

cgaagacgtc gtaggcggcg gccagtccgt cgcccatgcc gggacgttgt ga

#gccctgtc  31920

cggagaagag ccacacgagg cggcggtccg gttctgcggc gccggtgacc gt

#gtcggtgc  31980

cgatcagcgc ggcccggtgc gggaaggccg tgcgggcgag cagggccgcg gc

#caccgcgc  32040

gctcgtcctc ctcgccggtg gcgaggtggg cgcgcaggcg gtgtacctgt gc

#gtcgagtg  32100

cctgcggggt gcgtgccgag agcagcaggg gcagcggtcc ggtgtcgggt gc

#cggggcgg  32160

gttcgggggc cggtcggggg tggctttcga ggatgatgtg agcgttggtg cc

#gctaacgc  32220

cgaaggagga caccccggcg cgccgtgggc ggtcggtttc gggccagggg cg

#ggcgtcgg  32280

tgaggagttc gacggcgccg gccgtccagt cgacgtgcga ggacggcgtg tc

#cacgtgca  32340

gggtgcgcgg cagggtgccg tgccgcatgg cgaggaccat cttgatgaca cc

#ggcgacgc  32400

ccgcggcggc ctgagtgtgg ccgatgttgg acttcagcga gcccagcagc ac

#cggggtgt  32460

cgcgatgctg cccgtaggtg gccagtaccg cctgcgcctc gatggggtcg cc

#cagcctgg  32520

tcccggtgcc atgcgcctcg acagcgtcca catccgccgg ggtgagcccg gc

#gttggcca  32580

gcgcctgccg gatcacccgc tcctgcgacg gcccgttcgg cgccgacaac cc

#gttggaag  32640

caccgtcctg gttgaccgcc gaaccacgca cgaccgccag gacattgtgg cc

#gtgccgct  32700

cggcgtcgga gagcctctcg acgatcagca caccggatcc ctcggcgaaa cc

#ggtgccat  32760

cagccgcatc cgcgaacgcc ttgcagcggc cgtccgggga gaggccccgc tg

#ctgggaga  32820

agtccacgaa gccggacggc gaggccatca ccgtgacgcc gccgaccacg gc

#gagcgagc  32880

actcccccga gcgcagcgac tgcccggcct ggtgcagcgc caccagcgac ga

#cgaacacg  32940

ccgtgtccac cgtgaccgcc ggaccctcca aaccgtagaa gtacgacagc cg

#accggaca  33000

gcacactggt ctgggtgctg gtggcaccga aaccgccgcg gtcggctcca gt

#gccgtacc  33060

cgtagaagta gccgcccatg aacacgccgg tgtcgcttcc gcgcagcgac tc

#cgggagga  33120

tcccggcgtg ttccagcgcc tcccacgagg tctccaggac cagacgctgc tg

#cgggtcca  33180

tcgccagcgc ctcacgcgga ctgatcccga agaacgccgc gtcgaagtcc gc

#caccccgg  33240

cgaggaagcc accatgacgc acggtcgacg tgcccggatg atccggatcg gg

#atcgtaca  33300

gcccgtccac gtcccaacca cggtccgtcg gaaacgccgt gatcccgtca cc

#acccgact  33360

ccagcagccg ccacaagtcc tccggcgacg cgaccccacc cggcagccgg ca

#ggccatcc  33420

ccacgatcgc caacggctcg tcctgccgga cggccgcggt cggggtacgc cg

#ccgggtgg  33480

tggcccgcgc gccggccagt tcgtccaggt gggcggcgag cgcctgcgcc gt

#ggggtggt  33540

cgaagacgag cgtagcgggc agcgtcaggc ccgtcgcgtc ggccagccgg tt

#gcgcagtt  33600

cgacgccggt cagcgagtcg aagcccactt ccctgaacgc gcgcgcgggt gc

#gatggcgt  33660

gggcgtcgcg gtggccgagc accgcggcag cgctggtacg gacgaggtcg ag

#catgtcgc  33720

gcgcggccgg aggtgcggac gtgcgccgga cggccggcac gagggtgcgt ag

#gaccggcg  33780

ggacccggtc ggacgcggcg acggcggcga ggtcgagccg gatcggcacg ag

#cgcgggcc  33840

ggtcggtgtg cagggccgcg tcgaacaggg cgagcccctg tgcggccgtc at

#cggggtca  33900

tgccgttgcg ggcgatgcgg gccaggtcgg tggcggtcag ccgcccgccc at

#cccgtccg  33960

ccgcgtccca cagtccccag gcgagcgaga cggcgggcag cccctggtgg tg

#ccggtggc  34020

gggcgagcgc gtcgaggaac gcgttgccgg tcgcgtagtt ggcctgaccc gc

#gccgccga  34080

acgtggcgga tatggacgag tacaggacga acgcggccag gtcgagatcg cg

#cgtcagct  34140

cgtgcaggtg ccaggcgacg tccgccttga cccgcagcac ggcgtcccac tg

#ctccggcc  34200

gcatggtcgt cacggccgcg tcgtcgacga tcccggccat gtgcacgacg gc

#gcgcagcc  34260

gctgggcgac gtcggcgacg actgcggcca gctcgtcgcg gtcgacgacg tc

#ggcggcca  34320

cgtaccgcac gcggtcgtcc tccggcgtgt cgccgggccg gccgttgcgg ga

#caccacga  34380

cgacctcggc ggcctcgtgc acggtgagca ggtggtccac gaggaggcgg cc

#gagcccgc  34440

cggtgccgcc ggtgacgagg acggtcccgc cggtcagcgg ggaggttccg gt

#ggccgcgg  34500

cgacacggcg cagacgggcc gcacgcgctg tgccgtcggc gacccggacg tg

#cggctcgt  34560

cgccggcggc gagcccggcc gctatggcgg cgggcgtgat ctcgtccgct tc

#gatcaggg  34620

cgacgcggcc gggatgctcc gtctccgccg tccggaccag gccgccgagc gc

#ttcctgcg  34680

cgggatcgcc ggtacgggtg gccacgatga gccgggatcg cgcccagcgc gg

#ctcggcga  34740

gccaggtctg cacggtggtg agcaggtcgc ggcccagctc ccgggtccgg gc

#gccgggcg  34800

aggtgcccgg gtcgccgggt tccacggcca ggaccacgac cggggggtgc tc

#gccgtcgg  34860

gcacgtcggc gaggtacgtc cagtcgggga cgggtgacgc gggcacgggc ac

#ccaggcga  34920

tctcgaacag cgcctcggca tcggggtcgg cggcccgcac ggtcaggctg tc

#gacgtcaa  34980

ggaccggtga gccgtgctcg tccgtggcga cgatgcggac catgtcgggg cc

#gacgcgtt  35040

ccagcagcac gcgcagcgcg gtcgcggcgc gcgcgtggat cctcacgccg ga

#ccaggaga  35100

acgccagccg gcgccgctcc gggtccgtga agaccgtccc gagggcgtgc ag

#ggccgcgt  35160

cgagcagcac ggggtgcagc ccgtaccggg cgtcggtgag ctgttcggcg ag

#gcggaccg  35220

acgcgtaggc gcggccctcc cccgtccaca tcgcggtcat ggcccggaac gc

#gggcccgt  35280

acgagagcgg cagcgcgtcg tagaagccgg tcaggtcggc cgggtcggcg tc

#ggcgggcg  35340

gccagtccac gggctccgcc ggaccgccag tgtccacgct cagcgctccg gt

#cgcactga  35400

gcgcccaggg gcccgtgccg gtacggctgt gcagactcac cgaccgccgt cc

#ggacacct  35460

cggttccgac ggtggcctgg atctccgtgt cgccgtcgcc gtcgaccacc ac

#cggcgcga  35520

cgatggtcag ctccgcgatc tccggcgtgc cgagccgggc tcccgcttcg gc

#gagcagtt  35580

ccacgagcgc cgagccgggc acgatgaccc ggccgtccac ctcgtggtcg gc

#gagccagg  35640

gctgacggcg taccgagaca ccgcggtggc cagcgcgccc tcgccgtcgg gc

#gaggtcga  35700

cccacgagcc gagcagcggg tggccggacg ttcccgccgg ttccgcgtcg at

#ccagtagc  35760

ggtcacggcg gaacgggtac gtgggcagcg gcaccacccg acgcgtcgcg aa

#cgaccagg  35820

tgacgggcac gccccggacc cagagcgcgg cgagcgaccg agtgaagcgg tc

#caggccgc  35880

cctcgcctcg ccgcagtgtg ccggtgacga ccgtatgcgc atgcccggcg ag

#cgtgtcct  35940

ccagtgcggt ggtgagcacg ggatgcgcgc tgacctcgac gaacgcgcgg ta

#tccgcggt  36000

ccgccaggtg gccggtcgcg gcggcgaacc gaacggtgcg gcgcaggttg tc

#gtaccagt  36060

aggcggcgtc cgcgggccgg tccagccacg cctcgtccac ggtggagaag aa

#cgggacgt  36120

ccggcgtgcg cggagtgatg ccggcgagag cgtcgagcag cgcgccgcgg at

#cgtttcga  36180

catgcgcggt gtgcgacgcg tagtcgacgg cgatccggcg ggcgcggggg gt

#ggcggcca  36240

gcagctcctc cacggcgtcg gccgcaccgg cgacaacgat cgacgcgggt cc

#gttgaccg  36300

cggcgacctc caggcgcccg gcccacacgg cggcgtcgaa gtcggcgggc gg

#caccgaga  36360

ccatgccgcc ctgcccggcc agttcggtgg cgacgagtcg gctgcgcacc gc

#gacgacct  36420

tcgcggcgtc gtccagggtg agcaccccgg cgacgcaggc cgcggcgact tc

#gccctggg  36480

agtggccgac gaccgcggcc ggggcgaccc cgtgcgcacg ccacagctcc gc

#cagcgcca  36540

ccatcaccgc gaacgacgcg ggctgcacga catcgacccg gtcgaacgcg gg

#cgctccgg  36600

gccgctgggc gatgacgtcc agcaggtccc atccggtgtg cggggcgagc gc

#cgtggcgc  36660

actcgcggag ccgccgggcg aacacgggct cggtggcgag cagttcggca cc

#catgccgg  36720

cccactggga gccctgcccg gggaacgcga acacgacacg tgtgtcggtg ac

#gtcggcgg  36780

ttcccgtcac ggcccccggc acttcggcac cacgggcgaa cgcctccgcc tc

#tcgggccg  36840

gcacgaccgc ccggtggcgc atggccgtcc gggtggtggc gagcgagtgg cc

#gaccgcgg  36900

ccgcggcgcc agtgagcggg gccagctgtc ccgcgacgtc ccgcagtccc tc

#cggggtcc  36960

gggccgacat cggccagacc acgtcctcgg gcaccggctc ggcttcgggt gc

#ggacacgg  37020

gtgcgggcgc ggcggggggc ccggcctcca ggacgacatg ggcgttggtg cc

#gctgatgc  37080

cgaacgacga gacacccgca cgccgggcgc gcccggtgac cggccacggc tc

#actgcggt  37140

gcagcagccg gatgtcgccg tcccagtcga cgtgccggga cggctcgtcg ac

#gtgcagcg  37200

tgcgcggcag gacgccgtgc cgcatcgcca tgaccatctt gatgacgccg gc

#gacgccgg  37260

ccgcggcctg ggtgtggccg atgttcgact tgagcgagcc gatcagcagc gg

#atgcacgc  37320

gttcgcgccc gtaggccact tgcagggcct gggcctcgac ggggtcgccg ag

#acgggtgc  37380

cggtgccgtg tgcctccacg gcgtcgacgt cacccggcgc caggccggcg tc

#ggcgagcg  37440

cacgctggat gacgcgctgc tgcgcaggcc cgttcggggc ggacagcccg tt

#cgacgcgc  37500

cgtcggagtt gaccgcggag ccgcgcacca gcgccagcac ggggtggccg tg

#gcgggtgg  37560

cgtcggagag ccgctccagc accaggacac cggcgccctc ggcgaagctc gt

#gccgtccg  37620

cggtgtccgc gaaggccttg gcacggccgt cgggggcgag cccgcgctgc cg

#ggagaact  37680

cgacgaaccc ggtcgtcgtc gccatcaccg tgacaccgcc gaccagggcg ag

#cgagcact  37740

cccccgagcg cagcgaccgc gcggcctggt gcagcgccac cagcgacgac ga

#acacgccg  37800

tgtcgacggt gaccgacggg ccctccagac cgaagtagta cgagagccgc cc

#ggagagaa  37860

cgctggtcgg cgtgccggtc gccccgaaac cgcccaggtc cacgcccgcg cc

#gtagccct  37920

gggtgaacgc gcccatgaat acgccggtgt cgctgccgcg gacgctttcg gg

#caggatgc  37980

ccgctcgttc gaacgcctcc cacgacgctt cgaggaccag acgctgctgc gg

#gtccatcg  38040

ccagcgcctc acgcgggctg atcccgaaga acgcggcgtc gaagtcggcg gc

#gccggtga  38100

ggaagccgcc gtgacgcacg gaaaccttgc cgaccgcgtc ggggttcggg tc

#gtagagcg  38160

cggcgaggtc ccagccgcgg tcggcgggga actcggtgat cgcgtccccg cc

#ggagtcga  38220

ccagccgcca caggtcctcc ggtgaccgca cgccaccggg catccggcac gc

#catggcca  38280

cgatcgccag cggctcgttc cccgccaccg tcggtgcggg cactgtcgcc gc

#cggagcgg  38340

caggggccgg ctcaccccgc cgttcctcat ccaggcgggc ggcgagcgcg gc

#cggtgtcg  38400

ggtggtcgaa gacggccgtc gcggagagcc gtacccccgt cgtctcggcg ag

#gctgttgc  38460

gcaaccggac accgctgagc gagtcgatgc cgaggtcctt gaacgccgtc gt

#gggcgtga  38520

tctcggaggc gtcggcgtgg ccgagcacgg cggccgtggc cgcacacacg at

#ggccagca  38580

ggtcacgatc gcggtcgcgg tcgcggtcgc ggttgtcctc cgcacgggcg gc

#gatgcggc  38640

gctcggtccg ctgccggacg ggctcggtgg gaatcgccgc gaccatgaac gg

#cacgtccg  38700

cggcgaggct cgcgtcgatg aagtgggtgc cctcggcctc ggtgagcggc cg

#gaacccgt  38760

cgcgcacccg ctgccggtcg gcgtcgtcaa gttgtccggt gagggtgctg gt

#ggtgtgcc  38820

acatgcccca ggcgatggag gtggcgggtt ggccgagggt gtggcggtgg gt

#ggcgaggg  38880

cgtcgaggaa ggcgttggcg gcggcgtagt ttccttgtcc ggggctgccg ag

#gacggcgg  38940

cggcgctgga gtagaggacg aagtgggtga ggggttggtt ttgggtgagg tg

#gtgcaggt  39000

gccaggcggc gttggctttg gggtggagga cggtggtgag gcggtcgggg gt

#gagggcgt  39060

cgaggatgcc gtcgtcgagg gtggcggcgg tgtggaagac ggcggtgagg gg

#ttggggga  39120

tgtgggcgag ggtggtggcg agttggtggg ggtcgccgac gtcgcagggg ag

#gtgggtgc  39180

cgggggtggt gtcggggggt ggggtgcggg agaggaggta ggtgtggggg tg

#gttcaggt  39240

ggcgggcgag gatgccggcg agggtgccgg agccgccggt gatgatgatg gc

#gtgttcgg  39300

ggttgagggg ggtggtggtg ggtggggtgg tggtgtggag gggggtgagg tg

#gggtcggt  39360

ggagggtgtg gtgggtgagg cggaggtggg ggtggtcgag ggtggcgagt tg

#ggccaggg  39420

ggaggggagt gtgggggtgg tcggtttcga tgaggcggat gcggtggggg tg

#ttcgttct  39480

gggcggtgcg ggtgaggccg gtgacggtgg cgccggcggg gtcggtggtg gt

#gtggacga  39540

tgagggtgtg gtcggtggtg gtgaggtggt gttgcagggc ggtcaggacg cg

#ggtggcgc  39600

gggtgtgggc gcgggtgggt atgtcctcgg ggtcgtcggg gtgggcggcg gt

#gatcagga  39660

cgtgtccctc gggcaggtca ccgtcgtaga ccgcctcggc gaccgcgagc ca

#ctccaacc  39720

ggagcgggtt cggccccgac ggggtgtcgg cccgctccct cagcaccagc ga

#gtccaccg  39780

acacgacagg acggccatcc gggtcggcca cgcgcacggc gacgccggcc tc

#cccccggg  39840

tgagggcgac gcgcaccgcg gcggccccgg tggcgttcag gcgcacgccc gt

#ccaggaga  39900

acggcagctc gatcccgccg cccgcgtcga ggcgcccggc gtgcagggcc gc

#gtcgagca  39960

gtgccggatg cacaccgaaa ccgtccgcct cggcggcctg ctcgtcgggc ag

#cgccacct  40020

cggcatacac ggtgtcacca tcacgccagg cagcccgcaa cccctggaac gc

#cgacccgt  40080

actcataacc ggcatcccgc agttcgtcat agaaccccga gacgtcgacg gc

#cgcggccg  40140

tggccggcgg ccactgcgag aacggctcac cggaagcgtt ggaggtatcc gg

#ggtgtcgg  40200

gggtcagggt gccgctggcg tgccgggtcc agctgcccgt gccctcggta cg

#cgcgtgga  40260

cggtcaccgg ccgccgtccg gcctcatcgg ccccttccac ggtcaccgac ac

#atccaccg  40320

ctgcggtcac cggcaccacg agcggggatt cgatgaccag ttcatccacc ac

#cccgcaac  40380

cggtctcgtc accggcccgg atgaccagct ccacaaacgc cgtacccggc ag

#cagaaccg  40440

tgccccgcac cgcgtgatca gccagccagg gatgcgtacg caatgagatc cg

#gccggtga  40500

gaacaacacc accaccgtcg tcggcgggca gtgctgtgac ggcggccagc at

#cggatgcg  40560

ccgccccggt cagcccggcc gcggacaggt cggtggcacc ggccgcctcc ag

#ccagtacc  40620

gcctgtgctc gaacgcgtag gtgggcagat ccagcagccg ccccggcacc gg

#ttcgacca  40680

ccgtgcccca gtccaccccc gcacccagag tccacgcctg cgccaacgcc cc

#cagccacc  40740

gctcccagcc accgtcacca gtccgcaacg acgccaccgt gcgggcctgt tc

#catcgccg  40800

gcagcagcac cggatgggca ctgcactcca cgaacaccga cccgtccagc tc

#cgccaccg  40860

ccgcatccag cgcgacaggg cgacgcaggt tccggtacca gtacccctca tc

#caccggct  40920

cggtcaccca ggcgctgtcc acggtcgacc accacgccac cgacccggtc cc

#gccggaaa  40980

ttcccttcag tacctcagcg agttcgtcct cgatggcctc cacgtgaggc gt

#gtgggagg  41040

cgtagtcgac cgcgatacga cgcacccgca ccccatcagc ctcataccgc gc

#caccacct  41100

cctccaccgc cgacgggtcc cccgccacca ccgtcgaagc cggaccatta cg

#cgccgcga  41160

tccacacacc ctcgaccaga cccacctcac cggccggcaa cgccaccgaa gc

#catcgccc  41220

cccggccggc cagccgcgcc gcgatcaccc gactgcgcaa cgccaccacg cg

#ggcggcgt  41280

cctccaggct gagggctccg gccacacacg ccgccgcgat ctccccctgc ga

#gtgtccga  41340

ccacagcgtc cggcacgacc ccatgcgcct gccacagcgc ggccaggctc ac

#cgcgaccg  41400

cccagctggc cggctggacc acctccaccc gctccgccac atccgaccgc ga

#caacatct  41460

cccgcacatc ccagcccgtg tgcggcaaca acgcccgcgc acactcctcc at

#acgagccg  41520

cgaacaccgc ggaacggtcc atgagttcca cgcccatgcc cacccactgg gc

#accctgcc  41580

cggggaagac gaacaccgta cgcggctgat ccaccgccac acccatcacc cg

#ggcatcac  41640

ccagcagcac cgcacggtga ccgaagacag cacgctcacg caccaacccc tg

#cgcgaccg  41700

cggccacatc caccccaccc ccgcgcagat acccctccag ccgctccacc tg

#cccccgca  41760

gactcacctc accacgagcc gacaccggca acggcaccaa cccatcacca cc

#cgactcca  41820

cacgcgacgg cccaggaaca ccctccagga tcacgtgcgc gttcgtaccg ct

#caccccga  41880

acgacgacac acccgcatgc ggtgcccgat ccgactcggg ccacggcctc gc

#ctcggtga  41940

gcagctccac cgcaccggcc gaccagtcca catgcgacga cggctcgtcc ac

#gtgcagcg  42000

tcttcggcgc gatcccatgc cgcatcgcca tgaccatctt gatgacaccg gc

#gacacccg  42060

cagccgcctg cgcatgaccg atgttcgact tgaccgaacc gaggtagagc gg

#cgtgtcgc  42120

ggtcctgccc gtaggccgcg aggacggcct gcgcctcgat cgggtcgccc ag

#ccgcgtgc  42180

cggtgccgtg cgcctccacc acgtccacat cggcggcgcg cagtccggcg tt

#gaccaacg  42240

cctgccggat cacgcgctgc tgggcgacgc cgttgggggc ggacagtccg tt

#ggaggcac  42300

cgtcctggtt caccgccgag ccgcggacga ccgcgagaac ggtgtgcccg tt

#gcgctcgg  42360

cgtcggagag ccgctccagc acgagaacgc cgacgccctc ggcgaagccg gt

#cccgtccg  42420

ccgcgtcggc gaacgccttg caccgtccgt ccggggagag tccgcgctgc cg

#ggagaact  42480

ccacgagctc tgcggtgttc gccatgacgg tgacaccgcc gaccagcgcc ag

#ggagcact  42540

ccccggcccg cagtgcctgt gccgcctggt gcagggcgac cagcgacgac ga

#gcacgccg  42600

tgtcgaccgt gaccgccggg ccctgaagtc cgtacacgta cgagaggcgc cc

#ggacagga  42660

cgctcgtctg cgtcgccgtg acaccgagcc cgcccaggtc ccggccgacg cc

#gtagccct  42720

ggttgaacgc gcccatgaac acgccggtgt cgctctcccg gagcctgtcc gg

#cacgatgc  42780

cggcgttctc gaacgcctcc caggaggtct ccaggatcag gcgctgctgg gg

#gtccatcg  42840

ccagcgcctc gttcggactg atgccgaaga acgcggcgtc gaacccggcg cc

#ggccagga  42900

atccgccgtg gcgtgtcgtg gagcggccgg ccgcgtccgg gtccgggtcg ta

#cagcgcgt  42960

cgacgtccca gccccggtcg gtggggaact cggtgatcgc ctcggtaccg gc

#ggcgacga  43020

gccgccacag gtcctccggc gaggcgaccc cgccgggcag tcggcacgcc at

#gccgacga  43080

tcgcgacggg gtcgccggag ccgagggtct gggcggtcgc gggtgccgct gt

#cgcggagc  43140

cggcgaggtg ggcggcgaac gcacgcggag tggggtggtc gaacgcggtt ga

#cgcgggca  43200

cccgcagacc cgtccgcgcg gcgacggtgt tggtgaactc gacggtggtg ag

#cgagtcga  43260

ggccgttctc gcggaacgtg cggtccgggg agcagtgtcc ggcgcccggc ag

#gcccagga  43320

cggtggcgac gctgtcgcgg accaggtcga gcagtacgtc ctcccggccc gc

#acgggccg  43380

cggcgaggcg gttcgcccac tcctgttccg tggcgtcggg ctcggccggt cc

#ggtcagtg  43440

cggtgaggat cggcggcgtg gcgcccgcca tcgtcgcggc ccgcgccccg gc

#ggaaccgg  43500

tccgggccac gatgtacgag ccgccgcccg cgatggcctt ctcgatcagg tc

#gccggtga  43560

gcgccggccg ttcgatgccg ggcagcgcgc ggacggtgac ggtggggagt cc

#ctccgcgg  43620

cccgtggccg ggtgtgggcg tcggcgccgg ccgggccgtc gagcaggacg tg

#cacgagcg  43680

cgccggggtt cgcggcttcc tcggctgcgg tggtcacgtg ggtgaggccg gt

#ctcgtcgc  43740

ggagcaggcc ggcgacggtg tcggcgtcct ccccggtgac caggaccggc gc

#gtccgggc  43800

cgatcggagg cggcacggtg aggaccatct tgccggtgtg ccgggcgtgg ct

#catccacg  43860

cgaacgcgtc ccgcgcacgg cggatgtccc acggctgcac cggcagcggg ca

#cagctcac  43920

cgcggtcgaa caggtcgagg agcagttcga ggatctcccg caggcgcgcg gg

#atccacgt  43980

cggccaggtc gaacggctgc tgggcggcgt ggcggatgtc ggtcttgccc at

#ctcgacga  44040

accggccgcc cggtgcgagc aggccgatgg acgcgtcgag gagttcaccg gt

#gagcgagt  44100

tgagcacgac gtcgaccggc gggaaggtgt cggcgaacgc ggcgctgcgg ga

#gttcgcca  44160

catggtcggt gtcgaagccg tcggcgtgca gcaggtgttg tttggcggga ct

#ggcggtgg  44220

cgtacacctc ggcgccgagg tggcgggcga tccgggtcgc cgccatgccg ac

#accgcccg  44280

tcgcggcgtg gaccaggacc ttctggccgg gtcgcagctc gcccgcgtcg ac

#gaggccgt  44340

accaggcggt ggcgaacacg atgggcacgg acgcggcgat ggggaacgac ca

#tccccgtg  44400

ggatccgtgc gaccagccgc cggtccgcga ccacgctgcg ccggaacgcg tc

#ctgcacga  44460

gaccgaacac gcggtcgccg ggggccaggt cgtcgacgcc gggtccgact tc

#ggtcacga  44520

tgcccgcggc ctccccgccc atctcgccct cgcccgggta ggtgccgagc gc

#gatcagca  44580

cgtcgcggaa gttcagcccc gcggcgcgga cgtcgatgcg gacctcgccg gc

#ggccaggg  44640

gcgcggcggg acgtcgagcg gggcgacgac gaggtcgcgg agcgttccgg ag

#gcgggcgg  44700

gcgcagcgcc cactggcgcg gtcggcaggg gggtggtgtc cgcgcgtacc ag

#ccggggca  44760

cgtaggccac gccggcccgc agcgcgatct ggggttcgcc gagcgaggcc gc

#ggcgggga  44820

cgaggtcgtc atcgccgtcc gtgtccacca gcacgaacga tccgggttcg gc

#ggcctggc  44880

ggcgcagcgc ctcgtcccag agccgggcct ggtccgcgtc cgggatctcg gc

#cgggccga  44940

cgcccaccgc gcggcgggtg acgaccgtcc ggcggggtga cggggtgccg gg

#caggtcgc  45000

gccgctccca gaccagttcg cacagcgtgg cctcgccact gccggtggcg ac

#cagatggg  45060

ccggcagccc cgcgagccgc gcgcgctgga ccttgcccga cgcggtgcgg gg

#gatcgtgg  45120

tgacgtgcca gatctcgtcg ggcaccttga agtaggcgag ccggcggcgg ca

#ctcggcga  45180

ggatcgcctc ggcggggacg cgggggccgt cggaaacgac gtagagcacg gg

#tatgtcgc  45240

cgaggacggg gtgcgggcgg cccgccgcgg cggcgtcccg gacaccggcc ac

#ctcctggg  45300

cgacggtctc gatctcccgg gggtggatgt tctccccgcc gcggatgatc ag

#ctccttga  45360

cccggccggt gatcgtcacg tgtccggtct cggcctgacg tgcgaggtcc cc

#ggtgcggt  45420

accagccgtc cacgagcacc tgggcggtcg cctccggctg ggcgtggtag cc

#gagcatga  45480

ggctcggccc gctcgcccac agctcgccct cctcgccggg tgccacgtcg gc

#gccggaca  45540

ccgggtcgac gaaccgcagc gacaggcccg gcacgggcag cccgcacgag cc

#gggaaccc  45600

gcgcatcctc cagggtgttg gcggtgagcg agccggtcgt ctcggtgcag cc

#gtacgtgt  45660

cgagcagggg cacgccgaac gtcgcctcga aatccctggt gagcgacgcc gg

#cgaggtgg  45720

atccggcgac cagcgccacg cgcagcgcgc gagcccgcgg ctcgccggac ac

#ggcgccga  45780

ggaggtagcg gtacatcgtc ggcacgccga cgagcacggt gctggagtgt tc

#ggccaggg  45840

cgtcgaggac gtcacgcgcg acgaagccgc ccaggatacg ggcggacgcg cc

#gaccgtga  45900

ggacggcgag caggcagagg tggtggccga ggctgtggaa cagcggggcg gg

#ccagagca  45960

gttcgtcgtc ctcggtcagc cgccaggacg gcacgtcgca gtgcatcgcg ga

#ccacaggc  46020

cgctgcgctg tgcggaaacc acgcccttgg gacggccggt ggtgccggag gt

#gtagagca  46080

tccaggcggg ttcgtccagg ccgaggtcgt cgcggggcgg gcacggcggc tc

#ggtcccgg  46140

cgaggtcctc gtaggagacg cagtccggtg cccggcgccc gacgagcacg ac

#ggtggcgt  46200

cggtgccggt gcggcgcacc tggtcgaggt gggtttcgtc ggtgaccagc ac

#ggtcgcgc  46260

cggagtccgt caggaagtgg gcgagttcgg cgtcggcggc gtccgggttg ag

#cgggacgg  46320

cgacggcggc ggcgcgggcg gcggcgaggt agacctcgat ggtctcgatc cg

#gttgccga  46380

gcagcatcgc gacccggtcg ccgcggtcga cgccggacgc ggcgaggtgt cc

#ggcgagcc  46440

ggccggcccg gagccggagt tgcgtgtacg tcacggcgcg ttgggaatcc gt

#gtaggcga  46500

tccggtcgcc gcgtcgctcg gcatggatgc ggagcaattc gtgcaacggc cg

#gattggtt  46560

ccacacgcgc catggaaaca cctttctctc gaccaaccgc acaacagcac gg

#aaccggcc  46620

acgagtagac gccggcgacg ctagcagcgt tttccggacc gccaccccct ga

#agatcccc  46680

