Patent application title: Method for Producing Recombinant 11-De-O-Methyltomaymycin
Inventors:
IPC8 Class: AC12N1576FI
USPC Class:
1 1
Class name:
Publication date: 2020-07-09
Patent application number: 20200216852
Abstract:
The present invention provides a tomaymycin biosynthetic gene cluster of
Streptomyces species FH6421, and its use for producing
11-de-O-methyltomaymycm.Claims:
1. A nucleic acid molecule comprising at least one nucleic acid selected
from the group consisting of: (a) a nucleic acid comprising at least one
of the Open Reading Frames (ORFs) 1 to 19 of SEQ ID NO: 2 that encodes
proteins of SEQ ID NOs: 4 to 22 or a variant or fragment thereof, whereby
the variant or fragment encodes a functionally active variant or fragment
of a protein of SEQ ID NOs: 4 to 22, (b) a nucleic acid encoding at least
one of the proteins of SEQ ID NOs: 4 to 22 or a functionally active
variant or fragment thereof, (c) a nucleic acid encoding a protein that
is at least 70%, 80%, 90%, 95% or 97% identical in amino acid sequence to
a protein or fragment thereof encoded by the nucleic acid of (a) or (b),
(d) a nucleic acid that hybridizes under stringent conditions with a
nucleic acid of (a) to (c), (e) a nucleic acid that is complementary to a
nucleic acid of (a) to (d).
2. The nucleic acid according to claim 1 comprising the tomaymycin biosynthetic gene cluster of SEQ ID NO: 2 or having a variant or fragment sequence of SEQ ID NO: 2 harboring a variant or fragment of at least one of ORFs 1 to 19, whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22.
3. An expression vector comprising a nucleic acid of claim 1.
4. A cell comprising the expression vector according to claim 3.
5. The cell of claim 4, wherein the cell is Streptomyces species FH6421.
6. A method for producing a cell that harbors a tomaymycin biosynthetic gene cluster or a functionally active variant or fragment thereof.
7. The method according to claim 6, wherein the cell harbors the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof.
8. The method according to claim 7, wherein the cell is a Streptomyces strain harboring the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof.
9. The method according to claim 8, wherein the Streptomyces strain is selected from the group consisting of Streptomyces achromogenes var. tomaymyceticus, Streptomyces species FH6421, and Streptomyces albus/pStW102tc.
10. The method according to claim 6, wherein the cell harbors at least one ORF of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or functionally active variant or fragment thereof, and is capable of producing 11-de-O-methyltomaymycin.
Description:
FIELD OF THE INVENTION
[0001] The present invention relates to a method for recombinantly producing the antibiotic 11-de-O-methyltomaymycin, as well as to nucleic acids coding for the tomaymycin biosynthetic gene cluster.
BACKGROUND
[0002] Tomaymycin is a naturally occurring antibiotic having, besides its anti-bacterial effectiveness, cytotoxic and antitumor activity. Tomaymycin belongs to the chemical group of pyrrolobenzodiazepines (PBD), which are monomeric compounds with weak antibiotic properties that specifically alkylate the minor groove of DNA at a 5'-Pu-G-Pu base sequence. Tomaymycin was isolated from the culture filtrate of Streptomyces achromogenes var. tomaymyceticus by K. Arima (Arima et al., 1972; Nishioka et al., 1972). It was found that tomaymycin is synthesized by a series of proteins that are encoded by a set of genes clustered within a gene cluster. The biosynthetic gene cluster for tomaymycin has been elucidated by Li et al. (Li et al., 2009). Tomaymycin has proven active against tumours in mice, such as sarcoma 180; the benzpyrene sarcoma; the mamma tumor; or leukosarcomatosis such as leukosarcomatosis AKR or O.sub.1498 or against leukemia, plasmacytoma and ovarian cancer cell lines. Tomaymycin has the structural formula presented in FIG. 1(A). 11-de-O-methyltomaymycin has the empirical formula C.sub.15H.sub.18N.sub.2O.sub.4 and a molecular weight of 290.32. The structural formula of 11-de-O-methyltomaymycin is shown in FIG. 1(B). The tomaymycin-urea adduct is shown in FIG. 1(C).
[0003] Whereas 11-de-O-methyltomaymycin is produced during natural biosynthesis, tomaymycin is produced artificially therefrom by addition of methanol during the isolation/purification process (cf. FIG. 1).
[0004] FR 1.516.743 discloses a method for the production of tomaymycin using chromatography of a solution comprising the antibiotic adsorbed to an adsorbens, countercurrent distribution, preparation of a complex with urea and regeneration of the antibiotic from the medium comprising the complex. The medium for culturing the strain producing tomaymycin contained usual nutrients, such as an assimilable carbon and nitrogen sources, minerals and optionally growth factors. Assimilable nitrogen sources comprise, for example, nitrate, anorganic or organic ammonium salts, urea or amino acids or substances comprising nitrogen in protidic form, such as casein, soybean flour or distillers' solubles. However, none of the media indicated in FR 1.516.743 resulted in the production of high, satisfactory yields of tomaymycin.
[0005] Therefore, there is a need in the art for a method for producing high amounts of tomaymycin. This problem is solved by the present invention.
SUMMARY OF THE INVENTION
[0006] Embodiments of the invention provide a nucleic acid molecule comprising at least one nucleic acid selected from the group consisting of: (a) a nucleic acid comprising at least one of the Open Reading Frames (ORFs) 1 to 19 of SEQ ID NO: 2 that encodes proteins of SEQ ID NOs: 4 to 22 or a variant or fragment thereof, whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22, (b) a nucleic acid encoding at least one of the proteins of SEQ ID NOs: 4 to 22 or a functionally active variant or fragment thereof, (c) a nucleic acid encoding a protein that is at least 70%, 80%, 90%, 95% or 97% identical in amino acid sequence to a protein or fragment thereof encoded by the nucleic acid of (a) or (b), (d) a nucleic acid that hybridizes under stringent conditions with a nucleic acid of (a) to (c), (e) a nucleic acid that is complementary to a nucleic acid of (a) to (d). In certain embodiments, the nucleic acid comprises the tomaymycin biosynthetic gene cluster of SEQ ID NO: 2 or a variant or fragment sequence of SEQ ID NO: 2 harboring a variant or fragment of at least one of ORFs 1 to 19, whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22.
[0007] Another embodiment of the invention provides an expression vector comprising any of the above nucleic acids.
[0008] Yet another embodiment of the invention provides a cell comprising the above expression vector. In a specific embodiment, the cell is Streptomyces species FH6421.
[0009] An embodiment of the invention provides a method for producing a cell that harbors a tomaymycin biosynthetic gene cluster or a functionally active variant or fragment thereof. In certain embodiments, the cell harbors the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof. In other specific embodiment, the cell is a Streptomyces strain harboring the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof. In yet other specific embodiments, the Streptomyces strain is selected from the group consisting of Streptomyces achromogenes var. tomaymyceticus, Streptomyces species FH6421, and Streptomyces albus/pStW102tc. In further specific embodiments, the cell harbors at least one ORF of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or functionally active variant or fragment thereof, and is capable of producing 11-de-O-methyltomaymycin.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] FIG. 1 is s schematic drawing of the chemical relationship between oxotomaymycin, 11-de-O-methyltomaymycin, tomaymycin, and the tomaymycin-urea adduct.
[0011] FIG. 2 is a schematic drawing of the tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421. The designations "orfX0" and "orfX1" and "A" to "Q" denote ORFs or genes, as listed in Table 1, whereby "A" to "Q" stand for "tomA" to "tomQ".
[0012] FIG. 3 is a presentation of pSTW102tc, which is the plasmid into which the tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421 is inserted. The ORFs or genes constituting the gene cluster as well as the respective putative proteins are indicated. Moreover, cleavage sites for restriction enzymes and their location are included.
[0013] FIG. 4 is a presentation of A) Biosynthetic pathway of tomaymycin, B) Structure of fed 2-amino-5-bromobenzoic acid and proposed structures for resulting mutasynthesis products, C) Extracted ion chromatogram (C.sub.14H.sub.16BrN.sub.2O.sub.2+: 323.03897 Da.+-.5 ppm; C.sub.14H.sub.14BrN.sub.2O.sub.2+: 321.02332 Da.+-.5 ppm) of Streptomyces albus J1074 pStW102tc.DELTA.CG culture without feeding (grey) and feeding with 2-amino-5-bromobenzoic acid (black). Mass spectra of the obtained substances and deviations to the theoretical mass are displayed below the respective structure. D) Structure of fed (S)-4-methylenepyrrolidine-2-carboxylic acid and proposed structures for resulting mutasynthesis products, E) Extracted ion chromatogram (C.sub.14H.sub.17N.sub.2O.sub.4+: 277.11828 Da.+-.5 ppm; C.sub.14H.sub.15N.sub.2O.sub.4+: 275.10263 Da.+-.5 ppm) of Streptomyces albus J1074 pStW102tc.DELTA.HI culture without feeding (grey) and feeding with (S)-4-methylenepyrrolidine-2-carboxylic acid (black). Mass spectra of the obtained substances and deviations to the theoretical mass are displayed below the respective structure.
[0014] FIG. 5 is a presentation of A) Structure of 9-chloro-11-de-O-methyl-8-deshydroxy-7-hydroxytomaymycin (CDHT); B) 1H,13C-HSQC-spectrum of CDHT.
DETAILED DESCRIPTION
[0015] In one embodiment, a nucleic acid is provided comprising at least one nucleic acid selected from:
[0016] (a) a nucleic acid comprising at least one of the Open Reading Frames (ORFs) 1 to 19 as comprised by SEQ ID NO: 2 that encodes proteins of SEQ ID NOs: 4 to 22 or a variant or fragment thereof, whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22,
[0017] (b) a nucleic acid encoding at least one of the proteins of SEQ ID NOs: 4 to 22 or a functionally active variant or fragment thereof,
[0018] (c) a nucleic acid encoding a protein that is at least 70%, 80%, 90%, 95% or 97% identical in amino acid sequence to a protein or fragment thereof encoded by the nucleic acid of (a) or (b),
[0019] (d) a nucleic acid that hybridizes under stringent conditions with a nucleic acid of (a) to (c),
[0020] (e) a nucleic acid that is complementary to a nucleic acid of (a) to (d).
[0021] In another embodiment, the nucleic acid comprises or consists of the tomaymycin biosynthetic gene cluster having the sequence of SEQ ID NO: 2 or a variant or fragmental sequence of SEQ ID NO: 2 harboring a variant or fragment of at least one of ORFs 1 to 19, whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22.
[0022] In a further embodiment an expression vector is provided comprising a nucleic acid of the embodiments mentioned above.
[0023] In yet another embodiment, a cell comprising a nucleic acid is provided according to the embodiments outlined above, or a cell transformed with the expression vector according to above embodiment. Preferably the cell is Streptomyces species FH6421.
[0024] Another embodiment is directed to the method for producing a cell wherein the cell harbors a tomaymycin biosynthetic gene cluster or a functionally active variant or fragment thereof.
[0025] In a further embodiment, the method produces a cell that harbors the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof.
[0026] In another embodiment, the method produces a cell that is a Streptomyces strain harboring the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof.
[0027] In yet another embodiment, the method produces a Streptomyces strain that is selected from Streptomyces achromogenes var. tomaymyceticus, Streptomyces species FH6421, and Streptomyces albus.
[0028] In yet a further embodiment, the method produces a cell that harbors at least one ORF of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or functionally active variants or fragments thereof, and is capable of producing 11-de-O-methyltomaymycin.
[0029] Thus, in sum, the present invention relates to a method for producing a cell, wherein the cell harbors a tomaymycin biosynthetic gene cluster or a functionally active variant or fragment thereof. Preferably the cell harbours the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof. Preferably, the cell is a Streptomyces strain harbouring the tomaymycin biosynthetic gene cluster of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a functionally active variant or fragment thereof, in particular Streptomyces achromogenes var. tomaymyceticus, Streptomyces species FH6421.
[0030] Alternatively, the cell harbours at least one, at least two, at least three, at least four, or at least five ORFs as comprised by SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO: 3, or functionally active variants or fragments thereof, and is capable of producing 11-de-O-methyltomaymycin.
[0031] The term "cell producing 11-de-O-methyltomaymycin" is, in principle, any cell that produces 11-de-O-methyltomaymycin. Herein, the terms "cell" and "cell producing 11-de-O-methyltomaymycin" are used interchangeably. 11-de-O-methyltomaymycin is a secondary metabolite of the class of pyrrolobenzodiazepines (PBD) and is naturally produced by the genus Streptomyces of the order Actinomycetales. In particular, 11-de-O-methyltomaymycin is naturally produced by microorganisms of the genus Streptomyces achromogenes, especially the strain Streptomyces achromogenes var. tomaymyceticus. All of these cells are comprised by the present invention.
[0032] Particularly included herein are cells that produce 11-de-O-methyltomaymycin and harbor a tomaymycin biosynthetic gene cluster, such as, e.g., the nucleic acid sequence of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO: 3, so that they effectively produce 11-de-O-methyltomaymycin. Included herein are cells that do not naturally produce 11-de-O-methyltomaymycin, but that have been transformed with a tomaymycin biosynthetic gene cluster, such as SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO: 3. The above mentioned embodiments may also be combined. For example, the cell may be transformed with a tomaymycin biosynthetic gene cluster, such as, e.g., comprised by SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO: 3, and the tomaymycin biosynthetic gene cluster is mutagenized, e.g., in order to enhance the productivity of 11-de-O-methyltomaymycin by the cell.
[0033] Moreover, the present inventors succeeded in identifying the tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421 that is comprised herein by the nucleic acid sequences of SEQ ID NO: 2 (tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421) or SEQ ID NO: 3 (plasmid pSTW102tc into which the tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421 is inserted). FIG. 2 schematically shows the tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421. FIG. 3 shows the plasmid pSTW102tc, into which the tomaymycin biosynthetic gene cluster of strain Streptomyces species FH6421 is inserted.
[0034] In the context of the present invention, the term "gene cluster" is a nucleic acid and refers to a set of several genes or ORFs that are located on a contiguous stretch of the genome and that participate in the synthesis of 11-de-O-methyltomaymycin. The encoded proteins are either enzymes that catalyse reactions of substrates into products, or are involved in regulation of the synthesis of 11-de-O-methyltomaymycin or intermediate products or the transport of 11-de-O-methyltomaymycin or intermediate products. Li et al. (2009) have assigned, by homology to known genes, specific functions to the proteins encoded by the ORFs. Altogether, the genes as comprised by the gene cluster encode proteins involved in the biosynthesis of 11-de-O-methyltomaymycin.
[0035] The term "tomaymycin biosynthetic gene cluster" refers to the tomaymycin biosynthetic gene cluster as comprised by Streptomyces species FH6421, which has been cloned and sequenced by the present inventors. The sequence is shown in SEQ ID NO: 2. The sequence of the tomaymycin biosynthetic gene cluster has been cloned into the vector pStW102 (derived from pOJ446), resulting in pSTW102tc, which is presented herein as SEQ ID NO: 3. A schematic drawing of pSTW102tc is shown in FIG. 3. The following ORFs were identified within the gene cluster: orfX0, orfX1, tomA, tomb, tomC, tomD, tome, tomF, tomG, toml, tomJ, tomK, tomL, tomM, tomN, tomO, tomP, and tomQ. These ORFs are assigned the ORF numbers ORF1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19, respectively. The genes that participate in the synthesis of 11-de-O-methyltomaymycin are tomA, tomb, tomC, tomD, tome, tomF, tomG, toml, tomJ, tomK, tomL, tomM, tomN, tomO, tomP, and tomQ. Table 1 shows a list of the specific putative ORFs, the designation of the corresponding genes, the start and stop nucleotides within SEQ ID NO: 2, the lengths of the genes in nucleotides (nt), and the strandedness.
TABLE-US-00001 TABLE 1 gene start nt stop nt gene ORF desig- within SEQ within SEQ length number nation ID NO: 2 ID NO: 2 (nt) strandedness 1 orfX0 201 779 579 forward 2 orfX1 1580 2068 489 reverse 3 tomA 2785 4617 1833 forward 4 tomB 4689 9296 4608 forward 5 tomC 9415 10635 1221 forward 6 tomD 10632 12614 1983 forward 7 tomE 12611 13210 600 forward 8 tomF 13215 14786 1572 forward 9 tomG 14867 15532 666 reverse 10 tomH 15785 16285 501 forward 11 tomI 16282 17085 804 forward 12 tomJ 17279 18178 900 forward 13 tomK 18175 19050 876 forward 14 tomL 19171 20961 1791 forward 15 tomM 21014 23323 2310 forward 16 tomN 23410 23613 204 forward 17 tomO 23648 24847 1200 reverse 18 tomP 24976 27390 2415 forward 19 tomQ 27422 28867 1446 reverse
[0036] Table 2 shows the ORF number, protein designation, the length in amino acids (aa) of the putative proteins, the SEQ ID numbers of the proteins as identified herein, and the putative function.
TABLE-US-00002 TABLE 2 Protein Protein SEQ ID ORF desig- length NO: of number nation (aa) protein Putative function 1 OrfX0 192 4 TetR transcriptional regulator family 2 OrfX1 162 5 MarR transcriptional regulator family 3 TomA 610 6 Nonribosomal peptide synthetase 4 TomB 1532 7 Nonribosomal peptide synthetase 5 TomC 406 8 Phenazine biosynthesis protein PhzC 6 TomD 660 9 Phenazine biosynthesis protein PhzE 7 TomE 199 10 Phenol hydroxylase, reductase component 8 TomF 523 11 Phenol-2-monoxygenase oxygenase component 9 TomG 222 12 O-Methyltransferase 10 TomH 166 13 ImbB1 protein, L-DOPA 2,3-dioxygenase 11 TomI 268 14 ImbB2 protein, L-tyrosine 3-hydroxylase 12 TomJ 299 15 ImbY protein, unknown, lincomycin biosynthesis 13 TomK 291 16 ImbX protein/PhzF protein 14 TomL 597 17 ImbA protein, unknown function 15 TomM 769 18 Putative drug resistance pump 16 TomN 67 19 4-oxalocrotoate tautomerase 17 TomO 400 20 NADH-dependent flavin oxidoreductase 18 TomP 804 21 Anthranilate synthase 19 TomQ 481 22 Flavin-containing amine oxidase
[0037] The present invention includes functionally active variants or functionally active fragments of a tomaymycin biosynthetic gene cluster. A functionally active variant of a tomaymycin biosynthetic gene cluster relates to a tomaymycin biosynthetic gene cluster having at least one variant ORF with respect to the ORFs as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. The variant ORF encodes a functionally active variant of the respective protein. Such functionally active variants have a sequence identity with the proteins encoded by ORFs comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 of more than 50%, of more than 60%, preferably more than 70%, more preferably of more than 80%, still more preferably more than 85%, even more preferably more than 90%, even more preferably more than 95%, most preferably more than 97%, and/or have an activity of more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 100%, e.g., more than 120%, 150%, 200%, 300%, 400%, or 500% of the activity of the respective proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. Preferably, the activity is at least 100%, more preferably at least 120%, most preferably at least 150%. Consequently, the nucleic acid encoding such variants contains deletions, insertions, substitutions, and/or additions within and/or at the 5' and/or 3' termini of the ORFs as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, and show an identity to the sequences of the ORFs as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 of more than 50%, more than 60%, more than 70%, preferably more than 80%, more preferably more than 85%, even more preferably more than 90%, even more preferably more than 95%, most preferably more than 97%. In the context of the present invention, a functionally active variant nucleic acid sequence relative to a nucleic acid sequence as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 encoding a functionally active variant protein with respect to a protein encoded by the tomaymycin biosynthetic gene cluster as comprised e.g. by SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3 means a sequence encoding a variant protein that is capable of participating in the synthesis of 11-de-O-methyltomaymycin and/or that can be substituted for the respective sequence to participate in the synthesis of 11-de-O-methyltomaymycin.
[0038] A functionally active fragment of a tomaymycin biosynthetic gene cluster relates to a tomaymycin biosynthetic gene cluster that comprises fragments of at least one ORF as comprised by a tomaymycin biosynthetic gene cluster, such as comprised by SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3. Such fragments of the ORFs encode fragments of the respective proteins. This may include fragment proteins with short internal and/or C- and/or N-terminal deletions whereby the activity of the resulting proteins as identified herein is maintained to an extent of more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 100%, e.g., more than 120%, 150%, 200%, 300%, 400%, or 500%, of the activity of the proteins encoded by a tomaymycin biosynthetic gene cluster, such as, e.g., comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. Preferably, the activity is at least 100%, more preferably at least 120%, and most preferably at least 150%. Consequently, the respective nucleic acid encoding such fragments may contain deletions within and/or at the 5' and/or 3' termini of the ORFs, e.g., deletions of at the most 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, or less. In the context of the present invention, a fragment of a nucleic acid sequence encoding a functionally active fragment of a protein as comprised herein means a sequence encoding a fragment that is capable of participating in the synthesis of 11-de-O-methyltomaymycin and/or that can be substituted for the respective sequence to participate in the synthesis of 11-de-O-methyltomaymycin. The term "fragment" may encompass full length ORFs in combination with fragment ORFs, as long as this combination results in the synthesis of 11-de-O-methyltomaymycin. Moreover, a fragment of a tomaymycin biosynthetic gene cluster also relates to a tomaymycin biosynthetic gene cluster with internal and/or 5'- and/or 3'-deletions, which may result in the deletion of parts of ORFs and/or in the deletion of whole ORFs, as long as the ability of the fragments of the tomaymycin biosynthetic gene cluster to produce 11-de-O-methyltomaymycin is maintained to an extent of more than 5%, more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, more than 97% or more than 100%, e.g., more than 150%, 200%, 300%, 400%, or 500%, of the activity of the proteins encoded by a tomaymycin biosynthetic gene cluster, such as, e.g., comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. A functionally active fragment of a tomaymycin biosynthetic gene cluster as comprised herein encodes proteins that, in their entirety, are capable of effecting the synthesis of 11-de-O-methyltomaymycin and/or proteins that can be substituted for the respective sequence to participate in the synthesis of 11-de-O-methyltomaymycin.
[0039] Included within the term "cell producing 11-de-O-methyltomaymycin" are cells that harbor one or more ORFs of a tomaymycin biosynthetic gene cluster, which one or more ORFs are suitable to effect production of 11-de-O-methyltomaymycin. Based on the information of the cluster of Li et al. (2009) and the information provided herein, the skilled person will be able to select those ORFs that are sufficient to effect synthesis of 11-de-O-methyltomaymycin. The at least one ORF may comprise one nucleic acid, or different ORFs may comprise different nucleic acids. Thus, the term "cell producing 11-de-O-methyltomaymycin" includes nucleic acids that comprise all of the ORFs as comprised by a tomaymycin biosynthetic gene cluster, such as ORFs 1 to 19 as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or nucleic acids that comprise an individual ORF or a combination of individual ORFs such as at least one, at least two, at least three, at least four, at least five or more ORFs, which individual ORF or combination of ORFs encode proteins that are capable of synthesising 11-de-O-methyltomaymycin in a cell.
[0040] The terms "comprise", "comprises", and "comprising", as used herein mean to "include or encompass" the desired feature and further features that must not be specifically mentioned. The terms also meant to "consist of" the desired feature and not to include further features except the desired feature. Thus, the nucleic acid or protein referred to herein may be defined by additional features in addition to the definition as indicated, e.g., in addition to the definition by an ORF or SEQ ID number, or may consist of such indicated feature only.
[0041] The nucleic acid as comprised herein may be any macromolecule composed of chains of monomeric nucleotides carrying genetic information or form structures within cells. The most common (and therefore preferred) nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). The nucleic acid can be a DNA molecule, such as a genomic DNA molecule, and may comprise the whole sequence or a fragment of a tomaymycin biosynthetic gene cluster, such as SEQ ID NO: 1 or 2, or a cDNA molecule which can be single- or double-stranded, such as a nucleic acid representing an ORF and encoding a protein, as well as a synthetic DNA, such as a synthesized single-stranded polynucleotide. The nucleic acid may also be an RNA molecule. Preferably, the term also relates to non-coding regions of a gene, wherein these sections are of a relevant size in order to be specific for that gene. Examples of those regions are regulatory elements, such as a promoter. More preferably, the term "nucleic acid" relates to a gene, ORF, promoter, DNA, cDNA, or mRNA. The nucleic acid encoding the desired genetic information, preferably DNA, may comprise the gene(s) of interest, a promoter region, a start codon and a stop codon, and possibly further regions that may be used for regulation of expression of the gene. The regulatory regions may be heterologous to the respective gene or may be associated therewith in nature. The genetic information may be expressed permanently or under the control of a repressor and/or a promoter region in a cell into which the nucleic acid of the present invention is introduced. The obtained cells may be either used directly or used for tissue cultures.
[0042] Also comprised by the present invention are nucleic acids that comprise functionally active variants or fragments of the ORFs as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. Functionally active variants or fragments are defined with respect to functionally active variants or fragments of the tomaymycin biosynthetic gene cluster.
[0043] It is noted that the above mentioned modifications may be combined. For example, a tomaymycin biosynthetic gene cluster or a nucleic acid as comprised by the present invention may be a fragment comprising one or more variations of the ORFs the invention. It should also be noted that fragments and/or variants include fragments and/or variants, as defined herein, of promoter or regulatory sequences with which the ORFs or fragments or variants thereof are associated in nature. The fragments and/or variants are functionally active in that they regulate the transcription or translation of the genes associated therewith. Moreover, the variant or fragment as referred to above may be an artificially produced nucleic acid.
[0044] The term "heterologous" as it relates to nucleic acid sequences, such as coding or control sequences denotes sequences that are normally not associated with a region of a recombinant construct and/or a particular cell. A "heterologous" region is an identifiable segment of a nucleic acid within or attached to another nucleic acid that is not found in association with the other molecule in nature. For example, a heterologous region of a construct could be a regulatory region not found to be associated with a gene as identified herein in nature. Similarly, a heterologous sequence could be a coding sequence that is itself not found in nature as it contains, e.g., synthetic sequences with codons different from the native gene. Moreover, a cell transformed with a construct that is not normally present in the cell would be considered heterologous for the purposes of the present invention. A homologous nucleic acid sequence is a variant sequence as defined herein. The term "homologous" may be used interchangeably with variant. The term "homologous" may also refer to an identical sequence.
[0045] An ORF is an open reading frame that is a DNA sequence that could potentially encode a protein. In the context of the present invention, the term "ORF" stands for open reading frame in the tomaymycin biosynthetic gene cluster as isolated from Streptomyces achromogenes var. tomaymyceticus, from Streptomyces species FH6421, or any other microorganism producing 11-de-O-methyltomaymycin. The tomaymycin biosynthetic gene cluster has been elucidated by Li et al. (2009) and the cluster and ORFs identified therein are comprised for the purposes of the present invention. Moreover, the present inventors succeeded in identifying the tomaymycin biosynthetic gene cluster of Streptomyces species FH6421 and identified 19 ORFs. Furthermore, any ORFs of tomaymycin biosynthetic gene clusters from strains other than Streptomyces achromogenes var. tomaymyceticus or Streptomyces species FH6421 that are known in the art or will be identified are included herein. Also functionally active variants or functionally active fragments of the ORFs of Streptomyces achromogenes var. tomaymyceticus or Streptomyces species FH6421 fall within the term ORFs as comprised herein, as long as such ORFs encode functionally active proteins.
[0046] The substitution of a variant or fragment nucleic acid for ORFs to participate in the synthesis of 11-de-O-methyltomaymycin means that this variant or fragment nucleic acid can be inserted into the genome of a microorganism harbouring a tomaymycin biosynthetic gene cluster instead of the ORF to which it is a variant or to which it is a fragment, thereby expressing a variant or fragment protein that takes over the function of the respective protein and participates in the synthesis of 11-de-O-methyltomaymycin. The extent to which the variant or fragment takes over the function is as defined herein.
[0047] In another embodiment, the nucleic acid may comprise the sequences of a tomaymycin biosynthetic gene cluster, such as, e.g., comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. The nucleic acid may also encode proteins with the same amino acids as the proteins encoded by a tomaymycin biosynthetic gene cluster, such as, e.g., comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but differs in its nucleotide composition due to the degeneracy of the genetic code.
[0048] In a further embodiment, the tomaymycin biosynthetic gene cluster or the nucleic acid hybridizes under stringent conditions to a nucleic acid that comprises the tomaymycin biosynthetic gene cluster as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. In the present invention, the term "hybridize(s)(ing) under stringent conditions" refers to the formation of a hybrid between two nucleic acid molecules under conditions that allow the formation of a so-called specific hybrid, while a non-specific hybrid is substantially not formed. An example of such conditions includes conditions under which a complementary strand of a highly identical nucleic acid, namely, a DNA composed of a nucleotide sequence having 70% or more, preferably 80% or more, more preferably 85% or more, still more preferably 90% or more and even more preferably 95% or more identity with the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 hybridizes, while a less complementary strand of a nucleic acid less identical than the above does not hybridize. More specifically, such conditions refer to conditions in which the sodium salt concentration is 15 to 750 mM, preferably 50 to 750 mM, and more preferably 300 to 750 mM; the temperature is 25 to 70.degree. C., preferably 50 to 70.degree. C., and more preferably 55 to 65.degree. C.; and the formamide concentration is 0 to 50%, preferably 20 to 50%, and more preferably 35 to 45%. Furthermore, under stringent conditions, conditions for washing a filter after hybridization normally comprises the following: the sodium salt concentration is 15 to 600 mM, preferably 50 to 600 mM, and more preferably 300 to 600 mM; and the temperature is 50 to 70.degree. C., preferably 55 to 70.degree. C., and more preferably 60.degree. C. Stringency, and thus specificity, can, e.g., be increased by increasing the reaction temperature and/or lowering the ion strength of the reaction buffer. For example, low stringent conditions comprise hybridization in 3.times.SSC at room temperature to 65.degree. C., and highly stringent conditions comprise hybridization in 0.1.times.SSC at 68.degree. C. Exemplary moderately stringent conditions (nucleic acids hybridize under moderately stringent conditions if they are maximally degenerate with respect to their codon composition) comprise 50% formamide, 5.times.SSC and 1% SDS at 42.degree. C. and washing in 1.times.SSC at 45.degree. C. Highly stringent conditions comprise incubation at 42.degree. C., 50% formamide, 5.times.SSC and 1% SDS (e.g., 50% formamide, 5.times.SSC and 1% SDS, 50 mM sodium phosphate, 5.times.Denhardt's solution, 10.times. dextran sulphate, 20 mg/ml sheared salmon sperm DNA) or 5.times.SSC and 1% SDS at 65.degree. C. and washing in 0.2.times.SSC and 0.1% SDS at about 65.degree. C. (1.times.SSC stands for 0.15 M sodium chloride and 0.015 M trisodium citrate buffer). Preferred in the present invention are moderately or highly stringent conditions, more preferred are highly stringent conditions. In the context of the present invention, a "hybridizing" sequence means a sequence that encodes a protein that participates in the synthesis of 11-de-O-methyltomaymycin and/or that can be substituted for the ORF to which it specifically hybridizes to participate in the synthesis of 11-de-O-methyltomaymycin.
[0049] The tomaymycin biosynthetic gene cluster or nucleic acid as comprised or referred to herein may be provided by any methods known in the art. Using the sequence information provided herein or in the prior art, primers suitable for amplification/isolation of one or more ORFs can be determined according to standard methods well known to those of skill in the art. Primers suitable for amplification/isolation of any one or more of the ORFs as defined herein are designed according to the nucleotide sequence information provided in the sequence listing. The procedure is as follows: a primer is selected that may consist of 10 to 40, preferably 15 to 25 nucleotides. It is advantageous to select primers containing C and G nucleotides in a proportion sufficient to ensure efficient hybridization; i.e., an amount of C and G nucleotides of at least 40%, preferably 50% of the total nucleotide content. Typically such amplifications will utilize the DNA or RNA of an organism containing the requisite genes (e.g., Streptomyces achromogenes, such as Streptomyces achromogenes var. tomaymyceticus, Streptomyces species FH6421, or any other strain producing 11-de-O-methyltomaymycin) as a template. A standard PCR reaction will be performed that typically contains 0.5 to 5 Units of Taq DNA polymerase per 100 .mu.l, 20 to 200 .mu.M deoxynucleotide each, preferably at equivalent concentrations, 0.5 to 2.5 mM magnesium over the total deoxynucleotide concentration, 105 to 106 target molecules, and about 20 pmol of each primer. About 25 to 50 PCR cycles are performed. A more stringent annealing temperature improves discrimination against incorrectly annealed primers and reduces incorporation of incorrect nucleotides at the 3' end of primers. A denaturation temperature of 95.degree. C. to 97.degree. C. is typical, although higher temperatures may be appropriate for denaturation of G+C-rich targets. The number of cycles performed depends on the starting concentration of target molecules, though typically more than 40 cycles are not recommended as non-specific background products tend to accumulate. An alternative method for retrieving polynucleotides encoding variant proteins defined herein is by hybridization screening of a DNA or RNA library using the primers and probes. A nucleotide probe has a sequence found in or derived by the degeneracy of the genetic code from a sequence within the tomaymycin biosynthetic gene cluster as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a variant thereof or encoding any of SEQ ID NOs: 4 to 22 or functionally active variants thereof. The term "probe" refers to DNA, preferably single-stranded, or RNA molecules or modifications or combinations thereof, that hybridize under stringent conditions, as defined herein, to nucleic acid molecules comprised within the tomaymycin biosynthetic gene cluster identified by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or variants thereof or encoding any of the proteins of SEQ ID Nos: 4 to 22 or functionally active variants thereof, or their complementary or sense sequences. Generally, probes are significantly shorter than full-length sequences. They may contain from 5 to 100, preferably 10 to 80 nucleotides, more preferably 10 to 50 nucleotides, still more preferably 10 to 40 nucleotides and still more preferably 15 to 25 nucleotides. In particular, such probes may have sequences that are at least 70%, at least 75%, preferably at least 85%, more preferably at least 95%, and most preferably 100% homologous to a coding (ORFs 1 to 19) or non-coding sequence as comprised by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or that are, to the above extents, complementary thereto. They may contain modified bases, such as inosine, methyl-5-deoxycytidine, deoxyuridine, dimethylamino-5-deoxyuridine or diamino-2,6-purine. Sugar or phosphate residues may also be modified or substituted as is known in the art. For example, a deoxyribose residue may be replaced by a polyamide, and a phosphate residue may be replaced by ester groups, such as diphosphate, alky, arylphosphonate or phosphorothioate esters. Alternatively or in addition, the 2'-hydroxyl group on ribonucleotides may be modified by including such groups as alkyl, O-alkyl or halogen groups. Probes of the invention are used in any conventional hybridization technique such as dot blot, Southern blot, northern blot, or sandwich technique, which is a technique using specific capture and/or detection probes with nucleotide sequences that at least differ partially from each other (Sambrook et al., Molecular cloning: A laboratory manual. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2001). Hybridization procedures are well-known and are described in the art and herein.
[0050] Alternatively or additionally to the above, the nucleic acid may be provided by cloning and thereby introducing it into and amplifying it in a cell. The procedure of introducing a gene into a recipient cell is called transformation. The genes can be introduced into the cells by a variety of means known in the art and adapted to each cell type. The term "cell" refers to the cell in which the gene is expressed irrespective of whether it is a prokaryotic cell or a eukaryotic cell and of whether the cell naturally expresses the respective genes or not. Thereby the cell may be a cell that naturally harbors the gene expressing the protein as comprised by the present invention, e.g., Streptomyces achromogenes, such as Streptomyces achromogenes var. tomaymyceticu, or Streptomyces species FH6421, or any other strain producing 11-de-O-methyltomaymycin. Recombinant DNA cloning techniques well known in the art for introducing and expressing a nucleic acid molecule can be used to introduce and express the gene that is either endogenous if the cell harbours the respective gene or is heterologous if the gene is not endogenous to the cell. Cells can be transformed using any appropriate means, including viral or bacteriophage based vectors, chemical agents, electroporation, calcium phosphate co-precipitation or direct diffusion of DNA. Vectors are agents that transport an endogenous or heterologous gene into the cell and may include appropriate transcriptional and translational control signals, such as a promoter. Vectors can be a plasmid, a virus (e.g. bacteriophage) or others known in the art. Vectors are able to autonomously replicate in a cell or can be incorporated into chromosomal DNA. The term "vectors" includes those that function primarily for insertion of a nucleic acid into a cell, those that function primarily for replication of a nucleic acid (replication vector) in a cell or those that function primarily for transcription and/or translation of DNA or RNA in a cell. Examples of vectors include pBTrp2, pBTac1, pBTac2 (all of which are manufactured by Boehringer Mannheim), pKK263-2 (manufactured by Pharmacia), pGEX (manufactured by Pharmacia), pSE280 (manufactured by Invitrogen), pGEMEX-1 (manufactured by Promega), pQE-8 (manufactured by Qiagene), pET-3 (manufactured by Novagen), pBluescriptll SK+(manufactured by Stratagene), pBluescript II SK (-) (manufactured by Stratagene), pTrS30 [prepared from Escherichia coli JM109/pTrS30 (FERM BP-5407)], pTrS32 [prepared from Escherichia coli JM109/pTrS32 (FERM BP-5408)], pSTV28 (manufactured by Takara Bio Inc.), pUC118 (manufactured by Takara Bio Inc.), pHW1520 (manufactured by MoBiTec), pSET152, pOJ436 and pOJ446 (Bierman M, et al., 1992), pSH19 (Herai S, et al., 2004), pUWL199, pUWL218 and pUWL219 (Wehmeier U. F., 1995) and pIJ6021 (Takano E. et al., 1995). A preferred vector is pOJ446 and derivatives thereof.
[0051] The promoter can be inducible or constitutive, general or cell specific, nuclear or cytoplasmic specific, heterologous or associated with the gene in nature. Any type of promoter can be used, as long as it functions in the cells producing 11-de-O-methyltomaymycin. Examples of the promoter include promoters derived from Escherichia coli or phage, such as a trp promoter (Ptrp), a lac promoter (Plac), a PL promoter, a PR promoter or a PSE promoter, a SPO1 promoter, a SPO2 promoter, and a penP promoter. In addition, artificially designed or modified promoters, such as a promoter formed by placing two Ptrp in series (Ptrp*2), a tac promoter, a lacT7 promoter or a let I promoter, can be used. Moreover, a xylA promoter for expression in the bacteria of the genus Bacillus, or a P54-6 promoter for expression in the bacteria of the genus Corynebacterium can be used. Additional useful promoters are PermE (Bibb et al., 1985, PermE* (Bibb et al., 1994), PtipA (Murakami et al., 1989), PnitA-NitR expression system (Herai et al., 2004) and actlI-ORF4/PactI activator-promoter system (Ferna'ndez-Moreno et al., 1991). Selection of promoters, vectors, and other elements are a matter of routine design. Many such elements are described in literature and are available through commercial suppliers. A single gene can be introduced into a cell.
[0052] Also, more than one gene can be introduced into a cell and expressed therein. Where large clusters are to be expressed, it is preferable that phagemids, cosmids, P1s, YACs, BACs, PACs, HACs, or similar cloning vectors are used. If more than one gene is introduced into a cell, then the genes may be under the regulation of the same promoter and/or regulatory elements. Alternatively, the genes may be under the regulation of different promoter and/or regulatory elements. Usually, the method of transfer includes transfer of a selectable marker to the cells. In general, a cell line is transformed by any of the means mentioned above wherein the transgene is operatively linked to a selectable marker. Following transformation, cells are grown for an adapted period of time. Transformed cells exhibit resistance to the selection and are able to grow, whereas non-transformed cells die in general. Examples for selective markers include puromycin, zeocin, neomycin and hygromycin B, which confer resistance to puromycin, zeocin, aminoglycoside G-418 and hygromycin B, respectively.
[0053] In principle, any cells capable of harboring and expressing a recombinant tomaymycin biosynthetic gene cluster or one or more genes of the tomaymycin biosynthetic gene cluster that are useful or sufficient to effect production of 11-de-O-methyltomaymycin can be used in the methods of the present invention. Examples include microorganisms such as bacteria, yeasts, filamentous fungi, animal cells, and plant cells, such as, without limitation, cells of E. coli strains, of the order Actinomycetales, such as a Streptomyces species, such as Streptomyces albus, of yeast strains such as Saccharomyces cerevisiae.
[0054] Preferred embodiments are bacterial cells, such as cells of the order Actinomycetales, such as Streptomyces species, such as Streptomyces species FH6421, or Streptomyces albus, such as Streptomyces albus pSTW102tc cells with a wildtype tomaymycin biosynthetic gene cluster that is mutagenized, such as Streptomyces species FH6421-1038, Streptomyces species FH6421-1069, or Streptomyces species FH6421-1334.
[0055] The object of the present invention is the provision of advantageous methods to enhance the production of 11-de-O-methyltomaymycin by cells that produce 11-de-O-methyltomaymycin. The cells may comprise a tomaymycin biosynthetic gene cluster that is identified by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or may comprise a tomaymycin biosynthetic gene cluster as identified by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, which comprises functionally active variant or fragments of ORFs or promoter or regulatory regions, as long as such a cell is capable of producing 11-de-O-methyltomaymycin. Moreover, comprised herein are cells that comprise only part of the ORFs, possibly comprising functionally active variants and/or fragments of ORF(s) as comprised herein, as long as the cells are capable of producing 11-de-O-methyltomaymycin. Variant or fragment ORFs or promoter or regulatory regions may be natural or may be artificial. Variant or fragment ORFs or promoter or regulatory regions may serve to enhance the productivity of 11-de-O-methyltomaymycin by cells harboring such variant or fragment ORFs. The tomaymycin biosynthetic gene cluster or ORFs or promoter or regulatory regions may be artificially modified to result in a tomaymycin biosynthetic gene cluster or ORFs or promoter or regulatory regions that result in the production of a higher yield of 11-de-O-methyltomaymycin versus the production of 11-de-O-methyltomaymycin by a strain harboring the tomaymycin biosynthetic gene cluster identified by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or versus the parent strain. Such mutated strains are inter alia disclosed herein as Streptomyces species FH6421-1038, Streptomyces species FH6421-1069, Streptomyces species FH6421-1334.
[0056] Consequently, in a further embodiment of the present invention, cells naturally producing 11-de-O-methyltomaymycin and/or comprising the tomaymycin biosynthetic gene cluster or cells that have been transformed with an individual ORF or a combination of ORFs that may comprise variant or fragment ORFs, promoter or regulatory regions, as referred to above, and producing 11-de-O-methyltomaymycin, are mutagenized in order to enhance the production rate of 11-de-O-methyltomaymycin.
[0057] Preferably, the production rate is enhanced by a factor of at least 1.3, at least 1.5, at least 1.8, at least 2.0, at least 2.5, at least 5.0, or a least 10.0. More preferably, the production rate is enhanced by the factor of 1.5 to 2.0, and most preferably by a factor of 1.5 to 1.8.
[0058] For this invention, Streptomyces albus J1074 was transformed with the vector pSTW102tc (SEQ ID NO: 3) resulting in Streptomyces albus J1074/pSTW102tc with a yield of 338.+-.18.8 mg/l in a coil fitted shake flask using production medium (20 g/l soy flour, 10 g/l corn steep solid, 20 g/l glycerol, 7.5 g/l NaCl, 2 g/l CaCO.sub.3), and strain Streptomyces species FH6421 with a productivity of about 50.+-.10 mg/l, and as compared to the standard strain with 20 mg/l under same conditions.
[0059] The proteins that are produced by a cell that produces 11-de-O-methyltomaymycin and participates in the synthesis of 11-de-O-methyltomaymycin encompasses proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, such as, e.g., proteins identified by SEQ ID Nos: 4 to 22, and encompass proteins as they occur in other organisms that produce 11-de-O-methyltomaymycin that are orthologs or homologs whereby these orthologs or homologs have the same function as the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. Preferably, orthologs or homologs thereof differ from the sequences of the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, e.g., by addition, deletion, substitution, and/or insertion of amino acids, and have a sequence identity with the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 of more than 50%, of more than 60%, more than 70%, preferably of more than 80%, more preferably more than 85%, even more preferably more than 90%, even more preferably more than 95%, most preferably more than 97%, and/or have an activity of more than 5%, more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, more than 97% or more than 100%, e.g. more than 150%, 200%, 300% 400% or 500% of the activity of the respective proteins of SEQ ID Nos: 4 to 22.
[0060] In the context of the present invention the naturally or non-naturally occurring variant of the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 is a functionally active protein in that it maintains the biological function of the reference protein, i.e. the involvement in a reaction in which the reference protein is involved under natural conditions (in case of a non-natural variant, the biological function of the reference protein).
[0061] Non-naturally occurring variants of the proteins of SEQ ID Nos: 4 to 22 or of naturally occurring variants thereof may be obtained by a limited number of amino acid deletions, insertions and/or substitutions, particularly deletions, insertions and/or substitutions of, e.g., at most 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid(s), thereby obtaining a sequence identity or activity of the respective wild-type proteins, e.g. with respect to SEQ ID Nos: 4 to 22, as mentioned above.
[0062] In another embodiment of the present invention, the variant of the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 could be a fragment, wherein the fragment is still functionally active. This may include proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or variants thereof as detailed above with short internal and/or C- and/or N-terminal deletions (e.g. deletions of at most 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6 5, 4, 3, 2, or 1 amino acids within the variant and/or at the C- and/or N-termini or total deletions of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% amino acids or any values in between these values). Additionally, the fragment may be further modified as detailed above with respect to variants.
[0063] Alternatively or additionally, the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 or variants thereof as described above may comprise one or more amino acid substitution(s). However, semi-conservative and especially conservative amino acid substitutions, wherein an amino acid is substituted with a chemically related amino acid, are preferred. Typical substitutions are among the aliphatic amino acids, among the amino acids having aliphatic hydroxyl side chain, among the amino acids having acidic residues, among the amide derivatives, among the amino acids with basic residues, or the amino acids having aromatic residues. Typical semi-conservative and conservative substitutions are:
TABLE-US-00003 TABLE 3 Amino acid Conservative substitution Semi-conservative substitution A G; S; T N; V; C C A; V; L M; I; F; G D E; N; Q A; S; T; K; R; H E D; Q; N A; S; T; K; R; H F W; Y; L; M; H I; V; A G A S; N; T; D; E; N; Q H Y; F; K; R L; M; A I V; L; M; A F; Y; W; G K R; H D; E; N; Q; S; T; A L M; I; V; A F; Y; W; H; C M L; I; V; A F; Y; W; C; N Q D; E; S; T; A; G; K; R P V; I L; A; M; W; Y; S; T; C; F Q N D; E; A; S; T; L; M; K; R R K; H N; Q; S; T; D; E; A S A; T; G; N D; E; R; K T A; S; G; N; V D; E; R; K; I V A; L; I M; T; C; N W F; Y; H L; M; I; V; C Y F; W; H L; M; I; V; C
[0064] Changing from A, F, H, I, L, M, P, V, W or Y to C is semi-conservative if the new cysteine remains as a free thiol. Furthermore, the skilled person will appreciate that glycines at sterically demanding positions should not be substituted and that P should not be introduced into parts of the protein that have an alpha-helical or a beta-sheet structure.
[0065] It is noted that the above modifications of the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 may be combined. The variants of the present invention may be e.g. a fragment of a protein encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 comprising one or more amino acid substitutions. It is furthermore noted that any of the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 may be combined with any of a variant or fragment of the proteins encoded by SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
[0066] In one aspect, the present invention provides a nucleic acid comprising at least one nucleic acid selected from:
[0067] (a) a nucleic acid comprising at least one of the Open Reading Frames (ORFs) 1 to 19 as comprised by SEQ ID NO: 2 encoding proteins of SEQ ID NOs: 4 to 22 or a variant or fragment thereof whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22,
[0068] (b) a nucleic acid encoding at least one of the proteins of SEQ ID NOs: 4 to 22 or a functionally active variant or fragment thereof,
[0069] (c) a nucleic acid encoding a protein that is at least 70%, 80%, 90%, 95% or 97% identical in amino acid sequence to a protein or fragment thereof encoded by the nucleic acid of (a) or (b),
[0070] (d) a nucleic acid that hybridizes under stringent conditions with a nucleic acid of (a) to (c),
[0071] (e) a nucleic acid that is complementary to a nucleic acid of (a) to (d).
[0072] In another aspect, the nucleic acid comprises or consists of the tomaymycin biosynthetic gene cluster having the sequence of SEQ ID NO: 2 or having a variant or fragment sequence of SEQ ID NO: 2 harboring a variant or fragment of at least one of ORFs 1 to 19, whereby the variant or fragment encodes a functionally active variant or fragment of a protein of SEQ ID NOs: 4 to 22.
[0073] In a further aspect, an expression vector comprising the nucleic acid as referred to above is provided.
[0074] In still a further aspect, a cell comprising the above nucleic acid, or a cell transformed with the above expression vector is provided. Preferably the cell is Streptomyces species FH6421.
[0075] With respect to the tomaymycin biosynthetic gene cluster, the ORFs or genes comprised therein, the proteins encoded thereby and variants and fragments of the tomaymycin biosynthetic gene cluster or of ORFs or proteins, these features are described above in the context of the method for producing 11-de-O-methyltomaymycin and it is referred to the definitions provided therein.
EXAMPLES
[0076] The invention is further exemplified by the following examples:
Example 1--Knock Out of Biosynthetic Genes from the Tomaymycin Biosynthetic Gene Cluster
[0077] To enable mutasynthesis of tomaymycin analogous structures genes providing the precursors of the biosynthesis were deleted in the heterologous expression system. The anthranilic acid derivative incorporated in tomaymycin is derived from 3-deoxy-D-arabino-heptulosonate-7-phosphate partially utilizing the intrinsic shikimate pathway of the native producer strain or the heterologous host. For exchange of the A-Ring the genes tomC, tomD, tomE, tomF and tomG, were deleted. S. albus J1074 strains carrying the resulting plasmid pStW102tc.DELTA.C-G are not producing tomaymycin but did incorporate 2-amino-5-bromobenzoic acid. Deletion of tomH and toml also eliminated tomaymycin production but allowed incorporation of (S)-4-methylenepyrrolidine-2-carboxylic acid.
[0078] Deletion of genes from pStW102tc involved in the supply of precursors for tomaymycin was performed by Red/ET as described by the supplier (GeneBridges). The zeocin resistance gene from pCK_T7A1_att was amplified by the primer pair pr130f, pr130r (for primers see below) and the PCR product used to delete the genes tomC, tomD, tomE, tomF and tomG--(without being bound to theory, which is believed to be involved in the supply of the anthranilic acid derived residue). The tetracycline resistance gene from pACYC184 was amplified by the primer pair pr156f, pr156r and the PCR product used to delete tomH and toml--(without being bound to theory, which is believed to be involved in the supply of the ethylidene proline residue). The deletions were verified by restriction digest followed by gel electrophoresis. Resistance genes were removed via XmaJI, XbaI digest and religation yielding pStW102tc.DELTA.tomC-G and pStW102tc.DELTA.tomHI respectively. Plasmids were then transferred into S. albus J1074.
[0079] PCR-primers used for deletion of biosynthetic genes via Red/ET. Restriction sites are underlined and the corresponding restriction enzyme given in parentheses.
TABLE-US-00004 pr130f (SEQ ID NO: 23) 5'-CCGACCATCCACCACACGGCAATCGCCGAAGCGGTCGCCGGACACCGA AAGCCTAGGGCGAGGAAGCGGTGATCACAC-3' (XmaJI) pr130r (SEQ ID NO: 24) 5'-GCAACCATGGAACAAGAGCGATGGAACAGTGTCGACGTCTACTTCAGC TCTCTAGATTGATAAGCTTGGCGTAATGGATCTG-3' (XbaI) pr156f (SEQ ID NO: 25) 5'-GAAAAAGCCTGTCCCGGATAGGAGTGTCATTTCATGCGAGAAGACTCG GCCGTCCCTAGGCCTGAAGTCAG0000ATACG-3' (XmaJI) pr156r (SEQ ID NO: 26) 5'-CCTCGGGCAGTGCGGCGTCCTCCTGCGCGGTCAGCCCGGGGTACAGCC CGTTTCTAGACTTCCATTCAGGTCGAGGTG-3' (XbaI)
Example 2--Phenotypical Verification of S. albus J1074/pStW102tc.DELTA.tomC-G and S. albus J1074/pStW102tc.DELTA.tomHI
[0080] Mutasynthesis was performed by cultivation of the mutagenized strains in 500 .mu.l production medium (20 g/l soy flour, 10 g/l corn steep solid, 20 g/l glycerol, 7.5 g/l NaCl, 2 g/l CaCO.sub.3) in a punctured 2 ml reaction tube at 30.degree. C. and 1000 rpm. After 24 h S. albus J1074/pStW102tc.DELTA.tomC-G cultures were complemented with 2-amino-5-bromobenzoic acid. S. albus J1074/pStW102tc.DELTA.tomHI cultures were supplemented with (S)-4-methylenepyrrolidine-2-carboxylic acid (chemicals provided by Sanofi-Aventis) to a final concentration of 500 .mu.M each. After 24 h samples were taken and analyzed by HPLC-MS.
Example 3--Production of 9-Chloro-11-De-O-Methyl-8-Deshydroxy-7-Hydroxy-tomaymycin (CDHT)
[0081] 200 ml of production medium (20 g/l soy flour, 10 g/l corn steep solid, 20 g/l glycerol, 7.5 g/l NaCl, 2 g/l CaCO.sub.3) complemented with 60 .mu.g Apramycin/l in 2.5 l buffled flasks with tissue caps were inoculated with 10 ml from a densely grown overnight culture of S. albus J1074/pStW102tc.DELTA.C-G. The culture was incubated at 30.degree. C. and 150 rpm overnight followed by feeding with 17.2 mg 2-amino-3-chlorobenzoic acid dissolved in 200 .mu.l DMSO giving a final concentration of 0.5 mM. Incubation was repeated overnight under said conditions. Cells were pelleted by centrifugation and discarded. pH of the supernatant was adjusted to 7.0 and it was washed two times with 1 volume of hexane. Extraction was performed twice with 1 volume of ethyl acetate, organic layers were pooled and dried by rotary evaporation. Crude extract was solved in 1.5 ml H2O/acetonitrile (1:1 v/v) and subjected to semipreprative HPLC for isolation of CDHT.
[0082] The biosynthetic pathway, fed amino acids, proposed structures and HPLC-MS measurements are shown in FIG. 4 (C and E).
Example 4--Purification of CDHT
[0083] Reversed phase chromatography was performed by a Dionex HPLC system (Famos autosampler, P680 pump, TCC100 thermostat, and PDA100 detector) equipped with a Phenomenex Luna C18, 250.times.4.6 mm, 5 .mu.m dp column. Separation was achieved by a linear gradient using (A) H.sub.2O+0.1% formic acid to (B) aceto nitrile+0.1% formic acid at a flow rate of 5 ml/min and 30.degree. C. The gradient started at 10% B and increased to 56% B in 18 min (2.56% B/min). UV data was acquired at 254 nm. The sample was injected by .mu.l-pick-up technology with a water/methanol (50:50 v/v) mixture as supporting solvent. Fractions were collected manually and analysed by LC-HRMS. Fractions containing a mass corresponding to CDHT were pooled, pH adjusted to 7.0, extracted two times with two volumes ethyl acetate, organic fractions pooled and dried by rotary evaporation. Obtained CDHT was analysed by LC-HRMS and NMR.
[0084] Obtained substances did match the mass for the proposed structures with deviations <1 ppm; showed the typical elimination of water for the hemiaminal form of the PBD; the mass reduction of 2 for the oxidized form characteristic for tomaymycin and in case of the 2-amino-5-bromobenzoic feeding the isotope distribution exhibited the M+: M+2 intensity ratio of 1:1 of brominated structures in MS.
Example 5--NMR-Spectroscopy
[0085] To further affirm the successful mutasynthesis 2-amino-3-chlorobenzoic acid was fed in larger scale, the product purified and its structure elucidated by NMR. Due to the remaining activity of tomO the obtained structure was hydroxylated at C-7 yielding 9-chloro-8-deshydroxy-7-hydroxytomaymycin. Structure data showed the presence of both diastereomers of the hemiaminal as well as the imine at the N-10, C-11 position.
[0086] NMR spectra were recorded at 298 K on a 500 MHz Avance III spectrometer by Bruker BioSpin GmbH equipped with a cryoplatform. CD3CN was used as solvent. Chemical shift values of 1H and 13C NMR spectra are reported in ppm relative to the residual solvent signal given as an internal standard. Multiplicities are described using the following abbreviations: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, b=broad; corrected coupling constants are reported in Hz (cf. Figure X).
[0087] 1H-NMR Data relating to 9-chloro-8-deshydroxy-7-hydroxytomaymycin: (500 MHz, MeCN-d4): .delta. 7.62 (bs, Ph-OH) 7.00 (m, 2H, 6-H, 8-H), 5.59 (m, 1H, 12H), 5.20 (d, J=8.9 Hz, 1H, H-11), 5.10 (bs, 1H, NH), 4.22 (m, 1H, 3-Ha), 4.07 (m, 1H, 3-Hb), 3.65 (t, J=9.0 Hz, 1H, 11a-H), 2.64 (m, 1H, 1-Ha), 2.51 (m, 1H, 1-Hb), 1.66 (m, 3H, 13-H) ppm; 13C-NMR (125 MHz, MeCN-d4): .delta. 167.5 (5-C), 153.4 (7-C), 134.2 (9 C or 5a-C), 133.9 (2-C), 131.8 (9a-C), 128.3 (5a-C or 9-C), 119.6 (6-C), 119.0 (13-C), 115.6 (8-C), 87.5 (11-C), 60.6 (11a-C), 51.8 (3-C), 31.5 (1-C), 15.0 (13-C) ppm; HR-MS (ESI): calculated for C14H16ClN2O3 [M+H]+: 295.0849, found 295.0844.
[0088] 1H-NMR relating to imine 9-chloro-8,11-dideshydroxy-7-hydroxytomaymycin: (500 MHz, MeCN-d4): .delta. 7.75 (d, J=4.6 Hz, 1H, 11-H), 7.62 (bs, Ph-OH) 7.26 (d, J=2.8 Hz, 6-H), 7.16 (d, J=2.8 Hz, 8-H), 5.59 (m, 1H, 12H), 4.16 (m, 1H, 3-Ha), 4.10 (m, 1H, 3-Hb), 3.90 (m, 1H, 11a-H), 3.03 (m, 1H, 1-Ha), 2.94 (m, 1H, 1-Hb), 1.72 (m, 3H, 13-H) ppm; 13C-NMR (125 MHz, MeCN-d4): .delta. 165.6 (11-C), 164.0 (5-C), 155.9 (7-C), 136.9 (9a C), 134.6 (2-C), 132.5 (9 C or 5a-C), 131.1 (5a-C or 9-C), 120.6 (8-C), 119.0 (13-C), 115.1 (6-C), 54.9 (11a-C), 52.1 (3-C), 31.3 (1-C), 14.6 (13-C) ppm; HR-MS (ESI): calculated for C14H14ClN2O2 [M+H]+: 277.0744, found 277.0741.