ctaccgtggc cggcctcccc ggacgctcat ctagggggtt gcacgcatac cg

#ccgtgcgt  46740

aattgccttc ctgatgaccg atgccggacg ccagggaagg gtggaggcgt tg

#tccatatc  46800

tgtcacggcg ccgtattgcc gcttcgagaa gaccggatca ccggacctcg ag

#ggtgacga  46860

gacggtgctc ggcctgatcg agcacggcac cggccacacc gacgtgtcgc tg

#gtggacgg  46920

tgctccccgg accgccgtgc acaccacgac ccgtgacgac gaggcgttca cc

#gaggtctg  46980

gcacgcacag cgccctgtcg agtccggcat ggacaacggc atcgcctggg cc

#cgcaccga  47040

cgcgtacctg ttcggtgtcg tgcgcaccgg cgagagcggc aggtacgccg at

#gccaccgc  47100

ggccctctac acgaacgtct tccagctcac ccggtcgctg gggtatcccc tg

#ctcgcccg  47160

gacctggaac tacgtcagcg gtatcaacac gacgaacgcg gacgggctgg ag

#gtgtaccg  47220

ggacttctgc gtgggccgcg cccaggcgct cgacgagggc gggatcgacc cg

#gccaccat  47280

gcccgcggcc accggtatcg gcgcccacgg gggcggcatc acctgcgtgt tc

#ctcgccgc  47340

ccggggcgga gtgcggatca acatcgagaa ccccgccgtc ctcacggccc ac

#cactaccc  47400

gacgacgtac ggtccgcggc ccccggtctt cgcacgggcc acctggctgg gc

#ccgccgga  47460

ggggggccgg ctgttcatct ccgcgacggc cggcatcctc ggacaccgaa cg

#gtgcacca  47520

cggtgatgtg accggccagt gcgaggtcgc cctcgacaac atggcccggg tc

#atcggcgc  47580

ggagaacctg cggcgccacg gcgtccagcg ggggcacgtc ctcgccgacg tg

#gaccacct  47640

caaggtctac gtccgccgcc gcgaggatct cgatacggtc cgccgggtct gc

#gccgcacg  47700

cctgtcgagc accgcggccg tcgccctttt gcacaccgac atagcccgcg ag

#gatctgct  47760

cgtcgaaatc gaaggcatgg tggcgtgaca atacccggta aaaggcccgc ga

#cgctgcgc  47820

ctcggcggat ccgcgaagag aaagaagagc gtcaccgcac agcgcggcag cc

#cggtcctt  47880

tcgtccttcg cacagcggcg gatctggttt ctccagcaat tggacccgga ga

#gcaacgcc  47940

tataatctcc cgctcgtgca acgcctgcgc ggtctattgg acgcgccggc cc

#tggagcgt  48000

gcgctggcgc tcgtcgtcgc gcgccacgag gcgttgcgga cggtgttcga ca

#ccgccgac  48060

ggcgagcccc tccagcgggt gcttcccgcc ccggaacacc tcctgcgcca cg

#cgcgggcg  48120

ggcagcgagg aggacgccgc ccggctcgtc cgcgacgaga tcgccgcgcc gt

#tcgacctc  48180

gccaccgggc cgttgatcag ggccctgctg atccgcctcg gtgacgacga cc

#acgttctc  48240

gcggtgaccg tgcaccatgt cgccggcgac ggctggtcgt tcgggctcct cc

#aacatgaa  48300

ctcgcagccc actacacggc gctgcgcgac actgcccgcc ctgccgaact gc

#cgccgttg  48360

ccggtgcagt acgccgactt cgccgcctgg gagcggcgcg aactcaccgg cg

#ccggactg  48420

gacaggcgtc tggcctactg gcgcgagcaa ctccggggcg ccccggcgcg gc

#tcgccctc  48480

cccaccgacc gtccccgccc gccggtcgcc gacgcggacg cgggcatggc cg

#agtggcgg  48540

ccgccggccg cgctggccac cgcggtcctc acgctcgcgc gcgactccgg tg

#cgtccgtg  48600

ttcatgaccc tgctggcggc cttccaagcg gtcctcgccc ggcaggcggg ca

#cgcgggac  48660

gtgctggtcg gcacgcccgt ggcgaaccgt acgcgggcgg cgtacgaggg cc

#tgatcggc  48720

atgttcgtca acacgctcgc gctgcgcggc gacctctcgg gcgatccgtc gt

#tccgggaa  48780

ctcctcgacc gctgccgggc cacgaccacg gacgcgttcg cccacgccga cc

#tgccgttc  48840

gagaacgtca tcgaactcgt cgcaccggaa cgcgacctgt cggtcaaccc gg

#tcgtccag  48900

gtgctgttgc aggtgctgcg gcgcgacgcg gcgacggccg cgctgcccgg ca

#tcgcggcc  48960

gaaccgttcc gcaccggacg ctggttcacc cgcttcgacc tcgaattcca tg

#tgtacgag  49020

gagccgggtg gcgcgctgac cggcgaactg ctctacagcc gtgcgctgtt cg

#acgagcca  49080

cggatcacgg ggttgctgga ggagttcacg gcggtgcttc aggcggtcac cg

#ccgacccg  49140

gacgtacggc tgtcgcggct gccggccggc gacgcgacgg cggcagcgcc cg

#tggtgccc  49200

tcgaacgaca cggcgcggga cctgcccgtc gacacgctgc cgggcctgct gg

#cccggtac  49260

gccgcacgca cccccggcgc cgtggccgtc accgacccgc acatctccct ca

#cctacgcg  49320

cagctggacc ggcgggcgaa ccgcctcgcg cacctgctcc gcgcgcgcgg ca

#ccgccacc  49380

ggcgacctgg tcgggatctg cgccgatcgc ggcgccgacc tgatcgtcgg ca

#tcgtgggg  49440

atcctcaagg cgggcgccgc ttatgtgccg ctggaccccg aacatcctcc gg

#agcgcacg  49500

gcgttcgtgc tggccgacgc gcagctgacc acggtggtgg cgcacgaggt ct

#accgttcc  49560

cggttccccg atgtgccgca cgtggtggcg ttggacgacc cggagctgga cc

#ggcagccg  49620

gacgacacgg cgccggacgt cgagctggac cgggacagcc tcgcctacgc ga

#tctacacg  49680

tccgggtcga ccggcaggcc gaaggccgtg ctcatgccgg gtgtcagcgc cg

#tcaacctg  49740

ctgctctggc aggagcgcac gatgggccgc gagccggcca gccgcaccgt cc

#agttcgtg  49800

acgcccacgt tcgactactc ggtgcaggag atcttttccg cgctgctggg cg

#gcacgctc  49860

gtcatcccgc cggacgaggt gcggttcgac ccgccgggac tcgcccggtg ga

#tggacgaa  49920

caggcgatta cccggatcta cgcgccgacg gccgtactgc gcgcgctgat cg

#agcacgtc  49980

gatccgcaca gcgaccagct cgccgccctg cggcacctgt gccagggcgg cg

#aggcgctg  50040

atcctcgacg cgcggttgcg cgagctgtgc cggcaccggc cccacctgcg cg

#tgcacaat  50100

cactacggtc cggccgaaag ccagctcatc accgggtaca cgctgcccgc cg

#accccgac  50160

gcgtggcccg ccaccgcacc gatcggcccg ccgatcgaca acacccgcat cc

#atctgctc  50220

gacgaggcga tgcggccggt tccggacggt atgccggggc agctctgcgt cg

#ccggcgtc  50280

ggcctcgccc gtgggtacct ggcccgtccc gagctgaccg ccgagcgctg gg

#tgccggga  50340

gatgcggtcg gcgaggagcg catgtacctc accggcgacc tggcccgccg cg

#cgcccgac  50400

ggcgacctgg aattcctcgg ccggatcgac gaccaggtca agatccgcgg ca

#tccgcgtc  50460

gaaccgggtg agatcgagag cctgctcgcc gaggacgccc gcgtcacgca gg

#cggcggtg  50520

tccgtgcgcg aggaccggcg gggcgagaag ttcctggccg cgtacgtcgt ac

#cggtggcc  50580

ggccggcacg gcgacgactt cgccgcgtcg ctgcgcgcgg gactggccgc cc

#ggctgccc  50640

gccgcgctcg tgccctccgc cgtcgtcctg gtggagcgac tgccgaggac ca

#cgagcggc  50700

aaggtggacc ggcgcgcgct gcccgacccg gagccgggcc cggcgtcgac cg

#gggcggtt  50760

acgccccgca ccgatgccga gcggacggtg tgccggatct tccaggaggt gc

#tcgacgtc  50820

ccgcgggtcg gtgccgacga cgacttcttc acgctcggcg ggcactccct gc

#tcgccacc  50880

cgggtcgtct cccgcatccg cgccgagctg ggtgccgatg tcccgctgcg ta

#cgctcttc  50940

gacgggcgga cgcccgccgc gctcgcccgt gcggcggacg aggccggccc gg

#ccgccctg  51000

cccccgatcg cgccctccgc ggagaacggg ccggcccccc tcaccgcggc ac

#aggaacag  51060

atgctgcact cgcacggctc gctgctcgcc gcgccctcct acacggtcgc cc

#cgtacggg  51120

ttccggctgc gcgggccact cgaccgcgaa gcgctcgacg cggcactgac cc

#ggatcgcc  51180

gcgcgccacg agccgctgcg gaccgggttc cgcgatcggg aacaggtcgt cc

#ggccgccc  51240

gctccggtgc gcgccgaggt ggttccggtg ccggtcggcg acgtcgacgc cg

#cggtccgg  51300

gtcgcccacc gggagctgac ccggccgttc gacctcgtga acgggtcgtt gc

#tgcgtgcc  51360

gtgctgctgc cgctgggcgc cgaggatcac gtgctgctgc tgatgctgca cc

#acctcgcc  51420

ggtgacggat ggtccttcga cctcctggtc cgggagttgt cggggacgca ac

#cggacctt  51480

ccggtgtcct acacggacgt ggcccggtgg gaacggagtc cggccgtgat cg

#cggccagg  51540

gagaacgacc gggcctactg gcgccggcgg ctggggggcg ccaccgcgcc gg

#agctgccc  51600

gcggtccggc ccggcggggc accgaccggg cgggcgttcc tgtggacgct ca

#aggacacc  51660

gccgtcctgg cggcacgccg ggtcgcggac gcccacgacg cgacgttgca cg

#aaaccgtg  51720

ctcggcgcct tcgccctggt cgtggcggag accgccgaca ccgacgacgt gc

#tcgtcgcg  51780

acgccgttcg cggaccgggg gtacgccggg accgaccacc tcatcggctt ct

#tcgcgaag  51840

gtcctcgcgc tgcgcctcga cctcggcggc acgccgtcgt tccccgaggt gc

#tgcgccgg  51900

gtgcacaccg cgatggtggg cgcgcacgcc caccaggcgg tgccctactc cg

#cgctgcgc  51960

gccgaggacc ccgcgctgcc gccggccccc gtgtcgttcc agctcatcag cg

#cgctcagc  52020

gcggaactgc ggctgcccgg catgcacacc gagccgttcc ccgtcgtcgc cg

#agaccgtc  52080

gacgagatga ccggcgaact gtcgatcaac ctcttcgacg acggtcgcac cg

#tctccggc  52140

gcggtggtcc acgatgccgc gctgctcgac cgtgccaccg tcgacgattt gc

#tcacccgg  52200

gtggaggcga cgctgcgtgc cgccgcgggc gacctcaccg tacgcgtcac cg

#gttacgtg  52260

gaaagcgagt agcc atg ccc gag cag gac aag aca g

#tc gag tac ctt cgc    52310

Met P

#ro Glu Gln Asp Lys Thr Val Glu Tyr Leu Ar

#g

1

#              5

#               10

tgg gcg acc gcg gaa ctc cag aag acc cgt gc

#g gaa ctc gcc gcg cac    52358

Trp Ala Thr Ala Glu Leu Gln Lys Thr Arg Al

#a Glu Leu Ala Ala His

15

#         20

#         25

agc gag ccg ttg gcg atc gtg ggg atg gcc tg

#c cgg ctg ccc ggc ggg    52406

Ser Glu Pro Leu Ala Ile Val Gly Met Ala Cy

#s Arg Leu Pro Gly Gly

30

#     35

#     40

gtc gcg tcg ccg gag gac ctg tgg cag ttg ct

#g gag tcc ggt ggc gac    52454

Val Ala Ser Pro Glu Asp Leu Trp Gln Leu Le

#u Glu Ser Gly Gly Asp

45

# 50

# 55

# 60

ggc atc acc gcg ttc ccc acg gac cgg ggc tg

#g gag acc acc gcc gac    52502

Gly Ile Thr Ala Phe Pro Thr Asp Arg Gly Tr

#p Glu Thr Thr Ala Asp

65

#                 70

#                 75

ggt cgc ggc ggc ttc ctc acc ggg gcg gcc gg

#c ttc gac gcg gcg ttc    52550

Gly Arg Gly Gly Phe Leu Thr Gly Ala Ala Gl

#y Phe Asp Ala Ala Phe

80

#             85

#             90

ttc ggc atc agc ccg cgc gag gcg ctg gcg at

#g gac ccg cag cag cgc    52598

Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Me

#t Asp Pro Gln Gln Arg

95

#        100

#        105

ctg gcc ctg gag acc tcg tgg gag gcg ttc ga

#g cac gcg ggc atc gat    52646

Leu Ala Leu Glu Thr Ser Trp Glu Ala Phe Gl

#u His Ala Gly Ile Asp

110

#   115

#   120

ccg cag acg ctg cgg ggc agt gac acg ggg gt

#g ttc ctc ggc gcg ttc    52694

Pro Gln Thr Leu Arg Gly Ser Asp Thr Gly Va

#l Phe Leu Gly Ala Phe

125                 1

#30                 1

#35                 1

#40

ttc cag ggg tac ggc atc ggc gcc gac ttc ga

#c ggt tac ggc acc acg    52742

Phe Gln Gly Tyr Gly Ile Gly Ala Asp Phe As

#p Gly Tyr Gly Thr Thr

145

#               150

#               155

agc att cac acg agc gtg ctc tcc ggc cgc ct

#c gcg tac ttc tac ggt    52790

Ser Ile His Thr Ser Val Leu Ser Gly Arg Le

#u Ala Tyr Phe Tyr Gly

160

#           165

#           170

ctg gag ggt ccg gcg gtc acg gtc gac acg gc

#g tgt tcg tcg tcg ctg    52838

Leu Glu Gly Pro Ala Val Thr Val Asp Thr Al

#a Cys Ser Ser Ser Leu

175

#       180

#       185

gtg gcg ctg cac cag gcc ggg cag tcg ctg cg

#c tcc ggc gaa tgc tcg    52886

Val Ala Leu His Gln Ala Gly Gln Ser Leu Ar

#g Ser Gly Glu Cys Ser

190

#   195

#   200

ctc gcc ctg gtc ggc ggc gtc acg gtg atg gc

#c tcg ccg gcg ggg ttc    52934

Leu Ala Leu Val Gly Gly Val Thr Val Met Al

#a Ser Pro Ala Gly Phe

205                 2

#10                 2

#15                 2

#20

gcg gac ttc tcc gag cag ggc ggc ctg gcc cc

#c gac gcg cgc tgc aag    52982

Ala Asp Phe Ser Glu Gln Gly Gly Leu Ala Pr

#o Asp Ala Arg Cys Lys

225

#               230

#               235

gcc ttc gcg gaa gcg gct gac ggc acc ggt tt

#c gcc gag ggg tcc ggc    53030

Ala Phe Ala Glu Ala Ala Asp Gly Thr Gly Ph

#e Ala Glu Gly Ser Gly

240

#           245

#           250

gtc ctg atc gtc gag aag ctc tcc gac gcc ga

#g cgc aac ggc cac cgc    53078

Val Leu Ile Val Glu Lys Leu Ser Asp Ala Gl

#u Arg Asn Gly His Arg

255

#       260

#       265

gtg ctg gcg gtc gtc cgg ggt tcc gcc gtc aa

#c cag gac ggt gcc tcc    53126

Val Leu Ala Val Val Arg Gly Ser Ala Val As

#n Gln Asp Gly Ala Ser

270

#   275

#   280

aac ggg ctg tcc gcg ccg aac ggg ccg tcg ca

#g gag cgg gtg atc cgg    53174

Asn Gly Leu Ser Ala Pro Asn Gly Pro Ser Gl

#n Glu Arg Val Ile Arg

285                 2

#90                 2

#95                 3

#00

cag gcc ctg gcc aac gcc gga ctc acc ccg gc

#g gac gtg gac gcc gtc    53222

Gln Ala Leu Ala Asn Ala Gly Leu Thr Pro Al

#a Asp Val Asp Ala Val

305

#               310

#               315

gag gcc cac ggc acc ggc acc agg ctg ggc ga

#c ccc atc gag gca cag    53270

Glu Ala His Gly Thr Gly Thr Arg Leu Gly As

#p Pro Ile Glu Ala Gln

320

#           325

#           330

gcc gtg ctg gcc acc tac ggg cag ggg cgc ga

#c acc cct gtg ctg ctg    53318

Ala Val Leu Ala Thr Tyr Gly Gln Gly Arg As

#p Thr Pro Val Leu Leu

335

#       340

#       345

ggc tcg ctg aag tcc aac atc ggc cac acc ca

#g gcc gcc gcg ggc gtc    53366

Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gl

#n Ala Ala Ala Gly Val

350

#   355

#   360

gcc ggt gtc atc aag atg gtc ctc gcc atg cg

#g cac ggc acc ctg ccc    53414

Ala Gly Val Ile Lys Met Val Leu Ala Met Ar

#g His Gly Thr Leu Pro

365                 3

#70                 3

#75                 3

#80

cgc acc ctg cac gtg gac acg ccg tcc tcg ca

#c gtc gac tgg acg gcc    53462

Arg Thr Leu His Val Asp Thr Pro Ser Ser Hi

#s Val Asp Trp Thr Ala

385

#               390

#               395

ggc gcc gtc gaa ctc ctc acc gac gcc cgg cc

#c tgg ccc gaa acc gac    53510

Gly Ala Val Glu Leu Leu Thr Asp Ala Arg Pr

#o Trp Pro Glu Thr Asp

400

#           405

#           410

cgc cca cgg cgc gcc ggt gtc tcc tcc ttc gg

#c gtc agc ggc acc aac    53558

Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gl

#y Val Ser Gly Thr Asn

415

#       420

#       425

gcc cac atc atc ctc gaa agc cac ccc cga cc

#g gcc ccc gaa ccc gcc    53606

Ala His Ile Ile Leu Glu Ser His Pro Arg Pr

#o Ala Pro Glu Pro Ala

430

#   435

#   440

ccg gca ccc gac acc gga ccg ctg ccg ctg ct

#g ctc tcg gcc cgc acc    53654

Pro Ala Pro Asp Thr Gly Pro Leu Pro Leu Le

#u Leu Ser Ala Arg Thr

445                 4

#50                 4

#55                 4

#60

ccg cag gca ctc gac gca cag gta cac cgc ct

#g cgc gcg ttc ctc gac    53702

Pro Gln Ala Leu Asp Ala Gln Val His Arg Le

#u Arg Ala Phe Leu Asp

465

#               470

#               475

gac aac ccc ggc gcg gac cgg gtc gcc gtc gc

#g cag aca ctc gcc cgg    53750

Asp Asn Pro Gly Ala Asp Arg Val Ala Val Al

#a Gln Thr Leu Ala Arg

480

#           485

#           490

cgc acc cag ttc gag cac cgc gcc gtg ctg ct

#c ggc gac acg ctc atc    53798

Arg Thr Gln Phe Glu His Arg Ala Val Leu Le

#u Gly Asp Thr Leu Ile

495

#       500

#       505

acc gtg agc ccg aac gcc ggc cgc gga ccg gt

#g gtc ttc gtc tac tcg    53846

Thr Val Ser Pro Asn Ala Gly Arg Gly Pro Va

#l Val Phe Val Tyr Ser

510

#   515

#   520

ggg caa agc acg ctg cac ccg cac acc ggg cg

#g caa ctc gcg tcc acc    53894

Gly Gln Ser Thr Leu His Pro His Thr Gly Ar

#g Gln Leu Ala Ser Thr

525                 5

#30                 5

#35                 5

#40

tac ccc gtg ttc gcc gaa gcg tgg cgc gag gc

#c ctc gac cac ctc gac    53942

Tyr Pro Val Phe Ala Glu Ala Trp Arg Glu Al

#a Leu Asp His Leu Asp

545

#               550

#               555

ccc acc cag ggc ccg gcc acg cac ttc gcc ca

#c cag acc gcg ctc acc    53990

Pro Thr Gln Gly Pro Ala Thr His Phe Ala Hi

#s Gln Thr Ala Leu Thr

560

#           565

#           570

gcg ctc ctg cgg tcc tgg ggc atc acc ccg ca

#c gcg gtc atc ggc cac    54038

Ala Leu Leu Arg Ser Trp Gly Ile Thr Pro Hi

#s Ala Val Ile Gly His

575

#       580

#       585

tcc ctc ggt gag atc acc gcc gcg cac gcc gc

#c ggt gtc ctg tcc ctg    54086

Ser Leu Gly Glu Ile Thr Ala Ala His Ala Al

#a Gly Val Leu Ser Leu

590

#   595

#   600

agg gac gcg ggc gcg ctc ctc acc acc cgc ac

#c cgc ctg atg gac caa    54134

Arg Asp Ala Gly Ala Leu Leu Thr Thr Arg Th

#r Arg Leu Met Asp Gln

605                 6

#10                 6

#15                 6

#20

ctg ccg tcg ggc ggc gcg atg gtc acc gtc ct

#g acc agc gag gaa aag    54182

Leu Pro Ser Gly Gly Ala Met Val Thr Val Le

#u Thr Ser Glu Glu Lys

625

#               630

#               635

gca cgc cag gtg ctg cgg ccg ggc gtg gag at

#c gcc gcc gtc aac ggc    54230

Ala Arg Gln Val Leu Arg Pro Gly Val Glu Il

#e Ala Ala Val Asn Gly

640

#           645

#           650

ccc cac tcc ctc gtg ctg tcc ggg gac gag ga

#a gcc gta ctc gaa gcc    54278

Pro His Ser Leu Val Leu Ser Gly Asp Glu Gl

#u Ala Val Leu Glu Ala

655

#       660

#       665

gcc cgg cag ctc ggc atc cac cac cgc ctg cc

#g acc cgc cac gcc ggc    54326

Ala Arg Gln Leu Gly Ile His His Arg Leu Pr

#o Thr Arg His Ala Gly

670

#   675

#   680

cac tcc gag cgc atg cag cca ctc gtc gcc cc

#c ctc ctc gac gtc gcc    54374

His Ser Glu Arg Met Gln Pro Leu Val Ala Pr

#o Leu Leu Asp Val Ala

685                 6

#90                 6

#95                 7

#00

cgg acc ctg acg tac cac cag ccc cac acc gc

#c atc ccc ggc gac ccc    54422

Arg Thr Leu Thr Tyr His Gln Pro His Thr Al

#a Ile Pro Gly Asp Pro

705

#               710

#               715

acc acc gcc gaa tac tgg gcg cac cag gtc cg

#c gac caa gta cgt ttc    54470

Thr Thr Ala Glu Tyr Trp Ala His Gln Val Ar

#g Asp Gln Val Arg Phe

720

#           725

#           730

cag gcg cac acc gag cag tac ccg ggc gcg ac

#g ttc ctc gag atc ggc    54518

Gln Ala His Thr Glu Gln Tyr Pro Gly Ala Th

#r Phe Leu Glu Ile Gly

735

#       740

#       745

ccc aac cag gac ctc tcg ccg ctc gtc gac gg

#c gtt gcc gcc cag acc    54566

Pro Asn Gln Asp Leu Ser Pro Leu Val Asp Gl

#y Val Ala Ala Gln Thr

750

#   755

#   760

ggt acg ccc gac gag gtg cgg gcg ctg cac ac

#c gcg ctc gcg cag ctc    54614

Gly Thr Pro Asp Glu Val Arg Ala Leu His Th

#r Ala Leu Ala Gln Leu

765                 7

#70                 7

#75                 7

#80

cac gtc cgc ggc gtc gcg atc gac tgg acg ct

#c gtc ctc ggc ggg gac    54662

His Val Arg Gly Val Ala Ile Asp Trp Thr Le

#u Val Leu Gly Gly Asp

785

#               790

#               795

cgc gcg ccc gtc acg ctg ccc acg tat ccg tt

#c cag cac aag gac tac    54710

Arg Ala Pro Val Thr Leu Pro Thr Tyr Pro Ph

#e Gln His Lys Asp Tyr

800

#           805

#           810

tgg ctg cgg ccc acc tcc cgg gcc gat gtg ac

#c ggc gcg ggg cag gag    54758

Trp Leu Arg Pro Thr Ser Arg Ala Asp Val Th

#r Gly Ala Gly Gln Glu

815

#       820

#       825

cag gtg gcg cac ccg ctg ctc ggc gcc gcg gt

#c gcg ctg ccc ggc acg    54806

Gln Val Ala His Pro Leu Leu Gly Ala Ala Va

#l Ala Leu Pro Gly Thr

830

#   835

#   840

ggc gga gtc gtc ctg acc ggc cgc ctg tcg ct

#g gcc tcc cat ccg tgg    54854

Gly Gly Val Val Leu Thr Gly Arg Leu Ser Le

#u Ala Ser His Pro Trp

845                 8

#50                 8

#55                 8

#60

ctc ggc gag cac gcg gtc gac ggc acc gtg ct

#c ctg ccc ggc gcg gcc    54902

Leu Gly Glu His Ala Val Asp Gly Thr Val Le

#u Leu Pro Gly Ala Ala

865

#               870

#               875

ttc ctc gaa ctc gcg gcg cgc gcc ggc gac ga

#g gtc ggc tgc gac ctg    54950

Phe Leu Glu Leu Ala Ala Arg Ala Gly Asp Gl

#u Val Gly Cys Asp Leu

880

#           885

#           890

ctg cac gaa ctc gtc atc gag acg ccg ctc gt

#g ctg ccc gcg acc ggc    54998

Leu His Glu Leu Val Ile Glu Thr Pro Leu Va

#l Leu Pro Ala Thr Gly

895

#       900

#       905

ggt gtg gcg gtc tcc gtc gag atc gcc gaa cc

#c gac gac acg ggg cgg    55046

Gly Val Ala Val Ser Val Glu Ile Ala Glu Pr

#o Asp Asp Thr Gly Arg

910

#   915

#   920

cgg gcg gtc acc gtc cac gcg cgg gcc gac gg

#c tcg ggc ctg tgg acc    55094

Arg Ala Val Thr Val His Ala Arg Ala Asp Gl

#y Ser Gly Leu Trp Thr

925                 9

#30                 9

#35                 9

#40

cga cac gcc ggc gga ttc ctc ggc acg gca cc

#g gca ccg gcc acg gcc    55142

Arg His Ala Gly Gly Phe Leu Gly Thr Ala Pr

#o Ala Pro Ala Thr Ala

945

#               950

#               955

acg gac ccg gca ccc tgg ccg ccc gcg gaa gc

#c gga ccg gtc gac gtc    55190

Thr Asp Pro Ala Pro Trp Pro Pro Ala Glu Al

#a Gly Pro Val Asp Val

960

#           965

#           970

gcc gac gtc tac gac cgg ttc gag gac atc gg

#g tac tcc tac gga ccg    55238

Ala Asp Val Tyr Asp Arg Phe Glu Asp Ile Gl

#y Tyr Ser Tyr Gly Pro

975

#       980

#       985

ggc ttc cgg ggg ctg cgg gcc gcc tgg cgc gc

#c ggc gac acc gtg tac    55286

Gly Phe Arg Gly Leu Arg Ala Ala Trp Arg Al

#a Gly Asp Thr Val Tyr

990

#    995

#   1000

gcc gag gtc gcg ctc ccc gac gag cag agc gc

#c gac gcc gcc cgt ttc    55334

Ala Glu Val Ala Leu Pro Asp Glu Gln Ser Al