Sequence CWU
1
1
22125954DNAStreptomyces achromogenes 1atggaaagca attccccgtc attatcgacg
gaagagtcga cggaaaagct ggaagccgca 60ttgcgggcgg gaagtccacc gacccaggac
ccccctctcg tcctgacgac cctcttcgca 120cggacggtcc gcctgcacgg cggggccgtc
gccctcgcgg acggcgcgga acgcttcacg 180tacgaccggc tcaaccggcg ggccaaccgc
atggcccggc acctccaggg tcgcggcgtc 240ggcagaggcg accgtgtcgg agtgcacatg
caccgctctc ccgacctcta cgcggtcatt 300ctcgcggtcc tcaaggccgg aggctgcgtg
gtcccgctcg accccgccca tcccacgcag 360tacctcgctc gcatcctccg cgaggcccgc
ccgtcgccgg tgatctgcga cgaccccgac 420gacctgccct ccgaggcccg cgcgggcgcg
ctctcccggg acgacctggt ccgaggcagc 480gagggactcc ccgacggcga ccccgatccc
ggcaccggcc cggaggacac cgcgttcctc 540atgtacacct ccggctccac cggccggccc
aagggcgtac ggatcgcgca ccgcgggctg 600gcccggctcg gcccccacag cggccccctc
gacatcggcc ccccggacgg cctgacccag 660tcggccgcgt tctcgttcgc cgcctcgacc
atcgagatct ggctcgcgtt cctgcacggc 720gccaccctgc tgccgatgcc cccgggactc
ccctcgctgc cggtgctgcg cgaggccgtc 780gaggagcgcg gcgcgaccgt gctcagcctc
ccctgcggcc tgttcaacgc cctggtcgac 840caggagccgg agtgtctgcg ctcggtgcgc
atcgtgctgc tcagcggcga cttcccttcc 900cccgaccact tgcgccgcgc cctcgcgcac
accgacgcgg tcgtctacaa cggctacggg 960tgcacggaga actcctccat caccgctctg
cacccgctga cctcgcccga cgacgtggac 1020gggacgggcg tcgtcccgat cggccgcccg
ctgccgacgg tcaccctgga ggtctacgac 1080ccgtcgatgc gcccctgcga cgccggcgag
gtgggcgaac tctgcgtggg cggcgccggc 1140gtcgccctcg gctacgccga ccagccggag
ctgaccgccg agaagttcac ccgggacccg 1200gacggcgacg gactgctcta ccgcaccggc
gacctcgcgc ggcggaacga caacggcgac 1260atcgtgctgg tggggcgggg cgacagcatg
gtgaagatcc gcgggttccg ggtggagacc 1320agcgccgtga cgctggccgt gcgcgccctc
gacggggtcg cggacgccgc cgtcaaggcc 1380ttcgaggacg aggacacaca cgagaagcga
ctcgtcgcct tctacaccac cggcgacggc 1440tgcccggccg accccgcgga cctggtccgg
cggctctccg cggacctgcc ctcccacatg 1500gtcccctccg cgttccggca cctggagaaa
atgccgacga acgtgaacgg gaagatcgac 1560cgcaccgcgc tcaccgtgga atcccggaag
aacaggaaaa tccggaacaa caggaacgag 1620aaatccgaga aaggcgagaa gaccatgcag
aaccccctcg aagctgtcgt cctccagtcc 1680tggatcgaga tctccggaat ggacgacttc
tcgaccaccg actccttcct gggacacggc 1740ggtaattccc tccacttcgt gcagctggcc
tcccgtctcc agaagatatt cgggatcgag 1800atcaccaccg agtcggtctt ccggcacggc
acggtcgagc agcttgcgcg tttcatcgag 1860gagtcccgcg accaggcggc caccacctcg
tcgtcgcacg gctgacggag acccgacagc 1920gcccggcacg cgcgccgcgc cggtcgcccg
acgtctccgg agggggcgga ccgtgaacaa 1980ccctttgacc tcggctgttc tggacctggc
caggagaaca ctcggttccc ccggtctcag 2040gcccgacgag gccttctccg accactgcgg
cgaccccgcg ctgctggagc agctgggcgc 2100ggtgctgcgg gcggtgttcg ccgtcgaccc
gccggccggg gaacggctgg gcgcgcagag 2160cgcggcggcc gtggccgagc ggctcgcgcg
gggccgctcc ggctccggcg gacaggagct 2220gacggccggg tccgcgccgc gcgacggcgc
tacgacggag gcctcgttcg gccagagcgg 2280catctggctc atcgaccagt acctgccgac
cccgcaggcc tacaacgggc cgttcctgac 2340gcggctgccg ttccggctcg acggggaacg
gctgcgccgg gccgtcgccg gggtgctgcg 2400gcgccaggaa gtcctgcgca ccacctacgc
gttgcgcgac gcccggctcc tccaggtcgt 2460ctccgacgac gacacggtct tccactacgg
cgtcgagcac tacggcgacg acaaggaact 2520cgacgcgctc gcccggcggg tggccaacac
ccgcctcgac ctggagcacg ggcccgtggt 2580ctcggtgacc tgtgcgctcg gtccgggtgc
ggagagcgcg atcgtctgca acatccacca 2640catcgcctcc gatgccgcct cggccgggat
cttcctgcgc gaactcctcg acgcctacga 2700ccgcaccggg cggggcctgc cggtggaggc
cgtgccggga cgcccgcagt acgccgactt 2760cgcggagtgg caccgcaggc acctcacgcc
cgagcggacc acccggttgc tcgacgagtg 2820ggccgaccgg ctcgccgggg acctgcccgt
gctggccctg ccctccgacc ggccccggcc 2880cgccgcgcag gaacaccggg gcggcaccgt
cccgttacgt gtcccggccg gggtgaccga 2940gaagctggaa cggctcgcgg agcgcgaggg
cgtcacgctg ttcatggcac tcctcgcggt 3000gtacggcacg ttcctcgcca agctgagccg
tcaggagcgg gtgctgatcg gctcgcccgt 3060ctccctgcgg gacgacccgc agaccaagga
cctcgtcggc tacttcgtca acatggtcgt 3120gctgcgtcag gacgtgacgg gcgcgatgac
cgtgcgcgag gtgctggggc gggcgcgcga 3180ggaggtggcc ggcgcactgc ggctgaagtg
ggcccccttc gacaaggtcg tggagcggct 3240gcggcccgac cgcaccggcg ctcacacgcc
ggtcgtgcag acgatgctcg tcctgaccga 3300tcccggctcc gcccaagtca tgcacggagc
aacgtcattg accatccggc gtgacatggc 3360gcacggcgcg aagtacgacc tgtcggtggt
gttcgcacgg gaggcggccg ggctgctcgg 3420gagcctggag tacgacgcgc acctcttcga
cgaggcgacg gcgcggagca tgggcgaacg 3480gctgggccgg ctgctgaccc ggttcgccga
ggcggcgccg gacaccccgg tccgcgatct 3540cggtctgctt gccgccgacg aggagcggga
ggtgctggcc cacgacgacc gtgtggaccg 3600ctcggcgcct ccggtgccgg tggccgagct
gttcgagcgg caggcggcgg cgaccccgga 3660cgccgtggcc gtgacggacg gcgcccacgg
gtggacgtac cgcgaactgg acgagcgggc 3720cgaacggctc gcgcgggttc tgcgcgagga
gggggcggcg gcgggccgcc gggtggcgat 3780cgccctgccc cgctcggtcg agatggtcgc
gggactgctc gccgtcgtga agtccggggc 3840gtcctacgtg ccgctggacc cgtcgcaccc
gccggagcgg gtggcgtacg tcctggacga 3900cgccgaggcc ctcctcgtcc tcaccgacag
cgccacggcc gatcaacttc cgcctggtaa 3960aaggctgttg gagatcgacc gggaggcgga
ccggatcgcc gccgccgacc ccgcgccgct 4020cggcccgacc cggacgcccg acgacgagat
ctacgtcatc cacacctccg ggtcgaccgg 4080cgcccccaag ggcgtcgtca tccgggaccg
caccgtcggc aacctggtcg ccgcccagca 4140ccggatcagt ccctgcgggg cgacgggcac
caccctccag tacatgacgc tctccttcga 4200cgtctccgtc atggagatcc tggggacgct
ctgcgtcggc gggacgctcg ccctgatccc 4260cggtgaactg cagaaggacc tgcaccggct
ggccgcgttc gtcgccgagc acgacgtcac 4320ccgcctgtac ctgccgtaca tcgcgctcca
gcggttcgcg gcgctcgccg tcgcggagga 4380cctgcgctgc gacgcgctgc gcgaggtgac
ctcggtgggc gaggcgctgg tcgtctcccc 4440gcagatccgg gagttcttcg cccggcaccc
ccgcgcccgg ctgctcaaca tgtacggccc 4500gtccgagacc cacctcgcca cctggcacga
gctgtcgggc gaccccgccg actggcccga 4560ggccgcgccg atcgggcggg cggtcgacgg
gatgcggctg cgcgtcctcg gcccggaccg 4620cgaactcctc ccgccgggcg tcacgggcga
gctgtacatc ggcggcccgt acctgtcgcc 4680gggctaccgg ggccggctcg aggagacggc
acggcgcttc ctgcccgacc cgtacggcgg 4740cgacggcgag gtgatctacc gtacgggcga
cctcgtccgg tggaacgccc gcggcgacct 4800ggagtacctg ggccgcgccg acgaccagat
caagattcgc ggctaccgcg tggagcccag 4860cgaggtcgag gccgccctcg acgccctcga
cggggtgcgc gactcggcgg tggtcgcggt 4920cgagttcggg cccggcgacc ggcggctggt
ggccgccgtc acgggcgacg gcgccgacga 4980caccgtacgg ctgcgggccg cgctcgccga
gcggctgccg gagtacctgg tgcccgcgca 5040cctggtccgg ctggaccggc tgccgaccac
gcccagcggg aagatcgacc ggggcgggct 5100ggccggacgg ctggccgcag aggtgcgcga
ccggggcgcc gacgcgggcg ccaagacggc 5160acaaccgccg cggaccggga ccgagcgcgc
gatcgccgat gcctgggcgg acatcctggg 5220cggcggtgtc cccggacgcg acggcgactt
cttcaccctc ggcggcaaca gcatcatcgc 5280caccgaactc gtctaccggc tgcggacgtt
gttcgaggtc gacatcccgc tgcgaaccct 5340gttcgacaac ccgacggtcg ccgggatggc
cgcgcggatc gacgagcggc gggcaggcga 5400caccaccgcc ttcacggagc ccacggccga
cctgcgcgcc gacgtgaccc tgccggacac 5460cgtccacccc ggcgaccgga caccggtccc
gccggacgcc gcgacccggt tcctgctgac 5520cggcgcgacc ggcttcctcg gctgctggct
gctgcgggag ctggcgaccg tccccgggca 5580caccgtcacc tgtctggtgc gcgccgacga
cgcggacgcc gccctggccc ggctgcacac 5640cacggccgac cggtacggca tcaccggtgg
gatcgactgg gaacgggtgc gcgccgtccc 5700cggcgacctg gcccggcccc ggatgggcct
gtccgacgcg gaccacgacg agctggccga 5760gtcggtcgag gcggtccacc acgcggcagc
gcacatcaac ttcgtgctgc cgtacgcctc 5820ggtgaagccg gccaacgtcg acggcctgcg
ctccgtactg gagttcgccg ccaccggacg 5880gctcaagcac gtgcaccaca tgtcgaccgt
cgcggtgttc gcccccggcc gcgagggtgg 5940gacgatcacc gaggaagcgg tgcccgacag
gtgtgagggc ctcggcatcg gctacacgca 6000gagcaagtgg gtcgccgagg gcatcgcccg
gctggcgcgg gagcgcggga tgccggtgac 6060cgtctaccgc atcggccgga tctccggcga
cagcgttacc ggcgcctgcc aggccgacga 6120cttcctgtgg cggcagatca agagcttcat
cgaactcggc tcggccccgc ccgccgagga 6180actcaccacc gacctgctgc ccgtggactt
cgtcggccgg gccgtcgccg ccctctcccg 6240cgagccgtcc gccgacggcg ccacgtacca
cctgttccac ccgcgcggct ccgacttcac 6300ccccgtgcac gccgccgtcc gcgactgcgg
gcacccgctg gacacggtgc ccgccgagga 6360gtggctgaca cgcctggagg agtcggcccg
ccggcccggg ggcaacgccc tcgccgcggc 6420agtccccctg ttccgggagg gcgcgctcga
actcggcgag aacacctacg gcaacgagcg 6480gacgacccgg ctcctcgacc gtctcgggct
gcgctggccc gacatcgacc gccggtccct 6540cgcccgcatg atccgctggt tcgaagcggc
cggtgagctg gacgaccggg cccttgcgcg 6600acagggcgcg tagaccgggc cgcccagcgg
ctcccccacc cagacatccg caggcggtca 6660ccctccctgg ccgctcataa cgaaggccga
cagcatgtct cgatacccct ttccgggcgc 6720cggacacgac ccgttgatcg cctctatggt
ggccgctctc cccgcccgcc agcagcccca 6780atggcccgag ccctcccgcc tggcccccgt
gcacgagcgg ctggcccgcg agtcgccgct 6840ggtgtcgtac acgagcgtcc gcgagctgcg
cgggctcctc gcgcgggccg ccgagggcga 6900gttctgcctc atccaggccg gtgactgcgt
ggagctgacc accgagtgcg aaccctgcga 6960cgtcgaacgc aaggtggaga tgctcgacgt
cctcggcgac gtcatgcgca ccggctccgg 7020tctgcccgtc gtgcgcgtcg ggcggatggc
cgggcagttc gccaagcccc gctccgacga 7080ctgggagacc gtccccggcg gacggctccc
ggtgttccgg ggcccggtcg tcaacgcacc 7140ggacgcctgc gaggaggccc gcacacccga
cccgtcccgc gtcatcaccg gactggaggc 7200cgcccgctgc gccgtcaccg ccctcgaccg
gctcggccgc ggcgagggcg ccgctcccga 7260ggagcgggtg tggaccagcc acgaggcgct
cctgctcgac tacgaactcc ctcaagtccg 7320gcggcacacg gacggcggca gctatctggc
ctccacgcac tggccctgga tcggcgagcg 7380cacccggcag gccgacttcg cacacgtgcg
tctgatgacc gagctggaca acccggtggc 7440gtgcaaagtc gggcccgagg cgaccgtcga
cgaggtgctg gacctgtgcg cggtgctcga 7500cccgcaccgc accccgggcc ggctcaccct
catcgcccgg ttcggcgcgg accgggtcgc 7560cgcactggcc ccgctggtac gggcggtgcg
gcaggccggg cacccggtgc tgtggatgtg 7620cgacccgatg cacggcaaca ccgtgaagac
gcccgacggg ctgaagacgc gacgcctcga 7680cacgatcatg tccgagatcc ggcagtgcgt
ggacgtcctg gcggagaacg gggagcggtg 7740cgcggggctg cacctggagg cctcccccga
cgacatctgc gagtgcgagg gcgcgggtcg 7800cgtccccgtc cgcgggccgg gctaccgcag
cctgtgcgac ccccggctca gcctcgtcca 7860ggccgtcgcc gccgtcgcgc actggcggct
gcccgtgggg gcggcggccg tatgaccgcg 7920ccccccgcca ccaccgccgc tccggcgcag
gcccacgacg cgctgctgcc gccgccgggc 7980caccccttcg cggtgctgca ccgctccggc
gccggccgcg cggcaccggt ggagatcctc 8040agcggcccgg tacggcgcga ggacgacctg
gacgccctcg gcctggaccg gcccggcgga 8100gcgaagcggt ccacggacgg cggcgcggac
gccccggaca ccctcgtcct cgccccctac 8160cggcagatcc gggagcgcgg cttcgcgcac
cccgacgacg gcgagccgct gctcgccatg 8220gaggtgcgcg agcaccggag ggtccccctc
gaccggctgc tggcctgcct gccggaccgg 8280gcgccgcggg tggaggacac ccgcttcgac
gtcgacgacg acggctacgc ggcggccgtg 8340gacggcatcg tccaccgcga gatccaccgc
ggcgagggct ccaacttcgt gctggcccgc 8400agcctgcacg gccgcatccg cgccttcgac
cgcacggccg ccctcgcggt gctgcggggg 8460ctgatgacgg ccgagaccgg cgcctactgg
acgttcctga tcttcaccgg cgaccggtac 8520ctgatcggct cgaccccgga gcagcaggtg
cgggtgcggg gcgacctggt cgagatgaac 8580ccgatcagcg ggacctaccg ctacccgcgg
accggcgccg acctgccggg gctgctgcgc 8640ttcctgcgcg accccaagga gaccgacgag
ctgtacatgg tcgtcgacga ggaactcaag 8700atgatggcgg ccctgtgcgg cgacgaggtc
cgggtcagcg ggccgtccct gaagtggatg 8760tcccggctcg cgcacaccga gtactacctc
tccggccgct cggaccgccc gctcaccgag 8820atcctccgca ccacgatgcc cgcgccgacc
gtgacgggca gcccggtgga gaacgcctgc 8880cgggtgatcg agcggtacga gccgcgcggc
cggggctact acagcggggt catcgccctg 8940ctgggccgcg agggcgggca gcggcggttg
gacgcggcgc tgatgctgcg catggcggac 9000ctcggggcgg acggcaccgt acggctgacg
gcgggcgcca ccgtcgtacg ggagtcggta 9060ccggcgcacg agacggcgga gaccaccgcc
aagctgtcgg gtctgctgga cgcgctgtcc 9120ggtcggcggt cggcgaaggc gccggtgccg
gcgtccgccg cggggctggc cgaggccggt 9180ccggtgcgcg gcgcgctggc cgcccgcaac
gacgggctgg cggcgttctg gctgcgcggg 9240accggcgacg tccgcgccgc ccgggccgtg
gcgggcgccg acgtcacgat cgtcgacgcc 9300gaggacgggt tcacgtcgat gctcgcctac
caactgcggt cgctgggctg ctcggtgcgg 9360gtggtgccct ggcaccggac ggcgggcggg
gtcgccgggg cgccgggcgt ggtcctgctc 9420ggaccggggc ccggcgatcc gcgggacacg
gccgatcccc gggtgcgggc cctgcgggcc 9480acggcccgcg aacggctcac ggccaggctg
ccgttggcgg cggtctgcct gggacaccag 9540gtggtgtgca cgctgctggg gctgcgggtg
gcccggctgc ccgacccggc gcagggccga 9600cggctgcggg tcccgctgtg gggcaggccg
cgccgggccg gcttctacaa cagctacgcc 9660gcgcgggccg acggcgaccg gctgcacccg
ccgctgggcg acgggccggt gcaggtgcag 9720cgcgtcggcg acgaggtcat cgcgctgcgc
ggcccggccc tgtccaccat ccagttccac 9780gcggagtcgt tcctgaccga ggacggcccc
ggcatcctgg ccgacctgct gaccggcgcc 9840ctggcaccgg ccgcggcacc ccggccgggc
gtccacgaca gcgacggggc cggtgtccac 9900gacaccggac tccgggagtc cagggggacc
catgagcatg cctgagcgag ggggcgtgga 9960caccgcgccc cgtccgatcg acccgcgcgc
cctgcgcagt tgcatgggcc agttcgcgac 10020gggggtcacg gtgatcacct gccgccgcgg
cgacctcgtg cacggcacga cggtcaacgc 10080gttcacctcg gtgtcactgg acccgccgct
ggccctggtg gcgttggacc ggcgcagtcg 10140ggcgggcgcg ctgctgcggg agaacggcga
ctacgtcgtc aacatcctcg acgcctcgca 10200gcgcgatctg gcgatgcact tcgcaggtcg
gcccatggcg gagccggtgc cgtgggtgga 10260cgaggacggg ccgcacccgc ggctggcggg
cacggtggct catctggtct gccggccctg 10320gcagatccac gacggcggcg accacacgct
gcacatcggg agcgtcgagg agttcgagag 10380ccgtccggga cggccgctgc tgttccacgg
cggcgccttc ccggagctgg cgccggacga 10440ctccgccgtc gcctggtcgc tgtgcctgga
cggcatgaac ccggtgacgc acctcccggt 10500acccgaagag cccgacgagc ccgacaggac
agacaggaca gacaggacag gggactgacg 10560tacatgacgg acacaaggca cagcacgggc
gcgacgggcg cccccggcac cgggaccacc 10620ggctccgcgg gcgccaccgg tgatgcggcg
ccgcgccgca cgacccgccc gatgaccggt 10680gacgagtacc tggagagcat ccgcgacggc
cgggagatct gggcgtacgg ggagcgggtc 10740gacgacgtca ccaagcaccc ggcgttccgc
aacacggcgc ggatgacggc tcggctgtac 10800gacgccctgc acgacccgga gcaccacgac
acgctgacca cgccgaccga caccggcagc 10860gacggctaca cgcacaagtt ctaccgggtg
ccgcgcagtg tggaggacct ggtcggcgac 10920cgggacgcga tcgcttgctg ggcccggatg
acgtacggct ggatgggccg cagccccgac 10980tacaaggcga gcttcctcgt cactctcgga
gcggacccgg actactacgg ggagttcgcg 11040gacaacgccc gacgctggta cgcggaggcg
caggagcggg tgctgttctg gaaccacgcg 11100gtgatcaacc cgcccgtgga ccgcaaccgg
gcgccggacg aggtcggcga cgtgttcgtg 11160cacgtcgaga aggagtgcga cgacggcctg
gtggtcagcg gcgcgaaggt ggtggcgacg 11220gggtcggcgc tgacccactt caacttcgtt
gcgcactacg gactgccggt gaagaagaag 11280gagttcgcgc tcgtcgcgac gctgccgatg
gacgccccgg gggtgaagct gatctgccgc 11340cagtcgtacg agctggccgc gaaccggatg
ggcagcccct tcgactaccc gctgtcgagc 11400cgcctcgacg agaacgacac cgtcttcatc
ctggacaagg tgaagatccc ctgggagaac 11460gtcttcatct acggcgacac cgcgaaggcc
gggaccttcc tgaacacgtc cgggttcacg 11520caccggctca ccttccacgg cgtgacccgg
ctcgccgtga agctggactt cctggccggt 11580ctgctcctca aggggctgga cgtgaccggc
acgaaggatt tccggggcat tcagaccagg 11640gtcggcgagg tgctcgcctg gcggaacatg
ttctggggtc tcagcgacgc catggcgcac 11700aatccgaacg catggcacga cggcggactg
ctgccgaacc tggattacgg catggcatac 11760cgctggttca tgacgatcgg ctatccgcgg
gtccgcgaga tcatcctcca ggacctcagc 11820agcggcctca tctacttgaa ttcgcacgcc
aaggatttcc agaaccccga acttcggccg 11880catctcgacc gctacatgcg agggtcgaac
ggatacgact cggtggagcg ggtcaagctg 11940atgaagctga tctgggactc ggtgggcacg
gagttcggcg gccggcacga gctgtacgag 12000cgcaactact cgggcaacca cgagaacgtc
cgtatcgaac tgctgatggc gcagaccgcg 12060tccggcctgg tcgacggcta ccggggcttc
gccgaggagt gcatgtccga gtacgacctg 12120gacggctgga cggcgccgga cctcatcggg
cccgaccagg catgagcggc ggggcggcac 12180gacgacgggg gcggcggacg gccggtgtcc
ggtccgccgc cgcccccggg gggccgccct 12240cccgccgggc gtcagcgcgg ggcgggcgcc
tcgggccggt cgtccggggc gggcaggatc 12300acggcaaggg tgaagccgtc gtagcccttg
gcgcccaccg tctggatcgt ggtggcgctg 12360accctgggct ccgcggcgac cttctcgtgg
aagcgccgca tcccgcggac gccgccgtcg 12420gggtcgtccg agtcggcgac ggcgccgccg
agcaccacgt tgtcgacgac gatgacgccg 12480ccgggccggg acagttcgag cgcccagtcg
aagtactccg gggtgtcggg cttgttggcg 12540tcgatgaaga ccaggtcgaa cggctgctcc
ccggtcttcg cgaggtcggg caggatgtcg 12600agcgcccggc cgacccgctg gtcgacgcgg
tcggcgacac cggcccgcgc cagatggccg 12660gcggcggtct cggcgaacga gcgctcccac
tcgatggtga ccagccgtcc gccgggcggc 12720agggcgcggg cgagccagat ggtgctgtac
ccgccgaagg tgcctatctc cagaatgcgc 12780tcggcacgct ggacgagggc cagcaggtgc
agcagctttc cctgggagcg ggagaccccg 12840atatcgggga ggtcgaatcc gatgtgtgcc
tgggcggcgg cggccagggc ggggtcctcg 12900tcaccgagaa ccgtgtcgaa gtagccgtcc
accgcgctcc actgatgctg tgtcatgact 12960ccagtgttcc ccgtgaggag gaaattcggc
accctgaagg aaggaattcc agaaaaacgg 13020cggattcctg tcgtccactt atccgcgtcg
gttagtggag cggattttcg ggaattcgcc 13080tgcctgaacg agaaaaagaa cgtctttcgt
ttcccaccag cgacccaccc ctcgatctct 13140cctggcacga cttcaatgag aggagcgcag
tatcttgcag cgaaccggga ccacgacaga 13200gcgaccgccg aacccccacc tcggccctga
tcccgccccc gatctccacc tcgtaccgga 13260gctgcaccac gtgggcgtgc agacggacga
cctcgacaac tgcgccgcct ggtacgagga 13320gttcttcggc tgccggacca actggacgct
ggacaccttc tccgacctga cgctgagccg 13380cctgcccggc atcacccggc tgaccgaggt
gacggtggcc ggactgcgct tccacctctt 13440cgagcgcacc gggcacgacg gcgccgtccc
cggtgccaac acccgccagt tccagcacgt 13500ctgcctcgcc accggctccc ccgaggagct
gcgggcctgg cgcgagcgct ggttccggct 13560gtacgagtcc ggccggttca cgttcgccgg
tgacgaggag cccaccgaca tcgtcatcga 13620cgacgacggc gtccagagct gctacgtcct
ggacgtcaac ggcctggagt tcgagttcac 13680ctacgtgccg ccaggaggag accgatgagc
atgcgcgccc cccgcaccgt gaccgaactg 13740ccggtgcccg acggctggga cttcggggac
ttcccctacg gcctggaacc gctgacactg 13800cccgagccgc cgacgccggg aacggagacc
gcgatctcgg acgtgctccc cgccgacacc 13860tcccgcgtgc ggagcgccag gccgtgcccg
cgcacggggc cggcgctcgc cgccgaggag 13920atctcccacc agctgttctg gttccgctgg
atcacgggac accaagccac cttcgccatc 13980tggcagttga cggcgcacgc cctgcaccag
gcccgctccc gttccgaccc cgcgccgtcc 14040cttcgggcga tgacggacct caccgacgcc
tacaccgcga tgctgctgta caccagttcc 14100tgccccaccg acgtctacgg cacggtgatc
cggccgagca tgtacctcca gcaccgcagc 14160ttcagcggca cctgggcccc ggacttcgtg
cccgtgcgct cgctgctgcg gggcaagaag 14220accgagtggg aggggacgcc ggaggccgaa
cggctgaaga aggcggtgca gatgtatcac 14280agggtgcacg cgggggtggc ggccaagctg
gttccgggcg gccgttcgct gctccaggag 14340tcggccgcgg aggtcgcacc gacccgcccc
gagacccagg cgctcatcta cgaccactac 14400ttcctgaccc tgcgggcccc ggtcgacgcg
acggagctgg tcggccagtt gcggagccgg 14460ctgcgggcca tcacgcagga cgtggcgacc
aacggcctct atcccggcct cagtccgcag 14520gaggacgtcg ccttccccga ggaactgcgg
ggcgacgagg tacgccaggg gtacgaggag 14580ggcttcgcgt cggtcctggg ccggatcgac
gccgccgccg ggcagctcag gccgcgggtg 14640ctgcaccaca gcgccccctg acgggacggc
gtccggcacg acgaggccgg acaacgtccg 14700gcacgtcgga gcacgacggc atccggtacg
acgaaggagg agtacgcgtg cggcacggag 14760tggtgatcct tcccgagcgg cgctggtcgc
gggcgcgcga gcagtgggtc ctggccgagc 14820ggctgggctt cgaccacgcc tggacctacg
accagttgat gtggcggtgg ctgcgggacg 14880agccctggtt cgcctgcgtc cccaccttcg
ccgccgccgc ggccgtcacc tcgcggatca 14940cgctgggcac gatggtcgcc acgcccacct
accgccatcc ggtgacgctg gccaaggagg 15000tgatgtcgct ggaggacgtc gccggaggcc
gcttcgtctg cggactgggc gcgggggcgg 15060gcggcctgga cgaccgggtg gtggacccca
ccgagcggac cccgcgcgag cgcgccgacc 15120ggttcgccga gttcgtcgag gcgctggacc
tcctgctcac cgagcggagc gcctcgtacg 15180cgggcgagca ctacgccttc gacgacgtgc
cggtgaaccc cggttgcctg acacggcccc 15240gcgtgccgtt cgccgtcgcc gcgaccgggc
cgcgcgggat gcggctcgcg gcgcggcggg 15300cggacacctg ggtgacggcc ggtccgcccg
gccgtttcga cgccctgccc tacgagaagg 15360cgctgcccga catcgcccgg cagctggacg
agctggacgc cgcctgcgag gcgaccggcc 15420gcgacccggc cacgctggac cggctgctgc
tgaccggcgc cctggtcggc ggggtgctcg 15480actccgtcga gtcgttccgg gacgcggtgg
gccgcttcgg cgagctgggc gtcaccgacc 15540tggtcgtgca ctggccccgc gcgtcgttcc
cgtacgaggg cgacccgcgt gtgctggagg 15600ccgtcgcgga ggaggtgctg ctccccggac
gggaggcggc cccgcagacc gggcccgtgt 15660caccccggcc gggcagcgcc cccagtggcg
aaggagagcg acgttgatcg cgtacgagat 15720cgtcgacatg ttcacggaca ccccgtacgg
cggctgcgcc ctcggcgtcg tccccgacgc 15780ggcggcgctc tccacggcgg acatgctggc
cgtcgcgcgc gagaccgccc tgacggagac 15840ggcgttcgtg gtgccgccgg ccctgccggg
gtcgacgtac ggagtgcggg tgatgacccc 15900ggacggggag tcgccgtacg gcgggcacag
cgcggtgggc accgcgagcg cgctcgtgcg 15960cgccgggcgg ctcgcggccg gggaggcggt
gcaggagtgc ggcggccggc agctcgccgt 16020gaccgccggg gccgacggct ccaccctgtc
cgtcgcgggc gagccgctgc tgcggcccga 16080gtgggacccc ggcccgctgc tcacggcgtg
cggcctgacg gacaccgacc tgacgggcac 16140tccgcgtctg accggcttcg gaccggcgtt
ccacgtcctg ccggtgggcg cgaaggccct 16200cacgcgcgcc gcggccgacc tgaccgaccc
ggtctgggcg gactgcccgg acgcggtgct 16260cgtcgcctgg gaccaggccg gccgcaccgc
ccgcgtccgc gtgttcgctc ccggttacgg 16320catgcccgag gacccggcct gcgcctccgc
cgcgctgggt ctcggcgcct ggctcgccga 16380ggagaaggcg ctccccggcg cggacggccg
ccacgcctac caggtccgcc aaggcgaggg 16440cctcggccgc ccggccaccc tctcctgcac
cgtcgacctc accgggggcc gggccaccgc 16500cgccacggtc cacggccggg tcaccctcac
cgcctccggc cggatgacac cccccggacg 16560cccctgaccc cccgctcccc ccctgcggtc
cgaccgtacg accagaaagg atcttcccag 16620gtgctcttcc gaccggaact gcacggcacc
cgcggtgccg tcgcctccac ccactggctg 16680gcctcccaga cgggcatgtc catgctggac
cgcggcggca acgccttcga cgccgcgacc 16740gccgccgcct tcgtcatcca ggtggtggaa
ccgcatctca acgggccggg cggcgacgtc 16800ccgatcctcg cccatcgctc agggaccggc
tcggtcgacg tcgtctgcgg gcagggcccg 16860atgccggccg cggcgacgcc ggaggccttc
accgagcggg gcctggccgt cgtccccggt 16920tccggacttc tgcccgccgt cgtccccggc
gccttcggtg cctggctccg cctgctcaag 16980gagtacggca cgctgcccct ggaggacgtg
ctggaacccg ccatcggcta cgccgagcac 17040ggctacccgg tgctgcccaa gtgcgccgcc
atgatcggcg cgctcacgga gctgttccgc 17100gacacctgga cggagtcggg ccgcacctac
ctgcgcaacg gggtcgcccc gcgcgccggg 17160gagcgggtca ccaacccggc gctcgcgcgc
acctaccggc ggctggtcgc cgaggcgaag 17220gcgtccgccc gggaccgcga gggacagatc
gacgcggcga cggaggcgtt ctacaccggc 17280ttcgtcgcgg aggccgtcga cacctatctc
gccaaggccg aggagatgga ctccaccggc 17340cgccgcaaca gcggcctgct caccggcgcg
gacctcgcgg cgtggcgacc gcgtgtggag 17400cgtcctctga gcctcgacca ccgggacctc
accgtgcaca aggcgggccc ctggtcgcag 17460ggtcccgtct tcctccagca gctcgccctg
ctggccgacg ccgacctcgc cgcgaccggc 17520ccggagagct cggagtacgt ccacacggtg
gtcgaggccg ccaagctcgc cttcgccgac 17580cgggaggcct ggtacggcga cccggaccac
gcagacgtcc ccgtcggcgc gctgctggac 17640ccggcgtaca ccgccacgcg gcgggcgctg
ctcggcgcgg aagcctccta cgagctgcgg 17700ccggggcggc ccggcgggcg cgagccgctg
ctgccgccgc tgcacgacga gagcgtcgcg 17760cccgccgacc cggcctggct gggcgagctg
gagcagggca tcccggcgat catgaagctg 17820accgcggcga ccggcgacac ctgctgcgtg
agcgtcaccg acgcggcggg caacatggtg 17880gtggcgacgc ccagcggcgg ctggctgaag
agttccccgg tggtgccggg gctgggcttc 17940ccgctgggta cccggggaca gatggcgacc
ctcacccccg gccaccccaa cacggtggcg 18000cccggcaagc gtccgcggaa ccacgctgag
cccgaccttg gtcctgcggg acggccggcc 18060gtacctggcg ttcgggacgc cggggcggcg
accagcagga ccagtggacg ctcagcttct 18120tcctgcacca caccgagcac gggaagggcc
cgcaggcggc gggggggatc gaggcgcgca 18180ccttccacac cgatcacgtg cccacctcct
tcgccccgcg ccggttccgg ccgggcaccg 18240tcgtcctgga gggcggctgg cccgcggcgg
aggtcgaggc gctccggcgg cgcgggcacc 18300aggtggacct ggtgcccgac tactcgctca
gcaaggtgtg cgccaccggg ctgaccgccg 18360acgggctggt tctcggcgcc gcgagtccac
gcggcgcgca ggcgtacgcg gtcgcggggt 18420aggagaggag gacaggaaat ggtttccatc
aggaaaccac ccctccgagc gcggaaggtg 18480ctgacaatgg cggctcaccg gaagaaccac
agtggggact tcatcgtgct gagcgaggcc 18540cggcagaaca atctcaaggg cgtcgacctg
cgcatcccca aggggaagct gaccgtcttc 18600accggggtct ccgggtcggg gaagtcctcg
gtggtcttcg acacgctggc gatcgagtcg 18660cagcggcagc tcaacgagac cctgccggcc
ttcgtccgcc accggatgcc caagtacgaa 18720cggccggacg cgggggtgat ggagaacctc
tccacggcga tcgtcgtcga ccagcggcag 18780gtcggcggca acgcccgctc cacggtcggc
acgatgaccg agatcctgcc catgctgcgg 18840gtgctgttct cccgcgcggg cacgcccagc
gcgggcccct cgcacatgta ctcgccgaac 18900gacccgcgcg gcatgtgcga ggagtgccag
ggcctcggca cgacggtccg tctggagctc 18960gacaagctgc tcgacgagga caagagcctc
aacgaggggg ccatccgctt cccgtccttc 19020gccgtcggca ccttccagtg gcagctgtac
ggcgagtccg ggctcttcga ccccgacaaa 19080cggttacgcg acttctccgc caaggaccgc
gaactgctgc tgcacggcaa ggggttcacc 19140gtcgaccggg cgggtcgcaa cggtgtctac
aagaacgagt acgagggcgt cgtcgagcgc 19200ttcacccgcc ggtacatcac ccccggcctc
gaccacgcca agagcaagga gcgcgcggag 19260atcgagcggg tggtgaccca gggcccctgc
cccgtctgcc acggcggacg gctcaacaag 19320gccgccctcg cctcccgcat cgacggcgac
aacatcgcgg acttctcggc catggagatc 19380accgacctga tcgaacggct ggcccggatc
gacgcgcccg ccgtcaagcc ggtggtggcg 19440ggcgcgcggg ccgcgctcga gcgcatccag
gcggtcgggc tcggctatct gagcctcgga 19500cggcagacca ggaccctgtc cggcggcgag
gcccagcggc tcaagatggt gcgccacctg 19560ggcagcagcc tgaccgggct gacgtacatc
ttcgacgagc ccagcgtcgg cctgcacccg 19620cgggacgtgc gacggatgaa cgacatcctg
ctcgccctgc gcgacaaggg gaacaccgtg 19680ctggtcgtcg agcacgaccg ggacgtcatc
gcgatcgccg accacgtcgt cgacatgggc 19740ccgggggccg gccgcgacgg cggcgaggtg
accttcgagg gcaccccggc ggggctgcgc 19800cgctccggca ctgtcaccgg caagcagctg
cggagcgtga ccgggctcaa ggaggaggtc 19860cgctccccca ccggcgaact gaccgtgcgg
gcggcgaagc tgcacaacct gcgggacgtc 19920acggtcggct tccccaccgg ggtgctcacc
gcggtgaccg gagtcgccgg ctccggcaag 19980agcaccctcg tgtcgaaggt gttcaccgcc
cagtaccccg acgccgtcgt catcgaccag 20040tcctcggtgg ggatctccgc gcggtccaac
ccggcgacgt acaccgacat catgggcagt 20100ctgcggcagc ggttcgcgcg cgcctcgggc
gagaagccgg gactgttcag cttcaactcc 20160gagggcgcct gcccgcagtg cggcggcaag
ggcgtcatcg agaccgattt ggccttcatg 20220gacccggtca ccacggtgtg cgaggcgtgc
gacggccgcc ggttcaacga cgaggcgctg 20280cggcacaccg tgggcggccg gacggtcgtc
gacgtcctcg ccatgaccgc cgaggaggcg 20340tcggacttct tcgacgacgc gcccgtacgc
cgcaagctgg ccctcctcac cgaggtcggg 20400ctgggctacc tcaccctggg ccagcccctg
tccacgctct ccggaggcga gcggcagcgg 20460ctcaagctgg cgcaccgtct gaaggagacc
ggcaccgtct acgtcttcga cgagccgacc 20520accggtctgc acatgagcga cacgcgccgg
ctgctggacc tgttcgaccg tctcgtggac 20580ggcggcaaca cggtgctggt gatcgagcac
gacctcgacg tcgtcaagca cgccgactgg 20640gtcgtggacc tcggcccgga ggcgggcagc
cagggcggcc gggtggtgtt cgagggcacc 20700ccggccgagc tggcccgggc gaagggctcg
cacaccggcc gcagcctcgc ggcggacctg 20760cgcaccgccc gtgagcacag ctcgtcctga
gacacggtcc cagagcgcaa cccacgacac 20820agcccgaggg cacaacccag ggacacggta
ccggcccacg gtcgggacgc gtcccggggc 20880gctgtcccgg tcaggaccca aggagtcgga
gatgcccctc atccgcgtca ccttgctgga 20940gggccgctcg ccgcaggagg tggccgccct
cggagaggcg ctcaccgcgg cggcgcacga 21000gaccctcggc accccggtgg aggccgtccg
ggtgatcgtg gaggagacgc cgccggagcg 21060ctggttcgtc ggcgggcggt cggtggccga
gcggcgtgcc tccccctcgt gacccgctcc 21120gtcaccgccg cgggccgccc cggagccgcc
ggggcggccc gcacgtccgg gagcgccggt 21180tcagcggacg gcggccccgc gcgggctgcg
gtcggcgcgc gcgaagcgct cccggacccg 21240ctcgacgcgg tccggggacc aggacgcgag
gtcgtcgcgg gtgaggcggc cgctgcgggt 21300gatcagatgg gcgttgaacg cctccggttc
gaggtcggcg tactggtcca gttcctcgaa 21360ccactcctgg ctggtgcgcg cggtgcgctg
cacggccgcg acctccggca ggcgttcggc 21420ctcgtaggcg gcgagcgcct ccggcaccga
cccgtggtcg tccagggcca gggcgagcgc 21480gagggcgtcc tccatggcga gccgggtgcc
cgagccgacg gagaagtggg tggtgtgcgc 21540gctgtcgccg aggagggcga tgcggccgtg
gctccagcgc ccgttgcgga cggagacgaa 21600gcggccccag cgcgaccggt tggtgcgcag
gtcgtggccg tccaggacgt cggcgaagag 21660gtccgcgagc cggtccgtgt cgctcctgcc
ggaggcgtcc ggctcaccgg cgtcgccctc 21720ggtgaagccg gccgcctccc acacccggtc
gtcgacctcg gcgatgaagg tgctgaggcc 21780gggggcgtag gggtaggcgt gcgcgtgcag
caagccgtgg tcggtctcgg cgacggcgaa 21840ggtcagggcg gtgaaggccc gctcggtgcc
gagccacaga tagcgcgagc cgcgccgggt 21900ggtctcggtg ccgaactcct cccgacgggc
atcgcgggtc gcggagttga cgccgtcggc 21960tgccaccacc aggtcgtacg tgcgggccaa
ctcgtccgcg tcgggggcgc gggtgccgtg 22020gcggacggtg acgccgaggt cggcacagcg
ggcgcgcagg agggcgagca gggcgcggcg 22080gccgatcgcc gagaagcggt aaccgccgtt
gcgcgaggtg cggccccggt ggtggacgtc 22140gatcctcgac cagtgggcgc accgggcgcc
gagcgcggcg aacagcgggg ggtcggccgc 22200ctcgatgccg ccgagcgccc cctcggagaa
gaccacgccg aagccgaagg tcacgtcggg 22260ggcgttcgcc tcccagacgt cgatctcgct
tccggggcgc agccgtttga cgagcgcggc 22320cgtgtagagg ccgccgggtc cggctccgat
gatcgcgatt cgcatgacac ctcacgggga 22380ctcggtcgag tgacggggtg gcgcgcgcgg
gggacggcaa gtggcccctc caacggacta 22440cgtattgccc atgagagcaa aagtattatc
cgcagggaaa ccaatcgcct cgggcggtcc 22500gccgccgcgc ggccgaccgg gagggggcgc
ggccgatgag gccgacgacg gacgcggagg 22560tgctggagcg gctgtacggg gcggacgtgc
cgcccttcgc gctgctgcgc agacagcgtc 22620ccgacgcgcc cggaccagca ccggtggagc
tgctgttcgg ggacgtggac accgggaagt 22680cgatggggtc ttcggggtct ttggacaacg
cggagaccgc cgggtcgctg gacggggccg 22740gtccgcccgg tgccgcgccg gggccgggcc
tgctggccct gctgcccttc cggcacctgg 22800ccgaacacgg cctgccctgc cgtgacgacg
gcacgccggt gcgggtcctg cgggtgcgcg 22860agcaccacgt cctgccggcg gacgcgttca
ccgaccctcg ccttcccggc gcggggccgc 22920tggagtggcg ggacgcgggg ttcacgacga
ccgacaccca gtacgaggac ctggtgcgag 22980ggttcgtcgc ctcggacgcg gcccgcggcg
gactgaacgt cctgctgcgc cgcgactgga 23040ccgcgcggct gccggacttc cggcccgcgg
cggcggtgga cctgttccgg cggttgctga 23100ccgccgagtc gggcgcgtac tggaccttcg
tcctgcacac cgggggcgcg cggccgctca 23160ccctggtggg ggccacgccg caggggcacg
tgggcgtcga gggcggccgg gtggtgatga 23220acccgctgtg cggcacgtac cggctcccgc
cggaaggacc ggacgccggg gacctgctcg 23280ccttcctcac cgaccgcaag gagtcggagg
aactggcgac cacggtggac gccgaactgg 23340cggtgctgtg cgggctgtcc gggccgggag
tgcgggtgga ggggccgttc ctgcgccgga 23400tgtcgcatct gctgcacacg cagtgccgca
tcagcgggcc cgcggcagcc ggggcccggg 23460agaccctcgc gcgcgccctg ttcgcgtcga
cggtcgtggg cagcccgtac gccgacgcct 23520gccgggcgat ccaccgccac gagacgacgg
ggcgcggcta ctacggcggc gtcctcgcgc 23580tgatcggccg cgacgccgcg ggggcggagg
agctggactc ggcggcggtg atccggaccc 23640tggtcgtgga ccaccggggt gaagcgcggc
tgtcggtggg ggcgaccgtg ggcggccggt 23700cctcgccgga ggcggaggcg gccgagaccc
gcaccaaggc gcggaccgtg ctgaccgccc 23760tcaccgcgca accgcccgca gccgcgcaga
cgcccgcgcc ccgcgcgccg gtcctcgacg 23820cggccgtacg ggacgcgctg gaccggcgcc
acgcggcgct gtcgtcgttc tggacggagg 23880gcaccgtcgc cccgccagtg ggcgccgccc
cggtgctggt gctggacccg ggcggcgacc 23940cggtcgccgc actggcggcc ctgctgcacc
tggtcaccgc cgtgcacggc cccgccgtgg 24000tggtccggta cgacgccccc ggcgccgccg
agaccctggt ccggcacgcg gggcccgtac 24060tgctggcgtc cgggccgggc cgcccggacg
accccgggtg cgtacggatg gcggcgctgc 24120gccggacggc cgcgtccctg gtacgggaac
ggccgcccgg cgggcccccg gtggccgggg 24180tgggcctcgg cttccagttg ctggcgctgg
ccgccctgcc ggaggcccgg gtcgtgccca 24240ggaccggcgg cacaggactg gcgcgggagg
tcgaggtctt cggcaggccg gtcaccgccg 24300cgttccgcaa cgcccacgcg atcgtgtccg
gtccggacgg gcacgaggtg gtcgcgctcg 24360gcgaacagac cgtgcggggc gtgcagttcc
gcccggagtc ggtgctgacg accgacgggg 24420ccggggtgct gcgccgactc ctcggctgac
tcccccggcc cgtcgtcgtc cggtgccgtc 24480gtctcccggc cccgccgacg caggctcagg
acgcgtcggc cgccgcgccg agttccacga 24540gggtgcgggc cgcggactcc agggcgaagg
tcatcgagcc gccgttgggc tcgaacccgg 24600cgtggtcacc ggcgaagtgg atacgcccct
cgggcctgcg gatcaccggc atcagggcgc 24660tgtgcccacg ctcgggaagg atgtacgccc
cttccgcgta cgggcgttgg tcccagcaga 24720cggacgtgct ggtctcgtag tgctcgcggg
cgcccgggag catccgttcg acctcgtcga 24780gcacgaagtc caggcgctcc tcctcggtca
tgaccgacac cgcccgcgcc cgccaccccg 24840tcatcaggca ctccaggatc ttgcgcgggc
ccgggagccg gggggtggcg tcgcgtatcc 24900accgtatggg caggtcggtg gagaagctgg
cgttgcgctg cggccagaac tggcggcgca 24960tctggaggta gacgcggacg acggacgagt
acttcacccg gcgcatcacg tcgtgcttgg 25020cgtcggtcag gcccgcgtcg gtgaagtcga
tgtgccggat cgcgctgaac gggacggtga 25080cgacgactcg ttcggcctcc agggtctgca
tgcgaccgtg ggcgaggacg gtgacccggg 25140cccgctccgc gtcctgttcg acgcggacca
ccggacggcg gtagtggata cggctcttga 25200ggcgctcggc gaacgcctgc gggaagcggt
cggtgccgtg cttgaccttc gcccagcggg 25260gatcggcgtt gccgagcgag tgcgggctcg
cctcgtggcg cagccaggac agcgcggacg 25320ccgtcttcag gtcgccgccg cgcatctcca
ggaagagggg ttccatgagg tcgatggccg 25380cctgggaggc gccccgttcg gtgaggacct
cgtgcacgga caggtggtcg tagggggcga 25440ggcgctgggt gaccgcccag tcgggggcgt
tcaggtcggg ctggaggccg tcggcgaccg 25500ccgagacgta ccgctcgatc atcccgccga
cgccgagatc cttctcgtgc gggtgcaggc 25560cgagcccggc ctcggcgacc gactccgggt
cgggggtgaa gaaacggccg cgcgcgaagt 25620aggagaacgt ggtgtcgaag aggtcggcgg
tctccgtctc gacgccgagt tcgcgcaggt 25680agtgcatggc gtagtggcag tgcggggtga
gcgtcatcgc gccggcctcg gcatgcaggc 25740cgtcggcgaa cggctccctt aaggtgtacg
cgcgcccgcc ggcccggtcg gacgcctcca 25800ggacggtcac ctcgcagtcc ctcgcggcga
gttcgtgggc gacggccagc ccggcgagtc 25860cggctccgac gacgatgacg gatcgcgggg
gccgcagccc cggcagtccg ctgtcgaagg 25920cacgtcgtac ctcggtctga gtggcttccc
gcat 25954229067DNAStreptomyces species
FH6421 2ggagcgcgca agatccgacg ggaaccgagc ccccggcccc ctcgtcggcg ttgtcattcg
60aaagaaatac gtaatgaccg ctcaagtgtg gccaattccg ctccgggcgt acgctggcgc
120actaaattca gggtcggcgt aagaatcgag cgagggcgga cggagcgttg caggaacggg
180cgaaagcaac ccgtagatca ctgctggaag cggctgccca gctcttcgcc gaacagggat
240acgcggccac cagcgtcaac gacatgagcg cccggtcggg ccggaccagc ggcgccgtct
300acttccacta cgccggcaag gaggccgtcg ccgtcgccgt cgtccaggac cggttcgcca
360cctggccaca gctcgccgca cgctacgcgg acgaggcggt ccccccggtc gaccggctcg
420tcgccctcag ctacgacatc gcccacgctc tcgccgagga cccggtgacg cgtgccggcg
480cccgcctgtg ggccgaacgc gccaccatca acgttcccct tccccacccc ttcgcgctgt
540ggaccaccgc cgccacacga ctgctcgcga aggcccggct cgccggccac ctccacccgc
600atgtccgccc cgctcgcgcg gcccggaccc tggtccctgc cttcttcggc ctgtgcgcgc
660tcaccgagga actcgaaggc acggccgccc tcaccgaccg cctgaccgac tggtggcagc
720tgacgctgcc ctgcctccgc ccgcacccgg tgcgcgagga cctgccgcgc gggcggtgac
780gcgcggcagg tcccgtagct ccggccgctg tcggagcacg ggcggcgcgt tacggggacc
840gcgccgggta ggccccgtgg ctcaccgctc gcggacaccc tgaacgggga gatgaggggc
900gcggcttccg ccgtgacgcc gcggtccggg aagccgtggt gttcgaggaa gcccgcgatt
960gtcgagcccg cgcggcagcg ccggcccctc ggccggacag cgcgtcgcac cgcccgtaca
1020gcggccacca gcaggtccgg tgggtgcgga taccgggtgg tgcggccgcg cacgcggtgg
1080gtgcggacgt cgggtggtgc ggcggcgtac gccgtgggga cgggacgggg ggcgtcgcgc
1140atgcggtggc tgcggatgcc gggcggtgcc gccgtgcatg ccgcgggtac ggacgctggg
1200tggtgcggcg gcgcacgccg tgggggtggg gcgggggtgc catgggtgcg gacgccgggc
1260gctgcggcgg cgtacgccgt gggccggggc ggaggtgccg tcacgcatgc cgtggctgcg
1320gacgtcgggt ggtgccgccg tgcatgccgt gggtgcggac gtcggcggcg gcgtacgccg
1380tgggccggga ccgggggctg tcacgcatgc cgtggctgcg gacgtccggt ggtgccgccc
1440tgcatgccgt gaccgcggac gccgggcagt gccgtcgcgc accccatggg gccggggcgg
1500gtggggcctg ccggtatccg ggtgggggcg gtaccgccca cgccgaccgc gccgcccgcc
1560cggcaccccc gcccgcggct cagggacgcg cctccaccgg ttcgaggtgc ggttccaggg
1620cgccggcgac cttgtggcac agctccacga acatcgcgcg ctcacccggg ctcagtgcgg
1680cctccacgat gcgggcgttc tcccgccgtt gctcggcgac gcgggtgagg aaggcctcgc
1740cggccggtgt gagcgaggtg gtgagttcac gccggtcctc gccgagcgtc cgctggtcga
1800cgagctccag gtccttgagg acgcgcagtg ctttggacgt ggtcgcccgc gacacaccga
1860gcgaggcact gagcaccgag ggagtcatgg ggccgcggat gcgcagcagc tccaggacgt
1920cgtactgctg ccaggtgacg ccttccggat tcgagcgagt gcggcgggcg acgaggatgc
1980actgcaagtg ggaaagcgcg tcctcgagtt cgcctggcac agccgaggtc tcggaggcgc
2040ttcgtgggct gtcggttcgg gtcatcaacg ccctcaggtg atcaggtcgg ggacgaggcg
2100ccacggccgg ctcggggcgg gagcgggccg acggcgacct ccgattggtt acccattatc
2160aacgatcctg gggcggcaac gtatacctct ccacacggtc atgtgatcca tcgcccaaca
2220gttgtttcca tcgggaaaat ttctacgcga tgatctctga tgagtggccc tccaggccgg
2280acacgcgccg cgcctcgctc cggcttttcc cggcccgcga gcgggaggcg cccctcgggc
2340gggcccccgt ggacggcagg gcgccgtcgg ccgggcgctc tggtttgctc cggtcgaaca
2400atggctgatg catcgtcagt gactcggttg ctttcctggc ggtaaacaag tggctgaaaa
2460cgcgccctgc gacgctgcgc tccaccgtgt cggcctgggg ttgtccgtgg cggcgaaggg
2520gcctccggtc ggggcccgac agcccaggca tgggagcgaa gagcgggggt agtggcagct
2580ccagcgtggg tggcgcagcg gtctcggcgc ctcaagatcg gggttgacac gtcgaagagc
2640gccgacctaa cgtgatttcc gaacagcaac catcaccttt cggttgattt gtgcgtattc
2700cttgagaagc cggtcactgg acgtcggatg tgctcgtcgc tgtgggcgcc ccgtggccat
2760ccgccagcga ggcaaggagc acgtgtgctc acggagaatg catcaggcga ggcgcgttcc
2820gctgttcccc tgaccctcac cgagggtttc gaccgcgtgg tgcgggccgc cggccaccag
2880gtcgccctcg tctccggcac ggagaccgtg acctaccggc aactgaacga acgagccgag
2940cgcgtggccc gcgggctggg cgcccgcaag gtggcgcccg gtgaccgggt cggcgtgtat
3000ctgcgccggt ctcccgacct ctacgcggtc atgctcggcg tcctcaaggc gggcgcctgt
3060gtggtgccgg tcaacccgga ccaccctgcg ccgttcgtct cccgcgtggt ggccgaatca
3120gcgccgcggg ccgtcgtgca cgacgcggga acaccggccg tcgcgcccgc tgctccgggc
3180gcaccgctgt gggtaccggt cgaggagctc accacggccg cggaaccgga cgacggcgtc
3240gcgctgcccg ccgtgaacga tccggacagc accgcgttcc tgatgttcac ctccggatcc
3300accggccggc cgaagggcgt ccgcatcgcc caccgcgggc tggcacgact cggcccgtac
3360agcggggaac tgcgcatggg cccgcaggac tgcctggtcc agtccgcggc gttctccttc
3420gccgcgtcca ccatcgagat ctggctcgcc ctcctgcacg gcgcacgact cgtcgtgatg
3480ccccaggggc tgcccagtct cccggccctc aaggacgccg tcgtccggca cggcgtcacc
3540gcgctgtccc tgccctgcgg tctgttcaac ctcctggtgg acgaggagcc ggaatgcctg
3600cggggcctga gggtgatcct cctcagcggc gacttcccgt cgccggagca cctcagccgt
3660gcggcacggg cgacgcgcgc ggtcatctac aacggctacg gctgcaccga gaactcctcc
3720atcaccgccc tgtaccccat ccgcgatgcc ggggacgtca cccgcgagaa ccgggtgccg
3780gtgggccgcc ccctgcccgg ggtcaccctg gaggtactcg acgactcgct gcggccctgc
3840ccgcccggaa cgcccggaca gctcgtcgtc ggcgggctcg gactcgccca gggctatctg
3900aacgacccgg aactcacgaa ccgcaagttc gtcaccgggc cggacggacg gccgcgctac
3960ctgaccggcg atctggcccg cgccaccgag gacggtgaca tcgtcctcat aggccgcgcc
4020gacagcatgg tcaagatccg cggctaccgc gtcgaactga ccgcggtgac cctcgccctg
4080cgcgccctcg acgggatcgg cgacgccgtc gtcaaggcgt tcccggaggg cgccggggag
4140aagtcgctca ccgccttcta caccaccgtc gacggacgac cgctggacgg tgccgacctg
4200gcacgccgca tgggagacca actgccctcc tacatggtcc cctccacgtt ccaccacctc
4260ggcgacctgc cgagaaacgc caacggaaag atcgaccggt ccgccctcac ggacccgtcg
4320gacaccaacc gcgatccgaa gaaaggtcac acagccgtgc agaacccgct cgagaccgtc
4380gtactccagg cgtggaagga catctccggc gccgacgact tcaccaccac cgactccttc
4440ctcggccacg gcgggaactc cctgcacttc gtccagctcg cctccaggct gcagaagatc
4500ttcggtgtgg aggtcagcac cgaggacgtc ttccggcacg gcacggtgga gcagctggcg
4560cgcttcgtcg agcagtcgcg ggacaccgga cgcaaccccg ccgcacagac ccagtaggcg
4620tcacccggcc gtggccgtgc ggcgcccgtc gccggcggcc ggggcatctc tgcagaggac
4680tacgaccggt gaactccccc ctccgaacca ccgtgctcga ccttgcacgg accaccctcg
4740gcagcgccga cctcaccgcg cacgaaccgt tggccgaccg gtgcgaacac ccggccctgc
4800tcgacgacct cgccaccacg ctgaccgccg tcttcgcggt cgagatcacc ggcgcggacc
4860tggcggccgg tgccaccgtc gccgacgtgg ccgcgcgaat ggacgaccgg cgcgacgccc
4920cccggatccc ggaactgcgc gccgggctcg ctccccgcga cggccgggcg gtggaggcgt
4980ccttcgggca gagcggcatc tggctgatcg accagtacct gcccaacccg gccgcctaca
5040acggcccctt cttcgtccgg ctgccgttct cagccgatcc cgaccgcctg cacgcggccg
5100tgcgcggagt gctgcgccgc caggaggtcc tgcgcaccac ctacgccctg agcgacggca
5160cgctccggca gaacgtctcg cgggacgatg acgcggtcgt cttcgaggta gcccgctacg
5220gcgacgacaa ggaactcgac gccctcgtcc accgggtggc caatctccgc ctcgacctgg
5280cccgcgggcc ggtcatcgcc gtgacctgcg cgctcggccc cgcgaaccgg tccgccgtca
5340tctgcaacat ccaccacatc gcctccgacg ccgcctccgc cggtgtcttc ctgcgggaac
5400tcctcgacgc ctacgaccgc ctcggccgcg gtctgcccgt cgaggccgac ccgctgcggc
5460ccacctacgg ggacttcagc cagtggtacc gggaactgat gaaccccgag gccctcaccc
5520gctccctcga ccacttcgcc gcccggctcg ccggggaact cccggtgctc gacctgccca
5580ccgaccggcc ccgcccgccg gtgaagcaac accggggcgg caccctcccg ttgcacctgc
5640cggccgccgc ggccgacgac ttcgaggcgc tcgcccggac cgagggggtg accctgttca
5700tggccctcgt cgccgcgtac gcggtcttcc tctcccgcca caccggtcag cggcgcgtgc
5760tgatcggcag ccccgtctcg ctccgcgacg acccggccac ccacgaactg atcggctact
5820tcgtcaacct ggtcgtcctt cagcaggaga tcgacgaccg gatgaccgtc cgggacgtgc
5880tccgccgggc gcgggaggag gtgagcgagg cgctgcggca caagtgggcg cccttcgaca
5940aggttgtcga gcgtctgcag ccaccgcgca gcagcggcta caccccgctc gtgcagacca
6000tgctcgtgct cacccagggc gacgccggac ggatatccca cgacgacacg gaactgcgca
6060tcgagcgcgg ggccgcgcac ggcgccaagt acgacctgtc cctcgttttc gagcgggact
6120ccgaaggcct gcacggtctg atcgagtacg acgcggacct cttcgacgag ccgacggtac
6180gggccatggg cgaccggctg cggcacctga tggagcagtt cgcccgacgt cccgacgcac
6240ccctgcacga actggaggcg ctcggtgcgc aggagcggcg gtcggtgctg gtccgcgggg
6300accggaccgc gcacgccgtg cacgacgcac ccgtcatgga actgttcgag gcccaggccc
6360gggcgacccc cgacgcggtg gcgctggagg acggcgacac caccctgtcc taccgcgaac
6420tcgacgagcg cgccaaccgg ctcgcccacg tgctgcgcgc ctccggcgct gcggccggca
6480cccgggtcgg gatctgcctg ccccgctccc acgacatggt cgtcgccctg ttcgccatcc
6540tgaagaccgg ggcggcgtac gtaccgctcg acccgtccta ccccaggcag cggatcaccc
6600acacgctgcg cgacgccggg gtcttcctga ccgtgacgga cagctcactg gccgacgaac
6660tccccccgag ggagccgctg ttcgtgctgg accggcacga cggaccgatc gccgcggccc
6720ccgccaccgg cctcggccgg gtgaagacac ccgacgacga gatctacgtc gtgcacacct
6780cgggctccac cggcctgccc aagggggtgg tcatcgccga ccggaccgtc gccaacctcg
6840tccgggccca gcaccgttgc tcgccggccg gagcgaccgg gcggacgctc cagtacatgt
6900cgctgtcgtt cgacgtgtcc gtgatggaga tcctcggcac cctgtgcgtc ggcggcaccc
6960tcgtgctggt ctccgaggaa ctgcgcaagg atctgcacgc gctcgccgga ttcctcgccg
7020aacgccgcgt cacccgggtg tacctgccct acatcgcgct ccagcagctg gcctccctgg
7080ccaccgacgc cggtgtgcgc ctggacgacc tgcgcgagat cacctccgtc ggcgaggccc
7140tcgtggtctc cccgcagatc cgggagttcg ccacccgtca cccggcggtc cggctggtga
7200acatgtacgg gccgtcggaa acgcacctgg ccagctggta cccgctcacc ggctcgcccg
7260cgacctggcc cgacaggccg ccgatcggcc gcccggtgga cggcgtgcgg ctggtggtcc
7320tggacgccca catgcggctc gtccccccgg gtgtccccgg cgagctgtac atcggagggc
7380ccgtgctgtc ccccggatac cgcaaccgtc cggacgagac ggcccgccgg ttcctcccgg
7440accccttcgg cggccccgcc gaccggctct accgcaccgg cgacctggtg cgctggaaca
7500gcgagggcga cctggagtac ctgggccgga ccgacgacca gatcaagatc cgtggctatc
7560ggatcgagcc cgccgagatc gaggccgcac tcgacgacct ggacggcgtc gcctcctccg
7620cggtcgccgc cgtggacgtc gcccccggcg accgcagact cgtggccgtc ctggagacct
7680cccgcacctg ggagaccgcg gagctgcgcc gcgccctgtc cggcacgctg cccgactaca
7740tggtgcccgc gctggtggtc gcggtggagc acatgccgac gaccccgagc gggaagatcg
7800accgccgggc cgtcgccggc ctggccgcgg cacaggcgac cgcggcacgg accgcgcccg
7860cgccacccgg ccggccgccc aggccgggcc tggagcagcg gatcgcgcgg gagtgggcgg
7920atgtgctgaa ggtgcccgcg gtgggcaggg acgaggactt cttctccgtc ggagggaact
7980cgatcatcgc cacggaactg gtctatcggc tgcgccgggc gttcgaccag gacctctcgc
8040tgcgcgccct gctggagaat ccgacggtcg cgggcatggc cgcccggctg cgttccggcc
8100ccggcgctcc caccaccgcc cccgccgcgc tgcgggagga cgcgacgctt cccgacgacc
8160tgcccgccgt caccggcacc ccggtaccgg tcgcccgggc ccgtgaggtc ctgctcaccg
8220gcgcgaccgg gttcctcggc agctacctgc tgcgggagct gaccggaacc accggcggcc
8280gggtgcactg tctggtgcgg gcggcggacg aacgggccgg catggagcgg ctgcgggcca
8340ccgccgagcg ctaccggctg gacgggcgga tcgactggaa ccgggtgcgc gccgtgcccg
8400gcgacctgag ccggcccggg ttcggtctgc ccggtgcgga gtacgacgcg ctggccggca