#a Asp Ala Ala Arg Phe

1005                1010

#                1015

#              1020

acg ctg cac ccc gcg ctg ctc gac gcc gcg tt

#c cag gcc ggc gcg ctg    55382

Thr Leu His Pro Ala Leu Leu Asp Ala Ala Ph

#e Gln Ala Gly Ala Leu

1025

#               1030

#              1035

gcc gcg ctc gac gca ccc ggc ggg gcg gcc cg

#a ctg ccg ttc tcg ttc    55430

Ala Ala Leu Asp Ala Pro Gly Gly Ala Ala Ar

#g Leu Pro Phe Ser Phe

1040

#           1045

#          1050

cag gac gtc cgc atc cac gcg gcc ggg gcg ac

#g cgg ctg cgg gtc acg    55478

Gln Asp Val Arg Ile His Ala Ala Gly Ala Th

#r Arg Leu Arg Val Thr

1055

#       1060

#      1065

gtc ggc cgc gac ggc gag cgc agc acc gtc cg

#c atg acc ggc ccg gac    55526

Val Gly Arg Asp Gly Glu Arg Ser Thr Val Ar

#g Met Thr Gly Pro Asp

1070

#   1075

#  1080

ggg cag ctg gtg gcc gtg gtc ggt gcc gtg ct

#g tcg cgc ccg tac gcg    55574

Gly Gln Leu Val Ala Val Val Gly Ala Val Le

#u Ser Arg Pro Tyr Ala

1085                1090

#                1095

#              1100

gaa ggc tcc ggt gac ggc ctg ctg cgc ccg gt

#c tgg acc gag ctg ccg    55622

Glu Gly Ser Gly Asp Gly Leu Leu Arg Pro Va

#l Trp Thr Glu Leu Pro

1105

#               1110

#              1115

atg ccc gtc ccg tcc gcg gac gat ccg cgc gt

#g gag gtc ctc ggc gcc    55670

Met Pro Val Pro Ser Ala Asp Asp Pro Arg Va

#l Glu Val Leu Gly Ala

1120

#           1125

#          1130

gac ccg ggc gac ggc gac gtt ccg gcg gcc ac

#c cgg gag ctg acc gcc    55718

Asp Pro Gly Asp Gly Asp Val Pro Ala Ala Th

#r Arg Glu Leu Thr Ala

1135

#       1140

#      1145

cgc gtc ctc ggc gcg ctc cag cgc cac ctg tc

#c gcc gcc gag gac acc    55766

Arg Val Leu Gly Ala Leu Gln Arg His Leu Se

#r Ala Ala Glu Asp Thr

1150

#   1155

#  1160

acc ttg gtg gta cgg acc ggc acc ggc ccg gc

#c gct gcc gcc gcc gcg    55814

Thr Leu Val Val Arg Thr Gly Thr Gly Pro Al

#a Ala Ala Ala Ala Ala

1165                1170

#                1175

#              1180

ggt ctg gtc cgc tcg gcg cag gcg gag aac cc

#c ggc cgc gtc gtg ctc    55862

Gly Leu Val Arg Ser Ala Gln Ala Glu Asn Pr

#o Gly Arg Val Val Leu

1185

#               1190

#              1195

gtc gag gcg tcc ccg gac acc tcg gtg gag ct

#g ctc gcc gcg tgc gcc    55910

Val Glu Ala Ser Pro Asp Thr Ser Val Glu Le

#u Leu Ala Ala Cys Ala

1200

#           1205

#          1210

gcg ctg gac gaa ccg cag ctg gcc gtc cgg ga

#c ggc gtg ctc ttc gcg    55958

Ala Leu Asp Glu Pro Gln Leu Ala Val Arg As

#p Gly Val Leu Phe Ala

1215

#       1220

#      1225

ccg cgg ctg gtc cgg atg tcc gac ccc gcg ca

#c ggc ccg ctg tcc ctg    56006

Pro Arg Leu Val Arg Met Ser Asp Pro Ala Hi

#s Gly Pro Leu Ser Leu

1230

#   1235

#  1240

ccg gac ggc gac tgg ctg ctc acc cgg tcc gc

#c tcc ggc acg ttg cac    56054

Pro Asp Gly Asp Trp Leu Leu Thr Arg Ser Al

#a Ser Gly Thr Leu His

1245                1250

#                1255

#              1260

gac gtc gcg ctc ata gcc gac gac acg ccc cg

#g cgg gcg ctc gaa gcc    56102

Asp Val Ala Leu Ile Ala Asp Asp Thr Pro Ar

#g Arg Ala Leu Glu Ala

1265

#               1270

#              1275

ggc gag gtc cgc atc gac gtc cgc gcg gcc gg

#a ctg aac ttc cgc gat    56150

Gly Glu Val Arg Ile Asp Val Arg Ala Ala Gl

#y Leu Asn Phe Arg Asp

1280

#           1285

#          1290

gtg ctg atc gcg ctc ggg acg tac acc ggg gc

#c acg gcc atg ggc ggc    56198

Val Leu Ile Ala Leu Gly Thr Tyr Thr Gly Al

#a Thr Ala Met Gly Gly

1295

#       1300

#      1305

gag gcc gcg ggc gtc gtg gtg gag acc ggg cc

#c ggc gtg gac gac ctg    56246

Glu Ala Ala Gly Val Val Val Glu Thr Gly Pr

#o Gly Val Asp Asp Leu

1310

#   1315

#  1320

tcc ccc ggc gac cgg gtg ttc ggc ctg acc cg

#g ggc ggc atc ggc ccg    56294

Ser Pro Gly Asp Arg Val Phe Gly Leu Thr Ar

#g Gly Gly Ile Gly Pro

1325                1330

#                1335

#              1340

acg gcc gtc acc gac cgg cgc tgg ctg gcc cg

#g atc ccc gac ggc tgg    56342

Thr Ala Val Thr Asp Arg Arg Trp Leu Ala Ar

#g Ile Pro Asp Gly Trp

1345

#               1350

#              1355

agc ttc acc acg gcg gcg tcc gtc ccg atc gt

#g ttc gcg acc gcg tgg    56390

Ser Phe Thr Thr Ala Ala Ser Val Pro Ile Va

#l Phe Ala Thr Ala Trp

1360

#           1365

#          1370

tac ggc ctg gtc gac ctc ggc aca ctg cgc gc

#c ggc gag aag gtc ctc    56438

Tyr Gly Leu Val Asp Leu Gly Thr Leu Arg Al

#a Gly Glu Lys Val Leu

1375

#       1380

#      1385

gtc cac gcg gcc acc ggc ggt gtc ggc atg gc

#c gcc gca cag atc gcc    56486

Val His Ala Ala Thr Gly Gly Val Gly Met Al

#a Ala Ala Gln Ile Ala

1390

#   1395

#  1400

cgc cac ctg ggc gcc gag ctc tac gcc acc gc

#c agt acc ggc aag cag    56534

Arg His Leu Gly Ala Glu Leu Tyr Ala Thr Al

#a Ser Thr Gly Lys Gln

1405                1410

#                1415

#              1420

cac gtc ctg cgc gcc gcc ggg ctg ccc gac ac

#g cac atc gcc gac tct    56582

His Val Leu Arg Ala Ala Gly Leu Pro Asp Th

#r His Ile Ala Asp Ser

1425

#               1430

#              1435

cgg acg acc gcg ttc cgg acc gct ttc ccg cg

#c atg gac gtc gtc ctg    56630

Arg Thr Thr Ala Phe Arg Thr Ala Phe Pro Ar

#g Met Asp Val Val Leu

1440

#           1445

#          1450

aac gcg ctg acc ggc gag ttc atc gac gcg tc

#g ctc gac ctg ctg gac    56678

Asn Ala Leu Thr Gly Glu Phe Ile Asp Ala Se

#r Leu Asp Leu Leu Asp

1455

#       1460

#      1465

gcc gac ggc cgg ttc gtc gag atg ggc cgc ac

#c gag ctg cgc gac ccg    56726

Ala Asp Gly Arg Phe Val Glu Met Gly Arg Th

#r Glu Leu Arg Asp Pro

1470

#   1475

#  1480

gcc gcg atc gtc ccc gcc tac ctg ccg ttc ga

#c ctg ctg gac gcg ggc    56774

Ala Ala Ile Val Pro Ala Tyr Leu Pro Phe As

#p Leu Leu Asp Ala Gly

1485                1490

#                1495

#              1500

gcc gac cgc atc ggc gag atc ctg ggc gaa ct

#g ctc cgg ctg ttc gac    56822

Ala Asp Arg Ile Gly Glu Ile Leu Gly Glu Le

#u Leu Arg Leu Phe Asp

1505

#               1510

#              1515

gcg ggc gcg ctg gag ccg ctg ccg gtc cgt gc

#c tgg gac gtc cgg cag    56870

Ala Gly Ala Leu Glu Pro Leu Pro Val Arg Al

#a Trp Asp Val Arg Gln

1520

#           1525

#          1530

gca cgc gac gcg ctc ggc tgg atg agc cgc gc

#c cgc cac atc ggc aag    56918

Ala Arg Asp Ala Leu Gly Trp Met Ser Arg Al

#a Arg His Ile Gly Lys

1535

#       1540

#      1545

aac gtc ctg acg ctg ccc cgg ccg ctc gac cc

#g gag ggc gcc gtc gtc    56966

Asn Val Leu Thr Leu Pro Arg Pro Leu Asp Pr

#o Glu Gly Ala Val Val

1550

#   1555

#  1560

ctc acc ggc ggc tcc ggc acg ctc gcc ggc at

#c ctc gcc cgc cac ctg    57014

Leu Thr Gly Gly Ser Gly Thr Leu Ala Gly Il

#e Leu Ala Arg His Leu

1565                1570

#                1575

#              1580

cgc gaa cgg cat gtc tac ctg ctg tcc cgg ac

#g gca ccg ccc gag ggg    57062

Arg Glu Arg His Val Tyr Leu Leu Ser Arg Th

#r Ala Pro Pro Glu Gly

1585

#               1590

#              1595

acg ccc ggc gtc cac ctg ccc tgc gac gtc gg

#t gac cgg gac cag ctg    57110

Thr Pro Gly Val His Leu Pro Cys Asp Val Gl

#y Asp Arg Asp Gln Leu

1600

#           1605

#          1610

gcg gcg gcc ctg gag cgg gtg gac cgg ccg at

#c acc gcc gtg gtg cac    57158

Ala Ala Ala Leu Glu Arg Val Asp Arg Pro Il

#e Thr Ala Val Val His

1615

#       1620

#      1625

ctc gcc ggt gcg ctg gac gac ggc acc gtc gc

#g tcg ctc acc ccc gag    57206

Leu Ala Gly Ala Leu Asp Asp Gly Thr Val Al

#a Ser Leu Thr Pro Glu

1630

#   1635

#  1640

cgt ttc gac acg gtg ctg cgc ccg aag gcc ga

#c ggc gcc tgg tac ctg    57254

Arg Phe Asp Thr Val Leu Arg Pro Lys Ala As

#p Gly Ala Trp Tyr Leu

1645                1650

#                1655

#              1660

cac gag ctg acg aag gag cag gac ctc gcc gc

#g ttc gtg ctc tac tcg    57302

His Glu Leu Thr Lys Glu Gln Asp Leu Ala Al

#a Phe Val Leu Tyr Ser

1665

#               1670

#              1675

tcg gcc gcc ggc gtg ctc ggc aac gcc ggc ca

#g ggc aac tac gtc gcc    57350

Ser Ala Ala Gly Val Leu Gly Asn Ala Gly Gl

#n Gly Asn Tyr Val Ala

1680

#           1685

#          1690

gcg aac gcg ttc ctc gac gcg ctc gcc gag ct

#g cgc cac ggt tcc ggg    57398

Ala Asn Ala Phe Leu Asp Ala Leu Ala Glu Le

#u Arg His Gly Ser Gly

1695

#       1700

#      1705

ctg ccg gcc ctc tcc atc gcc tgg ggg ctc tg

#g gag gac gtg agc ggg    57446

Leu Pro Ala Leu Ser Ile Ala Trp Gly Leu Tr

#p Glu Asp Val Ser Gly

1710

#   1715

#  1720

ctc acc gcg gcg ctc ggc gaa gcc gac cgg ga

#c cgg atg cgg cgc agc    57494

Leu Thr Ala Ala Leu Gly Glu Ala Asp Arg As

#p Arg Met Arg Arg Ser

1725                1730

#                1735

#              1740

ggt ttc cgg gcc atc acc gcg caa cag ggc at

#g cac ctg tac gag gcg    57542

Gly Phe Arg Ala Ile Thr Ala Gln Gln Gly Me

#t His Leu Tyr Glu Ala

1745

#               1750

#              1755

gcc ggc cgc acc gga agt ccc gtg gtg gtc gc

#g gcg gcg ctc gac gac    57590

Ala Gly Arg Thr Gly Ser Pro Val Val Val Al

#a Ala Ala Leu Asp Asp

1760

#           1765

#          1770

gcg ccg gac gtg ccg ctg ctg cgc ggc ctg cg

#g cgg acg acc gtc cgg    57638

Ala Pro Asp Val Pro Leu Leu Arg Gly Leu Ar

#g Arg Thr Thr Val Arg

1775

#       1780

#      1785

cgg gcc gcc gtc cgg gag tgt tcg tcc gcc ga

#c cgg ctc gcc gcg ctg    57686

Arg Ala Ala Val Arg Glu Cys Ser Ser Ala As

#p Arg Leu Ala Ala Leu

1790

#   1795

#  1800

acc ggc gac gag ctc gcc gaa gcg ctg ctg ac

#g ctc gtc cgg gag agc    57734

Thr Gly Asp Glu Leu Ala Glu Ala Leu Leu Th

#r Leu Val Arg Glu Ser

1805                1810

#                1815

#              1820

acc gcc gcc gtg ctc ggc cac gtg ggt ggc ga

#g gac atc ccc gcg acg    57782

Thr Ala Ala Val Leu Gly His Val Gly Gly Gl

#u Asp Ile Pro Ala Thr

1825

#               1830

#              1835

gcg gcg ttc aag gac ctc ggc atc gac tcg ct

#c acc gcg gtc cag ctg    57830

Ala Ala Phe Lys Asp Leu Gly Ile Asp Ser Le

#u Thr Ala Val Gln Leu

1840

#           1845

#          1850

cgc aac gcc ctc acc gag gcg acc ggt gtg cg

#g ctg aac gcc acg gcg    57878

Arg Asn Ala Leu Thr Glu Ala Thr Gly Val Ar

#g Leu Asn Ala Thr Ala

1855

#       1860

#      1865

gtc ttc gac ttc ccg acc ccg cac gtg ctc gc

#c ggg aag ctc ggc gac    57926

Val Phe Asp Phe Pro Thr Pro His Val Leu Al

#a Gly Lys Leu Gly Asp

1870

#   1875

#  1880

gaa ctg acc ggc acc cgc gcg ccc gtc gtg cc

#c cgg acc gcg gcc acg    57974

Glu Leu Thr Gly Thr Arg Ala Pro Val Val Pr

#o Arg Thr Ala Ala Thr

1885                1890

#                1895

#              1900

gcc ggt gcg cac gac gag ccg ctg gcg atc gt

#g gga atg gcc tgc cgg    58022

Ala Gly Ala His Asp Glu Pro Leu Ala Ile Va

#l Gly Met Ala Cys Arg

1905

#               1910

#              1915

ctg ccc ggc ggg gtc gcg tca ccc gag gag ct

#g tgg cac ctc gtg gca    58070

Leu Pro Gly Gly Val Ala Ser Pro Glu Glu Le

#u Trp His Leu Val Ala

1920

#           1925

#          1930

tcc ggc acc gac gcc atc acg gag ttc ccg ac

#g gac cgc ggc tgg gac    58118

Ser Gly Thr Asp Ala Ile Thr Glu Phe Pro Th

#r Asp Arg Gly Trp Asp

1935

#       1940

#      1945

gtc gac gcg atc tac gac ccg gac ccc gac gc

#g atc ggc aag acc ttc    58166

Val Asp Ala Ile Tyr Asp Pro Asp Pro Asp Al

#a Ile Gly Lys Thr Phe

1950

#   1955

#  1960

gtc cgg cac ggt ggc ttc ctc acc ggc gcg ac

#a ggc ttc gac gcg gcg    58214

Val Arg His Gly Gly Phe Leu Thr Gly Ala Th

#r Gly Phe Asp Ala Ala

1965                1970

#                1975

#              1980

ttc ttc ggc atc agc ccg cgc gag gcc ctc gc

#g atg gac ccg cag cag    58262

Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Al

#a Met Asp Pro Gln Gln

1985

#               1990

#              1995

cgg gtg ctc ctg gag acg tcg tgg gag gcg tt

#c gaa agc gcc ggc atc    58310

Arg Val Leu Leu Glu Thr Ser Trp Glu Ala Ph

#e Glu Ser Ala Gly Ile

2000

#           2005

#          2010

acc ccg gac tcg acc cgc ggc agc gac acc gg

#c gtg ttc gtc ggc gcc    58358

Thr Pro Asp Ser Thr Arg Gly Ser Asp Thr Gl

#y Val Phe Val Gly Ala

2015

#       2020

#      2025

ttc tcc tac ggt tac ggc acc ggt gcg gac ac

#c gac ggc ttc ggc gcg    58406

Phe Ser Tyr Gly Tyr Gly Thr Gly Ala Asp Th

#r Asp Gly Phe Gly Ala

2030

#   2035

#  2040

acc ggc tcg cag acc agt gtg ctc tcc ggc cg

#g ctg tcg tac ttc tac    58454

Thr Gly Ser Gln Thr Ser Val Leu Ser Gly Ar

#g Leu Ser Tyr Phe Tyr

2045                2050

#                2055

#              2060

ggt ctg gag ggt ccg gcg gtc acg gtc gac ac

#g gcg tgt tcg tcg tcg    58502

Gly Leu Glu Gly Pro Ala Val Thr Val Asp Th

#r Ala Cys Ser Ser Ser

2065

#               2070

#              2075

ctg gtg gcg ctg cac cag gcc ggg cag tcg ct

#g cgc tcc ggc gaa tgc    58550

Leu Val Ala Leu His Gln Ala Gly Gln Ser Le

#u Arg Ser Gly Glu Cys

2080

#           2085

#          2090

tcg ctc gcc ctg gtc ggc ggc gtc acg gtg at

#g gcg tct ccc ggc ggc    58598

Ser Leu Ala Leu Val Gly Gly Val Thr Val Me

#t Ala Ser Pro Gly Gly

2095

#       2100

#      2105

ttc gtg gag ttc tcc cgg cag cgc ggc ctc gc

#g ccg gac ggc cgg gcg    58646

Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Al

#a Pro Asp Gly Arg Ala

2110

#   2115

#  2120

aag gcg ttc ggc gcg ggt gcg gac ggc acg ag

#c ttc gcc gag ggt gcc    58694

Lys Ala Phe Gly Ala Gly Ala Asp Gly Thr Se

#r Phe Ala Glu Gly Ala

2125                2130

#                2135

#              2140

ggt gtg ctg atc gtc gag agg ctc tcc gac gc

#c gaa cgc aac ggt cac    58742

Gly Val Leu Ile Val Glu Arg Leu Ser Asp Al

#a Glu Arg Asn Gly His

2145

#               2150

#              2155

acc gtc ctg gcg gtc gtc cgt ggt tcg gcg gt

#c aac cag gat ggt gcc    58790

Thr Val Leu Ala Val Val Arg Gly Ser Ala Va

#l Asn Gln Asp Gly Ala

2160

#           2165

#          2170

tcc aac ggg ctg tcg gcg ccg aac ggg ccg tc

#g cag gag cgg gtg atc    58838

Ser Asn Gly Leu Ser Ala Pro Asn Gly Pro Se

#r Gln Glu Arg Val Ile

2175

#       2180

#      2185

cgg cag gcc ctg gcc aac gcc ggg ctc acc cc

#g gcg gac gtg gac gcc    58886

Arg Gln Ala Leu Ala Asn Ala Gly Leu Thr Pr

#o Ala Asp Val Asp Ala

2190

#   2195

#  2200

gtc gag gcc cac ggc acc ggc acc agg ctg gg

#c gac ccc atc gag gca    58934

Val Glu Ala His Gly Thr Gly Thr Arg Leu Gl

#y Asp Pro Ile Glu Ala

2205                2210

#                2215

#              2220

cag gcg gta ctg gcc acc tac gga cag gag cg

#c gcc acc ccc ctg ctg    58982

Gln Ala Val Leu Ala Thr Tyr Gly Gln Glu Ar

#g Ala Thr Pro Leu Leu

2225

#               2230

#              2235

ctg ggc tcg ctg aag tcc aac atc ggc cac gc

#c cag gcc gcg tcc ggc    59030

Leu Gly Ser Leu Lys Ser Asn Ile Gly His Al

#a Gln Ala Ala Ser Gly

2240

#           2245

#          2250

gtc gcc ggc atc atc aag atg gtg cag gcc ct

#c cgg cac ggg gag ctg    59078

Val Ala Gly Ile Ile Lys Met Val Gln Ala Le

#u Arg His Gly Glu Leu

2255

#       2260

#      2265

ccg ccg acg ctg cac gcc gac gag ccg tcg cc

#g cac gtc gac tgg acg    59126

Pro Pro Thr Leu His Ala Asp Glu Pro Ser Pr

#o His Val Asp Trp Thr

2270

#   2275

#  2280

gcc ggc gcc gtc gaa ctg ctg acg tcg gcc cg

#g ccg tgg ccc gag acc    59174

Ala Gly Ala Val Glu Leu Leu Thr Ser Ala Ar

#g Pro Trp Pro Glu Thr

2285                2290

#                2295

#              2300

gac cgg cca cgg cgt gcc gcc gtc tcc tcg tt

#c ggg gtg agc ggc acc    59222

Asp Arg Pro Arg Arg Ala Ala Val Ser Ser Ph

#e Gly Val Ser Gly Thr

2305

#               2310

#              2315

aac gcc cac gtc atc ctg gag gcc gga ccg gt

#a acg gag acg ccc gcg    59270

Asn Ala His Val Ile Leu Glu Ala Gly Pro Va

#l Thr Glu Thr Pro Ala

2320

#           2325

#          2330

gca tcg cct tcc ggt gac ctt ccc ctg ctg gt

#g tcg gca cgc tca ccg    59318

Ala Ser Pro Ser Gly Asp Leu Pro Leu Leu Va

#l Ser Ala Arg Ser Pro

2335

#       2340

#      2345

gaa gcg ctc gac gag cag atc cgc cga ctg cg

#c gcc tac ctg gac acc    59366

Glu Ala Leu Asp Glu Gln Ile Arg Arg Leu Ar

#g Ala Tyr Leu Asp Thr

2350

#   2355

#  2360

acc ccg gac gtc gac cgg gtg gcc gtg gca ca

#g acg ctg gcc cgg cgc    59414

Thr Pro Asp Val Asp Arg Val Ala Val Ala Gl

#n Thr Leu Ala Arg Arg

2365                2370

#                2375

#              2380

aca cac ttc gcc cac cgc gcc gtg ctg ctc gg

#t gac acc gtc atc acc    59462

Thr His Phe Ala His Arg Ala Val Leu Leu Gl

#y Asp Thr Val Ile Thr

2385

#               2390

#              2395

aca ccc ccc gcg gac cgg ccc gac gaa ctc gt

#c ttc gtc tac tcc ggc    59510

Thr Pro Pro Ala Asp Arg Pro Asp Glu Leu Va

#l Phe Val Tyr Ser Gly

2400

#           2405

#          2410

cag ggc acc cag cat ccc gcg atg ggc gag ca

#g ctc gcc gcc gcc cat    59558

Gln Gly Thr Gln His Pro Ala Met Gly Glu Gl

#n Leu Ala Ala Ala His

2415

#       2420

#      2425

ccc gtg ttc gcc gac gcc tgg cat gaa gcg ct

#c cgc cgc ctt gac aac    59606

Pro Val Phe Ala Asp Ala Trp His Glu Ala Le

#u Arg Arg Leu Asp Asn

2430

#   2435

#  2440

ccc gac ccc cac gac ccc acg cac agc cag ca

#t gtg ctc ttc gcc cac    59654

Pro Asp Pro His Asp Pro Thr His Ser Gln Hi

#s Val Leu Phe Ala His

2445                2450

#                2455

#              2460

cag gcg gcg ttc acc gcc ctc ctg cgg tcc tg

#g ggc atc acc ccg cac    59702

Gln Ala Ala Phe Thr Ala Leu Leu Arg Ser Tr

#p Gly Ile Thr Pro His

2465

#               2470

#              2475

gcg gtc atc ggc cac tcg ctg ggc gag atc ac

#c gcg gcg cac gcc gcc    59750

Ala Val Ile Gly His Ser Leu Gly Glu Ile Th

#r Ala Ala His Ala Ala

2480

#           2485

#          2490

ggc atc ctg tcg ctg gac gac gcg tgc acc ct

#g atc acc acg cgc gcc    59798

Gly Ile Leu Ser Leu Asp Asp Ala Cys Thr Le

#u Ile Thr Thr Arg Ala

2495

#       2500

#      2505

cgc ctc atg cac acg ctc ccg cca ccc ggt gc

#c atg gtc acc gta ctg    59846

Arg Leu Met His Thr Leu Pro Pro Pro Gly Al

#a Met Val Thr Val Leu

2510

#   2515

#  2520

acc agc gaa gag aag gca cgc cag gcg ttg cg

#g ccg ggc gtg gag atc    59894

Thr Ser Glu Glu Lys Ala Arg Gln Ala Leu Ar

#g Pro Gly Val Glu Ile

2525                2530

#                2535

#              2540

gcc gcc gtc aac ggg ccc cac tcc atc gtg ct

#g tcc ggg gac gag gac    59942

Ala Ala Val Asn Gly Pro His Ser Ile Val Le

#u Ser Gly Asp Glu Asp

2545

#               2550

#              2555

gcc gtg ctc acc gtc gcc ggg cag ctc ggc at

#c cac cac cgc ctg ccc    59990

Ala Val Leu Thr Val Ala Gly Gln Leu Gly Il

#e His His Arg Leu Pro

2560

#           2565

#          2570

gcc ccg cac gcc ggg cac tcc gcg cac atg ga

#g ccc gtg gcc gcc gag    60038

Ala Pro His Ala Gly His Ser Ala His Met Gl

#u Pro Val Ala Ala Glu

2575

#       2580

#      2585

ctg ctc gcc acc acc cgc ggg ctc cgc tac ca

#c cct ccc cac acc tcc    60086

Leu Leu Ala Thr Thr Arg Gly Leu Arg Tyr Hi

#s Pro Pro His Thr Ser

2590

#   2595

#  2600

att ccg aac gac ccc acc acc gct gag tac tg

#g gcc gag cag gtc cgc    60134

Ile Pro Asn Asp Pro Thr Thr Ala Glu Tyr Tr

#p Ala Glu Gln Val Arg

2605                2610

#                2615

#              2620

aag ccc gtg ctg ttc cac gcc cac gcg cag ca