8460ccgtcgacgt cgtctaccac gcggccgcgc acatcaactt cgtgctgccg tacgcctcgg
8520tgaaaccgac gaacgtggac ggcttccgcc acgtggtccg tttcgccgcg acggaccgcc
8580ccaagcacgt gcagtacatg tccaccatcg ccgtgttccc tccgggcgag gcgcccgacg
8640gcacggtcct caccgaggac gacgtgcccg aggcgtgcga acgactgggc atcggctaca
8700cccagagcaa gtgggtcgcc gagcgcatcg cactcgcggc ccgcgcgcac ggcgtgccgg
8760tcaccatcca ccgcatcggg cgcatctcgg gcgacagcgt cacaggcgcc tgccagagcg
8820acgacttcct gtggcggcag atcaagagct tcatcgaact cggctcggcc ccgccggccg
8880aggacctcac caccgatctg ctgcccgtcg atttcgtcgc ccgcgccgtc gtcgccctct
8940cccgccaccc cgccacccac aaccgcaccc tgcacgtctt ccacccgagc ggatcggact
9000tcaccccggt ccacgcggcc ctgcgcgcgg acggccaccg cctggagatc gtcccggccg
9060acacctggct tgcccggctg gaggagtccg cacggcggcc cggcggcaac gccctggcgg
9120cagccgtgcc cctcttccgc gagggcgccc tggaactggg cgacaacacc tacggcaaca
9180ccgccaccac ccgcctgctg atggacctcg gactgccctg gcccgccatc gacgagcagg
9240cgatcacgcg gatgctccgc tacttccgct ccgtcggaga actggccgac gactgagggg
9300acttccctgt cccgggccct cccggcagcg cggcgcagcc gccggcgccg atccgaccat
9360ccaccacacg gcaatcgccg aagcggtcgc cggacaccga aagcaccagc agccatgcca
9420acgagctcct gccccgacac cgcatacgac accctcatac cctccgtcgt cgcggctctc
9480ccggccgcac agcagccgga gtggccggac cccgggcgac tcgccctcgt ccacaccgaa
9540ctggcccgcg cggacccgct ggtgacgtac gacagcgtgc gtgccctgcg ccggctgctg
9600tcccgtgccg ccgaaggcga actgtgcgtc ctccaggccg gcgactgcgc ggaggacccc
9660gccgagtgcg gcccggcccc gctggcccgc aaggccgaga tgctggacgt cctcagcgac
9720atcgtgcgaa cgggcgccgg acggccggtc gtccgggtgg gccgtgtcgc cgggcagtac
9780gccaaacccc gctcccaccc ggaggagctg cacgacggcg tccggctgcc ggtctaccgc
9840ggtcccatgg tgaacgcccc ccaccccgac gccgacgcgc gccggcccga tcccgcccgc
9900atcctgagct gctaccgggc cgcccgccgg gccgtggagt ccctggaccg gctgggccgc
9960ggcgagggtt cgcccgccga gacccgggtg tggaccagcc acgaggcact gctgctcgac
10020tacgagctgc ccctcgtgcg ccggcaccgc tcgggccgca gttacctcgc cagcacccac
10080tggccgtggg tcggcgaacg gacccggcaa ccggacggtg cccacgtacg gctgctggcg
10140gaagtggaca acccggtggc gtgcaaggtc ggcccgacca cgaccgtgga gcaggtgctc
10200gccctgtgca ccgcgctgga cccggagcgc tcaccgggcc ggctgtcgct ggtcgcccgg
10260ttcggcgcct cccgcatcga cggcctggcc cccctggtcc gcgcggtacg gcgggccggt
10320cacccggtgc tgtggctgtg cgacccgatg cacggcaacg gtgaacgcac cgcgcacgga
10380ctgaagacgc gccggctcag cgccgtgatg gcggagatca gccggttcgt ggacatcgtc
10440tccgccgagg gcggccgcag cgccggcctg cacctggagg cctcaccgga cgacatcgcc
10500gagtgcaccg gcgccggatt caccccggcc cccgggccgg cctaccgcac cctgtgcgat
10560ccccgcctga acctggtgca agccgtcgcg gcgaccgcct actggcggct gccggccctg
10620gaggccgtcg catgagcgag tctgcgcgga acgcgcgcgg cctggcggcc ctgctgcccc
10680cgccgggcac ccccttcgcc gtcctgcacc ggccgggtgc cgggcaccct ggcaccgtgg
10740acgtcgtcag cgggcccctc cgcaccgccg ccaccctcgc cgagctgagc ctcgacgacg
10800agtccgcgcc ggcctcccag ggccccggac cggcgcacag ggtcctcgcc ctggtgcccc
10860accggcagat cgccgaacga ggcttcgcgg ctcccgacga cggcaccccg ttgctggcca
10920tggacatcgg cacccagcac accgtgcccc tggagcggat gctggcactc ctgcccgacc
10980gcgaactgca cgtcgaggaa accgggttcg acctcgacga cgaccggtac gcggccggcg
11040tcgacgcgct cacccgccag gagatccagc gagggcaggg cgcgaacttc gtcctcgccc
11100gcagcctgca cggccggatc cgcgacttcg accggacccg ggccctggcc gcgctgcggc
11160ggctgctgat cgcggagagc ggcgcctact ggacctacct ggtctgcacc ggcgaccggt
11220acctcatcgg cagttccccc gagcagcacg tacgggtggc cggctcgcgg gtgtcgatga
11280acccgatcag cggcacctac cgctaccccg aggggggccg cccggaccgc gaaagcctcc
11340tccggttcct cgccgacccc aaggagatcc acgagctgta catggtcgtg gacgaggaac
11400tgaagatgat gaccgaactg tgcggttccc gcgtgcgcgt gtcgggcccc acgctcgcgt
11460ggatgtcgcg tctcgcccac acgcagtacc acctgcacgg cgagtccccg ctgcccctga
11520ccgacatcct gcgcgggaca ctgcccgcgc ccacggtgac gggcagcccg gtggagaacg
11580cctgccgggt catcgcccgc cacgaaccgg caggccgggg ctactacagc ggcgtgctgg
11640cgctggccgg ccaggagggg ggacggcgcg ccctggacgc ggtcatcgtg ctgcgcaccg
11700ccgacatcac cgcggacgga tccgtgcggc tgaccaccgg cgcgaccgtg gtgcgtgact
11760cggtgccccg cgaggaggcg gcggagacca cggcgaaggc cgcggggctc ctcaccgcgc
11820tgacccgcgg cccggccggc cggtccgcgg ccccggcgca cgcggcaccc gacgtctccc
11880tgggagccga tccggcggtg cgcgcggcgc tgcgctcccg caacgacggc atcgccgcct
11940tctggctcgg cggcggggcg cgcctgccgg cgccctcgcc acacggacca cgggtggcgg
12000tgatcgacgc ggaggaccgg ttcaccagca tgctcgcgca gcagctccgc gcggtgggct
12060gccacgtcac cctgcacccc tggtggtcgg ttccggaggc agccgacgac cccggcaccg
12120tgctgctgct ggggccggga cccggggacc cccgggacgt cggcgacccg cgggtggcgc
12180ggctgcgctc cctggccggc cgccggctcg cccggcggtt gccgctggcc gcggtgtgcc
12240tcggtcacca ggccgtctgc ggggtgctgg gccttcccct ggtccggctc gcgcggcccc
12300gccagggcgc ccggatgcgg gtcggcctgt ggggacgcga ccggcacgtc ggcttctaca
12360acagcttcac cgcgcgctcc gacaccgatc gctgtccgct gcccggccgg gacgccacgg
12420cccgggtgtg gcgccgggac gggggagacg tggtcgcgct ggacggcccg gggctggcca
12480ccgtccagtt ccacgccgag tcgctgctca ccgaggacgg cccggacatc ctgcgcgagc
12540tggtggaccg ggcggcccgt accgagcggc gcaccgaggc cctgatgtcc cgccgagcca
12600aggagcacgc gtgaacccgc ccggaaccgt cgtcgccaac gccgccctgg acccccggga
12660gctgcgccgg accatggggc acttcgcgac cggcgtcacc gtgctgacct gccggcgcgg
12720cgcccggctg cacggggcga cggtgaactc cttcacctcg gtgtcgctcg atccgccgct
12780cgccctggtc gccctggacc gtcgcacccg cgccgccgcc ctcctggacg acggcccctt
12840cgtcgtcaat ctgctcggcg agcaccagca ggacctggcc ctgcactttg ccggcggctc
12900gccggccgat tccgtgccgt gggtggacgg cgacggcgac cggccccggc tggcgggaac
12960cctcgggcac ctggtgtgcc gaccctggcg cacctacgac ggaggcgacc acacgctgca
13020tgtcggccgt gtcgaggagt tcgccgccgg agggggacgg ccgctgctct tctaccgagg
13080cgtcttcccc cgcctcatgc cggacggagg aggagacccg gagggacccg aggaggtgtg
13140gtcgctctgt ctggacggcc caggaccggc cacggatcag ttcgtcaccg atcatgagac
13200acggaagtag ggacatggca cccgacaacg gacagtccgc agcacccggc acctccgggg
13260cgtccaccgg caaggcccgg gtcacccggc cgctgaccgg ggacgagtac atcgagagca
13320tccgcgacgg acgggagatc tgggcgtacg gcgagaaggt cgacgacgtc accaagcacc
13380cggcgttccg caacaccgtg cggatgacgg cccgcttgta cgacgccctg cacgatcccg
13440agcaccacga caccctgacc gcgcccaccg acaccggcag cgacggcttc acccacaagt
13500tctaccgggt gccgcgcagc gtgcaggacc tcgtggggga cagggacgcc atcgccgact
13560gggccaggct gacctacggc tggatgggac gcagccccga ctacaaggcc agcttcctgg
13620tgaccctcgg cgcgaacccc gactactacg gcgacttcgc ggacaacgcc cgccggtggt
13680acgccaccgc ccaggagaac gtgctgttct ggaaccacgc ggtgatcaac cctccggtcg
13740accggcaccg gcccgccgac gaggtggacg acgtcttcgt gcacgtggag aaggagtgcg
13800acgacgggct ggtggtgagc ggggcgaagg tggtggcgac cgggtccgcg ctcacccact
13860tcaacttcgt ggcgcactac ggactgcccg tgaagaagaa ggagttcgcc ctcgtcgcca
13920ccctgcccct ggcggcaccc ggcgtgaagc tcatctgccg ccagtcatac gaactggccg
13980cgagccgcac gggcagcccg ttcgactacc cgctgtcgag ccggctcgac gagaacgaca
14040ccatcttcat cctggacaag gtgaagatcc cctgggagaa cgtcctcatc tacggggaca
14100ccgccagggc cggcaccttc ctgcagacct ccggcttcac ccaccggctc accttccacg
14160gggtgacccg gctggccgtg aaactggact tcctggcggg cctgctgctg aagggcgtgg
14220aggtcaccgg caccaaggac ttccggggca tccagacccg ggtcggcgag gtcctcgcct
14280ggcgcaacat gttctgggca ctgagcgacg cgatggcgca caaccccgat ccgtggcacg
14340acggagccct gctgcccaac ctcgactacg gcatggccta ccggtggttc atgaccgtcg
14400gctacccgag ggtccgggag atcatcatgc aggacctcag cagcgggctg atctacctca
14460cgtcgcacgc caaggacttc aacgaacctg aactccgtcc ccatctcgac cgcttcatgc
14520ggggttccaa cggttacgag gcggtggagc gcgccaagct gatgaagctc atctgggact
14580cggtgggcac cgagttcgcg ggccggcacg agctgtacga gcggaactac tccggcaacc
14640acgagagcgt gcggatcgaa ctgctgcacg cccagacggc ttccggtctc gtcgaccagt
14700accggggctt cgccgaacag tgcatggcgg aatacgacct ggacggctgg acggcaccgg
14760acctggtgcc gcccgacgtc gactgagcgc cgcggccatc cggccggacg ggcccgccgg
14820gcggtacggc aggggcggga ggagcccccc ccgcccctgc cgctcagccg gtgaccaggg
14880cgagcgtgaa accgtcgtac cccttggtgc cgacggtctg gatcgaggtg gcggtgacgt
14940ccgagcggcc ggccagcatc tcgtggaacc ggcgcacgcc ctggacgccg gcgtcgggat
15000ggtccgggtc ggtcaccgcg ccgcccagga cgacgttgtc gacgaccacc accgcgcccg
15060gccgggacag tttcagcgcc caggtgaagt actcggggat gtcgggcttg ttggcgtcga
15120cgaacaccat gtcgaagggg gcggtaccgg gccggtcgag ggtcggcagg atgtccagcg
15180cccggccgac gtgctgttcg acgaggtggg cgacccccgc ctcggcgagg cgcgacgcgg
15240ccgactcggc gaaggaccgc tcccactcga tggtgacgag gcggccgtcc ggcggcagcg
15300cgcgggccag ccagatgctg ctgtacccgc cgaacgtgcc gatctccagg atgcgccgcg
15360cctgccgcag acgcgccagc agatgaagga gtttgccctg gggggcgctg acggcgaggt
15420ccggcaggtc gaattcccta tgggcctgag cggccttcga aagtgcttcg tcctccttca
15480cgagaaggga gctgaagtag acgtcgacac tgttccatcg ctcttgttcc atggttgcaa
15540taattcacga taaaccctcg gatggcaaga gcagaattcg atatccatgg tgccgtgaac
15600ctgagaatgc ccaggtcagc ggcctattca aggctgtcca gtcaccttcg ccaaagaaat
15660taacgggcgg tcgataatgt tcggaaattc cttcggtgcg gcgttgactg tcgtccttgc
15720ggtcacctag cctccctttc tggaaccttg tgaaaaagcc tgtcccggat aggagtgtca
15780tttcatgcga gaagactcgg ccgtcacaac ggccgcaccc ccggtgcacc tggtgccggc
15840gatgcaccac ctgggcgtcc aaacccgcga cctggacaac tccctggcgt ggtacaagga
15900cttcttcggc tgcgccgaga cctggacgct caccacgttc tcggacctga cccgcagcag
15960gctccccggc atcacccggc tcaccgagat cagcgtcgcc gacgtccgct tccacctctt
16020cgaacgcgcc gggcacgacc cggccctgcc cggcggcaac aaggcccagt tccagcacgt
16080ctgcctcgcc accggttccc cggaggaact gcgcgcctgg cgcgatcgct ggatcgagct
16140ctaccgctcg ggccgctacg acttcgcgac cgatgagcag cccacggaca tcgtggtcga
16200cgccgacgga gtgcacagct gctacctgtt cgaccccaac ggcctcgagt tcgagttcac
16260ctacgttccg ggcggtgcgg catgagcgcg ggcccgcacc ggaccgtcac cgaactgccg
16320gtcgccgaag gctgggactt cggggacttc ccctacggcc tggagccgct gaccctgccc
16380gagcccccgc acgagcccgc ggccgacgtt ccggacgtgc tgtgcgccga gcccgccccc
16440ggcggtgcgc ggacgtcctg cccgcgcacc ggaccggcgc ccggcctccc ggagctggcc
16500caccagctct tctggttccg ctggatcacc ggacaccagc tgaccttcgc catctggcag
16560ttactcggcc acgcgctgca ccaggcgcac gcccggcccg accccggccc gtcgctgcga
16620gccatgacgg acctgacacg ggcgtacacc gcgatgctgc tctacaccgg ctcctgtccc
16680aaggacgtct acagcgacgt gatccggccc agcatgttcc tgcagcaccg cggcttcagc
16740gggacctggg cgccggactt cgtccccgtc cgccggctgc tgcggggcag gaagacgccg
16800tggcacgaga ccccggaggg cggccggctg gccgacgagg tccgtctcta ccacctggtg
16860cactcggggg tcgccgcgaa actcgtaccc ggcggcaggt ccctgctcca ggacaccgcc
16920cccacggccc ggccgcacga cccccggatg caggcgctgg tctacgacaa ctacttcctc
16980accctgcgcg ccgacgtccc gaccgccgag gtcgtcgagc agctccggcg ccgactggcc
17040gcggtgcgcc tggacgtctc ggtcaacggg ctgtaccccg ggctgaccgc gcaggaggac
17100gccgcactgc ccgaggagtt gcgcagcgag gacacacagg cctgcgagcg ggacttcgac
17160gccgtcctgc ggcgcgtcga cggccttgcc gccgcactcg accggcggtt gctcgacggc
17220acgatcgccc gctgagcctg agccacccga tcacgcgaca ggaaaggagc ggtggaccgt
17280gcggtacgga gtcgtcgtcc tgcccgaacg ccggtgggcg caggcccgcg aacagtgggt
17340ccgtgccgag gagttcggat tcgaccacgc ctggacctat gaccagctga tgtggcgttg
17400gctgcgcgac gagccctggt tcggcgccgt gcccaccctg gcggcggcgg ccgaggccac
17460ctcgaccctg accgtgggca ccatggtggc cacacccacc tatcggcacc cggtgacgct
17520ggccaaggag gtgatgaccc tcgaggacat cgcgggcggc cggttcgtct gcgggctggg
17580agccggggcc ggcggcctcg acgaccgcgt cgtcgatccg gccgcctact ccccacggca
17640acgcgccgac cgcttcacgg agttcgtcga cctgctcgac aagctgctga gccgccgcag
17700caccacacac accggcacct actacgacgt ccgggaggtg cccgtgcacc cgggctgcct
17760ggccacgccc cgggtgccgt tcgccatcgc ggcgaccggg ccgcgcggca tgcggctggc
17820ggcccgccac gccgacatgt ggatcaccgc ggggcggccc ggcgacttcg acgcccttcc
17880gtacgaggag accctgccgg tgatcaagga gcagctggcg cgcctcgacg aggcgtgcga
17940gcggaccggg cgggatcccg ccaccctgcg ccggctgctg ctgaccggcg ccatggtggg
18000cggcaccctg gactccgtcg aggcgtaccg cgacgccgcc ggccgcttcg gcgaactcgg
18060catcaccgac ttcgtcgtcc actggccccg gccctccttc ccctaccagg gcagggtgga
18120agtgctggag cagatcgcgc gggacgtgct gaccgtccgg ggcggggagc ggccgtgatc
18180gcctacgaga tcgtcgacat gttcaccggc acgcccttcc agggctgcgc gctcggggtg
18240gtcccggacg cgaccgcact cgacgacgac ggcatgcggg cggtggcccg cgagatcggc
18300ctcaccgaga cggcgttcgt cctgccaccc gagtcgcccg acgccaccca ccgggtacgg
18360gtcttcaccc cggagcggga gtcaccgtac ggcgggcact ccgccatcgg cacggccacc
18420accctggtgc ggctgggccg tctgcgcgcg ggggagctgg tgcaggagtg cgggggccgc
18480ctgatgaccg tgcgcgccag tgcccgacgg gccacgctcg gcgtccgggg ggagcccgtg
18540ccacccggcg cctgggatcc cgtgccgttg ctggaggcgt gcggcctcac cgaggacgac
18600ctggtcgccg ggccccgcgt gaccgggttc ggaccggcct tccacgtgct gccggtcgga
18660cccgaggcgg tcgcccgtgc cgcacacgac ccggcgcacc ccgtgtggtc cacctgcccg
18720gacgcggtgg tggtcgccta cgacaggcgc ggacacctgg ccgacgtcag ggtcttcgcc
18780cccggctacg gcatgccgga ggacccggcg tgcgcctccg ccgccctggc actgggcgcc
18840tggctcaccg gcgcgggcct ggtgccggcg acggacggta cccgccttta ccgggtccgg
18900caggggcacg ggctgggccg ccccgcccgg ctcgactgcg ccgtgaccgt acgcgacggc
18960cgagcggtcg cagccgaggt gaccggggag gtggcggcca ccgccgccgg ccggatgcac
19020ctgccccgca cggcggccgt cgcgcgctga gccggggccc gggccgtgta ccaggccgag
19080ccgcggaagg cacccggtcc ggtcgacgca cacccagagc acacgacgcg aatccctgta
19140tcgcagaacg aagaggagag gaacccgact gtgttgttcc gtccagagct gcgcggcacc
19200cggggcgcgg tcgcctcgac ccactggctg gcctcggccg cgggcttccg catgtacgac
19260aagggcggca acgcgttcga cgcggccgtc gccgccgcgt tcgtcatcca ggtcgtggag
19320ccccacctca acgggcccgg gggagacgtg cccgtcctcg tccaccgggc cgggagcggc
19380cgggtcgacg tcgtctgcgg ccagggcccc atgccccggg ccgcgaccat cgagaggttc
19440gaacagctcg gcctgtccgt ggtccccggc tccggcctgc tgcccgcggt ggtgcccggc
19500gccttcggcg cgtggctgcg ggtcctcgcc gagtacggca ccctgcgtct ggaggacgtc
19560ctggagccgg cgatcggcta cgccgaacgc ggctatccgc tgcttcccaa ggcggcggcg
19620atgatcgagg cgctccagga actcttccgc gacgagtgga ccgagtccgc ccgcacgtac
19680ctggtgggcg gggccgcgcc gcggcccggt cagcgcatga ccaaccccga cctggcccgg
19740acctaccggc gcgtcctcga cgaggccagg gcggcgggcg ccgatcgcga caagcagatc
19800gacgcggccc tgcgcgcctt ctacgagggc ttcgtggccg aggccatcga cggctatctc
19860gccaaggcgg aggagatcga tgccaccggc cgccgccacc gcggcctgct gaccggcgcc
19920gacctggcag gctggcgggc gacggtggag ccctcgctct ccttcgacca ccgcggcctc
19980accgtgcaca aggcgggacc ctggtcccag ggcccggtct tcctgcaaca gctcgcgctg
20040ctgcgggagt tcgacctcgc cggtatggga ccgcacagcg cggagttcgt gcacaccgtc
20100accgaggcgg cgaaactggc gttcgcggac cgcgaggcct ggtacggcga tcccgcgcac
20160gcggaggtgc cggtcggcga cctgctggac ccggcctaca ccgcggcccg ccgcgagctg
20220atcggcagcg aggcgtccac ggagctgcgg ccgggctcac cgggaggccg gacaccggtg
20280ctgccgcccg tccacgacga gtccgccggt ccggccggtc cctcctggct cggcgagctc
20340gaggagggca tcccggcggt ggtgcgctcc acggccgccc ggggcgacac ctgctgcgtc
20400accgccaccg acgcccacgg gaacatggtg gtcgcgacgc cgagcggcgg ctggctgaag
20460agttcgccgg tggtgcccgg tctgggcttc ccgctcggta cccgtgggca gatggccacg
20520ctcacccggg ggcacgccaa cgcgctggct cccggcaagc gcccccgcac caccctcagc
20580ccgaccctgg tgctccggga gggcaggccc gccctggcgt tcggcacacc gggcggcgac
20640cagcaggacc agtggacgct gcagttcttc ctgaggcaca ccgaacacgg catggggctt
20700caggaggccg tcgaggcacg gaccttccac accgaccacg tcccgacgtc cttcaccccc
20760cggcgtttcg ctcccgggac ggtgaccgtc gaaagcggca tgccggagga aaccatccag
20820gagctcaggc ggcgcggcca ccaggtccgc acggtcgccg actacagcct gagcaaggtg
20880tgtgtcaccg gcctggccag cgacgacatg gtcatcgcgg cggccagtcc gcgcggcgcg
20940caggcgtacg ccgtcgcgga ttgaggatgc cgaccggatg cgataagttt ccaaacggaa
21000atcgtctcgg cgaatggaag gggaacgcat ggtgccccac ccgtcactgg acccgggtga
21060ccacatcgtt ctgggagaag cacggcagaa caacctcaag ggcgtcagcc tgcgcatccc
21120caagggacgg ttgaccgtct tcaccggcgt ttcgggatcc ggcaagtcct ccctggtctt
21180cgggacgatc gccgtcgagt cgcagcggca gatgaacgag acctaccccg cgttcatccg
21240caaccgcctc cccaagttcg agcggcccga cgcggaggtc atcgagaacc tctccaccgc
21300catcgtgatc gaccagcgcc cggtcggcgg caacgcgcgc tccacggtcg gcaccatgac
21360cgagatccac gccatgctgc gggtgctgtt ctcccggcac ggcaggccca gcgcgggtcc
21420ctcacacatg tactccttca acgatccgcg cggcatgtgc ccggagtgcg agggactcgg
21480atccagggtg cggctggatt tgaaccgcct tctggacgag gacaagagcc tcaacgaggg
21540cgccatccgc ttccagccct tcgcggtggg caccttcccg tggcagctgt acgcggagtc
21600cgggctgttc gatcccgacc tgccgctgcg ggagttctcc gcggacgacc gcgaactgct
21660gctgcacggt tccgggttca aggtcgaccg ggccggccgg cacggcgtct acaagaacga
21720gtacgagggc atcgtgctgc gcttcacccg gcgctacctc aaggcgggcc tcgacaccct
21780caagccgaag gaacgggcgg cggtgcagga ggtcgtgacg gaggggccct gcgaggcctg
21840cggaggcgcc cggctgggac cggccgcgct cgcgtcgcgg atcgccgggg agaacatcgc
21900cgactactcc gccctggagg tcaccgatct gatcggccgc ctggagcgca acgacgcccc
21960accggtcaag ccggtggtcc aggcggcgct ggccgcactg cgcaggatcg aggccgtcgg
22020actcggctac ctcagcctcg accgccagac cgccacgctc tccggcggcg aggcgcagcg
22080gctgaagacg gtacgccacc tgggcagcag cctgaccggg ctgacgtaca tcttcgacga
22140gccgagcgtg ggcctgcacc cgcgtgacgt gcgccgtctg aacgagctgt tgctcgccct
22200gcgcgacaag ggcaacaccg tgctcgtggt ggagcacgac cgggacgtga tcgccatcgc
22260cgaccacgtc gtcgacatgg gcccgggcgc gggcagccag ggcggcgagg tggtctacga
22320gggatcgccg accgggttac ggggctcgga cagcccgacc ggacgcggcc tgcgttcggt
22380gccgggactg aagcgccgac tgcgcgcccc cgacggcagg ctgacggtcc gcggcgcgcg
22440gctgcacaac ctcaaggacg tcacggtcga cgtgcccacc ggtgtgctgg tggcgctgag
22500cggtgtcgcc ggctcgggca agagctccct cgcccgggag ctggcagcgc ggcacccgga
22560ggaaacggtc gtggtcgacc agtcctccat cgggatctcc tcccgatcca cccccgcgac
22620gtacaccgac atcatggaca ccgtccggcg gctgttcgcc cgcgcatccg gaaccgaccc
22680cggcctgttc agcttcaact ccgcgggcgc ctgcccggag tgccagggcc gcggtgtgat
22740cgagaccgac ctcgcgttca tggacccggt caccaccgtc tgcgagcgct gcgaggggcg
22800ccgcttcaac gacgaggcgc tgagccacac cctgtccggc cggaacatcg ccgacgtcct
22860cgccatgacg gccgaggagg cgatcgggtt cttcgcggag gactccgtcc gccgcaaact
22920ggccctgctg acggaggtcg gcctcggcta cctgacgctt ggccgctccc tgtccaccct
22980gtccggcggc gaacgccaac ggctgaagct ggcgcaccgg ctgcacgcct ccggcagcgt
23040ctacatcttc gacgaaccgt ccaccggcct gcacatgacg gacgtgggca agctgctcac
23100cctgttcgac cgcctcgtcg acggcggcaa cacggtggtg gtcatcgaac acgacctcga
23160cgtgctcaag tacgcggact ggatcatcga tctcggcccg gaggccggcc ggcacggcgg
23220ccgggtggtc ttcgagggca ccccggcgga cctggcgcgg gtgcgggaat cgcacaccgg
23280ccggtgtctg gccgaggacc tcgccgcaca cggtcacctc tgacggcccg gagcaccggc
23340tcgccgccgg gtccgggccg gggtcgcacc ccgtcctccc gcacacctcc cgtccgacaa
23400ggagtccgta tgcccctcat ccacgtcacc ctgctgagcg gtcgcggcga ggaggagatc
23460gccgccctcg gccgggccgt cacggaggcc gtacacacca cgctgggcac cccccgggag
23520gcgatccggg tgacggtgga cgcatgcccg cccgagcact ggttcgtggg cggcgtctcg
23580atggcggaga agaaggcggc ccggggcggc tgagcggtgc tccgttcgcc ccgggcgccg
23640cggttcaggc cgcgtccgcg aaccgccggc ggatccgccc gacgtgctcc gggtccacgg
23700ccgggaggtc gtcgcgggtg agccggccgc tgcgcgtcag caggtgcgcg ttgaaggcct
23760ccggcgccag ccccacgcag cggtcgatgt tctcgaacca ctccaggctg gtcagcgcgg
23820tccgctgcat ccgctccacg gcgggccgcc gctcggcctc gtaggccgcc agagcctcgg
23880ggaccgtccc gtggtcgtgc agggccgtgg cgagggacag accgtcctcc atcgccagtt
23940tggtacccga accgatcgag aagtgcgtcg tgtgggcgct gtcaccgagg agaacgaggt
24000tgccgtggct ccagtcgcgg ttgcgtacgg cagcgaagcg accccagtac gaccggttgc
24060cccacagccc gtgtccgtcc agcagaccgg tgaagtactc cctcaccctc ccgatgctct
24120cccggtcgcc caccgggtca tcggaccggc gctccgcggc gcggaaaccg gccgcccgcc
24180acaccgcgtc accgatctcc acgatgaacg tgctgcgacc gggggcgtac gggtaggcgt
24240gggcctgcac cgggccgtgg tcggtctcga ccacggcgaa ggtgagcgcg tcgaacggcc
24300ggtccgtgcc gagccacatg taacgcgacc cggcctgttc ccgctccgtg ccgaacgcgg
24360cctcgtaccg cgcgcgggtg cgcgatccca cgccgtccgc ggcgacgacg aggtcgtggg
24420acgcgcgcaa ccgggacacc tcgggcgccg gggagccgaa gtgcagacgc accccgaggt
24480cggcgcagcg ctcctgaagg agccgcagca gggtgtggcg gccgatcgcg gcgaacccgt
24540acccctcgtt gcgctggacc cggccccggt agcagacgtc gatccgcgtc caccgggcga
24600actcggcctc gacagcctcg aacagagccg ggtcggcggc ctcgatgccg ccgagggcgc
24660cgtcggagaa gacgacgccg aacccgaacg tgtcgtgggc cgcgttggcc tcccacacct
24720ccacgacgtc gtggggccgc aggcgcttga ccaggcaggc cgtgtacagc ccgccgggac
24780cggctccgat caccgctatc cgcatgacac ctcgctgggg ctccgagaga gatggatgga
24840cgtgcagaga ccagtaaatg ctatccgaag agaaatgatc tggtggtcaa tctcctttcc
24900agctccgcga gttccgggac tgaccggagc agatcgttgc ccacacgtaa tcaagtctga
24960cggaaggatg cccacatgcc ggacgtcctc acccccaccc cgctgccggc ggccgacctg
25020gcgggcctct tccgcgccct ggacccgccg ccgttcgccc tggtgcgtcg tgccgcaccg
25080gacgggacca gcaccggccc gttcgacgtg ttcatcggca ccatggacac cgtgcgacgg
25140gtgacggacc tgccgtcggg cccggccgtg ccgggcgggg gaccccacac actggcccta
25200ctgccgtacc ggtgcctggc cgaaagaggg ctggactgcc acgacgacgg cacaccactg
25260agggtcctgc ggatccgccg gcggcacacg gccgaccacg ccgcgctcac cgcggccctc
25320gccgcggtgc ggcccgcggg agacctcctc ggggaaggcg ccggcttcga cggctccgac
25380gaggactacg ccgacctggt ccgcgacctc atggccgacg aggtggcacg caccggtctg
25440cacgtcctga tccgcaggga cttcaccgcc cggttgccag gacacggacc cgtggtggtg
25500ggcgaactgt tccgccggct gctggccgtg gagcacggcg cgtactggac gttcgcggtg
25560tacaccggag gccccgacgg tgccgcactg gccggagcct caccgcaggg tcacgtcaca
25620ctgcggaacg gccgggtcgt gatgcgcccg atgtgcggca cgctccgcct tccacccggt
25680ggccggccga gcgccgccga cctggtggcc ttcctgcgtg acggcaagga gtccgaagaa
25740ctgggggccg tggtcgacgc ggaactcgcc atgctgtgcc ggatcagcga gggggacgta
25800cgcctggaag gaccgcgcct gcgaccgatg gcccgcgtac tgcacaccga gtgccgcatc
25860agcgccaccg ccgcgctgcc ggcccggcac acgctcgccg gctccctgtt cgcggcgacc
25920gccgtgggcc gccccttcgc ggacgcgtgc cgcgtcatca cccgccgcga accaaccggg
25980cgcggttact acggcggcct gatcgcgctg ctgggccacg acgacgcggg aaacgaggaa
26040ctggacaccg ccgtgctcat ccgcaccttc gaggtgtccg ggcagggccg gctgaagctg
26100tcggtcggag ccaccctcgg gccccgctcc gtggccgccg acgagacggc cgagacgcgc
26160gccaaggcct cggccctggt gtcggcgctc gcaagcggag gaccgactgc ggagggcggt
26220gccgggcgcc acgcgcgggc tggtcttggt cgcggcccgg aggcggccgg cggcccggcc
26280accggtgagc gaagcggagt gccgggtgac cggacgcggc accagcaggc cgccggccgg
26340cagcccacgt cccccgccga cccggcgtgg cgcccgtcgg tgaccgcgga gggcggtgcc
26400ggggaccacg cgcgggctgg tcttggtcgc ggcccggagg cggccggcgg cccggccacc
26460ggtgagggag gcggagtgcc gggtgaccgg acgcggcacc agcaggccgc cggccggcag
26520cccacgtccc ccgccgaccc ggcgtggcgc ccgtcggtga ccgcggaggg cggtgccggg
26580ggccacgcgc gggctggtct tggtcgcggc ccggaggcgg ccggcggccc ggccaccggt
26640gagggaggcg gagtgccggg tgaccggacg cggcaccagc aggccgccgg ccggcggccc
26700acgtcctccg ccgacccggc gtggtgcccg tcggtgaccg cggaactgga ccggcgccgc
26760gcacgtctgt ccgcctactg gcaacgcccc cgtcggccgg gcagccggcc cgctccgcgg
26820ccaccggtgc tgctcgtcga cacgggtggc gaggagacgg cgccgctggc cgccatgctg
26880cgcggactgg gccgcaccgt cgacgtgcgt cccgcgtacc ccgcggcggc cgcgccacgg
26940accgtcgcgc ccggaaccac ggtcgtcctc ggccccggcc cgggtgaccc gttggcccac
27000ggcgacgacc gcatcaccgc gctgcgggcc atgacgtccg ccctcctgtc cagcggagca
27060cccacgttcg gggtcgggct cggcttccac ctcctgctcg ccgtgctggg tctggccggg
27120gccgcgcgag cgtgggacgg ggccaccggc cagcgggaga tcgaggtctt cggcagacgc
27180gcgacggtgg ggtacggtgg cacgcacacc gtggtggccg gcccgcacac ggacaccctc
27240gcgcggcggc tgtccctgac gctctgctac ggcccggccc acggcgagct ggtggccatg
27300cgaggccctc gaaccggcgg cgtcgccttc ctcccggcat cggtgctgag cgtcgagggg
27360gcggagctgc tggatctcct gctgccctga gcgagcgggt gacgggccgg gccccggccg
27420ctcacccgtt cgccgtgccc ccgagctcga gcacggtccg cgcggccgac tccagcgcga
27480gtgtcatcga gccgccgttg ggctcgaacg aggtgtggtc accggcgaag tggatccggc
27540cctccggccg gcgcatggcc ggcatcaggg aactgtgccc catctcggga aggatgtagg
27600cgccttcgat gtacggctgc tggtcccaga ccaccgaggt gccggtctcg aagtgctcgc
27660gggcaccggg cagcatcgac tcgacgtgtt ccagggcgaa gcggatccgc tcctcggggc
27720tcagcacggc aagcgcccgg gcccgccagc cggttatcag gcactccagt atcttgcggg
27780gcccgggcaa ccggggagtg gcgtcccgga cccaccgcac cggcaggtcc gtggagaaac
27840tcgcgttgtc ctgggcccag aaccgccggc gcatctggag gtagacgcgc acgatcgacg
27900agtacttcac ccggcgcatc acggcctgtt tggcgtcgga cagcccggcg tccgtgaagt
27960cgatgtgccg gatcgcgctg aacggcaccg tcaccaccac gcggtccgcg tccacggagc
28020gcatgcgggt gccgtccagg aaggtgacac gagcgccctc gtcgtcctgg gcgacccgca
28080ccaccggagc gcggtagcgg atgcggtcct tcagccgctc ggcgaacgcc cgggggaagc
28140ggtcggttcc gcccttcacc ttcgaccacc gggggtcggc gttggccagc gagtgcgggc
28200tggactcgtg ccgcagccag gacagggcgg aggccgtctt gaggtccccg ccgcgcatct
28260ccaggaagtg cggttccacc aggtcgatcg ccgccggcga agcgccccgc ccggacagca
28320cctcgtacac cgagcgccgg tcgtacggtt cgagcagggg ggtcggcgcc cagtccgcag
28380ccgtgatgtc gggttccagc gcctcgtagg cgcggcgcac atagcggtcg atcatgtcgg
28440tgacgctcag ccccttctcg tgcggggcga gcgggagccc ggcgcggtcc agcgagtcgg
28500cgtcgggacc gaagaaccgg ttgccgacga agtacgagaa ctggctgccg accaggtcgg
28560ccgtctccag ttccaccccc agttcgcgca ggtagtgcat ggcgtagtgg cagtgcgggg
28620tcagcgtcat cgcgccggcc tcggcgtaca gaccgtccgt gaacggctcg cgcagtgtgt
28680acgcacgccc gcccggacgg ttcgacgcct ccaggaccgt cacgtcagtg ccgcgccggg
28740ccagttcgta ggccacggcg aggcccgaca ggcccgcacc tatgacgacg accctgccgg
28800gcacccggag tcccggcatt cccctgtcga actcgcgacg cacgtcggtc tgcgtgacct
28860cggccatcgg ctctccctcc cccgtgactg tgcacggttg ccgttcatat gattgacgtc
28920aggaaatcat attcggatgc tctcaacggt aaagccttgc atctcaggtg tgttgagccc
28980cgccccgact tggtagctgc tgaccagcaa tcgttgccct cgtcggagcc gtttccttcg
29040cacccggccg tcgttcggca gggcatg
29067337146DNAArtificial SequenceDescription of Artificial Sequence
Tomaymycin biosynthetic gene cluster of Streptomyces species FH6421
in plasmid 3ctttattgct agcgcccggg ccgccgccgc gtcgctcaag ccaggccagg
cgtaggccgg 60ccccgcccgg acacgagacc gccccccttc cgcagccacc ggaacggggg
cggtcctatg 120ccgcaccaga gccaacacac ccttggagga tcaccccgtg acccaatccc
ggaagccctc 180ggaagtcgtc tatctcttag gagccgaggg cctagacctg gtgaagatcg
gcaccacgac 240agacgtggag aggagagtcc gcacgatgca gaccggtctg ccgctgacgc
tgtcggtgct 300gtggacgtgc gagggcggcc gcgcccttga gggggcgctc caccaggagt
tccgtgagca 360caaccggcgg ggcgagtggt tcgacctgac cagccttggt gacccggtgg
ccgtcgtgag 420tgagacggtg cggaggcttg cgccaggtct tgggctcccg atcccgccgc
cgcgcgtctc 480gccggacctg gctctaacgc ctgtggcccc cacgattccg gggtgcgtcc
ggctggagcg 540caccctgacc gctcggatgc tcgcctggtt cgagtgcgaa gacttcggcc
ttccgccctc 600cgcgagcaag cctgtgatgc tgtaccggac cgtccagtac gacgcgaagg
gcgacgtcct 660ggccgtgacg cagacgctcg ttggcgatgg acacgagcag cctgaaggct
tcgagcttcc 720tcccgtacag ggcggcaact gaccgtctgg cacatagatc agcgcccccg
ctccgtggga 780gatcggagcg ggggcgctgt gttgtgcggt caagagacgg ggcggatcgt
ccggcggcgg 840cgagcgtcgg tgttgctgac cttgcggggc gcgcgggcac ggtcgtccgg
ggtgaccggg 900gggtccgtga gggcggcggt gatcatctcg cgcagctcgg ggtgcagcgt
gaagtagtac 960cgggcgttga tccggtagag gccggcggag gtccgcacga ggaggttcag
ggcctccagg 1020cgggcgagag cctcgtgcac cgtggagacg gagttgccgc agtcctcggc
gacggtttcg 1080accggtacgg ggatgacgtc aaacaggccg ccgtggtcct ccacccacgc
gagcgtgcag 1140tgctgagcgg cggacaggtt cagcgagtac agcgacgggg cggtcaggtc
gcgctccggg 1200ttgaagcctt ctacgggcat ggtcgtcatc gcgtctctca tgcggcgcgg
cggcgggtag 1260ccgggatggt cttgggcttg ctggcggacc tggtcatggc cttgtcgtcc
ggggccacac 1320gcggcggcgc catgcggcgg caggcgttga cctgcgtgct cccggctccc
tcccagtatc 1380ggagggggtt cagctggtaa gagccggatc gcgggcccgg cttgaccaca
agcccgagct 1440tgaccagccg gttaagggcc ttccggcagg tgtccttgct ggcgccgacg
tcctcgccca 1500gctcctcggg cgtgaccggg acgtatccgg ccgcgcctcc gtgttcgcgg
agccattcga 1560gcgtcgccca ctccgctgcg gccagcccga gggagtacag ggggacgtcg
ctgcctacgg 1620cctgcaaggt gtaccccttc ccgtcgaact ggtgcggagt ccgcggcctg
gcggacaccg 1680cctctccggt gtccatgttg accagggcca tgttcggccc gcgcttcctc
gtcggcactc 1740gcgcctctcc catcctcgcg ctaaggggcg gtttttgacc ccttagctcc
gatgaaccgc 1800cgcttacgag aggaaactaa caggtcacca ctccgaaaaa cagccgctta
gtccgcgtgt 1860cacacgcata ggtgacgcga cttcctggcg accgggcgcg acgagggacg
cacggtgggt 1920agctaagcgg cgatttttga cccagtaacc ggcggttttt agccgcttag
ccgcgcgttt 1980taccaggtca agagcatgcg ccccttcttc actagaaggc gcgcgccgtc
actcacgcac 2040cacggcgcac accgtggtga ccgccctgga ctcctacccg cccggcatac
cgggccgctc 2100ttagcttcta cggggaaggc cggcggtcct gcgcacctgg tcctcgtcgg
cgcctacggt 2160cggcgagtcc tcagccctgg cgggcttgcg ggtcccgact gatgccacct
gatcacgcct 2220ggcgaccgca cgctgtcctc ggccggccac aagcaggaag gaagcgtcgt
cctgccggtg 2280ggcggctgag gtacctccgt accgccgccg ttgtcgcccc cgccgcaggg
gttgcctttc 2340tgtggctgag gtcgattccc ccgccattga tcacccgcac cagggagggc
ccagccgacg 2400aggtggcgcg cgtgcgcacg tcgctgggtg ggggtgcggt aggtaccggg
cagcagctcg 2460gggaccaggc cccggcgggc gggggtccgg aggagcccgc cgggggtgtg
cgcaagggcg 2520tagcccgtag ccccggagcg cagcggaggg gtcgctagta ggaggccgct
atggcgcgta 2580ccaccagccc acagggtccg gcctccggag gcgacgcact gccggaccgg
accgcaaggt 2640ccggtgagcc cacaacggcg gatcaaccga taagacggcc accgggcagg
aagagggtgt 2700acgcgcccct tggtggcccc tctcgcgcgc gatcacctca cggcgcgtct
cccaggtagt 2760cagatccgtg cggggttctc tacgggccgc acacggcctt cgtcctaaga
ggctcttttc 2820cgccgcttag ctccgaggaa ccgcccctta tctcaatgaa agcgcaggtc
accactccag 2880gaaacggccg ggcgagcgac cagccaggag gggagccgcg gatgcgccgg
tcctcgacga 2940tgtagaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat
gcaaagtgcc 3000gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
aggacatatc 3060cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
agctgcatca 3120ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
gcggtgagtt 3180caggcttttt catatctcat tgcccccgga cgagcgtctg ctccgccatt
cgccgtccgc 3240cgtgccaatc ggatcagccg tccaaatgcg ggattttcgt tagtcggagg
ccaaacggca 3300ttgagcgtca gcatatcatc agcgagctga agaaagacaa tccccgatcc
gctccacgtg 3360ttgccccagc aatcagcgcg accttgcccc tccaacgtca tctcgttctc
cgctcatgag 3420ctcagccaat cgactggcga gcggcatcgc attcttcgca tcccgcctct
ggcggatgca 3480ggaagatcaa cggatctcgg cccagttgac ccagggctgt cgccacaatg
tcgcgggagc 3540ggatcaaccg agcaaaggca tgaccgactg gaccttcctt ctgaaggctc
ttctccttga 3600gccacctgtc cgccaaggca aagcgctcac agcagtggtc attctcgaga
taatcgacgc 3660gtaccaactt gccatcctga agaatggtgc agtgtctcgg caccccatag
ggaacctttg 3720ccatcaactc ggcaagatgc agcgtcgtgt tggcatcgtg tcccacgccg
aggagaagta 3780cctgcccatc gagttcatgg acacgggcga ccgggcttgc aggcgagtga
ggtggcaggg 3840gcaatggatc agagatgatc tgctctgcct gtggccccgc tgccgcaaag
gcaaatggat 3900gggcgctgcg ctttacattt ggcaggcgcc agaatgtgtc agagacaact
ccaaggtccg 3960gtgtaacggg cgacgtggca ggatcgaacg gctcgtcgtc cagacctgac
cacgagggca 4020tgacgagcgt ccctcccgga cccagcgcag cacgcagggc ctcgatcagt
ccaagtggcc 4080catcttcgag gggccggacg ctacggaagg agctgtggac cagcagcaca
ccgccggggg 4140taaccccaag gttgagaagc tgaccgatga gctcggcttt tcgccattcg
tattgcacga 4200cattgcactc caccgctgat gacatcagtc gatcatagca cgatcaacgg
cactgttgca 4260aatagtcggt ggtgataaac ttatcatccc cttttgctga tggagctgca
catgaaccca 4320ttcaaaggcc ggcattttca gcgtgacatc attctgtggg ccgtacgctg
gtactgcaaa 4380tacggcatca gttaccgtga gctgcaggtc gacggatctt ttccgctgca
taaccctgct 4440tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
tatacaggat 4500tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
ccgggcagga 4560taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
tattcgcacc 4620tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
ggcgtaacag 4680atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
ccacctatca 4740aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg
gcggccggca 4800tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg
ggcgtcgtgg 4860actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc
ctgggcggcc 4920tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat
gccacgatcc 4980tcgccctgct ggcgaagatc gaagagaagc aggacgagct tggcaaggtc
atgatgggcg 5040tggtccgccc gagggcagag ccatgacttt tttagccgct aaaacggccg
gggggtgcgc 5100gtgattgcca agcacgtccc catgcgctcc atcaagaaga gcgacttcgc
ggagctggtg 5160aagtacatca ccgacgagca aggcaagacc gatccccggg gacctgcagc
aatggcaaca 5220acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
acaattaata 5280gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
tccggctggc 5340tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
cattgcagca 5400ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
gagtcaggca 5460actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
taagcattgg 5520taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
tcatttttaa 5580tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
cccttaacgt 5640gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
ttcttgagat 5700cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
accagcggtg 5760gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
cttcagcaga 5820gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
cttcaagaac 5880tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
tgctgccagt 5940ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
taaggcgcag 6000cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
gacctacacc 6060gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
agggagaaag 6120gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
ggagcttcca 6180gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
acttgagcgt 6240cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
caacgcggcc 6300caattgaggc ctaagcttca tatgttaatt aaactaggtg tccgcacgca
tgagcgtgcg 6360acatttccga ctatcaaccg gaaacggaaa gatagtgacg gctacgttat
ttccgtcccg 6420gcaaatcggg caataactct attcagtgac tcgactgcgc cctgatcgtg
gcgtatggcc 6480gtgcgacggc ggcgagaggc cgtcggccga ccgtggcccg caacacagcg
gcgggaagaa 6540ccagttgggg cccaagcgcg agggagcgcg caagatccga cgggaaccga
gcccccggcc 6600ccctcgtcgg cgttgtcatt cgaaagaaat acgtaatgac cgctcaagtg
tggccaattc 6660cgctccgggc gtacgctggc gcactaaatt cagggtcggc gtaagaatcg
agcgagggcg 6720gacggagcgt tgcaggaacg ggcgaaagca acccgtagat cactgctgga
agcggctgcc 6780cagctcttcg ccgaacaggg atacgcggcc accagcgtca acgacatgag
cgcccggtcg 6840ggccggacca gcggcgccgt ctacttccac tacgccggca aggaggccgt
cgccgtcgcc 6900gtcgtccagg accggttcgc cacctggcca cagctcgccg cacgctacgc
ggacgaggcg 6960gtccccccgg tcgaccggct cgtcgccctc agctacgaca tcgcccacgc
tctcgccgag 7020gacccggtga cgcgtgccgg cgcccgcctg tgggccgaac gcgccaccat
caacgttccc 7080cttccccacc ccttcgcgct gtggaccacc gccgccacac gactgctcgc
gaaggcccgg 7140ctcgccggcc acctccaccc gcatgtccgc cccgctcgcg cggcccggac
cctggtccct 7200gccttcttcg gcctgtgcgc gctcaccgag gaactcgaag gcacggccgc
cctcaccgac 7260cgcctgaccg actggtggca gctgacgctg ccctgcctcc gcccgcaccc
ggtgcgcgag 7320gacctgccgc gcgggcggtg acgcgcggca ggtcccgtag ctccggccgc
tgtcggagca 7380cgggcggcgc gttacgggga ccgcgccggg taggccccgt ggctcaccgc
tcgcggacac 7440cctgaacggg gagatgaggg gcgcggcttc cgccgtgacg ccgcggtccg
ggaagccgtg 7500gtgttcgagg aagcccgcga ttgtcgagcc cgcgcggcag cgccggcccc
tcggccggac 7560agcgcgtcgc accgcccgta cagcggccac cagcaggtcc ggtgggtgcg
gataccgggt 7620ggtgcggccg cgcacgcggt gggtgcggac gtcgggtggt gcggcggcgt
acgccgtggg 7680gacgggacgg ggggcgtcgc gcatgcggtg gctgcggatg ccgggcggtg
ccgccgtgca 7740tgccgcgggt acggacgctg ggtggtgcgg cggcgcacgc cgtgggggtg
gggcgggggt 7800gccatgggtg cggacgccgg gcgctgcggc ggcgtacgcc gtgggccggg
gcggaggtgc 7860cgtcacgcat gccgtggctg cggacgtcgg gtggtgccgc cgtgcatgcc
gtgggtgcgg 7920acgtcggcgg cggcgtacgc cgtgggccgg gaccgggggc tgtcacgcat
gccgtggctg 7980cggacgtccg gtggtgccgc cctgcatgcc gtgaccgcgg acgccgggca
gtgccgtcgc 8040gcaccccatg gggccggggc gggtggggcc tgccggtatc cgggtggggg
cggtaccgcc 8100cacgccgacc gcgccgcccg cccggcaccc ccgcccgcgg ctcagggacg
cgcctccacc 8160ggttcgaggt gcggttccag ggcgccggcg accttgtggc acagctccac
gaacatcgcg 8220cgctcacccg ggctcagtgc ggcctccacg atgcgggcgt tctcccgccg
ttgctcggcg 8280acgcgggtga ggaaggcctc gccggccggt gtgagcgagg tggtgagttc
acgccggtcc 8340tcgccgagcg tccgctggtc gacgagctcc aggtccttga ggacgcgcag
tgctttggac 8400gtggtcgccc gcgacacacc gagcgaggca ctgagcaccg agggagtcat
ggggccgcgg 8460atgcgcagca gctccaggac gtcgtactgc tgccaggtga cgccttccgg
attcgagcga 8520gtgcggcggg cgacgaggat gcactgcaag tgggaaagcg cgtcctcgag
ttcgcctggc 8580acagccgagg tctcggaggc gcttcgtggg ctgtcggttc gggtcatcaa
cgccctcagg 8640tgatcaggtc ggggacgagg cgccacggcc ggctcggggc gggagcgggc
cgacggcgac 8700ctccgattgg ttacccatta tcaacgatcc tggggcggca acgtatacct
ctccacacgg 8760tcatgtgatc catcgcccaa cagttgtttc catcgggaaa atttctacgc
gatgatctct 8820gatgagtggc cctccaggcc ggacacgcgc cgcgcctcgc tccggctttt
cccggcccgc 8880gagcgggagg cgcccctcgg gcgggccccc gtggacggca gggcgccgtc
ggccgggcgc 8940tctggtttgc tccggtcgaa caatggctga tgcatcgtca gtgactcggt
tgctttcctg 9000gcggtaaaca agtggctgaa aacgcgccct gcgacgctgc gctccaccgt
gtcggcctgg 9060ggttgtccgt ggcggcgaag gggcctccgg tcggggcccg acagcccagg
catgggagcg 9120aagagcgggg gtagtggcag ctccagcgtg ggtggcgcag cggtctcggc
gcctcaagat 9180cggggttgac acgtcgaaga gcgccgacct aacgtgattt ccgaacagca
accatcacct 9240ttcggttgat ttgtgcgtat tccttgagaa gccggtcact ggacgtcgga
tgtgctcgtc 9300gctgtgggcg ccccgtggcc atccgccagc gaggcaagga gcacgtgtgc
tcacggagaa 9360tgcatcaggc gaggcgcgtt ccgctgttcc cctgaccctc accgagggtt
tcgaccgcgt 9420ggtgcgggcc gccggccacc aggtcgccct cgtctccggc acggagaccg
tgacctaccg 9480gcaactgaac gaacgagccg agcgcgtggc ccgcgggctg ggcgcccgca
aggtggcgcc 9540cggtgaccgg gtcggcgtgt atctgcgccg gtctcccgac ctctacgcgg
tcatgctcgg 9600cgtcctcaag gcgggcgcct gtgtggtgcc ggtcaacccg gaccaccctg
cgccgttcgt 9660ctcccgcgtg gtggccgaat cagcgccgcg ggccgtcgtg cacgacgcgg
gaacaccggc 9720cgtcgcgccc gctgctccgg gcgcaccgct gtgggtaccg gtcgaggagc
tcaccacggc 9780cgcggaaccg gacgacggcg tcgcgctgcc cgccgtgaac gatccggaca
gcaccgcgtt 9840cctgatgttc acctccggat ccaccggccg gccgaagggc gtccgcatcg
cccaccgcgg 9900gctggcacga ctcggcccgt acagcgggga actgcgcatg ggcccgcagg
actgcctggt 9960ccagtccgcg gcgttctcct tcgccgcgtc caccatcgag atctggctcg
ccctcctgca 10020cggcgcacga ctcgtcgtga tgccccaggg gctgcccagt ctcccggccc
tcaaggacgc 10080cgtcgtccgg cacggcgtca ccgcgctgtc cctgccctgc ggtctgttca
acctcctggt 10140ggacgaggag ccggaatgcc tgcggggcct gagggtgatc ctcctcagcg
gcgacttccc 10200gtcgccggag cacctcagcc gtgcggcacg ggcgacgcgc gcggtcatct
acaacggcta 10260cggctgcacc gagaactcct ccatcaccgc cctgtacccc atccgcgatg
ccggggacgt 10320cacccgcgag aaccgggtgc cggtgggccg ccccctgccc ggggtcaccc
tggaggtact 10380cgacgactcg ctgcggccct gcccgcccgg aacgcccgga cagctcgtcg
tcggcgggct 10440cggactcgcc cagggctatc tgaacgaccc ggaactcacg aaccgcaagt
tcgtcaccgg 10500gccggacgga cggccgcgct acctgaccgg cgatctggcc cgcgccaccg
aggacggtga 10560catcgtcctc ataggccgcg ccgacagcat ggtcaagatc cgcggctacc
gcgtcgaact 10620gaccgcggtg accctcgccc tgcgcgccct cgacgggatc ggcgacgccg
tcgtcaaggc 10680gttcccggag ggcgccgggg agaagtcgct caccgccttc tacaccaccg
tcgacggacg 10740accgctggac ggtgccgacc tggcacgccg catgggagac caactgccct
cctacatggt 10800cccctccacg ttccaccacc tcggcgacct gccgagaaac gccaacggaa
agatcgaccg 10860gtccgccctc acggacccgt cggacaccaa ccgcgatccg aagaaaggtc
acacagccgt 10920gcagaacccg ctcgagaccg tcgtactcca ggcgtggaag gacatctccg
gcgccgacga 10980cttcaccacc accgactcct tcctcggcca cggcgggaac tccctgcact
tcgtccagct 11040cgcctccagg ctgcagaaga tcttcggtgt ggaggtcagc accgaggacg
tcttccggca 11100cggcacggtg gagcagctgg cgcgcttcgt cgagcagtcg cgggacaccg
gacgcaaccc 11160cgccgcacag acccagtagg cgtcacccgg ccgtggccgt gcggcgcccg
tcgccggcgg 11220ccggggcatc tctgcagagg actacgaccg gtgaactccc ccctccgaac
caccgtgctc 11280gaccttgcac ggaccaccct cggcagcgcc gacctcaccg cgcacgaacc
gttggccgac 11340cggtgcgaac acccggccct gctcgacgac ctcgccacca cgctgaccgc
cgtcttcgcg 11400gtcgagatca ccggcgcgga cctggcggcc ggtgccaccg tcgccgacgt
ggccgcgcga 11460atggacgacc ggcgcgacgc cccccggatc ccggaactgc gcgccgggct
cgctccccgc 11520gacggccggg cggtggaggc gtccttcggg cagagcggca tctggctgat
cgaccagtac 11580ctgcccaacc cggccgccta caacggcccc ttcttcgtcc ggctgccgtt
ctcagccgat 11640cccgaccgcc tgcacgcggc cgtgcgcgga gtgctgcgcc gccaggaggt
cctgcgcacc 11700acctacgccc tgagcgacgg cacgctccgg cagaacgtct cgcgggacga
tgacgcggtc 11760gtcttcgagg tagcccgcta cggcgacgac aaggaactcg acgccctcgt
ccaccgggtg 11820gccaatctcc gcctcgacct ggcccgcggg ccggtcatcg ccgtgacctg
cgcgctcggc 11880cccgcgaacc ggtccgccgt catctgcaac atccaccaca tcgcctccga
cgccgcctcc 11940gccggtgtct tcctgcggga actcctcgac gcctacgacc gcctcggccg
cggtctgccc 12000gtcgaggccg acccgctgcg gcccacctac ggggacttca gccagtggta
ccgggaactg 12060atgaaccccg aggccctcac ccgctccctc gaccacttcg ccgcccggct
cgccggggaa 12120ctcccggtgc tcgacctgcc caccgaccgg ccccgcccgc cggtgaagca
acaccggggc 12180ggcaccctcc cgttgcacct gccggccgcc gcggccgacg acttcgaggc
gctcgcccgg 12240accgaggggg tgaccctgtt catggccctc gtcgccgcgt acgcggtctt
cctctcccgc 12300cacaccggtc agcggcgcgt gctgatcggc agccccgtct cgctccgcga
cgacccggcc 12360acccacgaac tgatcggcta cttcgtcaac ctggtcgtcc ttcagcagga
gatcgacgac 12420cggatgaccg tccgggacgt gctccgccgg gcgcgggagg aggtgagcga
ggcgctgcgg 12480cacaagtggg cgcccttcga caaggttgtc gagcgtctgc agccaccgcg
cagcagcggc 12540tacaccccgc tcgtgcagac catgctcgtg ctcacccagg gcgacgccgg
acggatatcc 12600cacgacgaca cggaactgcg catcgagcgc ggggccgcgc acggcgccaa
gtacgacctg 12660tccctcgttt tcgagcggga ctccgaaggc ctgcacggtc tgatcgagta
cgacgcggac 12720ctcttcgacg agccgacggt acgggccatg ggcgaccggc tgcggcacct
gatggagcag 12780ttcgcccgac gtcccgacgc acccctgcac gaactggagg cgctcggtgc
gcaggagcgg 12840cggtcggtgc tggtccgcgg ggaccggacc gcgcacgccg tgcacgacgc
acccgtcatg 12900gaactgttcg aggcccaggc ccgggcgacc cccgacgcgg tggcgctgga
ggacggcgac 12960accaccctgt cctaccgcga actcgacgag cgcgccaacc ggctcgccca
cgtgctgcgc 13020gcctccggcg ctgcggccgg cacccgggtc gggatctgcc tgccccgctc
ccacgacatg 13080gtcgtcgccc tgttcgccat cctgaagacc ggggcggcgt acgtaccgct
cgacccgtcc 13140taccccaggc agcggatcac ccacacgctg cgcgacgccg gggtcttcct
gaccgtgacg 13200gacagctcac tggccgacga actccccccg agggagccgc tgttcgtgct
ggaccggcac 13260gacggaccga tcgccgcggc ccccgccacc ggcctcggcc gggtgaagac
acccgacgac 13320gagatctacg tcgtgcacac ctcgggctcc accggcctgc ccaagggggt
ggtcatcgcc 13380gaccggaccg tcgccaacct cgtccgggcc cagcaccgtt gctcgccggc
cggagcgacc 13440gggcggacgc tccagtacat gtcgctgtcg ttcgacgtgt ccgtgatgga
gatcctcggc 13500accctgtgcg tcggcggcac cctcgtgctg gtctccgagg aactgcgcaa
ggatctgcac 13560gcgctcgccg gattcctcgc cgaacgccgc gtcacccggg tgtacctgcc