#g tac ccg gac gcc gtg    60182

Lys Pro Val Leu Phe His Ala His Ala Gln Gl

#n Tyr Pro Asp Ala Val

2625

#               2630

#              2635

ttc gtg gag atc ggc ccc gcc cag gac ctc tc

#c ccg ctc gtc gac ggg    60230

Phe Val Glu Ile Gly Pro Ala Gln Asp Leu Se

#r Pro Leu Val Asp Gly

2640

#           2645

#          2650

atc ccg ctg cag aac ggc acc gcg gac gag gt

#g cac gcg ctg cac acc    60278

Ile Pro Leu Gln Asn Gly Thr Ala Asp Glu Va

#l His Ala Leu His Thr

2655

#       2660

#      2665

gcg ctc gcg cac ctc tac gcg cgc ggt gcc ac

#g ctc gac tgg ccc cgc    60326

Ala Leu Ala His Leu Tyr Ala Arg Gly Ala Th

#r Leu Asp Trp Pro Arg

2670

#   2675

#  2680

atc ctc ggg gct ggg tca cgg cac gac gcg ga

#t gtg ccc gcg tac gcg    60374

Ile Leu Gly Ala Gly Ser Arg His Asp Ala As

#p Val Pro Ala Tyr Ala

2685                2690

#                2695

#              2700

ttc caa cgg cgg cac tac tgg atc gag tcg gc

#a cgc ccg gcc gca tcc    60422

Phe Gln Arg Arg His Tyr Trp Ile Glu Ser Al

#a Arg Pro Ala Ala Ser

2705

#               2710

#              2715

gac gcg ggc cac ccc gtg ctg ggc tcc ggt at

#c gcc ctc gcc ggg tcg    60470

Asp Ala Gly His Pro Val Leu Gly Ser Gly Il

#e Ala Leu Ala Gly Ser

2720

#           2725

#          2730

ccg ggc cgg gtg ttc acg ggt tcc gtg ccg ac

#c ggt gcg gac cgc gcg    60518

Pro Gly Arg Val Phe Thr Gly Ser Val Pro Th

#r Gly Ala Asp Arg Ala

2735

#       2740

#      2745

gtg ttc gtc gcc gag ctg gcg ctg gcc gcc gc

#g gac gcg gtc gac tgc    60566

Val Phe Val Ala Glu Leu Ala Leu Ala Ala Al

#a Asp Ala Val Asp Cys

2750

#   2755

#  2760

gcc acg gtc gag cgg ctc gac atc gcc tcc gt

#g ccc ggc cgg ccg ggc    60614

Ala Thr Val Glu Arg Leu Asp Ile Ala Ser Va

#l Pro Gly Arg Pro Gly

2765                2770

#                2775

#              2780

cat ggc cgg acg acc gta cag acc tgg gtc ga

#c gag ccg gcg gac gac    60662

His Gly Arg Thr Thr Val Gln Thr Trp Val As

#p Glu Pro Ala Asp Asp

2785

#               2790

#              2795

ggc cgg cgc cgg ttc acc gtg cac acc cgc ac

#c ggc gac gcc ccg tgg    60710

Gly Arg Arg Arg Phe Thr Val His Thr Arg Th

#r Gly Asp Ala Pro Trp

2800

#           2805

#          2810

acg ctg cac gcc gag ggg gtg ctg cgc ccc ca

#t ggc acg gcc ctg ccc    60758

Thr Leu His Ala Glu Gly Val Leu Arg Pro Hi

#s Gly Thr Ala Leu Pro

2815

#       2820

#      2825

gat gcg gcc gac gcc gag tgg ccc cca ccg gg

#c gcg gtg ccc gcg gac    60806

Asp Ala Ala Asp Ala Glu Trp Pro Pro Pro Gl

#y Ala Val Pro Ala Asp

2830

#   2835

#  2840

ggg ctg ccg ggt gtg tgg cgc cgg ggg gac ca

#g gtc ttc gcc gag gcc    60854

Gly Leu Pro Gly Val Trp Arg Arg Gly Asp Gl

#n Val Phe Ala Glu Ala

2845                2850

#                2855

#              2860

gag gtg gac gga ccg gac ggt ttc gtg gtg ca

#c ccc gac ctg ctc gac    60902

Glu Val Asp Gly Pro Asp Gly Phe Val Val Hi

#s Pro Asp Leu Leu Asp

2865

#               2870

#              2875

gcg gtc ttc tcc gcg gtc ggc gac gga agc cg

#c cag ccg gcc gga tgg    60950

Ala Val Phe Ser Ala Val Gly Asp Gly Ser Ar

#g Gln Pro Ala Gly Trp

2880

#           2885

#          2890

cgc gac ctg acg gtg cac gcg tcg gac gcc ac

#c gta ctg cgc gcc tgc    60998

Arg Asp Leu Thr Val His Ala Ser Asp Ala Th

#r Val Leu Arg Ala Cys

2895

#       2900

#      2905

ctc acc cgg cgc acc gac gga gcc atg gga tt

#c gcc gcc ttc gac ggc    61046

Leu Thr Arg Arg Thr Asp Gly Ala Met Gly Ph

#e Ala Ala Phe Asp Gly

2910

#   2915

#  2920

gcc ggc ctg ccg gta ctc acc gcg gag gcg gt

#g acg ctg cgg gag gtg    61094

Ala Gly Leu Pro Val Leu Thr Ala Glu Ala Va

#l Thr Leu Arg Glu Val

2925                2930

#                2935

#              2940

gcg tca ccg tcc ggc tcc gag gag tcg gac gg

#c ctg cac cgg ttg gag    61142

Ala Ser Pro Ser Gly Ser Glu Glu Ser Asp Gl

#y Leu His Arg Leu Glu

2945

#               2950

#              2955

tgg ctc gcg gtc gcc gag gcg gtc tac gac gg

#t gac ctg ccc gag gga    61190

Trp Leu Ala Val Ala Glu Ala Val Tyr Asp Gl

#y Asp Leu Pro Glu Gly

2960

#           2965

#          2970

cat gtc ctg atc acc gcc gcc cac ccc gac ga

#c ccc gag gac ata ccc    61238

His Val Leu Ile Thr Ala Ala His Pro Asp As

#p Pro Glu Asp Ile Pro

2975

#       2980

#      2985

acc cgc gcc cac acc cgc gcc acc cgc gtc ct

#g acc gcc ctg caa cac    61286

Thr Arg Ala His Thr Arg Ala Thr Arg Val Le

#u Thr Ala Leu Gln His

2990

#   2995

#  3000

cac ctc acc acc acc gac cac acc ctc atc gt

#c cac acc acc acc gac    61334

His Leu Thr Thr Thr Asp His Thr Leu Ile Va

#l His Thr Thr Thr Asp

3005                3010

#                3015

#              3020

ccc gcc ggc gcc acc gtc acc ggc ctc acc cg

#c acc gcc cag aac gaa    61382

Pro Ala Gly Ala Thr Val Thr Gly Leu Thr Ar

#g Thr Ala Gln Asn Glu

3025

#               3030

#              3035

cac ccc cac cgc atc cgc ctc atc gaa acc ga

#c cac ccc cac acc ccc    61430

His Pro His Arg Ile Arg Leu Ile Glu Thr As

#p His Pro His Thr Pro

3040

#           3045

#          3050

ctc ccc ctg gcc caa ctc gcc acc ctc gac ca

#c ccc cac ctc cgc ctc    61478

Leu Pro Leu Ala Gln Leu Ala Thr Leu Asp Hi

#s Pro His Leu Arg Leu

3055

#       3060

#      3065

acc cac cac acc ctc cac cac ccc cac ctc ac

#c ccc ctc cac acc acc    61526

Thr His His Thr Leu His His Pro His Leu Th

#r Pro Leu His Thr Thr

3070

#   3075

#  3080

acc cca ccc acc acc acc ccc ctc aac ccc ga

#a cac gcc atc atc atc    61574

Thr Pro Pro Thr Thr Thr Pro Leu Asn Pro Gl

#u His Ala Ile Ile Ile

3085                3090

#                3095

#              3100

acc ggc ggc tcc ggc acc ctc gcc ggc atc ct

#c gcc cgc cac ctg aac    61622

Thr Gly Gly Ser Gly Thr Leu Ala Gly Ile Le

#u Ala Arg His Leu Asn

3105

#               3110

#              3115

cac ccc cac acc tac ctc ctc tcc cgc acc cc

#a ccc ccc gac gcc acc    61670

His Pro His Thr Tyr Leu Leu Ser Arg Thr Pr

#o Pro Pro Asp Ala Thr

3120

#           3125

#          3130

ccc ggc acc cac ctc ccc tgc gac gtc ggc ga

#c ccc cac caa ctc gcc    61718

Pro Gly Thr His Leu Pro Cys Asp Val Gly As

#p Pro His Gln Leu Ala

3135

#       3140

#      3145

acc acc ctc acc cac atc ccc caa ccc ctc ac

#c gcc atc ttc cac acc    61766

Thr Thr Leu Thr His Ile Pro Gln Pro Leu Th

#r Ala Ile Phe His Thr

3150

#   3155

#  3160

gcc gcc acc ctc gac gac ggc atc ctc cac gc

#c ctc acc ccc gac cgc    61814

Ala Ala Thr Leu Asp Asp Gly Ile Leu His Al

#a Leu Thr Pro Asp Arg

3165                3170

#                3175

#              3180

ctc acc acc gtc ctc cac ccc aaa gcc aac gc

#c gcc tgg cac ctg cac    61862

Leu Thr Thr Val Leu His Pro Lys Ala Asn Al

#a Ala Trp His Leu His

3185

#               3190

#              3195

cac ctc acc caa aac caa ccc ctc acc cac tt

#c gtc ctc tac tcc agc    61910

His Leu Thr Gln Asn Gln Pro Leu Thr His Ph

#e Val Leu Tyr Ser Ser

3200

#           3205

#          3210

gcc gcc gcc gtc ctc ggc agc ccc gga caa gg

#a aac tac gcc gcc gcc    61958

Ala Ala Ala Val Leu Gly Ser Pro Gly Gln Gl

#y Asn Tyr Ala Ala Ala

3215

#       3220

#      3225

aac gcc ttc ctc gac gcc ctc gcc acc cac cg

#c cac acc ctc ggc caa    62006

Asn Ala Phe Leu Asp Ala Leu Ala Thr His Ar

#g His Thr Leu Gly Gln

3230

#   3235

#  3240

ccc gcc acc tcc atc gcc tgg ggc atg tgg ca

#c acc acc agc acc ctc    62054

Pro Ala Thr Ser Ile Ala Trp Gly Met Trp Hi

#s Thr Thr Ser Thr Leu

3245                3250

#                3255

#              3260

acc gga caa ctc gac gac gcc gac cgg gac cg

#c atc cgc cgc ggc ggt    62102

Thr Gly Gln Leu Asp Asp Ala Asp Arg Asp Ar

#g Ile Arg Arg Gly Gly

3265

#               3270

#              3275

ttc ctc ccg atc acg gac gac gag ggc atg cg

#c ctc tac gag gcg gcc    62150

Phe Leu Pro Ile Thr Asp Asp Glu Gly Met Ar

#g Leu Tyr Glu Ala Ala

3280

#           3285

#          3290

gtc ggc tcc ggc gag gac ttc gtc atg gcc gc

#c gcg atg gac ccg gca    62198

Val Gly Ser Gly Glu Asp Phe Val Met Ala Al

#a Ala Met Asp Pro Ala

3295

#       3300

#      3305

cag ccg atg acc ggc tcc gta ccg ccc atc ct

#g agc ggc ctg cgc agg    62246

Gln Pro Met Thr Gly Ser Val Pro Pro Ile Le

#u Ser Gly Leu Arg Arg

3310

#   3315

#  3320

agc gcg cgg cgc gtc gcc cgt gcc ggg cag ac

#g ttc gcc cag cgg ctc    62294

Ser Ala Arg Arg Val Ala Arg Ala Gly Gln Th

#r Phe Ala Gln Arg Leu

3325                3330

#                3335

#              3340

gcc gag ctg ccc gac gcc gac cgc ggc gcg gc

#g ctg acc acc ctc gtc    62342

Ala Glu Leu Pro Asp Ala Asp Arg Gly Ala Al

#a Leu Thr Thr Leu Val

3345

#               3350

#              3355

tcg gac gcc acg gcc gcc gtg ctc ggc cac gc

#c gac gcc tcc gag atc    62390

Ser Asp Ala Thr Ala Ala Val Leu Gly His Al

#a Asp Ala Ser Glu Ile

3360

#           3365

#          3370

gcg ccg acc acg acg ttc aag gac ctc ggc at

#c gac tcg ctc acc gcg    62438

Ala Pro Thr Thr Thr Phe Lys Asp Leu Gly Il

#e Asp Ser Leu Thr Ala

3375

#       3380

#      3385

atc gag ctg cgc aac cgg ctc gcg gag gcg ac

#c ggg ctg cgg ctg agt    62486

Ile Glu Leu Arg Asn Arg Leu Ala Glu Ala Th

#r Gly Leu Arg Leu Ser

3390

#   3395

#  3400

gcc acg ctg gtg ttc gac cac ccg aca cct cg

#g gtc ctc gcc gcc aag    62534

Ala Thr Leu Val Phe Asp His Pro Thr Pro Ar

#g Val Leu Ala Ala Lys

3405                3410

#                3415

#              3420

ctc cgc acc gat ctg ttc ggc acg gcc gtg cc

#c acg ccc gcg cgg acg    62582

Leu Arg Thr Asp Leu Phe Gly Thr Ala Val Pr

#o Thr Pro Ala Arg Thr

3425

#               3430

#              3435

gca cgg acc cac cac gac gag cca ctc gcg at

#c gtc ggc atg gcg tgc    62630

Ala Arg Thr His His Asp Glu Pro Leu Ala Il

#e Val Gly Met Ala Cys

3440

#           3445

#          3450

cga ctg ccc ggc ggg gtc gcc tcg ccg gag ga

#c ctg tgg cag ctc gtg    62678

Arg Leu Pro Gly Gly Val Ala Ser Pro Glu As

#p Leu Trp Gln Leu Val

3455

#       3460

#      3465

gcg tcc ggc acc gac gcg atc acc gag ttc cc

#c acc gac cgc ggc tgg    62726

Ala Ser Gly Thr Asp Ala Ile Thr Glu Phe Pr

#o Thr Asp Arg Gly Trp

3470

#   3475

#  3480

gac atc gac cgg ctg ttc gac ccg gac ccg ga

#c gcc ccc ggc aag acc    62774

Asp Ile Asp Arg Leu Phe Asp Pro Asp Pro As

#p Ala Pro Gly Lys Thr

3485                3490

#                3495

#              3500

tac gtc cgg cac ggc ggc ttc ctc gcc gag gc

#c gcc ggc ttc gat gcc    62822

Tyr Val Arg His Gly Gly Phe Leu Ala Glu Al

#a Ala Gly Phe Asp Ala

3505

#               3510

#              3515

gcg ttc ttc ggc atc agc ccg cgc gag gca cg

#g gcc atg gac ccg cag    62870

Ala Phe Phe Gly Ile Ser Pro Arg Glu Ala Ar

#g Ala Met Asp Pro Gln

3520

#           3525

#          3530

cag cgc gtc atc ctc gaa acc tcc tgg gag gc

#g ttc gag aac gcg ggc    62918

Gln Arg Val Ile Leu Glu Thr Ser Trp Glu Al

#a Phe Glu Asn Ala Gly

3535

#       3540

#      3545

atc gtg ccg gac acg ctg cgc ggc agc gac ac

#c ggc gtg ttc atg ggc    62966

Ile Val Pro Asp Thr Leu Arg Gly Ser Asp Th

#r Gly Val Phe Met Gly

3550

#   3555

#  3560

gcg ttc tcc cat ggg tac ggc gcc ggc gtc ga

#c ctg ggc ggg ttc ggc    63014

Ala Phe Ser His Gly Tyr Gly Ala Gly Val As

#p Leu Gly Gly Phe Gly

3565                3570

#                3575

#              3580

gcc acc gcc acg cag aac agc gtg ctc tcc gg

#c cgg ttg tcg tac ttc    63062

Ala Thr Ala Thr Gln Asn Ser Val Leu Ser Gl

#y Arg Leu Ser Tyr Phe

3585

#               3590

#              3595

ttc ggc atg gag ggc ccg gcc gtc acc gtc ga

#c acc gcc tgc tcg tcg    63110

Phe Gly Met Glu Gly Pro Ala Val Thr Val As

#p Thr Ala Cys Ser Ser

3600

#           3605

#          3610

tcg ctg gtc gcc ctg cac cag gcg gca cag gc

#g ctg cgg act gga gaa    63158

Ser Leu Val Ala Leu His Gln Ala Ala Gln Al

#a Leu Arg Thr Gly Glu

3615

#       3620

#      3625

tgc tcg ctg gcg ctc gcc ggc ggt gtc acg gt

#g atg ccc acc ccg ctg    63206

Cys Ser Leu Ala Leu Ala Gly Gly Val Thr Va

#l Met Pro Thr Pro Leu

3630

#   3635

#  3640

ggc tac gtc gag ttc tgc cgc cag cgg gga ct

#c gcc ccc gac ggc cgt    63254

Gly Tyr Val Glu Phe Cys Arg Gln Arg Gly Le

#u Ala Pro Asp Gly Arg

3645                3650

#                3655

#              3660

tgc cag gcc ttc gcg gaa ggc gcc gac ggc ac

#g agc ttc tcg gag ggc    63302

Cys Gln Ala Phe Ala Glu Gly Ala Asp Gly Th

#r Ser Phe Ser Glu Gly

3665

#               3670

#              3675

gcc ggc gtt ctt gtg ctg gag cgg ctc tcc ga

#c gcc gag cgc aac gga    63350

Ala Gly Val Leu Val Leu Glu Arg Leu Ser As

#p Ala Glu Arg Asn Gly

3680

#           3685

#          3690

cac acc gtc ctc gcg gtc gtc cgc tcc tcc gc

#c gtc aac cag gac ggc    63398

His Thr Val Leu Ala Val Val Arg Ser Ser Al

#a Val Asn Gln Asp Gly

3695

#       3700

#      3705

gcc tcc aac ggc atc tcc gca ccc aac ggc cc

#c tcc cag cag cgc gtc    63446

Ala Ser Asn Gly Ile Ser Ala Pro Asn Gly Pr

#o Ser Gln Gln Arg Val

3710

#   3715

#  3720

atc cgc cag gcc ctc gac aag gcc ggg ctc gc

#c ccc gcc gac gtg gac    63494

Ile Arg Gln Ala Leu Asp Lys Ala Gly Leu Al

#a Pro Ala Asp Val Asp

3725                3730

#                3735

#              3740

gtg gtg gag gcc cac ggc acc gga acc ccg ct

#g ggc gac ccg atc gag    63542

Val Val Glu Ala His Gly Thr Gly Thr Pro Le

#u Gly Asp Pro Ile Glu

3745

#               3750

#              3755

gca cag gcc atc atc gcg acc tac ggc cag ga

#c cgc gac aca ccg ctc    63590

Ala Gln Ala Ile Ile Ala Thr Tyr Gly Gln As

#p Arg Asp Thr Pro Leu

3760

#           3765

#          3770

tac ctc ggt tcg gtc aag tcg aac atc gga ca

#c acc cag acc acc gcc    63638

Tyr Leu Gly Ser Val Lys Ser Asn Ile Gly Hi

#s Thr Gln Thr Thr Ala

3775

#       3780

#      3785

ggt gtc gcc ggc gtc atc aag atg gtc atg gc

#g atg cgc cac ggc atc    63686

Gly Val Ala Gly Val Ile Lys Met Val Met Al

#a Met Arg His Gly Ile

3790

#   3795

#  3800

gcg ccg aag aca ctg cac gtg gac gag ccg tc

#g tcg cat gtg gac tgg    63734

Ala Pro Lys Thr Leu His Val Asp Glu Pro Se

#r Ser His Val Asp Trp

3805                3810

#                3815

#              3820

acc gag ggt gcg gtg gaa ctg ctc acc gag gc

#g agg ccg tgg ccc gac    63782

Thr Glu Gly Ala Val Glu Leu Leu Thr Glu Al

#a Arg Pro Trp Pro Asp

3825

#               3830

#              3835

gcg gga cgc ccg cgc cgc gcg ggc gtg tcg tc

#g ctc ggt atc agc ggt    63830

Ala Gly Arg Pro Arg Arg Ala Gly Val Ser Se

#r Leu Gly Ile Ser Gly

3840

#           3845

#          3850

acg aac gcc cac gtg atc ctt gag ggt gtt cc

#c ggg ccg tcg cgt gtg    63878

Thr Asn Ala His Val Ile Leu Glu Gly Val Pr

#o Gly Pro Ser Arg Val

3855

#       3860

#      3865

gag ccg tct gtt gac ggg ttg gtg ccg ttg cc

#g gtg tcg gct cgg agt    63926

Glu Pro Ser Val Asp Gly Leu Val Pro Leu Pr

#o Val Ser Ala Arg Ser

3870

#   3875

#  3880

gag gcg agt ctg cgg ggg cag gtg gag cgg ct

#g gag ggg tat ctg cgc    63974

Glu Ala Ser Leu Arg Gly Gln Val Glu Arg Le

#u Glu Gly Tyr Leu Arg

3885                3890

#                3895

#              3900

ggg agt gtg gat gtg gcc gcg gtc gcg cag gg

#g ttg gtg cgt gag cgt    64022

Gly Ser Val Asp Val Ala Ala Val Ala Gln Gl

#y Leu Val Arg Glu Arg

3905

#               3910

#              3915

gct gtc ttc ggt cac cgt gcg gta ctg ctg gg

#t gat gcc cgg gtg atg    64070

Ala Val Phe Gly His Arg Ala Val Leu Leu Gl

#y Asp Ala Arg Val Met

3920

#           3925

#          3930

ggt gtg gcg gtg gat cag ccg cgt acg gtg tt

#c gtc ttt ccc ggg cag    64118

Gly Val Ala Val Asp Gln Pro Arg Thr Val Ph

#e Val Phe Pro Gly Gln

3935

#       3940

#      3945

ggt gct cag tgg gtg ggc atg ggt gtg gag tt

#g atg gac cgt tct gcg    64166

Gly Ala Gln Trp Val Gly Met Gly Val Glu Le

#u Met Asp Arg Ser Ala

3950

#   3955

#  3960

gtg ttc gcg gct cgt atg gag gag tgt gcg cg

#g gcg ttg ttg ccg cac    64214

Val Phe Ala Ala Arg Met Glu Glu Cys Ala Ar

#g Ala Leu Leu Pro His

3965                3970

#                3975

#              3980

acg ggc tgg gat gtg cgg gag atg ttg gcg cg

#g ccg gat gtg gcg gag    64262

Thr Gly Trp Asp Val Arg Glu Met Leu Ala Ar

#g Pro Asp Val Ala Glu

3985

#               3990

#              3995

cgg gtg gag gtg gtc cag ccg gcc agc tgg gc

#g gtc gcg gtc agc ctg    64310

Arg Val Glu Val Val Gln Pro Ala Ser Trp Al

#a Val Ala Val Ser Leu

4000

#           4005

#          4010

gcc gca ctg tgg cag gcc cac ggg gtc gta cc

#c gac gcg gtg atc gga    64358

Ala Ala Leu Trp Gln Ala His Gly Val Val Pr

#o Asp Ala Val Ile Gly

4015

#       4020

#      4025

cac tcc cag ggc gag atc gcg gcg gcg tgc gt

#g gcc ggg gcc ctc agc    64406

His Ser Gln Gly Glu Ile Ala Ala Ala Cys Va

#l Ala Gly Ala Leu Ser

4030

#   4035

#  4040

ctt gag gac gcc gcc cgc gtg gtg gcc ttg cg

#c agc cag gtc atc gcg    64454

Leu Glu Asp Ala Ala Arg Val Val Ala Leu Ar

#g Ser Gln Val Ile Ala

4045                4050

#                4055

#              4060

gcg cga ctg gcc ggg cgg gga gcg atg gct tc

#g gtg gca ttg ccg gcc    64502

Ala Arg Leu Ala Gly Arg Gly Ala Met Ala Se

#r Val Ala Leu Pro Ala

4065

#               4070

#              4075

ggt gag gtc ggt ctg gtc gag ggc gtg tgg at

#c gcg gcg cgt aac ggc    64550

Gly Glu Val Gly Leu Val Glu Gly Val Trp Il

#e Ala Ala Arg Asn Gly

4080

#           4085

#          4090

ccc gcc tcg aca gtc gtg gcc ggc gag ccg tc

#g gcg gtg gag gac gtg    64598

Pro Ala Ser Thr Val Val Ala Gly Glu Pro Se

#r Ala Val Glu Asp Val

4095

#       4100

#      4105

gtg acg cgg tat gag acc gaa ggc gtg cga gt

#g cgt cgt atc gcc gtc    64646

Val Thr Arg Tyr Glu Thr Glu Gly Val Arg Va

#l Arg Arg Ile Ala Val

4110

#   4115

#  4120

gac tac gcc tcc cac acg ccc cac gtg gaa gc

#c atc gag gac gaa ctc    64694

Asp Tyr Ala Ser His Thr Pro His Val Glu Al

#a Ile Glu Asp Glu Leu

4125                4130

#                4135

#              4140

gct gag gta ctg aag gga gtt gca ggg aag gc

#c gcg tcg gtg gcg tgg    64742

Ala Glu Val Leu Lys Gly Val Ala Gly Lys Al

#a Ala Ser Val Ala Trp

4145

#               4150

#              4155

tgg tcg acc gtg gac agc gcc tgg gtg acc ga

#g ccg gtg gat gag agt    64790

Trp Ser Thr Val Asp Ser Ala Trp Val Thr Gl

#u Pro Val Asp Glu Ser

4160

#           4165

#          4170

tac tgg tac cgg aac ctg cgt cgc ccc gtc gc

#g ctg gac gcg gcg gtg    64838

Tyr Trp Tyr Arg Asn Leu Arg Arg Pro Val Al

#a Leu Asp Ala Ala Val

4175

#       4180

#      4185

gcg gag ctg gac ggg tcc gtg ttc gtg gag tg

#c agc gcc cat ccg gtg    64886

Ala Glu Leu Asp Gly Ser Val Phe Val Glu Cy

#s Ser Ala His Pro Val

4190

#   4195

#  4200

ctg ctg ccg gcg atg gaa cag gcc cac acg gt

#g gcg tcg ttg cgc acc    64934

Leu Leu Pro Ala Met Glu Gln Ala His Thr Va

#l Ala Ser Leu Arg Thr

4205                4210

#                4215

#              4220

ggt gac ggc ggc tgg gag cga tgg ctg acg gc

#g ttg gcg cag gcg tgg    64982

Gly Asp Gly Gly Trp Glu Arg Trp Leu Thr Al

#a Leu Ala Gln Ala Trp

4225

#               4230

#              4235

acc ctg ggc gcg gca gtg gac tgg gac acg gt

#g gtc gaa ccg gtg cca    65030

Thr Leu Gly Ala Ala Val Asp Trp Asp Thr Va

#l Val Glu Pro Val Pro

4240

#           4245

#          4250

ggg cgg ctg ctc gat ctg ccc acc tac gcg tt

#c gag cgc cgg cgc tac    65078

Gly Arg Leu Leu Asp Leu Pro Thr Tyr Ala Ph

#e Glu Arg Arg Arg Tyr

4255

#       4260

#      4265

tgg ctg gaa gcg gcc ggt gcc acc gac ctg tc

#c gcg gcc ggg ctg aca    65126

Trp Leu Glu Ala Ala Gly Ala Thr Asp Leu Se

#r Ala Ala Gly Leu Thr

4270

#   4275

#  4280

ggg gca gca cat ccc atg ctg gcc gcc atc ac

#g gca cta ccc gcc gac    65174

Gly Ala Ala His Pro Met Leu Ala Ala Ile Th

#r Ala Leu Pro Ala Asp

4285                4290

#                4295

#              4300

gac ggt ggt gtt gtt ctc acc ggc cgg atc tc

#g ttg cgc acg cat ccc    65222

Asp Gly Gly Val Val Leu Thr Gly Arg Ile Se

#r Leu Arg Thr His Pro

4305

#               4310

#              4315

tgg ctg gct gat cac gcg gtg cgg ggc acg gt

#c ctg ctg ccg ggc acg    65270

Trp Leu Ala Asp His Ala Val Arg Gly Thr Va

#l Leu Leu Pro Gly Thr

4320

#           4325

#          4330

gcc ttt gtg gag ctg gtc atc cgg gcc ggt ga

#c gag acc ggt tgc ggg    65318

Ala Phe Val Glu Leu Val Ile Arg Ala Gly As

#p Glu Thr Gly Cys Gly

4335

#       4340

#      4345

ata gtg gat gaa ctg gtc atc gaa tcc ccc ct

#c gtg gtg ccg gcg acc    65366

Ile Val Asp Glu Leu Val Ile Glu Ser Pro Le

#u Val Val Pro Ala Thr

4350

#   4355

#  4360

gca gcc gtg gat ctg tcg gtg acc gtg gaa gg

#a gct gac gag gcc gga    65414

Ala Ala Val Asp Leu Ser Val Thr Val Glu Gl

#y Ala Asp Glu Ala Gly

4365                4370

#                4375

#              4380

cgg cgg cga gtg acc gtc cac gcc cgc acc ga

#a ggc acc ggc agc tgg    65462

Arg Arg Arg Val Thr Val His Ala Arg Thr Gl

#u Gly Thr Gly Ser Trp

4385

#               4390

#              4395

acc cgg cac gcc agc ggc acc ctg acc ccc ga

#c acc ccc gac acc ccc    65510

Thr Arg His Ala Ser Gly Thr Leu Thr Pro As

#p Thr Pro Asp Thr Pro

4400

#           4405

#          4410

aac gct tcc ggt gtt gtc ggt gcg gag ccg tt

#c tcg cag tgg cca cct    65558

Asn Ala Ser Gly Val Val Gly Ala Glu Pro Ph

#e Ser Gln Trp Pro Pro

4415

#       4420

#      4425

gcc act gcc gcg gcc gtc gac acc tcg gag tt

#c tac ttg cgc ctg gac    65606

Ala Thr Ala Ala Ala Val Asp Thr Ser Glu Ph

#e Tyr Leu Arg Leu Asp

4430

#   4435

#  4440

gcg ctg ggc tac cgg ttc gga ccc atg ttc cg

#c gga atg cgg gct gcc    65654

Ala Leu Gly Tyr Arg Phe Gly Pro Met Phe Ar

#g Gly Met Arg Ala Ala

4445                4450

#                4455

#              4460

tgg cgt gat ggt gac acc gtg tac gcc gag gt

#c gcg ctc ccc gag gac    65702

Trp Arg Asp Gly Asp Thr Val Tyr Ala Glu Va

#l Ala Leu Pro Glu Asp

4465

#               4470

#              4475

cgt gcc gcc gac gcg gac ggt ttc ggc atg ca

#c ccg gcg ctg ctc gac    65750

Arg Ala Ala Asp Ala Asp Gly Phe Gly Met Hi

#s Pro Ala Leu Leu Asp

4480

#           4485

#          4490

gcg gcc ttg cag agc ggc agc ctg ctc atg ct

#g gaa tcg gac ggc gag    65798

Ala Ala Leu Gln Ser Gly Ser Leu Leu Met Le

#u Glu Ser Asp Gly Glu

4495

#       4500

#      4505

cag agc gtg caa ctg ccg ttc tcc tgg cac gg

#c gtc cgg ttc cac gcg    65846

Gln Ser Val Gln Leu Pro Phe Ser Trp His Gl

#y Val Arg Phe His Ala

4510

#   4515

#  4520

acg ggc gcg acc atg ctg cgg gtg gcg gtc gt

#a ccg ggc ccg gac ggc    65894

Thr Gly Ala Thr Met Leu Arg Val Ala Val Va

#l Pro Gly Pro Asp Gly

4525                4530

#                4535

#              4540

ctc cgg ctg cat gcc gcg gac agc ggg aac cg

#t ccc gtc gcg acg atc    65942

Leu Arg Leu His Ala Ala Asp Ser Gly Asn Ar

#g Pro Val Ala Thr Ile

4545

#               4550

#              4555

gac gcg ctc gtg acc cgg tcc ccg gaa gcg ga

#c ctc gcg ccc gcc gat    65990

Asp Ala Leu Val Thr Arg Ser Pro Glu Ala As

#p Leu Ala Pro Ala Asp

4560

#           4565

#          4570

ccg atg ctg cgg gtc ggg tgg gcc ccg gtg cc

#c gta cct gcc ggg gcc    66038

Pro Met Leu Arg Val Gly Trp Ala Pro Val Pr

#o Val Pro Ala Gly Ala

4575

#       4580

#      4585

ggt ccg tcc gac gcg gac gtg ctg acg ctg cg

#c ggc gac gac gcc gac    66086

Gly Pro Ser Asp Ala Asp Val Leu Thr Leu Ar

#g Gly Asp Asp Ala Asp

4590

#   4595

#  4600

ccg ctc ggg gag acc cgg gac ctg acc acc cg

#t gtt ctc gac gcg ctg    66134

Pro Leu Gly Glu Thr Arg Asp Leu Thr Thr Ar

#g Val Leu Asp Ala Leu

4605                4610

#                4615

#              4620

ctc cgg gcc gac cgg ccg gtg atc ttc cag gt

#g acc ggt ggc ctc gcc    66182

Leu Arg Ala Asp Arg Pro Val Ile Phe Gln Va

#l Thr Gly Gly Leu Ala

4625

#               4630

#              4635

gcc aag gcg gcc gca ggc ctg gtc cgc acc gc

#t cag aac gag cag ccc    66230

Ala Lys Ala Ala Ala Gly Leu Val Arg Thr Al

#a Gln Asn Glu Gln Pro

4640

#           4645

#          4650

ggc cgc ttc ttc ctc gtc gaa acg gac ccg gg

#a gag gtc ctg gac ggc    66278

Gly Arg Phe Phe Leu Val Glu Thr Asp Pro Gl

#y Glu Val Leu Asp Gly

4655

#       4660

#      4665

gcg aag cgc gac gcg atc gcg gca ctc ggc ga

#g ccc cat gtg cgg ctg    66326

Ala Lys Arg Asp Ala Ile Ala Ala Leu Gly Gl

#u Pro His Val Arg Leu

4670

#   4675

#  4680

cgc gac ggc ctc ttc gag gca gcc cgg ctg at

#g cgg gcc acg ccg tcc    66374

Arg Asp Gly Leu Phe Glu Ala Ala Arg Leu Me

#t Arg Ala Thr Pro Ser

4685                4690

#                4695

#              4700

ctg acg ctc ccg gac acc ggg tcg tgg cag ct

#g cgg ccg tcc gcc acc    66422

Leu Thr Leu Pro Asp Thr Gly Ser Trp Gln Le

#u Arg Pro Ser Ala Thr

4705

#               4710

#              4715

ggt tcc ctc gac gac ctt gcc gtc gtc ccc ac

#c gac gcc ccg gac cgg    66470

Gly Ser Leu Asp Asp Leu Ala Val Val Pro Th

#r Asp Ala Pro Asp Arg

4720

#           4725

#          4730

ccg ctc gcg gcc ggc gag gtg cgg atc gcg gt

#a cgc gcg gcg ggc ctg    66518

Pro Leu Ala Ala Gly Glu Val Arg Ile Ala Va

#l Arg Ala Ala Gly Leu

4735

#       4740

#      4745

aac ttc cgg gat gtc acg gtc gcg ctc ggt gt

#g gtc gcc gat gcg cgt    66566

Asn Phe Arg Asp Val Thr Val Ala Leu Gly Va

#l Val Ala Asp Ala Arg

4750

#   4755

#  4760

ccg ctc ggc agc gag gcc gcg ggt gtc gtc ct

#g gag acc ggc ccc ggt    66614

Pro Leu Gly Ser Glu Ala Ala Gly Val Val Le

#u Glu Thr Gly Pro Gly

4765                4770

#                4775

#              4780

gtg cac gac ctg gcg ccc ggc gac cgg gtc ct

#g ggg atg ctc gcg ggc    66662

Val His Asp Leu Ala Pro Gly Asp Arg Val Le

#u Gly Met Leu Ala Gly

4785

#               4790

#              4795

gcc ttc gga ccg gtc gcg atc acc gac cgg cg

#g ctg ctc ggc cgg atg    66710

Ala Phe Gly Pro Val Ala Ile Thr Asp Arg Ar

#g Leu Leu Gly Arg Met

4800

#           4805

#          4810

ccg gac ggc tgg acg ttc ccg cag gcg gcg tc

#c gtg atg acc gcg ttc    66758

Pro Asp Gly Trp Thr Phe Pro Gln Ala Ala Se

#r Val Met Thr Ala Phe

4815

#       4820

#      4825

gcg acc gcg tgg tac ggc ctg gtc gac ctg gc

#c ggg ctg cgc ccc ggc    66806

Ala Thr Ala Trp Tyr Gly Leu Val Asp Leu Al

#a Gly Leu Arg Pro Gly

4830

#   4835

#  4840

gag aag gtc ctg atc cac gcg gcg gcg acc gg

#t gtc ggc gcg gcg gcc    66854

Glu Lys Val Leu Ile His Ala Ala Ala Thr Gl

#y Val Gly Ala Ala Ala

4845                4850

#                4855

#              4860

gtc cag atc gcg cgg cat ctg ggc gcg gag gt

#g tac gcg acc acc agc    66902

Val Gln Ile Ala Arg His Leu Gly Ala Glu Va

#l Tyr Ala Thr Thr Ser

4865

#               4870

#              4875

gcc gcg aag cgc cat ctg gtg gac ctg gac gg

#a gcg cat ctg gcc gat    66950

Ala Ala Lys Arg His Leu Val Asp Leu Asp Gl

#y Ala His Leu Ala Asp

4880

#           4885

#          4890

tcc cgc agc acc gcg ttc gcc gac gcg ttc cc

#g ccg gtc gat gtc gtg    66998

Ser Arg Ser Thr Ala Phe Ala Asp Ala Phe Pr

#o Pro Val Asp Val Val

4895

#       4900

#      4905

ctc aac tcg ctc acc ggt gaa ttc ctc gac gc

#g tcc gtc ggc ctg ctc    67046

Leu Asn Ser Leu Thr Gly Glu Phe Leu Asp Al

#a Ser Val Gly Leu Leu

4910

#   4915

#  4920

gcg gcg ggt ggc cgg ttc atc gag atg ggg aa

#g acg gac atc cgg cac    67094

Ala Ala Gly Gly Arg Phe Ile Glu Met Gly Ly

#s Thr Asp Ile Arg His

4925                4930

#                4935

#              4940

gcc gtc cag cag ccg ttc gac ctg atg gac gc

#c ggc ccc gac cgg atg    67142

Ala Val Gln Gln Pro Phe Asp Leu Met Asp Al

#a Gly Pro Asp Arg Met

4945

#               4950

#              4955

cag cgg atc atc gtc gag ctg ctc ggc ctg tt

#c gcg cgc gac gtg ctg    67190

Gln Arg Ile Ile Val Glu Leu Leu Gly Leu Ph

#e Ala Arg Asp Val Leu

4960

#           4965

#          4970

cac ccg ctg ccg gtc cac gcc tgg gac gtg cg

#g cag gcg cgg gag gcg    67238

His Pro Leu Pro Val His Ala Trp Asp Val Ar

#g Gln Ala Arg Glu Ala

4975

#       4980

#      4985

ttc ggc tgg atg agc agc ggg cgt cac acc gg

#c aag ctg gtg ctg acg    67286

Phe Gly Trp Met Ser Ser Gly Arg His Thr Gl

#y Lys Leu Val Leu Thr

4990

#   4995

#  5000

gtc ccg cgg ccg ctg gat ccc gag ggg gcc gt

#c gtc atc acc ggc ggc    67334

Val Pro Arg Pro Leu Asp Pro Glu Gly Ala Va

#l Val Ile Thr Gly Gly

5005                5010

#                5015

#              5020

tcc ggc acc ctc gcc ggc atc ctc gcc cgc ca

#c ctg ggc cac ccc cac    67382

Ser Gly Thr Leu Ala Gly Ile Leu Ala Arg Hi

#s Leu Gly His Pro His

5025

#               5030

#              5035

acc tac ctg ctc tcc cgc acc cca ccc ccc ga

#c acc acc ccc ggc acc    67430

Thr Tyr Leu Leu Ser Arg Thr Pro Pro Pro As

#p Thr Thr Pro Gly Thr

5040

#           5045

#          5050

cac ctc ccc tgc gac gtc ggc gac ccc cac ca

#a ctc gcc acc acc ctc    67478

His Leu Pro Cys Asp Val Gly Asp Pro His Gl

#n Leu Ala Thr Thr Leu

5055

#       5060

#      5065

gcc cgc atc ccc caa ccc ctc acc gcc gtc tt

#c cac acc gcc gga acc    67526

Ala Arg Ile Pro Gln Pro Leu Thr Ala Val Ph

#e His Thr Ala Gly Thr

5070

#   5075

#  5080

ctc gac gac gcc ctg ctc gac aac ctc acc cc

#c gac cgc gtc gac acc    67574

Leu Asp Asp Ala Leu Leu Asp Asn Leu Thr Pr

#o Asp Arg Val Asp Thr

5085                5090

#                5095

#              5100

gtc ctc aaa ccc aag gcc gac gcc gcc tgg ca

#c ctg cac cgg ctc acc    67622

Val Leu Lys Pro Lys Ala Asp Ala Ala Trp Hi

#s Leu His Arg Leu Thr

5105

#               5110

#              5115

cgc gac acc gac ctc gcc gcg ttc gtc gtc ta

#c tcc gcg gtc gcc ggc    67670

Arg Asp Thr Asp Leu Ala Ala Phe Val Val Ty

#r Ser Ala Val Ala Gly

5120

#           5125

#          5130

ctc atg ggc agc ccg ggg cag ggc aac tac gt

#c gcg gcg aac gcg ttc    67718

Leu Met Gly Ser Pro Gly Gln Gly Asn Tyr Va

#l Ala Ala Asn Ala Phe

5135

#       5140

#      5145

ctc gac gcg ctc gcc gaa cac cgc cgt gcg ca

#a ggg ctg ccc gcg cag    67766

Leu Asp Ala Leu Ala Glu His Arg Arg Ala Gl

#n Gly Leu Pro Ala Gln

5150

#   5155

#  5160

tcc ctc gca tgg ggc atg tgg gcg gac gtc ag

#c gcg ctc acc gcg aaa    67814

Ser Leu Ala Trp Gly Met Trp Ala Asp Val Se

#r Ala Leu Thr Ala Lys

5165                5170

#                5175

#              5180

ctc acc gac gcg gac cgc cag cgc atc cgg cg

#c agc gga ttc ccg ccg    67862

Leu Thr Asp Ala Asp Arg Gln Arg Ile Arg Ar

#g Ser Gly Phe Pro Pro

5185

#               5190

#              5195

ttg agc gcc gcg gac ggc atg cgg ctg ttc ga

#c gcg gcg acg cgt acc    67910

Leu Ser Ala Ala Asp Gly Met Arg Leu Phe As

#p Ala Ala Thr Arg Thr

5200

#           5205

#          5210

ccg gaa ccg gtc gtc gtc gcg acg acc gtc ga

#c ctc acc cag ctc gac    67958

Pro Glu Pro Val Val Val Ala Thr Thr Val As

#p Leu Thr Gln Leu Asp

5215

#       5220

#      5225

ggc gcc gtc gcg ccg ttg ctc cgc ggt ctg gc

#c gcg cac cgg gcc ggg    68006

Gly Ala Val Ala Pro Leu Leu Arg Gly Leu Al

#a Ala His Arg Ala Gly

5230

#   5235

#  5240

ccg gcg cgc acg gtc gcc cgc aac gcc ggc ga

#a gag ccc ctg gcc gtg    68054

Pro Ala Arg Thr Val Ala Arg Asn Ala Gly Gl

#u Glu Pro Leu Ala Val

5245                5250

#                5255

#              5260

cgt ctt gcc ggg cgt acc gcc gcc gag cag cg

#g cgc atc atg cag gag    68102

Arg Leu Ala Gly Arg Thr Ala Ala Glu Gln Ar

#g Arg Ile Met Gln Glu

5265

#               5270

#              5275

gtc gtg ctc cgc cac gcg gcc gcg gtc ctc gc

#g tac ggg ctg ggc gac    68150

Val Val Leu Arg His Ala Ala Ala Val Leu Al

#a Tyr Gly Leu Gly Asp

5280

#           5285

#          5290

cgc gtg gcg gcg gac cgt ccg ttc cgc gag ct

#c ggt ttc gat tcg ctg    68198

Arg Val Ala Ala Asp Arg Pro Phe Arg Glu Le

#u Gly Phe Asp Ser Leu

5295

#       5300

#      5305

acc gcg gtc gac ctg cgc aat cgg ctc gcg gc

#c gag acg ggg ctg cgg    68246

Thr Ala Val Asp Leu Arg Asn Arg Leu Ala Al

#a Glu Thr Gly Leu Arg

5310

#   5315

#  5320

ctg ccg acg acg ctg gtg ttc agc cac ccg ac

#g gcg gag gcg ctc acc    68294

Leu Pro Thr Thr Leu Val Phe Ser His Pro Th

#r Ala Glu Ala Leu Thr

5325                5330

#                5335

#              5340

gcc cac ctg ctc gac ctg atc gac gct ccc ac

#c gcc cgg atc gcc ggg    68342

Ala His Leu Leu Asp Leu Ile Asp Ala Pro Th

#r Ala Arg Ile Ala Gly

5345

#               5350

#              5355

gag tcc ctg ccc gcg gtg acg gcc gct ccc gt

#g gcg gcc gcg cgg gac    68390

Glu Ser Leu Pro Ala Val Thr Ala Ala Pro Va

#l Ala Ala Ala Arg Asp

5360

#           5365

#          5370

cag gac gag ccg atc gcc atc gtg gcg atg gc

#g tgc cgg ctg ccc ggt    68438

Gln Asp Glu Pro Ile Ala Ile Val Ala Met Al

#a Cys Arg Leu Pro Gly

5375

#       5380

#      5385

ggt gtg acg tcg ccc gag gac ctg tgg cgg ct

#c gtc gag tcc ggc acc    68486

Gly Val Thr Ser Pro Glu Asp Leu Trp Arg Le

#u Val Glu Ser Gly Thr

5390

#   5395

#  5400

gac gcg atc acc acg cct cct gac gac cgc gg

#c tgg gac gtc gac gcg    68534

Asp Ala Ile Thr Thr Pro Pro Asp Asp Arg Gl

#y Trp Asp Val Asp Ala

5405                5410

#                5415

#              5420

ctg tac gac gcg gac ccg gac gcg gcc ggc aa

#g gcg tac aac ctg cgg    68582

Leu Tyr Asp Ala Asp Pro Asp Ala Ala Gly Ly

#s Ala Tyr Asn Leu Arg

5425

#               5430

#              5435

ggc ggt tac ctg gcc ggg gcg gcg gag ttc ga

#c gcg gcg ttc ttc gac    68630

Gly Gly Tyr Leu Ala Gly Ala Ala Glu Phe As

#p Ala Ala Phe Phe Asp

5440

#           5445

#          5450

atc agt ccg cgc gaa gcg ctc ggc atg gac cc

#g cag caa cgc ctg ctg    68678

Ile Ser Pro Arg Glu Ala Leu Gly Met Asp Pr

#o Gln Gln Arg Leu Leu

5455

#       5460

#      5465

ctc gaa acg gcg tgg gag gcg atc gag cgc gg

#c cgg atc agt ccg gcg    68726

Leu Glu Thr Ala Trp Glu Ala Ile Glu Arg Gl

#y Arg Ile Ser Pro Ala

5470

#   5475

#  5480

tcg ctc cgc ggc cgg gag gtc ggc gtc tat gt

#c ggt gcg gcc gcg cag    68774

Ser Leu Arg Gly Arg Glu Val Gly Val Tyr Va

#l Gly Ala Ala Ala Gln

5485                5490

#                5495

#              5500

ggc tac ggg ctg ggc gcc gag gac acc gag gg

#c cac gcg atc acc ggt    68822

Gly Tyr Gly Leu Gly Ala Glu Asp Thr Glu Gl

#y His Ala Ile Thr Gly

5505

#               5510

#              5515

ggt tcc acg agc ctg ctg tcc gga cgg ctg gc

#g tac gtg ctc ggg ctg    68870

Gly Ser Thr Ser Leu Leu Ser Gly Arg Leu Al

#a Tyr Val Leu Gly Leu

5520

#           5525

#          5530

gag ggc ccg gcg gtc acc gtg gac acg gcg tg

#c tcg tcg tct ctg gtc    68918

Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cy

#s Ser Ser Ser Leu Val

5535

#       5540

#      5545

gcg ctg cat ctg gcg tgc cag ggg ctg cgc ct

#g ggc gag tgc gaa ctc    68966

Ala Leu His Leu Ala Cys Gln Gly Leu Arg Le

#u Gly Glu Cys Glu Leu

5550

#   5555

#  5560

gct ctg gcc gga ggg gtc tcc gta ctg agt tc

#g ccg gcc gcg ttc gtg    69014

Ala Leu Ala Gly Gly Val Ser Val Leu Ser Se

#r Pro Ala Ala Phe Val

5565                5570

#                5575

#              5580

gag ttc tcc cgc cag cgc ggg ctc gcg gcc ga

#c ggg cgc tgc aag tcg    69062

Glu Phe Ser Arg Gln Arg Gly Leu Ala Ala As

#p Gly Arg Cys Lys Ser

5585

#               5590

#              5595

ttc ggc gcg ggc gcg gac ggc acg acg tgg tc

#c gag ggc gtg ggc gtg    69110

Phe Gly Ala Gly Ala Asp Gly Thr Thr Trp Se

#r Glu Gly Val Gly Val

5600

#           5605

#          5610

ctc gta ctg gaa cgg ctc tcc gac gcc gag cg

#g ctc ggg cac acc gtg    69158

Leu Val Leu Glu Arg Leu Ser Asp Ala Glu Ar

#g Leu Gly His Thr Val

5615

#       5620

#      5625

ctc gcc gtc gtc cgc ggc agc gcc gtc acg tc

#c gac ggc gcc tcc aac    69206

Leu Ala Val Val Arg Gly Ser Ala Val Thr Se

#r Asp Gly Ala Ser Asn

5630

#   5635

#  5640

ggc ctc acc gcg ccg aac ggg ctc tcg cag ca

#g cgg gtc atc cgg aag    69254

Gly Leu Thr Ala Pro Asn Gly Leu Ser Gln Gl

#n Arg Val Ile Arg Lys

5645                5650

#                5655

#              5660

gcg ctc gcc gcg gcc ggg ctg acc ggc gcc ga

#c gtg gac gtc gtc gag    69302

Ala Leu Ala Ala Ala Gly Leu Thr Gly Ala As

#p Val Asp Val Val Glu

5665

#               5670

#              5675

ggg cac ggc acc ggc acc cgg ctc ggc gac cc

#g gtc gag gcg gac gcg    69350

Gly His Gly Thr Gly Thr Arg Leu Gly Asp Pr

#o Val Glu Ala Asp Ala

5680

#           5685

#          5690

ctg ctc gcg acg tac ggg cag gac cgt ccg gc

#a ccg gtc tgg ctg ggc    69398

Leu Leu Ala Thr Tyr Gly Gln Asp Arg Pro Al

#a Pro Val Trp Leu Gly

5695

#       5700

#      5705

tcg ctg aag tcg aac atc gga cat gcc acg gc

#c gcg gcc ggt gtc gcg    69446

Ser Leu Lys Ser Asn Ile Gly His Ala Thr Al

#a Ala Ala Gly Val Ala

5710

#   5715

#  5720

ggc gtc atc aag atg gtg cag gcg atc ggc gc

#g ggc acg atg ccg cgg    69494

Gly Val Ile Lys Met Val Gln Ala Ile Gly Al

#a Gly Thr Met Pro Arg

5725                5730

#                5735

#              5740

acg ctg cat gtg gag gag ccc tcg ccc gcc gt

#c gac tgg agc acc gga    69542

Thr Leu His Val Glu Glu Pro Ser Pro Ala Va

#l Asp Trp Ser Thr Gly

5745

#               5750

#              5755

cag gtg tcc ctg ctc ggc tcc aac cgg ccc tg

#g ccg gac gac gag cgt    69590

Gln Val Ser Leu Leu Gly Ser Asn Arg Pro Tr

#p Pro Asp Asp Glu Arg

5760

#           5765

#          5770

ccg cgc cgg gcg gcc gtc tcc gcg ttc ggg ct

#c agc ggg acg aac gcg    69638

Pro Arg Arg Ala Ala Val Ser Ala Phe Gly Le

#u Ser Gly Thr Asn Ala

5775

#       5780

#      5785

cac gtc atc ctg gaa cag cac cgt ccg gcg cc

#c gtg gcg tcc cag ccg    69686

His Val Ile Leu Glu Gln His Arg Pro Ala Pr

#o Val Ala Ser Gln Pro

5790

#   5795

#  5800

ccc cgg ccg ccc cgt gag gag tcc cag ccg ct

#g ccg tgg gtg ctc tcc    69734

Pro Arg Pro Pro Arg Glu Glu Ser Gln Pro Le

#u Pro Trp Val Leu Ser

5805                5810

#                5815

#              5820

gcg cgg act ccg gcc gcg ctg cgg gcc cag gc

#g gcc cgg ctg cgc gac    69782

Ala Arg Thr Pro Ala Ala Leu Arg Ala Gln Al

#a Ala Arg Leu Arg Asp

5825

#               5830

#              5835

cac ctc gcg gcg gca ccg gac gcg gat ccg tt

#g gac atc ggg tac gcg    69830

His Leu Ala Ala Ala Pro Asp Ala Asp Pro Le

#u Asp Ile Gly Tyr Ala

5840