ctacatcgcg 13620ctccagcagc tggcctccct ggccaccgac gccggtgtgc gcctggacga
cctgcgcgag 13680atcacctccg tcggcgaggc cctcgtggtc tccccgcaga tccgggagtt
cgccacccgt 13740cacccggcgg tccggctggt gaacatgtac gggccgtcgg aaacgcacct
ggccagctgg 13800tacccgctca ccggctcgcc cgcgacctgg cccgacaggc cgccgatcgg
ccgcccggtg 13860gacggcgtgc ggctggtggt cctggacgcc cacatgcggc tcgtcccccc
gggtgtcccc 13920ggcgagctgt acatcggagg gcccgtgctg tcccccggat accgcaaccg
tccggacgag 13980acggcccgcc ggttcctccc ggaccccttc ggcggccccg ccgaccggct
ctaccgcacc 14040ggcgacctgg tgcgctggaa cagcgagggc gacctggagt acctgggccg
gaccgacgac 14100cagatcaaga tccgtggcta tcggatcgag cccgccgaga tcgaggccgc
actcgacgac 14160ctggacggcg tcgcctcctc cgcggtcgcc gccgtggacg tcgcccccgg
cgaccgcaga 14220ctcgtggccg tcctggagac ctcccgcacc tgggagaccg cggagctgcg
ccgcgccctg 14280tccggcacgc tgcccgacta catggtgccc gcgctggtgg tcgcggtgga
gcacatgccg 14340acgaccccga gcgggaagat cgaccgccgg gccgtcgccg gcctggccgc
ggcacaggcg 14400accgcggcac ggaccgcgcc cgcgccaccc ggccggccgc ccaggccggg
cctggagcag 14460cggatcgcgc gggagtgggc ggatgtgctg aaggtgcccg cggtgggcag
ggacgaggac 14520ttcttctccg tcggagggaa ctcgatcatc gccacggaac tggtctatcg
gctgcgccgg 14580gcgttcgacc aggacctctc gctgcgcgcc ctgctggaga atccgacggt
cgcgggcatg 14640gccgcccggc tgcgttccgg ccccggcgct cccaccaccg cccccgccgc
gctgcgggag 14700gacgcgacgc ttcccgacga cctgcccgcc gtcaccggca ccccggtacc
ggtcgcccgg 14760gcccgtgagg tcctgctcac cggcgcgacc gggttcctcg gcagctacct
gctgcgggag 14820ctgaccggaa ccaccggcgg ccgggtgcac tgtctggtgc gggcggcgga
cgaacgggcc 14880ggcatggagc ggctgcgggc caccgccgag cgctaccggc tggacgggcg
gatcgactgg 14940aaccgggtgc gcgccgtgcc cggcgacctg agccggcccg ggttcggtct
gcccggtgcg 15000gagtacgacg cgctggccgg caccgtcgac gtcgtctacc acgcggccgc
gcacatcaac 15060ttcgtgctgc cgtacgcctc ggtgaaaccg acgaacgtgg acggcttccg
ccacgtggtc 15120cgtttcgccg cgacggaccg ccccaagcac gtgcagtaca tgtccaccat
cgccgtgttc 15180cctccgggcg aggcgcccga cggcacggtc ctcaccgagg acgacgtgcc
cgaggcgtgc 15240gaacgactgg gcatcggcta cacccagagc aagtgggtcg ccgagcgcat
cgcactcgcg 15300gcccgcgcgc acggcgtgcc ggtcaccatc caccgcatcg ggcgcatctc
gggcgacagc 15360gtcacaggcg cctgccagag cgacgacttc ctgtggcggc agatcaagag
cttcatcgaa 15420ctcggctcgg ccccgccggc cgaggacctc accaccgatc tgctgcccgt
cgatttcgtc 15480gcccgcgccg tcgtcgccct ctcccgccac cccgccaccc acaaccgcac
cctgcacgtc 15540ttccacccga gcggatcgga cttcaccccg gtccacgcgg ccctgcgcgc
ggacggccac 15600cgcctggaga tcgtcccggc cgacacctgg cttgcccggc tggaggagtc
cgcacggcgg 15660cccggcggca acgccctggc ggcagccgtg cccctcttcc gcgagggcgc
cctggaactg 15720ggcgacaaca cctacggcaa caccgccacc acccgcctgc tgatggacct
cggactgccc 15780tggcccgcca tcgacgagca ggcgatcacg cggatgctcc gctacttccg
ctccgtcgga 15840gaactggccg acgactgagg ggacttccct gtcccgggcc ctcccggcag
cgcggcgcag 15900ccgccggcgc cgatccgacc atccaccaca cggcaatcgc cgaagcggtc
gccggacacc 15960gaaagcacca gcagccatgc caacgagctc ctgccccgac accgcatacg
acaccctcat 16020accctccgtc gtcgcggctc tcccggccgc acagcagccg gagtggccgg
accccgggcg 16080actcgccctc gtccacaccg aactggcccg cgcggacccg ctggtgacgt
acgacagcgt 16140gcgtgccctg cgccggctgc tgtcccgtgc cgccgaaggc gaactgtgcg
tcctccaggc 16200cggcgactgc gcggaggacc ccgccgagtg cggcccggcc ccgctggccc
gcaaggccga 16260gatgctggac gtcctcagcg acatcgtgcg aacgggcgcc ggacggccgg
tcgtccgggt 16320gggccgtgtc gccgggcagt acgccaaacc ccgctcccac ccggaggagc
tgcacgacgg 16380cgtccggctg ccggtctacc gcggtcccat ggtgaacgcc ccccaccccg
acgccgacgc 16440gcgccggccc gatcccgccc gcatcctgag ctgctaccgg gccgcccgcc
gggccgtgga 16500gtccctggac cggctgggcc gcggcgaggg ttcgcccgcc gagacccggg
tgtggaccag 16560ccacgaggca ctgctgctcg actacgagct gcccctcgtg cgccggcacc
gctcgggccg 16620cagttacctc gccagcaccc actggccgtg ggtcggcgaa cggacccggc
aaccggacgg 16680tgcccacgta cggctgctgg cggaagtgga caacccggtg gcgtgcaagg
tcggcccgac 16740cacgaccgtg gagcaggtgc tcgccctgtg caccgcgctg gacccggagc
gctcaccggg 16800ccggctgtcg ctggtcgccc ggttcggcgc ctcccgcatc gacggcctgg
cccccctggt 16860ccgcgcggta cggcgggccg gtcacccggt gctgtggctg tgcgacccga
tgcacggcaa 16920cggtgaacgc accgcgcacg gactgaagac gcgccggctc agcgccgtga
tggcggagat 16980cagccggttc gtggacatcg tctccgccga gggcggccgc agcgccggcc
tgcacctgga 17040ggcctcaccg gacgacatcg ccgagtgcac cggcgccgga ttcaccccgg
cccccgggcc 17100ggcctaccgc accctgtgcg atccccgcct gaacctggtg caagccgtcg
cggcgaccgc 17160ctactggcgg ctgccggccc tggaggccgt cgcatgagcg agtctgcgcg
gaacgcgcgc 17220ggcctggcgg ccctgctgcc cccgccgggc acccccttcg ccgtcctgca
ccggccgggt 17280gccgggcacc ctggcaccgt ggacgtcgtc agcgggcccc tccgcaccgc
cgccaccctc 17340gccgagctga gcctcgacga cgagtccgcg ccggcctccc agggccccgg
accggcgcac 17400agggtcctcg ccctggtgcc ccaccggcag atcgccgaac gaggcttcgc
ggctcccgac 17460gacggcaccc cgttgctggc catggacatc ggcacccagc acaccgtgcc
cctggagcgg 17520atgctggcac tcctgcccga ccgcgaactg cacgtcgagg aaaccgggtt
cgacctcgac 17580gacgaccggt acgcggccgg cgtcgacgcg ctcacccgcc aggagatcca
gcgagggcag 17640ggcgcgaact tcgtcctcgc ccgcagcctg cacggccgga tccgcgactt
cgaccggacc 17700cgggccctgg ccgcgctgcg gcggctgctg atcgcggaga gcggcgccta
ctggacctac 17760ctggtctgca ccggcgaccg gtacctcatc ggcagttccc ccgagcagca
cgtacgggtg 17820gccggctcgc gggtgtcgat gaacccgatc agcggcacct accgctaccc
cgaggggggc 17880cgcccggacc gcgaaagcct cctccggttc ctcgccgacc ccaaggagat
ccacgagctg 17940tacatggtcg tggacgagga actgaagatg atgaccgaac tgtgcggttc
ccgcgtgcgc 18000gtgtcgggcc ccacgctcgc gtggatgtcg cgtctcgccc acacgcagta
ccacctgcac 18060ggcgagtccc cgctgcccct gaccgacatc ctgcgcggga cactgcccgc
gcccacggtg 18120acgggcagcc cggtggagaa cgcctgccgg gtcatcgccc gccacgaacc
ggcaggccgg 18180ggctactaca gcggcgtgct ggcgctggcc ggccaggagg ggggacggcg
cgccctggac 18240gcggtcatcg tgctgcgcac cgccgacatc accgcggacg gatccgtgcg
gctgaccacc 18300ggcgcgaccg tggtgcgtga ctcggtgccc cgcgaggagg cggcggagac
cacggcgaag 18360gccgcggggc tcctcaccgc gctgacccgc ggcccggccg gccggtccgc
ggccccggcg 18420cacgcggcac ccgacgtctc cctgggagcc gatccggcgg tgcgcgcggc
gctgcgctcc 18480cgcaacgacg gcatcgccgc cttctggctc ggcggcgggg cgcgcctgcc
ggcgccctcg 18540ccacacggac cacgggtggc ggtgatcgac gcggaggacc ggttcaccag
catgctcgcg 18600cagcagctcc gcgcggtggg ctgccacgtc accctgcacc cctggtggtc
ggttccggag 18660gcagccgacg accccggcac cgtgctgctg ctggggccgg gacccgggga
cccccgggac 18720gtcggcgacc cgcgggtggc gcggctgcgc tccctggccg gccgccggct
cgcccggcgg 18780ttgccgctgg ccgcggtgtg cctcggtcac caggccgtct gcggggtgct
gggccttccc 18840ctggtccggc tcgcgcggcc ccgccagggc gcccggatgc gggtcggcct
gtggggacgc 18900gaccggcacg tcggcttcta caacagcttc accgcgcgct ccgacaccga
tcgctgtccg 18960ctgcccggcc gggacgccac ggcccgggtg tggcgccggg acgggggaga
cgtggtcgcg 19020ctggacggcc cggggctggc caccgtccag ttccacgccg agtcgctgct
caccgaggac 19080ggcccggaca tcctgcgcga gctggtggac cgggcggccc gtaccgagcg
gcgcaccgag 19140gccctgatgt cccgccgagc caaggagcac gcgtgaaccc gcccggaacc
gtcgtcgcca 19200acgccgccct ggacccccgg gagctgcgcc ggaccatggg gcacttcgcg
accggcgtca 19260ccgtgctgac ctgccggcgc ggcgcccggc tgcacggggc gacggtgaac
tccttcacct 19320cggtgtcgct cgatccgccg ctcgccctgg tcgccctgga ccgtcgcacc
cgcgccgccg 19380ccctcctgga cgacggcccc ttcgtcgtca atctgctcgg cgagcaccag
caggacctgg 19440ccctgcactt tgccggcggc tcgccggccg attccgtgcc gtgggtggac
ggcgacggcg 19500accggccccg gctggcggga accctcgggc acctggtgtg ccgaccctgg
cgcacctacg 19560acggaggcga ccacacgctg catgtcggcc gtgtcgagga gttcgccgcc
ggagggggac 19620ggccgctgct cttctaccga ggcgtcttcc cccgcctcat gccggacgga
ggaggagacc 19680cggagggacc cgaggaggtg tggtcgctct gtctggacgg cccaggaccg
gccacggatc 19740agttcgtcac cgatcatgag acacggaagt agggacatgg cacccgacaa
cggacagtcc 19800gcagcacccg gcacctccgg ggcgtccacc ggcaaggccc gggtcacccg
gccgctgacc 19860ggggacgagt acatcgagag catccgcgac ggacgggaga tctgggcgta
cggcgagaag 19920gtcgacgacg tcaccaagca cccggcgttc cgcaacaccg tgcggatgac
ggcccgcttg 19980tacgacgccc tgcacgatcc cgagcaccac gacaccctga ccgcgcccac
cgacaccggc 20040agcgacggct tcacccacaa gttctaccgg gtgccgcgca gcgtgcagga
cctcgtgggg 20100gacagggacg ccatcgccga ctgggccagg ctgacctacg gctggatggg
acgcagcccc 20160gactacaagg ccagcttcct ggtgaccctc ggcgcgaacc ccgactacta
cggcgacttc 20220gcggacaacg cccgccggtg gtacgccacc gcccaggaga acgtgctgtt
ctggaaccac 20280gcggtgatca accctccggt cgaccggcac cggcccgccg acgaggtgga
cgacgtcttc 20340gtgcacgtgg agaaggagtg cgacgacggg ctggtggtga gcggggcgaa
ggtggtggcg 20400accgggtccg cgctcaccca cttcaacttc gtggcgcact acggactgcc
cgtgaagaag 20460aaggagttcg ccctcgtcgc caccctgccc ctggcggcac ccggcgtgaa
gctcatctgc 20520cgccagtcat acgaactggc cgcgagccgc acgggcagcc cgttcgacta
cccgctgtcg 20580agccggctcg acgagaacga caccatcttc atcctggaca aggtgaagat
cccctgggag 20640aacgtcctca tctacgggga caccgccagg gccggcacct tcctgcagac
ctccggcttc 20700acccaccggc tcaccttcca cggggtgacc cggctggccg tgaaactgga
cttcctggcg 20760ggcctgctgc tgaagggcgt ggaggtcacc ggcaccaagg acttccgggg
catccagacc 20820cgggtcggcg aggtcctcgc ctggcgcaac atgttctggg cactgagcga
cgcgatggcg 20880cacaaccccg atccgtggca cgacggagcc ctgctgccca acctcgacta
cggcatggcc 20940taccggtggt tcatgaccgt cggctacccg agggtccggg agatcatcat
gcaggacctc 21000agcagcgggc tgatctacct cacgtcgcac gccaaggact tcaacgaacc
tgaactccgt 21060ccccatctcg accgcttcat gcggggttcc aacggttacg aggcggtgga
gcgcgccaag 21120ctgatgaagc tcatctggga ctcggtgggc accgagttcg cgggccggca
cgagctgtac 21180gagcggaact actccggcaa ccacgagagc gtgcggatcg aactgctgca
cgcccagacg 21240gcttccggtc tcgtcgacca gtaccggggc ttcgccgaac agtgcatggc
ggaatacgac 21300ctggacggct ggacggcacc ggacctggtg ccgcccgacg tcgactgagc
gccgcggcca 21360tccggccgga cgggcccgcc gggcggtacg gcaggggcgg gaggagcccc
ccccgcccct 21420gccgctcagc cggtgaccag ggcgagcgtg aaaccgtcgt accccttggt
gccgacggtc 21480tggatcgagg tggcggtgac gtccgagcgg ccggccagca tctcgtggaa
ccggcgcacg 21540ccctggacgc cggcgtcggg atggtccggg tcggtcaccg cgccgcccag
gacgacgttg 21600tcgacgacca ccaccgcgcc cggccgggac agtttcagcg cccaggtgaa
gtactcgggg 21660atgtcgggct tgttggcgtc gacgaacacc atgtcgaagg gggcggtacc
gggccggtcg 21720agggtcggca ggatgtccag cgcccggccg acgtgctgtt cgacgaggtg
ggcgaccccc 21780gcctcggcga ggcgcgacgc ggccgactcg gcgaaggacc gctcccactc
gatggtgacg 21840aggcggccgt ccggcggcag cgcgcgggcc agccagatgc tgctgtaccc
gccgaacgtg 21900ccgatctcca ggatgcgccg cgcctgccgc agacgcgcca gcagatgaag
gagtttgccc 21960tggggggcgc tgacggcgag gtccggcagg tcgaattccc tatgggcctg
agcggccttc 22020gaaagtgctt cgtcctcctt cacgagaagg gagctgaagt agacgtcgac
actgttccat 22080cgctcttgtt ccatggttgc aataattcac gataaaccct cggatggcaa
gagcagaatt 22140cgatatccat ggtgccgtga acctgagaat gcccaggtca gcggcctatt
caaggctgtc 22200cagtcacctt cgccaaagaa attaacgggc ggtcgataat gttcggaaat
tccttcggtg 22260cggcgttgac tgtcgtcctt gcggtcacct agcctccctt tctggaacct
tgtgaaaaag 22320cctgtcccgg ataggagtgt catttcatgc gagaagactc ggccgtcaca
acggccgcac 22380ccccggtgca cctggtgccg gcgatgcacc acctgggcgt ccaaacccgc
gacctggaca 22440actccctggc gtggtacaag gacttcttcg gctgcgccga gacctggacg
ctcaccacgt 22500tctcggacct gacccgcagc aggctccccg gcatcacccg gctcaccgag
atcagcgtcg 22560ccgacgtccg cttccacctc ttcgaacgcg ccgggcacga cccggccctg
cccggcggca 22620acaaggccca gttccagcac gtctgcctcg ccaccggttc cccggaggaa
ctgcgcgcct 22680ggcgcgatcg ctggatcgag ctctaccgct cgggccgcta cgacttcgcg
accgatgagc 22740agcccacgga catcgtggtc gacgccgacg gagtgcacag ctgctacctg
ttcgacccca 22800acggcctcga gttcgagttc acctacgttc cgggcggtgc ggcatgagcg
cgggcccgca 22860ccggaccgtc accgaactgc cggtcgccga aggctgggac ttcggggact
tcccctacgg 22920cctggagccg ctgaccctgc ccgagccccc gcacgagccc gcggccgacg
ttccggacgt 22980gctgtgcgcc gagcccgccc ccggcggtgc gcggacgtcc tgcccgcgca
ccggaccggc 23040gcccggcctc ccggagctgg cccaccagct cttctggttc cgctggatca
ccggacacca 23100gctgaccttc gccatctggc agttactcgg ccacgcgctg caccaggcgc
acgcccggcc 23160cgaccccggc ccgtcgctgc gagccatgac ggacctgaca cgggcgtaca
ccgcgatgct 23220gctctacacc ggctcctgtc ccaaggacgt ctacagcgac gtgatccggc
ccagcatgtt 23280cctgcagcac cgcggcttca gcgggacctg ggcgccggac ttcgtccccg
tccgccggct 23340gctgcggggc aggaagacgc cgtggcacga gaccccggag ggcggccggc
tggccgacga 23400ggtccgtctc taccacctgg tgcactcggg ggtcgccgcg aaactcgtac
ccggcggcag 23460gtccctgctc caggacaccg cccccacggc ccggccgcac gacccccgga
tgcaggcgct 23520ggtctacgac aactacttcc tcaccctgcg cgccgacgtc ccgaccgccg
aggtcgtcga 23580gcagctccgg cgccgactgg ccgcggtgcg cctggacgtc tcggtcaacg
ggctgtaccc 23640cgggctgacc gcgcaggagg acgccgcact gcccgaggag ttgcgcagcg
aggacacaca 23700ggcctgcgag cgggacttcg acgccgtcct gcggcgcgtc gacggccttg
ccgccgcact 23760cgaccggcgg ttgctcgacg gcacgatcgc ccgctgagcc tgagccaccc
gatcacgcga 23820caggaaagga gcggtggacc gtgcggtacg gagtcgtcgt cctgcccgaa
cgccggtggg 23880cgcaggcccg cgaacagtgg gtccgtgccg aggagttcgg attcgaccac
gcctggacct 23940atgaccagct gatgtggcgt tggctgcgcg acgagccctg gttcggcgcc
gtgcccaccc 24000tggcggcggc ggccgaggcc acctcgaccc tgaccgtggg caccatggtg
gccacaccca 24060cctatcggca cccggtgacg ctggccaagg aggtgatgac cctcgaggac
atcgcgggcg 24120gccggttcgt ctgcgggctg ggagccgggg ccggcggcct cgacgaccgc
gtcgtcgatc 24180cggccgccta ctccccacgg caacgcgccg accgcttcac ggagttcgtc
gacctgctcg 24240acaagctgct gagccgccgc agcaccacac acaccggcac ctactacgac
gtccgggagg 24300tgcccgtgca cccgggctgc ctggccacgc cccgggtgcc gttcgccatc
gcggcgaccg 24360ggccgcgcgg catgcggctg gcggcccgcc acgccgacat gtggatcacc
gcggggcggc 24420ccggcgactt cgacgccctt ccgtacgagg agaccctgcc ggtgatcaag
gagcagctgg 24480cgcgcctcga cgaggcgtgc gagcggaccg ggcgggatcc cgccaccctg
cgccggctgc 24540tgctgaccgg cgccatggtg ggcggcaccc tggactccgt cgaggcgtac
cgcgacgccg 24600ccggccgctt cggcgaactc ggcatcaccg acttcgtcgt ccactggccc
cggccctcct 24660tcccctacca gggcagggtg gaagtgctgg agcagatcgc gcgggacgtg
ctgaccgtcc 24720ggggcgggga gcggccgtga tcgcctacga gatcgtcgac atgttcaccg
gcacgccctt 24780ccagggctgc gcgctcgggg tggtcccgga cgcgaccgca ctcgacgacg
acggcatgcg 24840ggcggtggcc cgcgagatcg gcctcaccga gacggcgttc gtcctgccac
ccgagtcgcc 24900cgacgccacc caccgggtac gggtcttcac cccggagcgg gagtcaccgt
acggcgggca 24960ctccgccatc ggcacggcca ccaccctggt gcggctgggc cgtctgcgcg
cgggggagct 25020ggtgcaggag tgcgggggcc gcctgatgac cgtgcgcgcc agtgcccgac
gggccacgct 25080cggcgtccgg ggggagcccg tgccacccgg cgcctgggat cccgtgccgt
tgctggaggc 25140gtgcggcctc accgaggacg acctggtcgc cgggccccgc gtgaccgggt
tcggaccggc 25200cttccacgtg ctgccggtcg gacccgaggc ggtcgcccgt gccgcacacg
acccggcgca 25260ccccgtgtgg tccacctgcc cggacgcggt ggtggtcgcc tacgacaggc
gcggacacct 25320ggccgacgtc agggtcttcg cccccggcta cggcatgccg gaggacccgg
cgtgcgcctc 25380cgccgccctg gcactgggcg cctggctcac cggcgcgggc ctggtgccgg
cgacggacgg 25440tacccgcctt taccgggtcc ggcaggggca cgggctgggc cgccccgccc
ggctcgactg 25500cgccgtgacc gtacgcgacg gccgagcggt cgcagccgag gtgaccgggg
aggtggcggc 25560caccgccgcc ggccggatgc acctgccccg cacggcggcc gtcgcgcgct
gagccggggc 25620ccgggccgtg taccaggccg agccgcggaa ggcacccggt ccggtcgacg
cacacccaga 25680gcacacgacg cgaatccctg tatcgcagaa cgaagaggag aggaacccga
ctgtgttgtt 25740ccgtccagag ctgcgcggca cccggggcgc ggtcgcctcg acccactggc
tggcctcggc 25800cgcgggcttc cgcatgtacg acaagggcgg caacgcgttc gacgcggccg
tcgccgccgc 25860gttcgtcatc caggtcgtgg agccccacct caacgggccc gggggagacg
tgcccgtcct 25920cgtccaccgg gccgggagcg gccgggtcga cgtcgtctgc ggccagggcc
ccatgccccg 25980ggccgcgacc atcgagaggt tcgaacagct cggcctgtcc gtggtccccg
gctccggcct 26040gctgcccgcg gtggtgcccg gcgccttcgg cgcgtggctg cgggtcctcg
ccgagtacgg 26100caccctgcgt ctggaggacg tcctggagcc ggcgatcggc tacgccgaac
gcggctatcc 26160gctgcttccc aaggcggcgg cgatgatcga ggcgctccag gaactcttcc
gcgacgagtg 26220gaccgagtcc gcccgcacgt acctggtggg cggggccgcg ccgcggcccg
gtcagcgcat 26280gaccaacccc gacctggccc ggacctaccg gcgcgtcctc gacgaggcca
gggcggcggg 26340cgccgatcgc gacaagcaga tcgacgcggc cctgcgcgcc ttctacgagg
gcttcgtggc 26400cgaggccatc gacggctatc tcgccaaggc ggaggagatc gatgccaccg
gccgccgcca 26460ccgcggcctg ctgaccggcg ccgacctggc aggctggcgg gcgacggtgg
agccctcgct 26520ctccttcgac caccgcggcc tcaccgtgca caaggcggga ccctggtccc
agggcccggt 26580cttcctgcaa cagctcgcgc tgctgcggga gttcgacctc gccggtatgg
gaccgcacag 26640cgcggagttc gtgcacaccg tcaccgaggc ggcgaaactg gcgttcgcgg
accgcgaggc 26700ctggtacggc gatcccgcgc acgcggaggt gccggtcggc gacctgctgg
acccggccta 26760caccgcggcc cgccgcgagc tgatcggcag cgaggcgtcc acggagctgc
ggccgggctc 26820accgggaggc cggacaccgg tgctgccgcc cgtccacgac gagtccgccg
gtccggccgg 26880tccctcctgg ctcggcgagc tcgaggaggg catcccggcg gtggtgcgct
ccacggccgc 26940ccggggcgac acctgctgcg tcaccgccac cgacgcccac gggaacatgg
tggtcgcgac 27000gccgagcggc ggctggctga agagttcgcc ggtggtgccc ggtctgggct
tcccgctcgg 27060tacccgtggg cagatggcca cgctcacccg ggggcacgcc aacgcgctgg
ctcccggcaa 27120gcgcccccgc accaccctca gcccgaccct ggtgctccgg gagggcaggc
ccgccctggc 27180gttcggcaca ccgggcggcg accagcagga ccagtggacg ctgcagttct
tcctgaggca 27240caccgaacac ggcatggggc ttcaggaggc cgtcgaggca cggaccttcc
acaccgacca 27300cgtcccgacg tccttcaccc cccggcgttt cgctcccggg acggtgaccg
tcgaaagcgg 27360catgccggag gaaaccatcc aggagctcag gcggcgcggc caccaggtcc
gcacggtcgc 27420cgactacagc ctgagcaagg tgtgtgtcac cggcctggcc agcgacgaca
tggtcatcgc 27480ggcggccagt ccgcgcggcg cgcaggcgta cgccgtcgcg gattgaggat
gccgaccgga 27540tgcgataagt ttccaaacgg aaatcgtctc ggcgaatgga aggggaacgc
atggtgcccc 27600acccgtcact ggacccgggt gaccacatcg ttctgggaga agcacggcag
aacaacctca 27660agggcgtcag cctgcgcatc cccaagggac ggttgaccgt cttcaccggc
gtttcgggat 27720ccggcaagtc ctccctggtc ttcgggacga tcgccgtcga gtcgcagcgg
cagatgaacg 27780agacctaccc cgcgttcatc cgcaaccgcc tccccaagtt cgagcggccc
gacgcggagg 27840tcatcgagaa cctctccacc gccatcgtga tcgaccagcg cccggtcggc
ggcaacgcgc 27900gctccacggt cggcaccatg accgagatcc acgccatgct gcgggtgctg
ttctcccggc 27960acggcaggcc cagcgcgggt ccctcacaca tgtactcctt caacgatccg
cgcggcatgt 28020gcccggagtg cgagggactc ggatccaggg tgcggctgga tttgaaccgc
cttctggacg 28080aggacaagag cctcaacgag ggcgccatcc gcttccagcc cttcgcggtg
ggcaccttcc 28140cgtggcagct gtacgcggag tccgggctgt tcgatcccga cctgccgctg
cgggagttct 28200ccgcggacga ccgcgaactg ctgctgcacg gttccgggtt caaggtcgac
cgggccggcc 28260ggcacggcgt ctacaagaac gagtacgagg gcatcgtgct gcgcttcacc
cggcgctacc 28320tcaaggcggg cctcgacacc ctcaagccga aggaacgggc ggcggtgcag
gaggtcgtga 28380cggaggggcc ctgcgaggcc tgcggaggcg cccggctggg accggccgcg
ctcgcgtcgc 28440ggatcgccgg ggagaacatc gccgactact ccgccctgga ggtcaccgat
ctgatcggcc 28500gcctggagcg caacgacgcc ccaccggtca agccggtggt ccaggcggcg
ctggccgcac 28560tgcgcaggat cgaggccgtc ggactcggct acctcagcct cgaccgccag
accgccacgc 28620tctccggcgg cgaggcgcag cggctgaaga cggtacgcca cctgggcagc
agcctgaccg 28680ggctgacgta catcttcgac gagccgagcg tgggcctgca cccgcgtgac
gtgcgccgtc 28740tgaacgagct gttgctcgcc ctgcgcgaca agggcaacac cgtgctcgtg
gtggagcacg 28800accgggacgt gatcgccatc gccgaccacg tcgtcgacat gggcccgggc
gcgggcagcc 28860agggcggcga ggtggtctac gagggatcgc cgaccgggtt acggggctcg
gacagcccga 28920ccggacgcgg cctgcgttcg gtgccgggac tgaagcgccg actgcgcgcc
cccgacggca 28980ggctgacggt ccgcggcgcg cggctgcaca acctcaagga cgtcacggtc
gacgtgccca 29040ccggtgtgct ggtggcgctg agcggtgtcg ccggctcggg caagagctcc
ctcgcccggg 29100agctggcagc gcggcacccg gaggaaacgg tcgtggtcga ccagtcctcc
atcgggatct 29160cctcccgatc cacccccgcg acgtacaccg acatcatgga caccgtccgg
cggctgttcg 29220cccgcgcatc cggaaccgac cccggcctgt tcagcttcaa ctccgcgggc
gcctgcccgg 29280agtgccaggg ccgcggtgtg atcgagaccg acctcgcgtt catggacccg
gtcaccaccg 29340tctgcgagcg ctgcgagggg cgccgcttca acgacgaggc gctgagccac
accctgtccg 29400gccggaacat cgccgacgtc ctcgccatga cggccgagga ggcgatcggg
ttcttcgcgg 29460aggactccgt ccgccgcaaa ctggccctgc tgacggaggt cggcctcggc
tacctgacgc 29520ttggccgctc cctgtccacc ctgtccggcg gcgaacgcca acggctgaag
ctggcgcacc 29580ggctgcacgc ctccggcagc gtctacatct tcgacgaacc gtccaccggc
ctgcacatga 29640cggacgtggg caagctgctc accctgttcg accgcctcgt cgacggcggc
aacacggtgg 29700tggtcatcga acacgacctc gacgtgctca agtacgcgga ctggatcatc
gatctcggcc 29760cggaggccgg ccggcacggc ggccgggtgg tcttcgaggg caccccggcg
gacctggcgc 29820gggtgcggga atcgcacacc ggccggtgtc tggccgagga cctcgccgca
cacggtcacc 29880tctgacggcc cggagcaccg gctcgccgcc gggtccgggc cggggtcgca
ccccgtcctc 29940ccgcacacct cccgtccgac aaggagtccg tatgcccctc atccacgtca
ccctgctgag 30000cggtcgcggc gaggaggaga tcgccgccct cggccgggcc gtcacggagg
ccgtacacac 30060cacgctgggc accccccggg aggcgatccg ggtgacggtg gacgcatgcc
cgcccgagca 30120ctggttcgtg ggcggcgtct cgatggcgga gaagaaggcg gcccggggcg
gctgagcggt 30180gctccgttcg ccccgggcgc cgcggttcag gccgcgtccg cgaaccgccg
gcggatccgc 30240ccgacgtgct ccgggtccac ggccgggagg tcgtcgcggg tgagccggcc
gctgcgcgtc 30300agcaggtgcg cgttgaaggc ctccggcgcc agccccacgc agcggtcgat
gttctcgaac 30360cactccaggc tggtcagcgc ggtccgctgc atccgctcca cggcgggccg
ccgctcggcc 30420tcgtaggccg ccagagcctc ggggaccgtc ccgtggtcgt gcagggccgt
ggcgagggac 30480agaccgtcct ccatcgccag tttggtaccc gaaccgatcg agaagtgcgt
cgtgtgggcg 30540ctgtcaccga ggagaacgag gttgccgtgg ctccagtcgc ggttgcgtac
ggcagcgaag 30600cgaccccagt acgaccggtt gccccacagc ccgtgtccgt ccagcagacc
ggtgaagtac 30660tccctcaccc tcccgatgct ctcccggtcg cccaccgggt catcggaccg