#           5845

#          5850

ctg gcc acc agc cgc gcc cag ttc gcc cac cg

#t gcc gcg gtc gtc gcc    69878

Leu Ala Thr Ser Arg Ala Gln Phe Ala His Ar

#g Ala Ala Val Val Ala

5855

#       5860

#      5865

acc acc ccg gac gga ttc cgt gcc gcg ctc ga

#c ggc ctc gcg gac ggc    69926

Thr Thr Pro Asp Gly Phe Arg Ala Ala Leu As

#p Gly Leu Ala Asp Gly

5870

#   5875

#  5880

gcg gag gcg ccc gga gtc gtc acc ggg acc gc

#t cag gag cgg cgc gtc    69974

Ala Glu Ala Pro Gly Val Val Thr Gly Thr Al

#a Gln Glu Arg Arg Val

5885                5890

#                5895

#              5900

gcc ttc ctc ttc gac ggc cag ggc gcc cag cg

#c gcc gga atg ggg cgc    70022

Ala Phe Leu Phe Asp Gly Gln Gly Ala Gln Ar

#g Ala Gly Met Gly Arg

5905

#               5910

#              5915

gag ctc cac cgc cgg ttc ccc gtc ttc gcc gc

#c gcg tgg gac gag gtc    70070

Glu Leu His Arg Arg Phe Pro Val Phe Ala Al

#a Ala Trp Asp Glu Val

5920

#           5925

#          5930

tcc gac gcg ttc ggc aag cac ctc aag cac tc

#c ccc acg gac gtc tac    70118

Ser Asp Ala Phe Gly Lys His Leu Lys His Se

#r Pro Thr Asp Val Tyr

5935

#       5940

#      5945

cac ggc gaa cac ggc gct ctc gcc cat gac ac

#c ctg tac gcc cag gcc    70166

His Gly Glu His Gly Ala Leu Ala His Asp Th

#r Leu Tyr Ala Gln Ala

5950

#   5955

#  5960

ggc ctg ttc acg ctc gaa gtg gcg ctg ctg cg

#g ctg ctg gag cac tgg    70214

Gly Leu Phe Thr Leu Glu Val Ala Leu Leu Ar

#g Leu Leu Glu His Trp

5965                5970

#                5975

#              5980

ggg gtg cgg ccg gac gtg ctc gtc ggg cac tc

#c gtc ggc gag gtg acc    70262

Gly Val Arg Pro Asp Val Leu Val Gly His Se

#r Val Gly Glu Val Thr

5985

#               5990

#              5995

gcg gcg tac gcg gcg ggg gtg ctc acc ctg gc

#g gac gcg acg gag ttg    70310

Ala Ala Tyr Ala Ala Gly Val Leu Thr Leu Al

#a Asp Ala Thr Glu Leu

6000

#           6005

#          6010

atc gtg gcc cgg ggg cgg gcg ctg cgg gcg ct

#g ccg ccc ggg gcg atg    70358

Ile Val Ala Arg Gly Arg Ala Leu Arg Ala Le

#u Pro Pro Gly Ala Met

6015

#       6020

#      6025

ctc gcc gtc gac gga agc ccg gcg gag gtc gg

#c gcc cgc acg gat ctg    70406

Leu Ala Val Asp Gly Ser Pro Ala Glu Val Gl

#y Ala Arg Thr Asp Leu

6030

#   6035

#  6040

gac atc gcc gcg gtc aac ggc ccg tcc gcc gt

#g gtg ctc gcc ggt tcg    70454

Asp Ile Ala Ala Val Asn Gly Pro Ser Ala Va

#l Val Leu Ala Gly Ser

6045                6050

#                6055

#              6060

ccg gac gat gtg gcg gcg ttc gaa cgg gag tg

#g tcg gcg gcc ggg cgg    70502

Pro Asp Asp Val Ala Ala Phe Glu Arg Glu Tr

#p Ser Ala Ala Gly Arg

6065

#               6070

#              6075

cgc acg aaa cgg ctc gac gtc ggg cac gcg tt

#c cac tcc cgg cac gtc    70550

Arg Thr Lys Arg Leu Asp Val Gly His Ala Ph

#e His Ser Arg His Val

6080

#           6085

#          6090

gac ggt gcg ctc gac ggc ttc cgt acg gtg ct

#g gag tcg ctc gcg ttc    70598

Asp Gly Ala Leu Asp Gly Phe Arg Thr Val Le

#u Glu Ser Leu Ala Phe

6095

#       6100

#      6105

ggc gcg gcg cgg ctg ccg gtg gtg tcc acg ac

#g acg ggc cgg gac gcc    70646

Gly Ala Ala Arg Leu Pro Val Val Ser Thr Th

#r Thr Gly Arg Asp Ala

6110

#   6115

#  6120

gcg gac gac ctc ata acg ccc gcg cac tgg ct

#g cgc cat gcg cgt cgg    70694

Ala Asp Asp Leu Ile Thr Pro Ala His Trp Le

#u Arg His Ala Arg Arg

6125                6130

#                6135

#              6140

ccg gtg ctg ttc tcg gat gcc gtc cgg gag ct

#g gcc gac cgc ggc gtc    70742

Pro Val Leu Phe Ser Asp Ala Val Arg Glu Le

#u Ala Asp Arg Gly Val

6145

#               6150

#              6155

acc acg ttc gtg gcc gtc ggc ccc tcc ggc tc

#c ctg gcg tcg gcc gcg    70790

Thr Thr Phe Val Ala Val Gly Pro Ser Gly Se

#r Leu Ala Ser Ala Ala

6160

#           6165

#          6170

gcg gag agc gcc ggg gag gac gcc ggg acc ta

#c cac gcg gtg ctg cgc    70838

Ala Glu Ser Ala Gly Glu Asp Ala Gly Thr Ty

#r His Ala Val Leu Arg

6175

#       6180

#      6185

gcc cgg acc ggt gag gag acc gcg gcg ctg ac

#c gcc ctc gcc gag ctg    70886

Ala Arg Thr Gly Glu Glu Thr Ala Ala Leu Th

#r Ala Leu Ala Glu Leu

6190

#   6195

#  6200

cac gcc cac ggc gtc ccg gtc gac ctg gcc gc

#g gta ctg gcc ggt ggc    70934

His Ala His Gly Val Pro Val Asp Leu Ala Al

#a Val Leu Ala Gly Gly

6205                6210

#                6215

#              6220

cgg cca gtg gac ctt ccc gtg tac gcg ttc ca

#g cac cgt tcc tac tgg    70982

Arg Pro Val Asp Leu Pro Val Tyr Ala Phe Gl

#n His Arg Ser Tyr Trp

6225

#               6230

#              6235

ctg gcc ccg gcc gtg gcg ggg gcg ccg gcc ac

#c gtg gcg gac acc ggg    71030

Leu Ala Pro Ala Val Ala Gly Ala Pro Ala Th

#r Val Ala Asp Thr Gly

6240

#           6245

#          6250

ggt ccg gcg gag tcc gag ccg gag gac ctc ac

#c gtc gcc gag atc gtc    71078

Gly Pro Ala Glu Ser Glu Pro Glu Asp Leu Th

#r Val Ala Glu Ile Val

6255

#       6260

#      6265

cgt cgg cgc acc gcg gcg ctg ctc ggc gtc ac

#g gac ccc gcc gac gtc    71126

Arg Arg Arg Thr Ala Ala Leu Leu Gly Val Th

#r Asp Pro Ala Asp Val

6270

#   6275

#  6280

gat gcg gaa gcg acg ttc ttc gcg ctc ggt tt

#c gac tca ctg gcg gtg    71174

Asp Ala Glu Ala Thr Phe Phe Ala Leu Gly Ph

#e Asp Ser Leu Ala Val

6285                6290

#                6295

#              6300

cag cgg ctg cgc aac cag ctc gcc tcg gca ac

#c ggg ctg gac ctg ccg    71222

Gln Arg Leu Arg Asn Gln Leu Ala Ser Ala Th

#r Gly Leu Asp Leu Pro

6305

#               6310

#              6315

gcg gcc gtc ctg ttc gac cac gac acc ccg gc

#c gcg ctc acc gcg ttc    71270

Ala Ala Val Leu Phe Asp His Asp Thr Pro Al

#a Ala Leu Thr Ala Phe

6320

#           6325

#          6330

ctc cag gac cgg atc gag gcc ggc cag gac cg

#g atc gag gcc ggc gag    71318

Leu Gln Asp Arg Ile Glu Ala Gly Gln Asp Ar

#g Ile Glu Ala Gly Glu

6335

#       6340

#      6345

gac gac gac gcg ccc acc gtg ctc tcg ctc ct

#g gag gag atg gag tcg    71366

Asp Asp Asp Ala Pro Thr Val Leu Ser Leu Le

#u Glu Glu Met Glu Ser

6350

#   6355

#  6360

ctc gac gcc gcg gac atc gcg gcg acg ccg gc

#c ccg gag cgt gcg gcc    71414

Leu Asp Ala Ala Asp Ile Ala Ala Thr Pro Al

#a Pro Glu Arg Ala Ala

6365                6370

#                6375

#              6380

atc gcc gat ctg ctc gac aag ctc gcc cat ac

#c tgg aag gac tac cga    71462

Ile Ala Asp Leu Leu Asp Lys Leu Ala His Th

#r Trp Lys Asp Tyr Arg

6385

#               6390

#              6395

tga gcaccgatac gcacgaggga acgccgcccg ccggccgctg cccattcgc

#g         71515

*

atccaggacg gtcaccgcgc catcctggag agcggcacgg tgggttcgtt cg

#acctgttc  71575

ggcgtcaagc actggctggt cgccgccgcc gaggacgtca agctggtcac ca

#acgatccg  71635

cggttcagct cggccgcgcc gtccgagatg ctgcccgacc ggcggcccgg ct

#ggttctcc  71695

gggatggact caccggagca caaccgctac cggcagaaga tcgcggggga ct

#tcacactg  71755

cgcgcggcgc gcaagcggga ggacttcgtc gccgaggccg ccgacgcctg cc

#tggacgac  71815

atcgaggccg cgggacccgg caccgacctc atccccgggt acgccaagcg gc

#tgccctcc  71875

ctcgtcatca acgcgctgta cgggctcacc cctgaggagg gggccgtgct gg

#aggcacgg  71935

atgcgcgaca tcaccggctc ggccgatctg gacagcgtca agacgctgac cg

#acgacttc  71995

ttcgggcacg cgctgcggct ggtccgcgcg aagcgtgacg agcggggcga gg

#acctgctg  72055

caccggctgg cctcggccga cgacggcgag atctcgctca gcgacgacga gg

#cgacgggc  72115

gtgttcgcga cgctgctgtt cgccggccac gactcggtgc agcagatggt cg

#gctactgc  72175

ctctacgcac tgctcagcca ccccgagcag caggcggcgc tgcgcgcgcg cc

#cggagctg  72235

gtcgacaacg cggtcgagga gatgctccgt ttcctgcccg tcaaccagat gg

#gcgtaccg  72295

cgcgtctgtg tcgaggacgt cgatgtgcgg ggcgtgcgca tccgtgcggg cg

#acaacgtg  72355

atcccgctct actcgacggc caaccgcgac cccgaggtgt tcccgcagcc cg

#acaccttc  72415

gatgtgacgc gcccgctgga gggcaacttc gcgttcggcc acggcattca ca

#agtgtccc  72475

ggccagcaca tcgcccgggt gctcatcaag gtcgcctgcc tgcggttgtt cg

#agcgtttc  72535

ccggacgtcc ggctggccgg cgacgtgccg atgaacgagg ggctcgggct gt

#tcagcccg  72595

gccgagctgc gggtcacctg gggggcggca tgagtcaccc ggtggagacg tt

#gcggttgc  72655

cgaacgggac gacggtcgcg cacatcaacg cgggcgaggc gcagttcctc ta

#ccgggaga  72715

tcttcaccca gcgctgctac ctgcgccacg gtgtcgacct gcgcccgggg ga

#cgtggtgt  72775

tcgacgtcgg cgcgaacatc ggcatgttca cgcttttcgc gcatctggag tg

#tcctggtg  72835

tgaccgtgca cgccttcgag cccgcgcccg tgccgttcgc ggcgctgcgg gc

#gaacgtga  72895

cgcggcacgg catcccgggc caggcggacc agtgcgcggt ctccgacagc tc

#cggcaccc  72955

ggaagatgac cttctatccc gacgccacgc tgatgtccgg tttccacgcg ga

#tgccgcgg  73015

cccggacgga gctgttgcgc acgctcggcc tcaacggcgg ctacaccgcc ga

#ggacgtcg  73075

acaccatgct cgcgcaactg cccgacgtca gcgaggagat cgaaacccct gt

#ggtccggc  73135

tctccgacgt catcgcggag cgcggtatcg aggccatcgg cctgctgaag gt

#cgacgtgg  73195

agaagagcga acggcaggtc ttcgccggcc tcgaggacac cgactggccc cg

#tatccgcc  73255

aggtcgtcgc ggaggtccac gacatcgacg gcgcgctcga ggaggtcgtc ac

#gctgctcc  73315

gcggccatgg cttcaccgtg gtcgccgagc aggaaccgct gttcgccggc ac

#gggcatcc  73375

accaggtcgc cgcgcggcgg gtggccggct gagcgccgtc ggggccgcgg cc

#gtccgcac  73435

cggcggccgc ggtgcggacg gcggctcagc cggcgtcgga cagttccttg gg

#cagttgct  73495

gacggccctt cacccccagc ttgcggaaca cgttggtgag gtgctgttcc ac

#cgtgctgg  73555

aggtgacgaa cagctggctg gcgatctcct tgttggtgcg cccgaccgcg gc

#gtgcgacg  73615

ccacccgccg ctccgcctcg gtcagcgatg tgatccgctg cgccggcgtc ac

#gtcctggg  73675

tgccgtccgc gtccgaggac tccccaccga gccgccggag gagcggcacg gc

#tccgcact  73735

gggtcgcgag gtgccgtgcg cggcggaaca gtccccgcgc acggctgtgc cg

#ccggagca  73795

tgccgcacgc ttcgcccatg tcggcgagga cgcgggccag ctcgtactgg tc

#gcggcaca  73855

tgatgagcag atcggcggcc tcgtcgagca gttcgatccg cttggccggc gg

#actgtagg  73915

ccgcctgcac ccgcagcgtc atcacccgcg cccgggaccc catcggccgg ga

#cagctgct  73975

cggagatgag cctcagcccc tcgtcacggc cgcggccgag cagcagaagc gc

#ttcggcgg  74035

cgtcgacccg ccacagggcc aggcccggca cgtcgacgga ccagcgtcgc at

#ccgctccc  74095

cgcagtcccg gaacgcgttg tacgccgccc ggtaccgccc ggccgcgaga tg

#gtgttgcc  74155

cacgggccca gaccatgtgc agtccgaaga ggctgtcgga ggtctcctcc gg

#caacggct  74215

cggcgagcca ccgctccgcc cggtccaggt cgcccagtcg gatcgcggcg gc

#cacggtgc  74275

tgctcagcgg caatgcggcg gccatccccc aggagggcac gacccggggg gc

#gagcgcgg  74335

cctcgccgca ttcgacggcg gcggtcaggt cgccgcggcg cagcgcggcc tc

#ggcgcgga  74395

accccgcgtg gaccgcctcg tcggccgggg tccgcatgtt gtcgtcaccg gc

#cagcttgt  74455

cgacccagga ctggacggca tcggtgtcct cggcgtagag cagggccagc aa

#cgccatca  74515

tggtcgtggt ccggtccgtc gtgacccggg agtgctggag cacgtactcg gc

#tttggcct  74575

cggcctgttc ggaccagccg cgcagcgcgt tgctcagggc cttgtcggcg ac

#ggcgcggt  74635

gccggacggc tccggaaaac gaggcgacct cgtcctcggc cggcggatcg gc

#cggacgcg  74695

gcggatcggc cgcgccggga tagatcagcg cgagggacag gtccgcgacg cg

#caggtgcg  74755

cccggccctg ctcgctcggg gcggcggagc gctgggccgc caggacctcg gc

#ggcctcgc  74815

ccggccgccc gtccatcgcc agccagcagg cgagcgacac ggcgtgctcg ct

#ggagagga  74875

gccgttcccg cgacgcggtg agcagctcgg gcacatgccg gccggatctg gc

#gggatcgc  74935

agagccgctc gatggcggcg gtgtcgacgc gcagtgcggc gtggacggcg gg

#gtcgtcgg  74995

aggcccggta ggcgaactcc aggtaggtga cggcctcgtc gagctcgccg cg

#caggtggt  75055

gctcgcgcgc ggcgtcggtg aacagcccgg cgacctcggc gccgtgcacc cg

#gccggtac  75115

ccatctggtg gcgggcgagc accttgctgg ccacgccgcg gtcccgcagc ag

#ttccagcg  75175

ccagctcgtg caggccacgc cgctcggcgg cggagaggtc gtcgagtacg ac

#ggagcggg  75235

ccgcggggtg cgggaaccgc ccttcccgca gcagccgccc ctcgaccagc tg

#ttcgtggg  75295

cctgctcgac cgcctcggtg tcgaggccgg tcatccgctg gacgagggtg ag

#ttcgacac  75355

tctcgccgag cacggcggaa gctcgggcga cgctcagcgc ggccgggccg ca

#acgataga  75415

gcgacccgag gtaggcgagc cggtacgccc gccccgcgac cacttccagg ca

#ccctgagg  75475

tccgtgtccg tgcctcccgg atgtcgtcga tcaggccgtg gccgaggagc ag

#gttgccgc  75535

cggtcgcccg gaacgcctgg gccaccacgt cgtcgtgcgc gtcctggccg ag

#gtgccggc  75595

gcacgagttc ggtggtctgc gcctcggtga gcgggcgcag cgcgatctcc tg

#gtagtggc  75655

gcagactcag cagtgccgcc cggaattggg agtgggcggg cgtcggccgg ag

#cagctcgg  75715

tcagcacgat ggcgacacgg gcccggctga tgcggcgcgc gaggtggagc ag

#gcagcgca  75775

gcgacggcgc gtcggcgtgg tgcacgtcgt cgatgccgat cagtacgggc cg

#ctccgcgg  75835

cgagcgtcag caccgtgcgg gtgagttcgg tccccaggcg gttgtcgacg tc

#ggccggca  75895

ggttttcgca cgatgccgtc agccggacca gctccggtgt ccgggcggcc ag

#ctcgggct  75955

ggtcgaggag ctggccgagc atgccgtacg gcagggcccg ctcctccatg ga

#gcacaccg  76015

cgcgaagggt gacgaagccg gccttggccg cggcggcgtc gaggagttcg gt

#cttgccgc  76075

aggcgatcgg cccggtgacg gcggcgacga cgccccgccc gccccccgct cg

#ggtgagcg  76135

cccggtggag ggaaccgaac tcgtcatcgc gggcgatcag gtctggggga ga

#taagcgcg  76195

ctatcacgaa tggaactacc tcgcgaccgt cgtggaaacc cataggcatc ac

#atggcttg  76255

ttgatctgta cggctgtgat tcagcctggc gggatgctgt gctacagatg gg

#aagatgtg  76315

atctagggcc gtgccgttcc ctcaggagcc gaccgccccc ggcgccaccc gc

#cgtacccc  76375

ctgggccacc agctcggcga cccgctcctg gtggtcgacg aggtagaagt gc

#ccgccggg  76435

gaagacctcc accgtggtcg gcgcggtcgt gtgcccggcc caggcgtggg cc

#tgctccac  76495

cgtcgtcttc ggatcgtcgt caccgatgca caccgtgatc ggcgtctcca gc

#ggcggcgc  76555

gggctcccac cggtacgtct ccgccgcgta gtagtccgcc cgcaacggcg cc

#aggatcag  76615

cgcgcgcatt tcgtcgtccg ccatcacatc ggcgctcgtc ccgccgaggc cg

#atgaccgc  76675

cgccagcagc tcgtcgtcgg acgcgaggtg gtcctggtcg gcgcgcggct gc

#gacggcgc  76735

ccgccggccc gagacgatca ggtgcgccac cgggagccgc tgggccagct cg

#aacgcgag  76795

tgtcgcgccc atgctgtggc cgaacagcac cagcggacgg tccagccccg gc

#ttcaacgc  76855

ctcggccacg aggccggcga gaacacgcag gtcgcgcacc gcctcctcgt cg

#cggcggtc  76915

ctggcggccg gggtactgca cggcgtacac gtccgccacc ggggcgagcg ca

#cgggccag  76975

cggaaggtag aacgtcgccg atccgccggc gtggggcagc agcaccaccc gt

#accggggc  77035

ctcgggcgtg gggaagaact gccgcagcca gagttccgag ctcaccgcac cc

#cctcggcc  77095

gcgacctggg gagcccggaa ccgggtgatc tcggccaagt gcttctcccg ca

#tctccggg  77155

tcggtcacgc cccatccctc ctccggcgcc agacagagga cgccgacttt gc

#cgttgtgc  77215

acattgcgat gcacatcgcg caccgccgac ccgacgtcgt cgagcgggta gg

#tcaccgac  77275

agcgtcgggt gcaccatccc cttgcagatc aggcggttcg cctcccacgc ct

#cacgatag  77335

ttcgcgaagt gggtaccgat gatccgcttc acggacatcc acaggtaccg at

#tgtcaaag  77395

gcgtgctcgt atcccgaggt tgacgcgcag gtgacgatcg tgccaccccg ac

#gtgtcacg  77455

tagacactcg cgccgaacgt cgcgcgcccc gggtgctcga acacgatgtc gg

#gatcgtca  77515

ccgccggtca gctcccggat c

#

#               77536

#

#

#             77536

<210> SEQ ID NO 2

<211> LENGTH: 360

<212> TYPE: PRT

<213> ORGANISM: Streptomyces sp.

<400> SEQUENCE: 2

Met Thr Ile Val Lys Cys Leu Val Trp Asp Le

#u Asp Asn Thr Leu Trp

1               5

#                 10

#                 15

Arg Gly Thr Val Leu Glu Asp Asp Glu Val Va

#l Leu Thr Asp Glu Ile

20

#             25

#             30

Arg Glu Val Ile Thr Thr Leu Asp Asp Arg Gl

#y Ile Leu Gln Ala Val

35

#         40

#         45

Ala Ser Lys Asn Asp His Asp Leu Ala Trp Gl

#u Arg Leu Glu Arg Leu

50

#     55

#     60

Gly Val Ala Glu Tyr Phe Val Leu Ala Arg Il

#e Gly Trp Gly Pro Lys

65

# 70

# 75

# 80

Ser Gln Ser Val Arg Glu Ile Ala Thr Glu Le

#u Asn Phe Ala Pro Thr

85

#                 90

#                 95

Thr Ile Ala Phe Ile Asp Asp Gln Pro Ala Gl

#u Arg Ala Glu Val Ala

100

#           105

#           110

Phe His Leu Pro Glu Val Arg Cys Tyr Pro Al

#a Glu Gln Ala Ala Thr

115

#       120

#       125

Leu Leu Ser Leu Pro Glu Phe Ser Pro Pro Va

#l Ser Thr Val Asp Ser

130

#   135

#   140

Arg Arg Arg Arg Leu Met Tyr Gln Ala Gly Ph

#e Ala Arg Asp Gln Ala

145                 1

#50                 1

#55                 1

#60

Arg Glu Ala Tyr Ser Gly Pro Asp Glu Asp Ph

#e Leu Arg Ser Leu Asp

165

#               170

#               175

Leu Ser Met Thr Ile Ala Pro Ala Gly Glu Gl

#u Glu Leu Ser Arg Val

180

#           185

#           190

Glu Glu Leu Thr Leu Arg Thr Ser Gln Met As

#n Ala Thr Gly Val His

195

#       200

#       205

Tyr Ser Asp Ala Asp Leu Arg Ala Leu Leu Th

#r Asp Pro Ala His Glu

210

#   215

#   220

Val Leu Val Val Thr Met Gly Asp Arg Phe Gl

#y Pro His Gly Ala Val

225                 2

#30                 2

#35                 2

#40

Gly Ile Ile Leu Leu Glu Lys Lys Pro Ser Th

#r Trp His Leu Lys Leu

245

#               250

#               255

Leu Ala Thr Ser Cys Arg Val Val Ser Phe Gl

#y Ala Gly Ala Thr Ile

260

#           265

#           270

Leu Asn Trp Leu Thr Asp Gln Gly Ala Arg Al

#a Gly Ala His Leu Val

275

#       280

#       285

Ala Asp Phe Arg Arg Thr Asp Arg Asn Arg Me

#t Met Glu Ile Ala Tyr

290

#   295

#   300

Arg Phe Ala Gly Phe Ala Asp Ser Asp Cys Pr

#o Cys Val Ser Glu Val

305                 3

#10                 3

#15                 3

#20

Ala Gly Ala Ser Ala Ala Gly Val Glu Arg Le

#u His Leu Glu Pro Ser

325

#               330

#               335

Ala Arg Pro Ala Pro Thr Thr Leu Thr Leu Th

#r Ala Ala Asp Ile Ala

340

#           345

#           350

Pro Val Thr Val Ser Ala Ala Gly

355

#       360

<210> SEQ ID NO 3

<211> LENGTH: 22

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: Linker

<400> SEQUENCE: 3

ctagtgggca gatctggcag ct

#

#                 22

<210> SEQ ID NO 4

<211> LENGTH: 14

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: Linker

<400> SEQUENCE: 4

gccagatctg ccca

#

#

#     14

<210> SEQ ID NO 5

<211> LENGTH: 12

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: Linker

<400> SEQUENCE: 5

gggatgcatg gc

#

#

#       12

<210> SEQ ID NO 6

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: Linker

<400> SEQUENCE: 6

ttaagccatg catccccatg

#

#

# 20

<210> SEQ ID NO 7

<211> LENGTH: 24

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 7

cgactcacta gtgggcagat ctgg

#

#                24

<210> SEQ ID NO 8

<211> LENGTH: 29

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 8

cacgcctagg ccggtcggtc tcgggccac

#

#            29

<210> SEQ ID NO 9

<211> LENGTH: 30

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 9

gcggctagct gctcgcccat cgcgggatgc

#

#           30

<210> SEQ ID NO 10

<211> LENGTH: 35

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 10

gatgtacagc tcgagtcggc acgcccggcc gcatc

#

#       35

<210> SEQ ID NO 11

<211> LENGTH: 23

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 11

cgactcactt aagccatgca tcc

#

#                23

<210> SEQ ID NO 12

<211> LENGTH: 31

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 12

atcctaggcg ggcrggygtg tcgtccttcg g

#

#          31

<210> SEQ ID NO 13

<211> LENGTH: 32

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 13

atgctagccg ccgcgttccc cgtcttcgcg cg

#

#          32

<210> SEQ ID NO 14

<211> LENGTH: 32

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 14

atgctagcgg attcgtcggt ggtgttcgcc ga

#

#          32

<210> SEQ ID NO 15

<211> LENGTH: 31

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: primer

<400> SEQUENCE: 15

atctcgagcc agtascgctg gtgytggaag g

#

#          31

<210> SEQ ID NO 16

<211> LENGTH: 4478

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: DNA

encoding synthetic PKS synthase frag

#ment

<220> FEATURE:

<221> NAME/KEY: CDS

<222> LOCATION: (3)..(4466)

<400> SEQUENCE: 16

ag atc tgg cag ctc gcc gaa gcg ctg ctg acg

# ctc gtc cgg gag agc        47

Ile Trp Gln Leu Ala Glu Ala Leu Leu

#Thr Leu Val Arg Glu Ser

1

#  5

#  10

#  15

acc gcc gcc gtg ctc ggc cac gtg ggt ggc ga

#g gac atc ccc gcg acg       95

Thr Ala Ala Val Leu Gly His Val Gly Gly Gl

#u Asp Ile Pro Ala Thr

20

#                 25

#                 30

gcg gcg ttc aag gac ctc ggc atc gac tcg ct

#c acc gcg gtc cag ctg      143

Ala Ala Phe Lys Asp Leu Gly Ile Asp Ser Le

#u Thr Ala Val Gln Leu

35

#             40

#             45

cgc aac gcc ctc acc gag gcg acc ggt gtg cg

#g ctg aac gcc acg gcg      191

Arg Asn Ala Leu Thr Glu Ala Thr Gly Val Ar

#g Leu Asn Ala Thr Ala

50

#         55

#         60

gtc ttc gac ttc ccg acc ccg cac gtg ctc gc

#c ggg aag ctc ggc gac      239

Val Phe Asp Phe Pro Thr Pro His Val Leu Al

#a Gly Lys Leu Gly Asp

65

#     70

#     75

gaa ctg acc ggc acc cgc gcg ccc gtc gtg cc

#c cgg acc gcg gcc acg      287

Glu Leu Thr Gly Thr Arg Ala Pro Val Val Pr

#o Arg Thr Ala Ala Thr

80

# 85

# 90

# 95

gcc ggt gcg cac gac gag ccg ctg gcg atc gt

#g gga atg gcc tgc cgg      335

Ala Gly Ala His Asp Glu Pro Leu Ala Ile Va

#l Gly Met Ala Cys Arg

100

#               105

#               110

ctg ccc ggc ggg gtc gcg tca ccc gag gag ct

#g tgg cac ctc gtg gca      383

Leu Pro Gly Gly Val Ala Ser Pro Glu Glu Le

#u Trp His Leu Val Ala

115

#           120

#           125

tcc ggc acc gac gcc atc acg gag ttc ccg ac

#g gac cgc ggc tgg gac      431

Ser Gly Thr Asp Ala Ile Thr Glu Phe Pro Th

#r Asp Arg Gly Trp Asp

130

#       135

#       140

gtc gac gcg atc tac gac ccg gac ccc gac gc

#g atc ggc aag acc ttc      479

Val Asp Ala Ile Tyr Asp Pro Asp Pro Asp Al

#a Ile Gly Lys Thr Phe

145

#   150

#   155

gtc cgg cac ggt ggc ttc ctc acc ggc gcg ac

#a ggc ttc gac gcg gcg      527

Val Arg His Gly Gly Phe Leu Thr Gly Ala Th

#r Gly Phe Asp Ala Ala

160                 1

#65                 1

#70                 1

#75

ttc ttc ggc atc agc ccg cgc gag gcc ctc gc

#g atg gac ccg cag cag      575

Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Al

#a Met Asp Pro Gln Gln

180

#               185

#               190

cgg gtg ctc ctg gag acg tcg tgg gag gcg tt

#c gaa agc gcc ggc atc      623

Arg Val Leu Leu Glu Thr Ser Trp Glu Ala Ph

#e Glu Ser Ala Gly Ile

195

#           200

#           205

acc ccg gac tcg acc cgc ggc agc gac acc gg

#c gtg ttc gtc ggc gcc      671

Thr Pro Asp Ser Thr Arg Gly Ser Asp Thr Gl

#y Val Phe Val Gly Ala

210

#       215

#       220

ttc tcc tac ggt tac ggc acc ggt gcg gac ac

#c gac ggc ttc ggc gcg      719

Phe Ser Tyr Gly Tyr Gly Thr Gly Ala Asp Th

#r Asp Gly Phe Gly Ala

225

#   230

#   235

acc ggc tcg cag acc agt gtg ctc tcc ggc cg

#g ctg tcg tac ttc tac      767

Thr Gly Ser Gln Thr Ser Val Leu Ser Gly Ar

#g Leu Ser Tyr Phe Tyr

240                 2

#45                 2

#50                 2

#55

ggt ctg gag ggt ccg gcg gtc acg gtc gac ac

#g gcg tgt tcg tcg tcg      815

Gly Leu Glu Gly Pro Ala Val Thr Val Asp Th

#r Ala Cys Ser Ser Ser

260

#               265

#               270

ctg gtg gcg ctg cac cag gcc ggg cag tcg ct

#g cgc tcc ggc gaa tgc      863

Leu Val Ala Leu His Gln Ala Gly Gln Ser Le

#u Arg Ser Gly Glu Cys

275

#           280

#           285

tcg ctc gcc ctg gtc ggc ggc gtc acg gtg at

#g gcg tct ccc ggc ggc      911

Ser Leu Ala Leu Val Gly Gly Val Thr Val Me

#t Ala Ser Pro Gly Gly

290

#       295

#       300

ttc gtg gag ttc tcc cgg cag cgc ggc ctc gc

#g ccg gac ggc cgg gcg      959

Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Al

#a Pro Asp Gly Arg Ala

305

#   310

#   315

aag gcg ttc ggc gcg ggt gcg gac ggc acg ag

#c ttc gcc gag ggt gcc     1007

Lys Ala Phe Gly Ala Gly Ala Asp Gly Thr Se

#r Phe Ala Glu Gly Ala

320                 3

#25                 3

#30                 3

#35

ggt gtg ctg atc gtc gag agg ctc tcc gac gc

#c gaa cgc aac ggt cac     1055

Gly Val Leu Ile Val Glu Arg Leu Ser Asp Al

#a Glu Arg Asn Gly His

340

#               345

#               350

acc gtc ctg gcg gtc gtc cgt ggt tcg gcg gt

#c aac cag gat ggt gcc     1103

Thr Val Leu Ala Val Val Arg Gly Ser Ala Va

#l Asn Gln Asp Gly Ala

355

#           360

#           365

tcc aac ggg ctg tcg gcg ccg aac ggg ccg tc

#g cag gag cgg gtg atc     1151

Ser Asn Gly Leu Ser Ala Pro Asn Gly Pro Se

#r Gln Glu Arg Val Ile

370

#       375

#       380

cgg cag gcc ctg gcc aac gcc ggg ctc acc cc

#g gcg gac gtg gac gcc     1199

Arg Gln Ala Leu Ala Asn Ala Gly Leu Thr Pr

#o Ala Asp Val Asp Ala

385

#   390

#   395

gtc gag gcc cac ggc acc ggc acc agg ctg gg

#c gac ccc atc gag gca     1247

Val Glu Ala His Gly Thr Gly Thr Arg Leu Gl

#y Asp Pro Ile Glu Ala

400                 4

#05                 4

#10                 4

#15

cag gcg gta ctg gcc acc tac gga cag gag cg

#c gcc acc ccc ctg ctg     1295

Gln Ala Val Leu Ala Thr Tyr Gly Gln Glu Ar

#g Ala Thr Pro Leu Leu

420

#               425

#               430

ctg ggc tcg ctg aag tcc aac atc ggc cac gc

#c cag gcc gcg tcc ggc     1343

Leu Gly Ser Leu Lys Ser Asn Ile Gly His Al

#a Gln Ala Ala Ser Gly

435

#           440

#           445

gtc gcc ggc atc atc aag atg gtg cag gcc ct

#c cgg cac ggg gag ctg     1391

Val Ala Gly Ile Ile Lys Met Val Gln Ala Le

#u Arg His Gly Glu Leu

450

#       455

#       460

ccg ccg acg ctg cac gcc gac gag ccg tcg cc

#g cac gtc gac tgg acg     1439

Pro Pro Thr Leu His Ala Asp Glu Pro Ser Pr

#o His Val Asp Trp Thr

465

#   470

#   475

gcc ggc gcc gtc gaa ctg ctg acg tcg gcc cg

#g ccg tgg ccc gag acc     1487

Ala Gly Ala Val Glu Leu Leu Thr Ser Ala Ar

#g Pro Trp Pro Glu Thr

480                 4

#85                 4

#90                 4

#95

gac cgg cct agg cgg gca ggc gtg tcg tcc tt

#c ggg atc agt ggc acc     1535

Asp Arg Pro Arg Arg Ala Gly Val Ser Ser Ph

#e Gly Ile Ser Gly Thr

500

#               505

#               510

aac gcc cac gtc atc ctg gaa agc gca ccc cc

#c act cag cct gcg gac     1583

Asn Ala His Val Ile Leu Glu Ser Ala Pro Pr

#o Thr Gln Pro Ala Asp

515

#           520

#           525

aac gcg gtg atc gag cgg gca ccg gag tgg gt

#g ccg ttg gtg att tcg     1631

Asn Ala Val Ile Glu Arg Ala Pro Glu Trp Va

#l Pro Leu Val Ile Ser

530

#       535

#       540

gcc agg acc cag tcg gct ttg act gag cac ga

#g ggc cgg ttg cgt gcg     1679

Ala Arg Thr Gln Ser Ala Leu Thr Glu His Gl

#u Gly Arg Leu Arg Ala

545

#   550

#   555

tat ctg gcg gcg tcg ccc ggg gtg gat atg cg

#g gct gtg gca tcg acg     1727

Tyr Leu Ala Ala Ser Pro Gly Val Asp Met Ar

#g Ala Val Ala Ser Thr

560                 5

#65                 5

#70                 5

#75

ctg gcg atg aca cgg tcg gtg ttc gag cac cg

#t gcc gtg ctg ctg gga     1775

Leu Ala Met Thr Arg Ser Val Phe Glu His Ar

#g Ala Val Leu Leu Gly

580

#               585

#               590

gat gac acc gtc acc ggc acc gct gtg tct ga

#c cct cgg gcg gtg ttc     1823

Asp Asp Thr Val Thr Gly Thr Ala Val Ser As

#p Pro Arg Ala Val Phe

595

#           600

#           605

gtc ttc ccg gga cag ggg tcg cag cgt gct gg

#c atg ggt gag gaa ctg     1871

Val Phe Pro Gly Gln Gly Ser Gln Arg Ala Gl

#y Met Gly Glu Glu Leu

610

#       615

#       620

gcc gcc gcg ttc ccc gtc ttc gcg cgg atc ca

#t cag cag gtg tgg gac     1919

Ala Ala Ala Phe Pro Val Phe Ala Arg Ile Hi

#s Gln Gln Val Trp Asp

625

#   630

#   635

ctg ctc gat gtg ccc gat ctg gag gtg aac ga

#g acc ggt tac gcc cag     1967

Leu Leu Asp Val Pro Asp Leu Glu Val Asn Gl

#u Thr Gly Tyr Ala Gln

640                 6

#45                 6

#50                 6

#55

ccg gcc ctg ttc gca atg cag gtg gct ctg tt

#c ggg ctg ctg gaa tcg     2015

Pro Ala Leu Phe Ala Met Gln Val Ala Leu Ph

#e Gly Leu Leu Glu Ser

660

#               665

#               670

tgg ggt gta cga ccg gac gcg gtg atc ggc ca

#t tcg gtg ggt gag ctt     2063

Trp Gly Val Arg Pro Asp Ala Val Ile Gly Hi

#s Ser Val Gly Glu Leu

675

#           680

#           685

gcg gct gcg tat gtg tcc ggg gtg tgg tcg tt

#g gag gat gcc tgc act     2111

Ala Ala Ala Tyr Val Ser Gly Val Trp Ser Le

#u Glu Asp Ala Cys Thr

690

#       695

#       700

ttg gtg tcg gcg cgg gct cgt ctg atg cag gc

#t ctg ccc gcg ggt ggg     2159

Leu Val Ser Ala Arg Ala Arg Leu Met Gln Al

#a Leu Pro Ala Gly Gly

705

#   710

#   715

gtg atg gtc gct gtc ccg gtc tcg gag gat ga

#g gcc cgg gcc gtg ctg     2207

Val Met Val Ala Val Pro Val Ser Glu Asp Gl

#u Ala Arg Ala Val Leu

720                 7

#25                 7

#30                 7

#35

ggt gag ggt gtg gag atc gcc gcg gtc aac gg

#c ccg tcg tcg gtg gtt     2255

Gly Glu Gly Val Glu Ile Ala Ala Val Asn Gl

#y Pro Ser Ser Val Val

740

#               745

#               750

ctc tcc ggt gat gag gcc gcc gtg ctg cag gc

#c gcg gag ggg ctg ggg     2303

Leu Ser Gly Asp Glu Ala Ala Val Leu Gln Al

#a Ala Glu Gly Leu Gly

755

#           760

#           765

aag tgg acg cgg ctg gcg acc agc cac gcg tt

#c cat tcc gcc cgt atg     2351

Lys Trp Thr Arg Leu Ala Thr Ser His Ala Ph

#e His Ser Ala Arg Met

770

#       775

#       780

gaa ccc atg ctg gag gag ttc cgg gcg gtc gc

#c gaa ggc ctg acc tac     2399

Glu Pro Met Leu Glu Glu Phe Arg Ala Val Al

#a Glu Gly Leu Thr Tyr

785

#   790

#   795

cgg acg ccg cag gtc tcc atg gcc gtt ggt ga

#t cag gtg acc acc gct     2447

Arg Thr Pro Gln Val Ser Met Ala Val Gly As

#p Gln Val Thr Thr Ala

800                 8

#05                 8

#10                 8

#15

gag tac tgg gtg cgg cag gtc cgg gac acg gt

#c cgg ttc ggc gag cag     2495

Glu Tyr Trp Val Arg Gln Val Arg Asp Thr Va

#l Arg Phe Gly Glu Gln

820

#               825

#               830

gtg gcc tcg tac gag gac gcc gtg ttc gtc ga

#g ctg ggt gcc gac cgg     2543

Val Ala Ser Tyr Glu Asp Ala Val Phe Val Gl

#u Leu Gly Ala Asp Arg

835

#           840

#           845

tca ctg gcc cgc ctg gtc gac ggt gtc gcg at

#g ctg cac ggc gac cac     2591

Ser Leu Ala Arg Leu Val Asp Gly Val Ala Me

#t Leu His Gly Asp His

850

#       855

#       860

gaa atc cag gcc gcg atc ggc gcc ctg gcc ca

#c ctg tat gtc aac ggc     2639

Glu Ile Gln Ala Ala Ile Gly Ala Leu Ala Hi

#s Leu Tyr Val Asn Gly

865

#   870

#   875

gtc acg gtc gac tgg ccc gcg ctc ctg ggc ga

#t gct ccg gca aca cgg     2687

Val Thr Val Asp Trp Pro Ala Leu Leu Gly As

#p Ala Pro Ala Thr Arg

880                 8

#85                 8

#90                 8

#95

gtg ctg gac ctt ccg aca tac gcc ttc cag ca

#c cag cgc tac tgg ctc     2735

Val Leu Asp Leu Pro Thr Tyr Ala Phe Gln Hi

#s Gln Arg Tyr Trp Leu

900

#               905

#               910

gag tcg gca cgc ccg gcc gca tcc gac gcg gg

#c cac ccc gtg ctg ggc     2783

Glu Ser Ala Arg Pro Ala Ala Ser Asp Ala Gl

#y His Pro Val Leu Gly

915

#           920

#           925

tcc ggt atc gcc ctc gcc ggg tcg ccg ggc cg

#g gtg ttc acg ggt tcc     2831

Ser Gly Ile Ala Leu Ala Gly Ser Pro Gly Ar

#g Val Phe Thr Gly Ser

930

#       935

#       940

gtg ccg acc ggt gcg gac cgc gcg gtg ttc gt

#c gcc gag ctg gcg ctg     2879

Val Pro Thr Gly Ala Asp Arg Ala Val Phe Va

#l Ala Glu Leu Ala Leu

945

#   950

#   955

gcc gcc gcg gac gcg gtc gac tgc gcc acg gt

#c gag cgg ctc gac atc     2927

Ala Ala Ala Asp Ala Val Asp Cys Ala Thr Va

#l Glu Arg Leu Asp Ile

960                 9

#65                 9

#70                 9

#75

gcc tcc gtg ccc ggc cgg ccg ggc cat ggc cg

#g acg acc gta cag acc     2975

Ala Ser Val Pro Gly Arg Pro Gly His Gly Ar

#g Thr Thr Val Gln Thr

980

#               985

#               990

tgg gtc gac gag ccg gcg gac gac ggc cgg cg

#c cgg ttc acc gtg cac     3023

Trp Val Asp Glu Pro Ala Asp Asp Gly Arg Ar

#g Arg Phe Thr Val His

995

#           1000

#          1005

acc cgc acc ggc gac gcc ccg tgg acg ctg ca

#c gcc gag ggg gtg ctg     3071

Thr Arg Thr Gly Asp Ala Pro Trp Thr Leu Hi

#s Ala Glu Gly Val Leu

1010

#       1015

#      1020

cgc ccc cat ggc acg gcc ctg ccc gat gcg gc

#c gac gcc gag tgg ccc     3119

Arg Pro His Gly Thr Ala Leu Pro Asp Ala Al

#a Asp Ala Glu Trp Pro

1025

#   1030

#  1035

cca ccg ggc gcg gtg ccc gcg gac ggg ctg cc

#g ggt gtg tgg cgc cgg     3167

Pro Pro Gly Ala Val Pro Ala Asp Gly Leu Pr

#o Gly Val Trp Arg Arg

1040                1045

#                1050

#              1055

ggg gac cag gtc ttc gcc gag gcc gag gtg ga

#c gga ccg gac ggt ttc     3215

Gly Asp Gln Val Phe Ala Glu Ala Glu Val As

#p Gly Pro Asp Gly Phe

1060

#               1065

#              1070

gtg gtg cac ccc gac ctg ctc gac gcg gtc tt

#c tcc gcg gtc ggc gac     3263

Val Val His Pro Asp Leu Leu Asp Ala Val Ph

#e Ser Ala Val Gly Asp

1075

#           1080

#          1085

gga agc cgc cag ccg gcc gga tgg cgc gac ct

#g acg gtg cac gcg tcg     3311

Gly Ser Arg Gln Pro Ala Gly Trp Arg Asp Le

#u Thr Val His Ala Ser

1090

#       1095

#      1100

gac gcc acc gta ctg cgc gcc tgc ctc acc cg

#g cgc acc gac gga gcc     3359

Asp Ala Thr Val Leu Arg Ala Cys Leu Thr Ar

#g Arg Thr Asp Gly Ala

1105

#   1110

#  1115

atg gga ttc gcc gcc ttc gac ggc gcc ggc ct

#g ccg gta ctc acc gcg     3407

Met Gly Phe Ala Ala Phe Asp Gly Ala Gly Le

#u Pro Val Leu Thr Ala

1120                1125

#                1130

#              1135

gag gcg gtg acg ctg cgg gag gtg gcg tca cc

#g tcc ggc tcc gag gag     3455

Glu Ala Val Thr Leu Arg Glu Val Ala Ser Pr

#o Ser Gly Ser Glu Glu

1140

#               1145

#              1150

tcg gac ggc ctg cac cgg ttg gag tgg ctc gc

#g gtc gcc gag gcg gtc     3503

Ser Asp Gly Leu His Arg Leu Glu Trp Leu Al

#a Val Ala Glu Ala Val

1155

#           1160

#          1165

tac gac ggt gac ctg ccc gag gga cat gtc ct

#g atc acc gcc gcc cac     3551

Tyr Asp Gly Asp Leu Pro Glu Gly His Val Le

#u Ile Thr Ala Ala His

1170

#       1175

#      1180

ccc gac gac ccc gag gac ata ccc acc cgc gc

#c cac acc cgc gcc acc     3599

Pro Asp Asp Pro Glu Asp Ile Pro Thr Arg Al

#a His Thr Arg Ala Thr

1185

#   1190

#  1195

cgc gtc ctg acc gcc ctg caa cac cac ctc ac

#c acc acc gac cac acc     3647

Arg Val Leu Thr Ala Leu Gln His His Leu Th

#r Thr Thr Asp His Thr

1200                1205

#                1210

#              1215

ctc atc gtc cac acc acc acc gac ccc gcc gg

#c gcc acc gtc acc ggc     3695

Leu Ile Val His Thr Thr Thr Asp Pro Ala Gl

#y Ala Thr Val Thr Gly

1220

#               1225

#              1230

ctc acc cgc acc gcc cag aac gaa cac ccc ca

#c cgc atc cgc ctc atc     3743

Leu Thr Arg Thr Ala Gln Asn Glu His Pro Hi

#s Arg Ile Arg Leu Ile

1235

#           1240

#          1245

gaa acc gac cac ccc cac acc ccc ctc ccc ct

#g gcc caa ctc gcc acc     3791

Glu Thr Asp His Pro His Thr Pro Leu Pro Le

#u Ala Gln Leu Ala Thr

1250

#       1255

#      1260

ctc gac cac ccc cac ctc cgc ctc acc cac ca

#c acc ctc cac cac ccc     3839

Leu Asp His Pro His Leu Arg Leu Thr His Hi

#s Thr Leu His His Pro

1265

#   1270

#  1275

cac ctc acc ccc ctc cac acc acc acc cca cc

#c acc acc acc ccc ctc     3887

His Leu Thr Pro Leu His Thr Thr Thr Pro Pr

#o Thr Thr Thr Pro Leu

1280                1285

#                1290

#              1295

aac ccc gaa cac gcc atc atc atc acc ggc gg

#c tcc ggc acc ctc gcc     3935

Asn Pro Glu His Ala Ile Ile Ile Thr Gly Gl

#y Ser Gly Thr Leu Ala

1300

#               1305

#              1310

ggc atc ctc gcc cgc cac ctg aac cac ccc ca

#c acc tac ctc ctc tcc     3983

Gly Ile Leu Ala Arg His Leu Asn His Pro Hi

#s Thr Tyr Leu Leu Ser

1315

#           1320

#          1325

cgc acc cca ccc ccc gac gcc acc ccc ggc ac

#c cac ctc ccc tgc gac     4031

Arg Thr Pro Pro Pro Asp Ala Thr Pro Gly Th

#r His Leu Pro Cys Asp

1330

#       1335

#      1340

gtc ggc gac ccc cac caa ctc gcc acc acc ct

#c acc cac atc ccc caa     4079

Val Gly Asp Pro His Gln Leu Ala Thr Thr Le

#u Thr His Ile Pro Gln

1345

#   1350

#  1355

ccc ctc acc gcc atc ttc cac acc gcc gcc ac

#c ctc gac gac ggc atc     4127

Pro Leu Thr Ala Ile Phe His Thr Ala Ala Th

#r Leu Asp Asp Gly Ile

1360                1365

#                1370

#              1375

ctc cac gcc ctc acc ccc gac cgc ctc acc ac

#c gtc ctc cac ccc aaa     4175

Leu His Ala Leu Thr Pro Asp Arg Leu Thr Th

#r Val Leu His Pro Lys

1380

#               1385

#              1390

gcc aac gcc gcc tgg cac ctg cac cac ctc ac

#c caa aac caa ccc ctc     4223

Ala Asn Ala Ala Trp His Leu His His Leu Th

#r Gln Asn Gln Pro Leu

1395

#           1400

#          1405

acc cac ttc gtc ctc tac tcc agc gcc gcc gc

#c gtc ctc ggc agc ccc     4271

Thr His Phe Val Leu Tyr Ser Ser Ala Ala Al

#a Val Leu Gly Ser Pro

1410

#       1415

#      1420

gga caa gga aac tac gcc gcc gcc aac gcc tt

#c ctc gac gcc ctc gcc     4319

Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ph

#e Leu Asp Ala Leu Ala

1425

#   1430

#  1435

acc cac cgc cac acc ctc ggc caa ccc gcc ac

#c tcc atc gcc tgg ggc     4367

Thr His Arg His Thr Leu Gly Gln Pro Ala Th

#r Ser Ile Ala Trp Gly

1440                1445

#                1450

#              1455

atg tgg cac acc acc agc acc ctc acc gga ca

#a ctc gac gac gcc gac     4415

Met Trp His Thr Thr Ser Thr Leu Thr Gly Gl

#n Leu Asp Asp Ala Asp

1460

#               1465

#              1470

cgg gac cgc atc cgc cgc ggc ggt ttc ctc cc

#g atc acg gac gac gag     4463

Arg Asp Arg Ile Arg Arg Gly Gly Phe Leu Pr

#o Ile Thr Asp Asp Glu

1475

#           1480

#          1485

ggc atggggatgc at

#

#

#  4478

Gly

<210> SEQ ID NO 17

<211> LENGTH: 1488

<212> TYPE: PRT

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: Synthetic

PKS synthase fragment

<400> SEQUENCE: 17

Ile Trp Gln Leu Ala Glu Ala Leu Leu Thr Le

#u Val Arg Glu Ser Thr

1               5

#                 10

#                 15

Ala Ala Val Leu Gly His Val Gly Gly Glu As

#p Ile Pro Ala Thr Ala

20

#             25

#             30

Ala Phe Lys Asp Leu Gly Ile Asp Ser Leu Th

#r Ala Val Gln Leu Arg

35

#         40

#         45

Asn Ala Leu Thr Glu Ala Thr Gly Val Arg Le

#u Asn Ala Thr Ala Val

50

#     55

#     60

Phe Asp Phe Pro Thr Pro His Val Leu Ala Gl

#y Lys Leu Gly Asp Glu

65

# 70

# 75

# 80

Leu Thr Gly Thr Arg Ala Pro Val Val Pro Ar

#g Thr Ala Ala Thr Ala

85

#                 90

#                 95

Gly Ala His Asp Glu Pro Leu Ala Ile Val Gl

#y Met Ala Cys Arg Leu

100

#           105

#           110

Pro Gly Gly Val Ala Ser Pro Glu Glu Leu Tr

#p His Leu Val Ala Ser

115

#       120

#       125

Gly Thr Asp Ala Ile Thr Glu Phe Pro Thr As

#p Arg Gly Trp Asp Val

130

#   135

#   140

Asp Ala Ile Tyr Asp Pro Asp Pro Asp Ala Il

#e Gly Lys Thr Phe Val

145                 1

#50                 1

#55                 1

#60

Arg His Gly Gly Phe Leu Thr Gly Ala Thr Gl

#y Phe Asp Ala Ala Phe

165

#               170

#               175

Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Me

#t Asp Pro Gln Gln Arg

180

#           185

#           190

Val Leu Leu Glu Thr Ser Trp Glu Ala Phe Gl

#u Ser Ala Gly Ile Thr

195

#       200

#       205

Pro Asp Ser Thr Arg Gly Ser Asp Thr Gly Va

#l Phe Val Gly Ala Phe

210

#   215

#   220

Ser Tyr Gly Tyr Gly Thr Gly Ala Asp Thr As

#p Gly Phe Gly Ala Thr

225                 2

#30                 2

#35                 2

#40

Gly Ser Gln Thr Ser Val Leu Ser Gly Arg Le

#u Ser Tyr Phe Tyr Gly

245

#               250

#               255

Leu Glu Gly Pro Ala Val Thr Val Asp Thr Al

#a Cys Ser Ser Ser Leu

260

#           265

#           270

Val Ala Leu His Gln Ala Gly Gln Ser Leu Ar

#g Ser Gly Glu Cys Ser

275

#       280

#       285

Leu Ala Leu Val Gly Gly Val Thr Val Met Al

#a Ser Pro Gly Gly Phe

290

#   295

#   300

Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Pr

#o Asp Gly Arg Ala Lys

305                 3

#10                 3

#15                 3

#20

Ala Phe Gly Ala Gly Ala Asp Gly Thr Ser Ph

#e Ala Glu Gly Ala Gly

325

#               330

#               335

Val Leu Ile Val Glu Arg Leu Ser Asp Ala Gl

#u Arg Asn Gly His Thr

340

#           345

#           350

Val Leu Ala Val Val Arg Gly Ser Ala Val As

#n Gln Asp Gly Ala Ser

355

#       360

#       365

Asn Gly Leu Ser Ala Pro Asn Gly Pro Ser Gl

#n Glu Arg Val Ile Arg

370

#   375

#   380

Gln Ala Leu Ala Asn Ala Gly Leu Thr Pro Al

#a Asp Val Asp Ala Val

385                 3

#90                 3

#95                 4

#00

Glu Ala His Gly Thr Gly Thr Arg Leu Gly As

#p Pro Ile Glu Ala Gln

405

#               410

#               415

Ala Val Leu Ala Thr Tyr Gly Gln Glu Arg Al

#a Thr Pro Leu Leu Leu

420

#           425

#           430

Gly Ser Leu Lys Ser Asn Ile Gly His Ala Gl

#n Ala Ala Ser Gly Val

435

#       440

#       445

Ala Gly Ile Ile Lys Met Val Gln Ala Leu Ar

#g His Gly Glu Leu Pro

450

#   455

#   460

Pro Thr Leu His Ala Asp Glu Pro Ser Pro Hi

#s Val Asp Trp Thr Ala

465                 4

#70                 4

#75                 4

#80

Gly Ala Val Glu Leu Leu Thr Ser Ala Arg Pr

#o Trp Pro Glu Thr Asp

485

#               490

#               495

Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gl

#y Ile Ser Gly Thr Asn

500

#           505

#           510

Ala His Val Ile Leu Glu Ser Ala Pro Pro Th

#r Gln Pro Ala Asp Asn

515

#       520

#       525

Ala Val Ile Glu Arg Ala Pro Glu Trp Val Pr

#o Leu Val Ile Ser Ala

530

#   535

#   540

Arg Thr Gln Ser Ala Leu Thr Glu His Glu Gl

#y Arg Leu Arg Ala Tyr

545                 5

#50                 5

#55                 5

#60

Leu Ala Ala Ser Pro Gly Val Asp Met Arg Al

#a Val Ala Ser Thr Leu

565

#               570

#               575

Ala Met Thr Arg Ser Val Phe Glu His Arg Al

#a Val Leu Leu Gly Asp

580

#           585

#           590

Asp Thr Val Thr Gly Thr Ala Val Ser Asp Pr

#o Arg Ala Val Phe Val

595

#       600

#       605

Phe Pro Gly Gln Gly Ser Gln Arg Ala Gly Me

#t Gly Glu Glu Leu Ala

610

#   615

#   620

Ala Ala Phe Pro Val Phe Ala Arg Ile His Gl

#n Gln Val Trp Asp Leu

625                 6

#30                 6

#35                 6

#40

Leu Asp Val Pro Asp Leu Glu Val Asn Glu Th

#r Gly Tyr Ala Gln Pro

645

#               650

#               655

Ala Leu Phe Ala Met Gln Val Ala Leu Phe Gl

#y Leu Leu Glu Ser Trp

660

#           665

#           670

Gly Val Arg Pro Asp Ala Val Ile Gly His Se

#r Val Gly Glu Leu Ala

675

#       680

#       685

Ala Ala Tyr Val Ser Gly Val Trp Ser Leu Gl

#u Asp Ala Cys Thr Leu

690

#   695

#   700

Val Ser Ala Arg Ala Arg Leu Met Gln Ala Le

#u Pro Ala Gly Gly Val

705                 7

#10                 7

#15                 7

#20

Met Val Ala Val Pro Val Ser Glu Asp Glu Al

#a Arg Ala Val Leu Gly

725

#               730

#               735

Glu Gly Val Glu Ile Ala Ala Val Asn Gly Pr

#o Ser Ser Val Val Leu

740

#           745

#           750

Ser Gly Asp Glu Ala Ala Val Leu Gln Ala Al

#a Glu Gly Leu Gly Lys

755

#       760

#       765

Trp Thr Arg Leu Ala Thr Ser His Ala Phe Hi

#s Ser Ala Arg Met Glu

770

#   775

#   780

Pro Met Leu Glu Glu Phe Arg Ala Val Ala Gl

#u Gly Leu Thr Tyr Arg

785                 7

#90                 7

#95                 8

#00

Thr Pro Gln Val Ser Met Ala Val Gly Asp Gl

#n Val Thr Thr Ala Glu

805

#               810

#               815

Tyr Trp Val Arg Gln Val Arg Asp Thr Val Ar

#g Phe Gly Glu Gln Val

820

#           825

#           830

Ala Ser Tyr Glu Asp Ala Val Phe Val Glu Le

#u Gly Ala Asp Arg Ser

835

#       840

#       845

Leu Ala Arg Leu Val Asp Gly Val Ala Met Le

#u His Gly Asp His Glu

850

#   855

#   860

Ile Gln Ala Ala Ile Gly Ala Leu Ala His Le

#u Tyr Val Asn Gly Val

865                 8

#70                 8

#75                 8

#80

Thr Val Asp Trp Pro Ala Leu Leu Gly Asp Al

#a Pro Ala Thr Arg Val

885

#               890

#               895

Leu Asp Leu Pro Thr Tyr Ala Phe Gln His Gl

#n Arg Tyr Trp Leu Glu

900

#           905

#           910

Ser Ala Arg Pro Ala Ala Ser Asp Ala Gly Hi

#s Pro Val Leu Gly Ser

915

#       920

#       925

Gly Ile Ala Leu Ala Gly Ser Pro Gly Arg Va

#l Phe Thr Gly Ser Val

930

#   935

#   940

Pro Thr Gly Ala Asp Arg Ala Val Phe Val Al

#a Glu Leu Ala Leu Ala

945                 9

#50                 9

#55                 9

#60

Ala Ala Asp Ala Val Asp Cys Ala Thr Val Gl

#u Arg Leu Asp Ile Ala

965

#               970

#               975

Ser Val Pro Gly Arg Pro Gly His Gly Arg Th

#r Thr Val Gln Thr Trp

980

#           985

#           990

Val Asp Glu Pro Ala Asp Asp Gly Arg Arg Ar

#g Phe Thr Val His Thr

995

#       1000

#      1005

Arg Thr Gly Asp Ala Pro Trp Thr Leu His Al

#a Glu Gly Val Leu Arg

1010

#   1015

#  1020

Pro His Gly Thr Ala Leu Pro Asp Ala Ala As

#p Ala Glu Trp Pro Pro

1025                1030

#                1035

#              1040

Pro Gly Ala Val Pro Ala Asp Gly Leu Pro Gl

#y Val Trp Arg Arg Gly

1045

#               1050

#              1055

Asp Gln Val Phe Ala Glu Ala Glu Val Asp Gl

#y Pro Asp Gly Phe Val

1060

#           1065

#          1070

Val His Pro Asp Leu Leu Asp Ala Val Phe Se

#r Ala Val Gly Asp Gly

1075

#       1080

#      1085

Ser Arg Gln Pro Ala Gly Trp Arg Asp Leu Th

#r Val His Ala Ser Asp

1090

#   1095

#  1100

Ala Thr Val Leu Arg Ala Cys Leu Thr Arg Ar

#g Thr Asp Gly Ala Met

1105                1110

#                1115

#              1120

Gly Phe Ala Ala Phe Asp Gly Ala Gly Leu Pr

#o Val Leu Thr Ala Glu

1125

#               1130

#              1135

Ala Val Thr Leu Arg Glu Val Ala Ser Pro Se

#r Gly Ser Glu Glu Ser

1140

#           1145

#          1150

Asp Gly Leu His Arg Leu Glu Trp Leu Ala Va

#l Ala Glu Ala Val Tyr

1155

#       1160

#      1165

Asp Gly Asp Leu Pro Glu Gly His Val Leu Il

#e Thr Ala Ala His Pro

1170

#   1175

#  1180

Asp Asp Pro Glu Asp Ile Pro Thr Arg Ala Hi

#s Thr Arg Ala Thr Arg

1185                1190

#                1195

#              1200

Val Leu Thr Ala Leu Gln His His Leu Thr Th

#r Thr Asp His Thr Leu

1205

#               1210

#              1215

Ile Val His Thr Thr Thr Asp Pro Ala Gly Al

#a Thr Val Thr Gly Leu

1220

#           1225

#          1230

Thr Arg Thr Ala Gln Asn Glu His Pro His Ar

#g Ile Arg Leu Ile Glu

1235

#       1240

#      1245

Thr Asp His Pro His Thr Pro Leu Pro Leu Al

#a Gln Leu Ala Thr Leu

1250

#   1255

#  1260

Asp His Pro His Leu Arg Leu Thr His His Th

#r Leu His His Pro His

1265                1270

#                1275

#              1280

Leu Thr Pro Leu His Thr Thr Thr Pro Pro Th

#r Thr Thr Pro Leu Asn

1285

#               1290

#              1295

Pro Glu His Ala Ile Ile Ile Thr Gly Gly Se

#r Gly Thr Leu Ala Gly

1300

#           1305

#          1310

Ile Leu Ala Arg His Leu Asn His Pro His Th

#r Tyr Leu Leu Ser Arg

1315

#       1320

#      1325

Thr Pro Pro Pro Asp Ala Thr Pro Gly Thr Hi

#s Leu Pro Cys Asp Val

1330

#   1335

#  1340

Gly Asp Pro His Gln Leu Ala Thr Thr Leu Th

#r His Ile Pro Gln Pro

1345                1350

#                1355

#              1360

Leu Thr Ala Ile Phe His Thr Ala Ala Thr Le

#u Asp Asp Gly Ile Leu

1365

#               1370

#              1375

His Ala Leu Thr Pro Asp Arg Leu Thr Thr Va

#l Leu His Pro Lys Ala

1380

#           1385

#          1390

Asn Ala Ala Trp His Leu His His Leu Thr Gl

#n Asn Gln Pro Leu Thr

1395

#       1400

#      1405

His Phe Val Leu Tyr Ser Ser Ala Ala Ala Va

#l Leu Gly Ser Pro Gly

1410

#   1415

#  1420

Gln Gly Asn Tyr Ala Ala Ala Asn Ala Phe Le

#u Asp Ala Leu Ala Thr

1425                1430

#                1435

#              1440

His Arg His Thr Leu Gly Gln Pro Ala Thr Se

#r Ile Ala Trp Gly Met

1445

#               1450

#              1455

Trp His Thr Thr Ser Thr Leu Thr Gly Gln Le

#u Asp Asp Ala Asp Arg

1460

#           1465

#          1470

Asp Arg Ile Arg Arg Gly Gly Phe Leu Pro Il

#e Thr Asp Asp Glu Gly

1475

#       1480

#      1485

<210> SEQ ID NO 18

<211> LENGTH: 4571

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Description of Artificial

#Sequence: DNA

encoding synthetic PKS synthase frag

#ment

<220> FEATURE:

<221> NAME/KEY: CDS

<222> LOCATION: (9)..(4559)

<400> SEQUENCE: 18

agatctgg cag ctc gcc gaa gcg ctg ctg acg ctc

#gtc cgg gag agc acc      50

Gln Leu Ala Glu Ala L

#eu Leu Thr Leu Val Arg Glu Ser Thr

1

#        5

#        10

gcc gcc gtg ctc ggc cac gtg ggt ggc gag ga

#c atc ccc gcg acg gcg       98

Ala Ala Val Leu Gly His Val Gly Gly Glu As

#p Ile Pro Ala Thr Ala

15

# 20

# 25

# 30

gcg ttc aag gac ctc ggc atc gac tcg ctc ac

#c gcg gtc cag ctg cgc      146

Ala Phe Lys Asp Leu Gly Ile Asp Ser Leu Th

#r Ala Val Gln Leu Arg

35

#                 40

#                 45

aac gcc ctc acc gag gcg acc ggt gtg cgg ct

#g aac gcc acg gcg gtc      194

Asn Ala Leu Thr Glu Ala Thr Gly Val Arg Le

#u Asn Ala Thr Ala Val

50

#             55

#             60

ttc gac ttc ccg acc ccg cac gtg ctc gcc gg

#g aag ctc ggc gac gaa      242

Phe Asp Phe Pro Thr Pro His Val Leu Ala Gl

#y Lys Leu Gly Asp Glu

65

#         70

#         75

ctg acc ggc acc cgc gcg ccc gtc gtg ccc cg

#g acc gcg gcc acg gcc      290

Leu Thr Gly Thr Arg Ala Pro Val Val Pro Ar

#g Thr Ala Ala Thr Ala

80

#     85

#     90

ggt gcg cac gac gag ccg ctg gcg atc gtg gg

#a atg gcc tgc cgg ctg      338

Gly Ala His Asp Glu Pro Leu Ala Ile Val Gl

#y Met Ala Cys Arg Leu

95

#100

#105

#110

ccc ggc ggg gtc gcg tca ccc gag gag ctg tg

#g cac ctc gtg gca tcc      386

Pro Gly Gly Val Ala Ser Pro Glu Glu Leu Tr

#p His Leu Val Ala Ser

115

#               120

#               125

ggc acc gac gcc atc acg gag ttc ccg acg ga

#c cgc ggc tgg gac gtc      434

Gly Thr Asp Ala Ile Thr Glu Phe Pro Thr As

#p Arg Gly Trp Asp Val

130

#           135

#           140

gac gcg atc tac gac ccg gac ccc gac gcg at

#c ggc aag acc ttc gtc      482

Asp Ala Ile Tyr Asp Pro Asp Pro Asp Ala Il

#e Gly Lys Thr Phe Val

145

#       150

#       155

cgg cac ggt ggc ttc ctc acc ggc gcg aca gg

#c ttc gac gcg gcg ttc      530

Arg His Gly Gly Phe Leu Thr Gly Ala Thr Gl

#y Phe Asp Ala Ala Phe

160

#   165

#   170

ttc ggc atc agc ccg cgc gag gcc ctc gcg at

#g gac ccg cag cag cgg      578

Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Me

#t Asp Pro Gln Gln Arg

175                 1

#80                 1

#85                 1

#90

gtg ctc ctg gag acg tcg tgg gag gcg ttc ga

#a agc gcc ggc atc acc      626

Val Leu Leu Glu Thr Ser Trp Glu Ala Phe Gl

#u Ser Ala Gly Ile Thr

195

#               200

#               205

ccg gac tcg acc cgc ggc agc gac acc ggc gt

#g ttc gtc ggc gcc ttc      674

Pro Asp Ser Thr Arg Gly Ser Asp Thr Gly Va

#l Phe Val Gly Ala Phe

210

#           215

#           220

tcc tac ggt tac ggc acc ggt gcg gac acc ga

#c ggc ttc ggc gcg acc      722

Ser Tyr Gly Tyr Gly Thr Gly Ala Asp Thr As

#p Gly Phe Gly Ala Thr

225

#       230

#       235

ggc tcg cag acc agt gtg ctc tcc ggc cgg ct

#g tcg tac ttc tac ggt      770

Gly Ser Gln Thr Ser Val Leu Ser Gly Arg Le

#u Ser Tyr Phe Tyr Gly

240

#   245

#   250

ctg gag ggt ccg gcg gtc acg gtc gac acg gc

#g tgt tcg tcg tcg ctg      818

Leu Glu Gly Pro Ala Val Thr Val Asp Thr Al

#a Cys Ser Ser Ser Leu

255                 2

#60                 2

#65                 2

#70

gtg gcg ctg cac cag gcc ggg cag tcg ctg cg

#c tcc ggc gaa tgc tcg      866

Val Ala Leu His Gln Ala Gly Gln Ser Leu Ar

#g Ser Gly Glu Cys Ser

275

#               280

#               285

ctc gcc ctg gtc ggc ggc gtc acg gtg atg gc

#g tct ccc ggc ggc ttc      914

Leu Ala Leu Val Gly Gly Val Thr Val Met Al

#a Ser Pro Gly Gly Phe

290

#           295

#           300

gtg gag ttc tcc cgg cag cgc ggc ctc gcg cc

#g gac ggc cgg gcg aag      962

Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Pr

#o Asp Gly Arg Ala Lys

305

#       310

#       315

gcg ttc ggc gcg ggt gcg gac ggc acg agc tt

#c gcc gag ggt gcc ggt     1010

Ala Phe Gly Ala Gly Ala Asp Gly Thr Ser Ph

#e Ala Glu Gly Ala Gly

320

#   325

#   330

gtg ctg atc gtc gag agg ctc tcc gac gcc ga

#a cgc aac ggt cac acc     1058

Val Leu Ile Val Glu Arg Leu Ser Asp Ala Gl

#u Arg Asn Gly His Thr

335                 3

#40                 3

#45                 3

#50

gtc ctg gcg gtc gtc cgt ggt tcg gcg gtc aa

#c cag gat ggt gcc tcc     1106

Val Leu Ala Val Val Arg Gly Ser Ala Val As

#n Gln Asp Gly Ala Ser

355

#               360

#               365

aac ggg ctg tcg gcg ccg aac ggg ccg tcg ca

#g gag cgg gtg atc cgg     1154

Asn Gly Leu Ser Ala Pro Asn Gly Pro Ser Gl

#n Glu Arg Val Ile Arg

370

#           375

#           380

cag gcc ctg gcc aac gcc ggg ctc acc ccg gc

#g gac gtg gac gcc gtc     1202

Gln Ala Leu Ala Asn Ala Gly Leu Thr Pro Al

#a Asp Val Asp Ala Val

385

#       390

#       395

gag gcc cac ggc acc ggc acc agg ctg ggc ga

#c ccc atc gag gca cag     1250

Glu Ala His Gly Thr Gly Thr Arg Leu Gly As

#p Pro Ile Glu Ala Gln

400

#   405

#   410

gcg gta ctg gcc acc tac gga cag gag cgc gc

#c acc ccc ctg ctg ctg     1298

Ala Val Leu Ala Thr Tyr Gly Gln Glu Arg Al

#a Thr Pro Leu Leu Leu

415                 4

#20                 4

#25                 4

#30

ggc tcg ctg aag tcc aac atc ggc cac gcc ca

#g gcc gcg tcc ggc gtc     1346

Gly Ser Leu Lys Ser Asn Ile Gly His Ala Gl

#n Ala Ala Ser Gly Val

435

#               440

#               445

gcc ggc atc atc aag atg gtg cag gcc ctc cg

#g cac ggg gag ctg ccg     1394

Ala Gly Ile Ile Lys Met Val Gln Ala Leu Ar

#g His Gly Glu Leu Pro

450

#           455

#           460

ccg acg ctg cac gcc gac gag ccg tcg ccg ca

#c gtc gac tgg acg gcc     1442

Pro Thr Leu His Ala Asp Glu Pro Ser Pro Hi

#s Val Asp Trp Thr Ala

465

#       470

#       475

ggc gcc gtc gaa ctg ctg acg tcg gcc cgg cc

#g tgg ccc gag acc gac     1490

Gly Ala Val Glu Leu Leu Thr Ser Ala Arg Pr

#o Trp Pro Glu Thr Asp

480

#   485

#   490

cgg cct agg cgg gcg ggc gtg tcg tcc ttc gg

#a gtc agc ggc acc aac     1538

Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gl

#y Val Ser Gly Thr Asn

495                 5

#00                 5

#05                 5

#10

gcc cac gtc atc ctg gag agc gca ccc ccc gc

#t cag ccc gcg gag gag     1586

Ala His Val Ile Leu Glu Ser Ala Pro Pro Al

#a Gln Pro Ala Glu Glu

515

#               520

#               525

gcg cag cct gtt gag acg ccg gtg gtg gcc tc

#g gat gtg ctg ccg ctg     1634

Ala Gln Pro Val Glu Thr Pro Val Val Ala Se

#r Asp Val Leu Pro Leu

530

#           535

#           540

gtg ata tcg gcc aag acc cag ccc gcc ctg ac

#c gaa cac gaa gac cgg     1682

Val Ile Ser Ala Lys Thr Gln Pro Ala Leu Th

#r Glu His Glu Asp Arg

545

#       550

#       555

ctg cgc gcc tac ctg gcg gcg tcg ccc ggg gc

#g gat ata cgg gct gtg     1730

Leu Arg Ala Tyr Leu Ala Ala Ser Pro Gly Al

#a Asp Ile Arg Ala Val

560

#   565

#   570

gca tcg acg ctg gcg gtg aca cgg tcg gtg tt

#c gag cac cgc gcc gta     1778

Ala Ser Thr Leu Ala Val Thr Arg Ser Val Ph

#e Glu His Arg Ala Val

575                 5

#80                 5

#85                 5

#90

ctc ctt gga gat gac acc gtc acc ggc acc gc

#g gtg acc gac ccc agg     1826

Leu Leu Gly Asp Asp Thr Val Thr Gly Thr Al

#a Val Thr Asp Pro Arg

595

#               600

#               605

atc gtg ttt gtc ttt ccc ggg cag ggg tgg ca

#g tgg ctg ggg atg ggc     1874

Ile Val Phe Val Phe Pro Gly Gln Gly Trp Gl

#n Trp Leu Gly Met Gly

610

#           615

#           620

agt gca ctg cgc gat tcg tcg gtg gtg ttc gc

#c gag cgg atg gcc gag     1922

Ser Ala Leu Arg Asp Ser Ser Val Val Phe Al

#a Glu Arg Met Ala Glu

625

#       630

#       635

tgt gcg gcg gcg ttg cgc gag ttc gtg gac tg

#g gat ctg ttc acg gtt     1970

Cys Ala Ala Ala Leu Arg Glu Phe Val Asp Tr

#p Asp Leu Phe Thr Val

640

#   645

#   650

ctg gat gat ccg gcg gtg gtg gac cgg gtt ga

#t gtg gtc cag ccc gct     2018

Leu Asp Asp Pro Ala Val Val Asp Arg Val As

#p Val Val Gln Pro Ala

655                 6

#60                 6

#65                 6

#70

tcc tgg gcg atg atg gtt tcc ctg gcc gcg gt

#g tgg cag gcg gcc ggt     2066

Ser Trp Ala Met Met Val Ser Leu Ala Ala Va

#l Trp Gln Ala Ala Gly

675

#               680

#               685

gtg cgg ccg gat gcg gtg atc ggc cat tcg ca

#g ggt gag atc gcc gca     2114

Val Arg Pro Asp Ala Val Ile Gly His Ser Gl

#n Gly Glu Ile Ala Ala

690

#           695

#           700

gct tgt gtg gcg ggt gcg gtg tca cta cgc ga

#t gcc gcc cgg atc gtg     2162

Ala Cys Val Ala Gly Ala Val Ser Leu Arg As

#p Ala Ala Arg Ile Val

705

#       710

#       715

acc ttg cgc agc cag gcg atc gcc cgg ggc ct

#g gcg ggc cgg ggc gcg     2210

Thr Leu Arg Ser Gln Ala Ile Ala Arg Gly Le

#u Ala Gly Arg Gly Ala

720

#   725

#   730

atg gca tcc gtc gcc ctg ccc gcg cag gat gt

#c gag ctg gtc gac ggg     2258

Met Ala Ser Val Ala Leu Pro Ala Gln Asp Va

#l Glu Leu Val Asp Gly

735                 7

#40                 7

#45                 7

#50

gcc tgg atc gcc gcc cac aac ggg ccc gcc tc

#c acc gtg atc gcg ggc     2306

Ala Trp Ile Ala Ala His Asn Gly Pro Ala Se

#r Thr Val Ile Ala Gly

755

#               760

#               765

acc ccg gaa gcg gtc gac cat gtc ctc acc gc

#t cat gag gca caa ggg     2354

Thr Pro Glu Ala Val Asp His Val Leu Thr Al

#a His Glu Ala Gln Gly

770

#           775

#           780

gtg cgg gtg cgg cgg atc acc gtc gac tat gc

#c tcg cac acc ccg cac     2402

Val Arg Val Arg Arg Ile Thr Val Asp Tyr Al

#a Ser His Thr Pro His

785

#       790

#       795

gtc gag ctg atc cgc gac gaa cta ctc gac at

#c act agc gac agc agc     2450

Val Glu Leu Ile Arg Asp Glu Leu Leu Asp Il

#e Thr Ser Asp Ser Ser

800

#   805

#   810

tcg cag acc ccg ctc gtg ccg tgg ctg tcg ac

#c gtg gac ggc acc tgg     2498

Ser Gln Thr Pro Leu Val Pro Trp Leu Ser Th

#r Val Asp Gly Thr Trp

815                 8

#20                 8

#25                 8

#30

gtc gac agc ccg ctg gac ggg gag tac tgg ta

#c cgg aac ctg cgt gaa     2546

Val Asp Ser Pro Leu Asp Gly Glu Tyr Trp Ty

#r Arg Asn Leu Arg Glu

835

#               840

#               845

ccg gtc ggt ttc cac ccc gcc gtc agc cag tt

#g cag gcc cag ggc gac     2594

Pro Val Gly Phe His Pro Ala Val Ser Gln Le

#u Gln Ala Gln Gly Asp

850

#           855

#           860

acc gtg ttc gtc gag gtc agc gcc agc ccg gt

#g ttg ttg cag gcg atg     2642

Thr Val Phe Val Glu Val Ser Ala Ser Pro Va

#l Leu Leu Gln Ala Met

865

#       870

#       875

gac gac gat gtc gtc acg gtt gcc acg ctg cg

#t cgt gac gac ggc gac     2690

Asp Asp Asp Val Val Thr Val Ala Thr Leu Ar

#g Arg Asp Asp Gly Asp

880

#   885

#   890

gcc acc cgg atg ctc acc gcc ctg gca cag gc

#c tat gtc cac ggc gtc     2738

Ala Thr Arg Met Leu Thr Ala Leu Ala Gln Al

#a Tyr Val His Gly Val

895                 9

#00                 9

#05                 9

#10

acc gtc gac tgg ccc gcc atc ctc ggc acc ac

#c aca acc cgg gta ctg     2786

Thr Val Asp Trp Pro Ala Ile Leu Gly Thr Th

#r Thr Thr Arg Val Leu

915

#               920

#               925

gac ctt ccg acc tac gcc ttc caa cac cag cg

#g tac tgg ctc gag tcg     2834

Asp Leu Pro Thr Tyr Ala Phe Gln His Gln Ar

#g Tyr Trp Leu Glu Ser

930

#           935

#           940

gca cgc ccg gcc gca tcc gac gcg ggc cac cc

#c gtg ctg ggc tcc ggt     2882

Ala Arg Pro Ala Ala Ser Asp Ala Gly His Pr

#o Val Leu Gly Ser Gly

945

#       950

#       955

atc gcc ctc gcc ggg tcg ccg ggc cgg gtg tt

#c acg ggt tcc gtg ccg     2930

Ile Ala Leu Ala Gly Ser Pro Gly Arg Val Ph

#e Thr Gly Ser Val Pro

960

#   965

#   970

acc ggt gcg gac cgc gcg gtg ttc gtc gcc ga

#g ctg gcg ctg gcc gcc     2978

Thr Gly Ala Asp Arg Ala Val Phe Val Ala Gl

#u Leu Ala Leu Ala Ala

975                 9

#80                 9

#85                 9

#90

gcg gac gcg gtc gac tgc gcc acg gtc gag cg

#g ctc gac atc gcc tcc     3026

Ala Asp Ala Val Asp Cys Ala Thr Val Glu Ar

#g Leu Asp Ile Ala Ser

995

#               1000

#              1005

gtg ccc ggc cgg ccg ggc cat ggc cgg acg ac

#c gta cag acc tgg gtc     3074

Val Pro Gly Arg Pro Gly His Gly Arg Thr Th

#r Val Gln Thr Trp Val

1010

#           1015

#          1020

gac gag ccg gcg gac gac ggc cgg cgc cgg tt

#c acc gtg cac acc cgc     3122

Asp Glu Pro Ala Asp Asp Gly Arg Arg Arg Ph

#e Thr Val His Thr Arg

1025

#       1030

#      1035

acc ggc gac gcc ccg tgg acg ctg cac gcc ga

#g ggg gtg ctg cgc ccc     3170

Thr Gly Asp Ala Pro Trp Thr Leu His Ala Gl

#u Gly Val Leu Arg Pro

1040

#   1045

#  1050

cat ggc acg gcc ctg ccc gat gcg gcc gac gc

#c gag tgg ccc cca ccg     3218

His Gly Thr Ala Leu Pro Asp Ala Ala Asp Al

#a Glu Trp Pro Pro Pro

1055                1060

#                1065

#              1070

ggc gcg gtg ccc gcg gac ggg ctg ccg ggt gt

#g tgg cgc cgg ggg gac     3266

Gly Ala Val Pro Ala Asp Gly Leu Pro Gly Va

#l Trp Arg Arg Gly Asp

1075

#               1080

#              1085

cag gtc ttc gcc gag gcc gag gtg gac gga cc

#g gac ggt ttc gtg gtg     3314

Gln Val Phe Ala Glu Ala Glu Val Asp Gly Pr

#o Asp Gly Phe Val Val

1090

#           1095

#          1100

cac ccc gac ctg ctc gac gcg gtc ttc tcc gc

#g gtc ggc gac gga agc     3362

His Pro Asp Leu Leu Asp Ala Val Phe Ser Al

#a Val Gly Asp Gly Ser

1105

#       1110

#      1115

cgc cag ccg gcc gga tgg cgc gac ctg acg gt

#g cac gcg tcg gac gcc     3410

Arg Gln Pro Ala Gly Trp Arg Asp Leu Thr Va

#l His Ala Ser Asp Ala

1120

#   1125

#  1130

acc gta ctg cgc gcc tgc ctc acc cgg cgc ac

#c gac gga gcc atg gga     3458

Thr Val Leu Arg Ala Cys Leu Thr Arg Arg Th

#r Asp Gly Ala Met Gly

1135                1140

#                1145

#              1150

ttc gcc gcc ttc gac ggc gcc ggc ctg ccg gt

#a ctc acc gcg gag gcg     3506

Phe Ala Ala Phe Asp Gly Ala Gly Leu Pro Va

#l Leu Thr Ala Glu Ala

1155

#               1160

#              1165

gtg acg ctg cgg gag gtg gcg tca ccg tcc gg

#c tcc gag gag tcg gac     3554

Val Thr Leu Arg Glu Val Ala Ser Pro Ser Gl

#y Ser Glu Glu Ser Asp

1170

#           1175

#          1180

ggc ctg cac cgg ttg gag tgg ctc gcg gtc gc

#c gag gcg gtc tac gac     3602

Gly Leu His Arg Leu Glu Trp Leu Ala Val Al

#a Glu Ala Val Tyr Asp

1185

#       1190

#      1195

ggt gac ctg ccc gag gga cat gtc ctg atc ac

#c gcc gcc cac ccc gac     3650

Gly Asp Leu Pro Glu Gly His Val Leu Ile Th

#r Ala Ala His Pro Asp

1200

#   1205

#  1210

gac ccc gag gac ata ccc acc cgc gcc cac ac

#c cgc gcc acc cgc gtc     3698

Asp Pro Glu Asp Ile Pro Thr Arg Ala His Th

#r Arg Ala Thr Arg Val

1215                1220

#                1225

#              1230

ctg acc gcc ctg caa cac cac ctc acc acc ac

#c gac cac acc ctc atc     3746

Leu Thr Ala Leu Gln His His Leu Thr Thr Th

#r Asp His Thr Leu Ile

1235

#               1240

#              1245

gtc cac acc acc acc gac ccc gcc ggc gcc ac

#c gtc acc ggc ctc acc     3794

Val His Thr Thr Thr Asp Pro Ala Gly Ala Th

#r Val Thr Gly Leu Thr

1250

#           1255

#          1260

cgc acc gcc cag aac gaa cac ccc cac cgc at

#c cgc ctc atc gaa acc     3842

Arg Thr Ala Gln Asn Glu His Pro His Arg Il

#e Arg Leu Ile Glu Thr

1265

#       1270

#      1275

gac cac ccc cac acc ccc ctc ccc ctg gcc ca

#a ctc gcc acc ctc gac     3890

Asp His Pro His Thr Pro Leu Pro Leu Ala Gl

#n Leu Ala Thr Leu Asp

1280

#   1285

#  1290

cac ccc cac ctc cgc ctc acc cac cac acc ct

#c cac cac ccc cac ctc     3938

His Pro His Leu Arg Leu Thr His His Thr Le

#u His His Pro His Leu

1295                1300

#                1305

#              1310

acc ccc ctc cac acc acc acc cca ccc acc ac

#c acc ccc ctc aac ccc     3986

Thr Pro Leu His Thr Thr Thr Pro Pro Thr Th

#r Thr Pro Leu Asn Pro

1315

#               1320

#              1325

gaa cac gcc atc atc atc acc ggc ggc tcc gg

#c acc ctc gcc ggc atc     4034

Glu His Ala Ile Ile Ile Thr Gly Gly Ser Gl

#y Thr Leu Ala Gly Ile

1330

#           1335

#          1340

ctc gcc cgc cac ctg aac cac ccc cac acc ta

#c ctc ctc tcc cgc acc     4082

Leu Ala Arg His Leu Asn His Pro His Thr Ty

#r Leu Leu Ser Arg Thr

1345

#       1350

#      1355