gcgctccgcg 30720gcgcggaaac cggccgcccg ccacaccgcg tcaccgatct ccacgatgaa
cgtgctgcga 30780ccgggggcgt acgggtaggc gtgggcctgc accgggccgt ggtcggtctc
gaccacggcg 30840aaggtgagcg cgtcgaacgg ccggtccgtg ccgagccaca tgtaacgcga
cccggcctgt 30900tcccgctccg tgccgaacgc ggcctcgtac cgcgcgcggg tgcgcgatcc
cacgccgtcc 30960gcggcgacga cgaggtcgtg ggacgcgcgc aaccgggaca cctcgggcgc
cggggagccg 31020aagtgcagac gcaccccgag gtcggcgcag cgctcctgaa ggagccgcag
cagggtgtgg 31080cggccgatcg cggcgaaccc gtacccctcg ttgcgctgga cccggccccg
gtagcagacg 31140tcgatccgcg tccaccgggc gaactcggcc tcgacagcct cgaacagagc
cgggtcggcg 31200gcctcgatgc cgccgagggc gccgtcggag aagacgacgc cgaacccgaa
cgtgtcgtgg 31260gccgcgttgg cctcccacac ctccacgacg tcgtggggcc gcaggcgctt
gaccaggcag 31320gccgtgtaca gcccgccggg accggctccg atcaccgcta tccgcatgac
acctcgctgg 31380ggctccgaga gagatggatg gacgtgcaga gaccagtaaa tgctatccga
agagaaatga 31440tctggtggtc aatctccttt ccagctccgc gagttccggg actgaccgga
gcagatcgtt 31500gcccacacgt aatcaagtct gacggaagga tgcccacatg ccggacgtcc
tcacccccac 31560cccgctgccg gcggccgacc tggcgggcct cttccgcgcc ctggacccgc
cgccgttcgc 31620cctggtgcgt cgtgccgcac cggacgggac cagcaccggc ccgttcgacg
tgttcatcgg 31680caccatggac accgtgcgac gggtgacgga cctgccgtcg ggcccggccg
tgccgggcgg 31740gggaccccac acactggccc tactgccgta ccggtgcctg gccgaaagag
ggctggactg 31800ccacgacgac ggcacaccac tgagggtcct gcggatccgc cggcggcaca
cggccgacca 31860cgccgcgctc accgcggccc tcgccgcggt gcggcccgcg ggagacctcc
tcggggaagg 31920cgccggcttc gacggctccg acgaggacta cgccgacctg gtccgcgacc
tcatggccga 31980cgaggtggca cgcaccggtc tgcacgtcct gatccgcagg gacttcaccg
cccggttgcc 32040aggacacgga cccgtggtgg tgggcgaact gttccgccgg ctgctggccg
tggagcacgg 32100cgcgtactgg acgttcgcgg tgtacaccgg aggccccgac ggtgccgcac
tggccggagc 32160ctcaccgcag ggtcacgtca cactgcggaa cggccgggtc gtgatgcgcc
cgatgtgcgg 32220cacgctccgc cttccacccg gtggccggcc gagcgccgcc gacctggtgg
ccttcctgcg 32280tgacggcaag gagtccgaag aactgggggc cgtggtcgac gcggaactcg
ccatgctgtg 32340ccggatcagc gagggggacg tacgcctgga aggaccgcgc ctgcgaccga
tggcccgcgt 32400actgcacacc gagtgccgca tcagcgccac cgccgcgctg ccggcccggc
acacgctcgc 32460cggctccctg ttcgcggcga ccgccgtggg ccgccccttc gcggacgcgt
gccgcgtcat 32520cacccgccgc gaaccaaccg ggcgcggtta ctacggcggc ctgatcgcgc
tgctgggcca 32580cgacgacgcg ggaaacgagg aactggacac cgccgtgctc atccgcacct
tcgaggtgtc 32640cgggcagggc cggctgaagc tgtcggtcgg agccaccctc gggccccgct
ccgtggccgc 32700cgacgagacg gccgagacgc gcgccaaggc ctcggccctg gtgtcggcgc
tcgcaagcgg 32760aggaccgact gcggagggcg gtgccgggcg ccacgcgcgg gctggtcttg
gtcgcggccc 32820ggaggcggcc ggcggcccgg ccaccggtga gcgaagcgga gtgccgggtg
accggacgcg 32880gcaccagcag gccgccggcc ggcagcccac gtcccccgcc gacccggcgt
ggcgcccgtc 32940ggtgaccgcg gagggcggtg ccggggacca cgcgcgggct ggtcttggtc
gcggcccgga 33000ggcggccggc ggcccggcca ccggtgaggg aggcggagtg ccgggtgacc
ggacgcggca 33060ccagcaggcc gccggccggc agcccacgtc ccccgccgac ccggcgtggc
gcccgtcggt 33120gaccgcggag ggcggtgccg ggggccacgc gcgggctggt cttggtcgcg
gcccggaggc 33180ggccggcggc ccggccaccg gtgagggagg cggagtgccg ggtgaccgga
cgcggcacca 33240gcaggccgcc ggccggcggc ccacgtcctc cgccgacccg gcgtggtgcc
cgtcggtgac 33300cgcggaactg gaccggcgcc gcgcacgtct gtccgcctac tggcaacgcc
cccgtcggcc 33360gggcagccgg cccgctccgc ggccaccggt gctgctcgtc gacacgggtg
gcgaggagac 33420ggcgccgctg gccgccatgc tgcgcggact gggccgcacc gtcgacgtgc
gtcccgcgta 33480ccccgcggcg gccgcgccac ggaccgtcgc gcccggaacc acggtcgtcc
tcggccccgg 33540cccgggtgac ccgttggccc acggcgacga ccgcatcacc gcgctgcggg
ccatgacgtc 33600cgccctcctg tccagcggag cacccacgtt cggggtcggg ctcggcttcc
acctcctgct 33660cgccgtgctg ggtctggccg gggccgcgcg agcgtgggac ggggccaccg
gccagcggga 33720gatcgaggtc ttcggcagac gcgcgacggt ggggtacggt ggcacgcaca
ccgtggtggc 33780cggcccgcac acggacaccc tcgcgcggcg gctgtccctg acgctctgct
acggcccggc 33840ccacggcgag ctggtggcca tgcgaggccc tcgaaccggc ggcgtcgcct
tcctcccggc 33900atcggtgctg agcgtcgagg gggcggagct gctggatctc ctgctgccct
gagcgagcgg 33960gtgacgggcc gggccccggc cgctcacccg ttcgccgtgc ccccgagctc
gagcacggtc 34020cgcgcggccg actccagcgc gagtgtcatc gagccgccgt tgggctcgaa
cgaggtgtgg 34080tcaccggcga agtggatccg gccctccggc cggcgcatgg ccggcatcag
ggaactgtgc 34140cccatctcgg gaaggatgta ggcgccttcg atgtacggct gctggtccca
gaccaccgag 34200gtgccggtct cgaagtgctc gcgggcaccg ggcagcatcg actcgacgtg
ttccagggcg 34260aagcggatcc gctcctcggg gctcagcacg gcaagcgccc gggcccgcca
gccggttatc 34320aggcactcca gtatcttgcg gggcccgggc aaccggggag tggcgtcccg
gacccaccgc 34380accggcaggt ccgtggagaa actcgcgttg tcctgggccc agaaccgccg
gcgcatctgg 34440aggtagacgc gcacgatcga cgagtacttc acccggcgca tcacggcctg
tttggcgtcg 34500gacagcccgg cgtccgtgaa gtcgatgtgc cggatcgcgc tgaacggcac
cgtcaccacc 34560acgcggtccg cgtccacgga gcgcatgcgg gtgccgtcca ggaaggtgac
acgagcgccc 34620tcgtcgtcct gggcgacccg caccaccgga gcgcggtagc ggatgcggtc
cttcagccgc 34680tcggcgaacg cccgggggaa gcggtcggtt ccgcccttca ccttcgacca
ccgggggtcg 34740gcgttggcca gcgagtgcgg gctggactcg tgccgcagcc aggacagggc
ggaggccgtc 34800ttgaggtccc cgccgcgcat ctccaggaag tgcggttcca ccaggtcgat
cgccgccggc 34860gaagcgcccc gcccggacag cacctcgtac accgagcgcc ggtcgtacgg
ttcgagcagg 34920ggggtcggcg cccagtccgc agccgtgatg tcgggttcca gcgcctcgta
ggcgcggcgc 34980acatagcggt cgatcatgtc ggtgacgctc agccccttct cgtgcggggc
gagcgggagc 35040ccggcgcggt ccagcgagtc ggcgtcggga ccgaagaacc ggttgccgac
gaagtacgag 35100aactggctgc cgaccaggtc ggccgtctcc agttccaccc ccagttcgcg
caggtagtgc 35160atggcgtagt ggcagtgcgg ggtcagcgtc atcgcgccgg cctcggcgta
cagaccgtcc 35220gtgaacggct cgcgcagtgt gtacgcacgc ccgcccggac ggttcgacgc
ctccaggacc 35280gtcacgtcag tgccgcgccg ggccagttcg taggccacgg cgaggcccga
caggcccgca 35340cctatgacga cgaccctgcc gggcacccgg agtcccggca ttcccctgtc
gaactcgcga 35400cgcacgtcgg tctgcgtgac ctcggccatc ggctctccct cccccgtgac
tgtgcacggt 35460tgccgttcat atgattgacg tcaggaaatc atattcggat gctctcaacg
gtaaagcctt 35520gcatctcagg tgtgttgagc cccgccccga cttggtagct gctgaccagc
aatcgttgcc 35580ctcgtcggag ccgtttcctt cgcacccggc cgtcgttcgg cagggcatgg
acaccgcacc 35640gatcctcgtc ctcggagcca ccggctccac gggccggcgt gtcaccgcac
acctgcgggc 35700tctgggagcg ccggtcagag gcgcctcccg gcacagcgcc atccgattcg
actggaacga 35760ccgcagtacg tgggaaccgg ccctgcgggg cgcgaaccgc atgttcctga
tggcccccga 35820cggcatcccc gtccacccgg acctcgtcgg cctggccgcg gacctcggcg
tcgaacgcat 35880cgtcctgctg tccagccgcg ccatcgaggt gataggagac cagaggctga
tcgcggccga 35940ggacaccgtc aagaagtccg gagccgcctg gacgatcctg cgcatcgact
ggttcgacca 36000gaacttcgac gagggcccct tccgcgacgc ggtccgcgcc ggtgagctgg
cgctgccgct 36060gggggattgc cgacaggggt tcgtggacct ggacgacgtg ggggccgctg
ccgcccggac 36120tctgaccgag gacggacaca ccggccggac gtacgaactg accgggccgc
aggcgctctc 36180cttcggcgag gcgtgcgccg tcatccagga cgtcaccggc cgcccgatga
ggttccacgg 36240cggcgacgac gcctaccgcg ccgcgcagac gtccttcggc cgctccgagg
agtccgtcga 36300acgcgacatc accgcctacg cggcactccg ctccctcggc gacaccgagc
ctctggacac 36360cgtgccccgc ctgaccgggc gccgggcccg cacgttccgg gagtacgtca
ccgaggccgc 36420gtccgggaac gactggcccc gcccctgaca ggctcacggt gggcaccggc
atgcccccgg 36480ctgtcgggac ggggggcgag ccgcggtcgt cagcggacgc cggtgccgga
ccgccgggag 36540acgagccccg tctcgtaggc gaggatcacc gcctggatcc ggtctctcag
cgccagcttc 36600ccgaggacgt tcgccacatg cgtcttgacg gtcgtttcgg cgaccgagaa
ctcggcggcg 36660atctcggcgt tggacagtcc acgggccatc gcgtggaggg tgtccctctc
acgggcggtg 36720agggccctca ggcccgctgg cggctgcggg ggcgccacct cgagtgcggc
gaatcggtcg 36780agaaggcgtc gggtgacgcg gggggcgatg acggcgtcgc ccgccgcgac
cgtgcggatg 36840gcgtcgacga gggactcggc cgggccgtcc ttgacgagga agccgctggc
gcctgctcgg 36900agggcgtcga gtacgtgctc gtccaggtcg aaggtggtca ggatgatgac
gcgggtgtcg 36960gaggtggagt cgaggagctg tcgcgtggcg tcgatgccgt tcgtgcccgg
catgcggatg 37020tccatgagga cgacgtcggg ctgcagggct cgggcgtctc ggaccgcggc
ccggccgtcc 37080gccgactccc cgacgacggt gaggcccggc tccgcctcca ggatgtagcg
gaagccggtt 37140cggatc
371464192PRTStreptomyces species FH6421 4Leu Leu Glu Ala Ala
Ala Gln Leu Phe Ala Glu Gln Gly Tyr Ala Ala1 5
10 15Thr Ser Val Asn Asp Met Ser Ala Arg Ser Gly
Arg Thr Ser Gly Ala 20 25
30Val Tyr Phe His Tyr Ala Gly Lys Glu Ala Val Ala Val Ala Val Val
35 40 45Gln Asp Arg Phe Ala Thr Trp Pro
Gln Leu Ala Ala Arg Tyr Ala Asp 50 55
60Glu Ala Val Pro Pro Val Asp Arg Leu Val Ala Leu Ser Tyr Asp Ile65
70 75 80Ala His Ala Leu Ala
Glu Asp Pro Val Thr Arg Ala Gly Ala Arg Leu 85
90 95Trp Ala Glu Arg Ala Thr Ile Asn Val Pro Leu
Pro His Pro Phe Ala 100 105
110Leu Trp Thr Thr Ala Ala Thr Arg Leu Leu Ala Lys Ala Arg Leu Ala
115 120 125Gly His Leu His Pro His Val
Arg Pro Ala Arg Ala Ala Arg Thr Leu 130 135
140Val Pro Ala Phe Phe Gly Leu Cys Ala Leu Thr Glu Glu Leu Glu
Gly145 150 155 160Thr Ala
Ala Leu Thr Asp Arg Leu Thr Asp Trp Trp Gln Leu Thr Leu
165 170 175Pro Cys Leu Arg Pro His Pro
Val Arg Glu Asp Leu Pro Arg Gly Arg 180 185
1905162PRTStreptomyces species FH6421 5Leu Met Thr Arg Thr
Asp Ser Pro Arg Ser Ala Ser Glu Thr Ser Ala1 5
10 15Val Pro Gly Glu Leu Glu Asp Ala Leu Ser His
Leu Gln Cys Ile Leu 20 25
30Val Ala Arg Arg Thr Arg Ser Asn Pro Glu Gly Val Thr Trp Gln Gln
35 40 45Tyr Asp Val Leu Glu Leu Leu Arg
Ile Arg Gly Pro Met Thr Pro Ser 50 55
60Val Leu Ser Ala Ser Leu Gly Val Ser Arg Ala Thr Thr Ser Lys Ala65
70 75 80Leu Arg Val Leu Lys
Asp Leu Glu Leu Val Asp Gln Arg Thr Leu Gly 85
90 95Glu Asp Arg Arg Glu Leu Thr Thr Ser Leu Thr
Pro Ala Gly Glu Ala 100 105
110Phe Leu Thr Arg Val Ala Glu Gln Arg Arg Glu Asn Ala Arg Ile Val
115 120 125Glu Ala Ala Leu Ser Pro Gly
Glu Arg Ala Met Phe Val Glu Leu Cys 130 135
140His Lys Val Ala Gly Ala Leu Glu Pro His Leu Glu Pro Val Glu
Ala145 150 155 160Arg
Pro6610PRTStreptomyces species FH6421 6Val Leu Thr Glu Asn Ala Ser Gly
Glu Ala Arg Ser Ala Val Pro Leu1 5 10
15Thr Leu Thr Glu Gly Phe Asp Arg Val Val Arg Ala Ala Gly
His Gln 20 25 30Val Ala Leu
Val Ser Gly Thr Glu Thr Val Thr Tyr Arg Gln Leu Asn 35
40 45Glu Arg Ala Glu Arg Val Ala Arg Gly Leu Gly
Ala Arg Lys Val Ala 50 55 60Pro Gly
Asp Arg Val Gly Val Tyr Leu Arg Arg Ser Pro Asp Leu Tyr65
70 75 80Ala Val Met Leu Gly Val Leu
Lys Ala Gly Ala Cys Val Val Pro Val 85 90
95Asn Pro Asp His Pro Ala Pro Phe Val Ser Arg Val Val
Ala Glu Ser 100 105 110Ala Pro
Arg Ala Val Val His Asp Ala Gly Thr Pro Ala Val Ala Pro 115
120 125Ala Ala Pro Gly Ala Pro Leu Trp Val Pro
Val Glu Glu Leu Thr Thr 130 135 140Ala
Ala Glu Pro Asp Asp Gly Val Ala Leu Pro Ala Val Asn Asp Pro145
150 155 160Asp Ser Thr Ala Phe Leu
Met Phe Thr Ser Gly Ser Thr Gly Arg Pro 165
170 175Lys Gly Val Arg Ile Ala His Arg Gly Leu Ala Arg
Leu Gly Pro Tyr 180 185 190Ser
Gly Glu Leu Arg Met Gly Pro Gln Asp Cys Leu Val Gln Ser Ala 195
200 205Ala Phe Ser Phe Ala Ala Ser Thr Ile
Glu Ile Trp Leu Ala Leu Leu 210 215
220His Gly Ala Arg Leu Val Val Met Pro Gln Gly Leu Pro Ser Leu Pro225
230 235 240Ala Leu Lys Asp
Ala Val Val Arg His Gly Val Thr Ala Leu Ser Leu 245
250 255Pro Cys Gly Leu Phe Asn Leu Leu Val Asp
Glu Glu Pro Glu Cys Leu 260 265
270Arg Gly Leu Arg Val Ile Leu Leu Ser Gly Asp Phe Pro Ser Pro Glu
275 280 285His Leu Ser Arg Ala Ala Arg
Ala Thr Arg Ala Val Ile Tyr Asn Gly 290 295
300Tyr Gly Cys Thr Glu Asn Ser Ser Ile Thr Ala Leu Tyr Pro Ile
Arg305 310 315 320Asp Ala
Gly Asp Val Thr Arg Glu Asn Arg Val Pro Val Gly Arg Pro
325 330 335Leu Pro Gly Val Thr Leu Glu
Val Leu Asp Asp Ser Leu Arg Pro Cys 340 345
350Pro Pro Gly Thr Pro Gly Gln Leu Val Val Gly Gly Leu Gly
Leu Ala 355 360 365Gln Gly Tyr Leu
Asn Asp Pro Glu Leu Thr Asn Arg Lys Phe Val Thr 370
375 380Gly Pro Asp Gly Arg Pro Arg Tyr Leu Thr Gly Asp
Leu Ala Arg Ala385 390 395
400Thr Glu Asp Gly Asp Ile Val Leu Ile Gly Arg Ala Asp Ser Met Val
405 410 415Lys Ile Arg Gly Tyr
Arg Val Glu Leu Thr Ala Val Thr Leu Ala Leu 420
425 430Arg Ala Leu Asp Gly Ile Gly Asp Ala Val Val Lys
Ala Phe Pro Glu 435 440 445Gly Ala
Gly Glu Lys Ser Leu Thr Ala Phe Tyr Thr Thr Val Asp Gly 450
455 460Arg Pro Leu Asp Gly Ala Asp Leu Ala Arg Arg
Met Gly Asp Gln Leu465 470 475
480Pro Ser Tyr Met Val Pro Ser Thr Phe His His Leu Gly Asp Leu Pro
485 490 495Arg Asn Ala Asn
Gly Lys Ile Asp Arg Ser Ala Leu Thr Asp Pro Ser 500
505 510Asp Thr Asn Arg Asp Pro Lys Lys Gly His Thr
Ala Val Gln Asn Pro 515 520 525Leu
Glu Thr Val Val Leu Gln Ala Trp Lys Asp Ile Ser Gly Ala Asp 530
535 540Asp Phe Thr Thr Thr Asp Ser Phe Leu Gly
His Gly Gly Asn Ser Leu545 550 555
560His Phe Val Gln Leu Ala Ser Arg Leu Gln Lys Ile Phe Gly Val
Glu 565 570 575Val Ser Thr
Glu Asp Val Phe Arg His Gly Thr Val Glu Gln Leu Ala 580
585 590Arg Phe Val Glu Gln Ser Arg Asp Thr Gly
Arg Asn Pro Ala Ala Gln 595 600
605Thr Gln 61071535PRTStreptomyces species FH6421 7Val Asn Ser Pro Leu
Arg Thr Thr Val Leu Asp Leu Ala Arg Thr Thr1 5
10 15Leu Gly Ser Ala Asp Leu Thr Ala His Glu Pro
Leu Ala Asp Arg Cys 20 25
30Glu His Pro Ala Leu Leu Asp Asp Leu Ala Thr Thr Leu Thr Ala Val
35 40 45Phe Ala Val Glu Ile Thr Gly Ala
Asp Leu Ala Ala Gly Ala Thr Val 50 55
60Ala Asp Val Ala Ala Arg Met Asp Asp Arg Arg Asp Ala Pro Arg Ile65
70 75 80Pro Glu Leu Arg Ala
Gly Leu Ala Pro Arg Asp Gly Arg Ala Val Glu 85
90 95Ala Ser Phe Gly Gln Ser Gly Ile Trp Leu Ile
Asp Gln Tyr Leu Pro 100 105
110Asn Pro Ala Ala Tyr Asn Gly Pro Phe Phe Val Arg Leu Pro Phe Ser
115 120 125Ala Asp Pro Asp Arg Leu His
Ala Ala Val Arg Gly Val Leu Arg Arg 130 135
140Gln Glu Val Leu Arg Thr Thr Tyr Ala Leu Ser Asp Gly Thr Leu
Arg145 150 155 160Gln Asn
Val Ser Arg Asp Asp Asp Ala Val Val Phe Glu Val Ala Arg
165 170 175Tyr Gly Asp Asp Lys Glu Leu
Asp Ala Leu Val His Arg Val Ala Asn 180 185
190Leu Arg Leu Asp Leu Ala Arg Gly Pro Val Ile Ala Val Thr
Cys Ala 195 200 205Leu Gly Pro Ala
Asn Arg Ser Ala Val Ile Cys Asn Ile His His Ile 210
215 220Ala Ser Asp Ala Ala Ser Ala Gly Val Phe Leu Arg
Glu Leu Leu Asp225 230 235
240Ala Tyr Asp Arg Leu Gly Arg Gly Leu Pro Val Glu Ala Asp Pro Leu
245 250 255Arg Pro Thr Tyr Gly
Asp Phe Ser Gln Trp Tyr Arg Glu Leu Met Asn 260
265 270Pro Glu Ala Leu Thr Arg Ser Leu Asp His Phe Ala
Ala Arg Leu Ala 275 280 285Gly Glu
Leu Pro Val Leu Asp Leu Pro Thr Asp Arg Pro Arg Pro Pro 290
295 300Val Lys Gln His Arg Gly Gly Thr Leu Pro Leu
His Leu Pro Ala Ala305 310 315
320Ala Ala Asp Asp Phe Glu Ala Leu Ala Arg Thr Glu Gly Val Thr Leu
325 330 335Phe Met Ala Leu
Val Ala Ala Tyr Ala Val Phe Leu Ser Arg His Thr 340
345 350Gly Gln Arg Arg Val Leu Ile Gly Ser Pro Val
Ser Leu Arg Asp Asp 355 360 365Pro
Ala Thr His Glu Leu Ile Gly Tyr Phe Val Asn Leu Val Val Leu 370
375 380Gln Gln Glu Ile Asp Asp Arg Met Thr Val
Arg Asp Val Leu Arg Arg385 390 395
400Ala Arg Glu Glu Val Ser Glu Ala Leu Arg His Lys Trp Ala Pro
Phe 405 410 415Asp Lys Val
Val Glu Arg Leu Gln Pro Pro Arg Ser Ser Gly Tyr Thr 420
425 430Pro Leu Val Gln Thr Met Leu Val Leu Thr
Gln Gly Asp Ala Gly Arg 435 440
445Ile Ser His Asp Asp Thr Glu Leu Arg Ile Glu Arg Gly Ala Ala His 450
455 460Gly Ala Lys Tyr Asp Leu Ser Leu
Val Phe Glu Arg Asp Ser Glu Gly465 470
475 480Leu His Gly Leu Ile Glu Tyr Asp Ala Asp Leu Phe
Asp Glu Pro Thr 485 490
495Val Arg Ala Met Gly Asp Arg Leu Arg His Leu Met Glu Gln Phe Ala
500 505 510Arg Arg Pro Asp Ala Pro
Leu His Glu Leu Glu Ala Leu Gly Ala Gln 515 520
525Glu Arg Arg Ser Val Leu Val Arg Gly Asp Arg Thr Ala His
Ala Val 530 535 540His Asp Ala Pro Val
Met Glu Leu Phe Glu Ala Gln Ala Arg Ala Thr545 550
555 560Pro Asp Ala Val Ala Leu Glu Asp Gly Asp
Thr Thr Leu Ser Tyr Arg 565 570
575Glu Leu Asp Glu Arg Ala Asn Arg Leu Ala His Val Leu Arg Ala Ser
580 585 590Gly Ala Ala Ala Gly
Thr Arg Val Gly Ile Cys Leu Pro Arg Ser His 595
600 605Asp Met Val Val Ala Leu Phe Ala Ile Leu Lys Thr
Gly Ala Ala Tyr 610 615 620Val Pro Leu
Asp Pro Ser Tyr Pro Arg Gln Arg Ile Thr His Thr Leu625
630 635 640Arg Asp Ala Gly Val Phe Leu
Thr Val Thr Asp Ser Ser Leu Ala Asp 645
650 655Glu Leu Pro Pro Arg Glu Pro Leu Phe Val Leu Asp
Arg His Asp Gly 660 665 670Pro
Ile Ala Ala Ala Pro Ala Thr Gly Leu Gly Arg Val Lys Thr Pro 675
680 685Asp Asp Glu Ile Tyr Val Val His Thr
Ser Gly Ser Thr Gly Leu Pro 690 695
700Lys Gly Val Val Ile Ala Asp Arg Thr Val Ala Asn Leu Val Arg Ala705
710 715 720Gln His Arg Cys
Ser Pro Ala Gly Ala Thr Gly Arg Thr Leu Gln Tyr 725
730 735Met Ser Leu Ser Phe Asp Val Ser Val Met
Glu Ile Leu Gly Thr Leu 740 745
750Cys Val Gly Gly Thr Leu Val Leu Val Ser Glu Glu Leu Arg Lys Asp
755 760 765Leu His Ala Leu Ala Gly Phe
Leu Ala Glu Arg Arg Val Thr Arg Val 770 775
780Tyr Leu Pro Tyr Ile Ala Leu Gln Gln Leu Ala Ser Leu Ala Thr
Asp785 790 795 800Ala Gly
Val Arg Leu Asp Asp Leu Arg Glu Ile Thr Ser Val Gly Glu
805 810 815Ala Leu Val Val Ser Pro Gln
Ile Arg Glu Phe Ala Thr Arg His Pro 820 825
830Ala Val Arg Leu Val Asn Met Tyr Gly Pro Ser Glu Thr His
Leu Ala 835 840 845Ser Trp Tyr Pro
Leu Thr Gly Ser Pro Ala Thr Trp Pro Asp Arg Pro 850
855 860Pro Ile Gly Arg Pro Val Asp Gly Val Arg Leu Val
Val Leu Asp Ala865 870 875
880His Met Arg Leu Val Pro Pro Gly Val Pro Gly Glu Leu Tyr Ile Gly
885 890 895Gly Pro Val Leu Ser
Pro Gly Tyr Arg Asn Arg Pro Asp Glu Thr Ala 900
905 910Arg Arg Phe Leu Pro Asp Pro Phe Gly Gly Pro Ala
Asp Arg Leu Tyr 915 920 925Arg Thr
Gly Asp Leu Val Arg Trp Asn Ser Glu Gly Asp Leu Glu Tyr 930
935 940Leu Gly Arg Thr Asp Asp Gln Ile Lys Ile Arg
Gly Tyr Arg Ile Glu945 950 955
960Pro Ala Glu Ile Glu Ala Ala Leu Asp Asp Leu Asp Gly Val Ala Ser
965 970 975Ser Ala Val Ala
Ala Val Asp Val Ala Pro Gly Asp Arg Arg Leu Val 980
985 990Ala Val Leu Glu Thr Ser Arg Thr Trp Glu Thr
Ala Glu Leu Arg Arg 995 1000
1005Ala Leu Ser Gly Thr Leu Pro Asp Tyr Met Val Pro Ala Leu Val
1010 1015 1020Val Ala Val Glu His Met
Pro Thr Thr Pro Ser Gly Lys Ile Asp 1025 1030
1035Arg Arg Ala Val Ala Gly Leu Ala Ala Ala Gln Ala Thr Ala
Ala 1040 1045 1050Arg Thr Ala Pro Ala
Pro Pro Gly Arg Pro Pro Arg Pro Gly Leu 1055 1060
1065Glu Gln Arg Ile Ala Arg Glu Trp Ala Asp Val Leu Lys
Val Pro 1070 1075 1080Ala Val Gly Arg
Asp Glu Asp Phe Phe Ser Val Gly Gly Asn Ser 1085
1090 1095Ile Ile Ala Thr Glu Leu Val Tyr Arg Leu Arg
Arg Ala Phe Asp 1100 1105 1110Gln Asp
Leu Ser Leu Arg Ala Leu Leu Glu Asn Pro Thr Val Ala 1115
1120 1125Gly Met Ala Ala Arg Leu Arg Ser Gly Pro
Gly Ala Pro Thr Thr 1130 1135 1140Ala
Pro Ala Ala Leu Arg Glu Asp Ala Thr Leu Pro Asp Asp Leu 1145
1150 1155Pro Ala Val Thr Gly Thr Pro Val Pro
Val Ala Arg Ala Arg Glu 1160 1165
1170Val Leu Leu Thr Gly Ala Thr Gly Phe Leu Gly Ser Tyr Leu Leu
1175 1180 1185Arg Glu Leu Thr Gly Thr
Thr Gly Gly Arg Val His Cys Leu Val 1190 1195
1200Arg Ala Ala Asp Glu Arg Ala Gly Met Glu Arg Leu Arg Ala
Thr 1205 1210 1215Ala Glu Arg Tyr Arg
Leu Asp Gly Arg Ile Asp Trp Asn Arg Val 1220 1225
1230Arg Ala Val Pro Gly Asp Leu Ser Arg Pro Gly Phe Gly
Leu Pro 1235 1240 1245Gly Ala Glu Tyr
Asp Ala Leu Ala Gly Thr Val Asp Val Val Tyr 1250
1255 1260His Ala Ala Ala His Ile Asn Phe Val Leu Pro
Tyr Ala Ser Val 1265 1270 1275Lys Pro
Thr Asn Val Asp Gly Phe Arg His Val Val Arg Phe Ala 1280
1285 1290Ala Thr Asp Arg Pro Lys His Val Gln Tyr
Met Ser Thr Ile Ala 1295 1300 1305Val
Phe Pro Pro Gly Glu Ala Pro Asp Gly Thr Val Leu Thr Glu 1310
1315 1320Asp Asp Val Pro Glu Ala Cys Glu Arg
Leu Gly Ile Gly Tyr Thr 1325 1330
1335Gln Ser Lys Trp Val Ala Glu Arg Ile Ala Leu Ala Ala Arg Ala
1340 1345 1350His Gly Val Pro Val Thr
Ile His Arg Ile Gly Arg Ile Ser Gly 1355 1360
1365Asp Ser Val Thr Gly Ala Cys Gln Ser Asp Asp Phe Leu Trp
Arg 1370 1375 1380Gln Ile Lys Ser Phe
Ile Glu Leu Gly Ser Ala Pro Pro Ala Glu 1385 1390
1395Asp Leu Thr Thr Asp Leu Leu Pro Val Asp Phe Val Ala
Arg Ala 1400 1405 1410Val Val Ala Leu
Ser Arg His Pro Ala Thr His Asn Arg Thr Leu 1415
1420 1425His Val Phe His Pro Ser Gly Ser Asp Phe Thr
Pro Val His Ala 1430 1435 1440Ala Leu
Arg Ala Asp Gly His Arg Leu Glu Ile Val Pro Ala Asp 1445
1450 1455Thr Trp Leu Ala Arg Leu Glu Glu Ser Ala
Arg Arg Pro Gly Gly 1460 1465 1470Asn
Ala Leu Ala Ala Ala Val Pro Leu Phe Arg Glu Gly Ala Leu 1475
1480 1485Glu Leu Gly Asp Asn Thr Tyr Gly Asn
Thr Ala Thr Thr Arg Leu 1490 1495
1500Leu Met Asp Leu Gly Leu Pro Trp Pro Ala Ile Asp Glu Gln Ala
1505 1510 1515Ile Thr Arg Met Leu Arg
Tyr Phe Arg Ser Val Gly Glu Leu Ala 1520 1525
1530Asp Asp 15358406PRTStreptomyces species FH6421 8Met Pro
Thr Ser Ser Cys Pro Asp Thr Ala Tyr Asp Thr Leu Ile Pro1 5
10 15Ser Val Val Ala Ala Leu Pro Ala
Ala Gln Gln Pro Glu Trp Pro Asp 20 25
30Pro Gly Arg Leu Ala Leu Val His Thr Glu Leu Ala Arg Ala Asp
Pro 35 40 45Leu Val Thr Tyr Asp
Ser Val Arg Ala Leu Arg Arg Leu Leu Ser Arg 50 55
60Ala Ala Glu Gly Glu Leu Cys Val Leu Gln Ala Gly Asp Cys
Ala Glu65 70 75 80Asp
Pro Ala Glu Cys Gly Pro Ala Pro Leu Ala Arg Lys Ala Glu Met
85 90 95Leu Asp Val Leu Ser Asp Ile
Val Arg Thr Gly Ala Gly Arg Pro Val 100 105
110Val Arg Val Gly Arg Val Ala Gly Gln Tyr Ala Lys Pro Arg
Ser His 115 120 125Pro Glu Glu Leu
His Asp Gly Val Arg Leu Pro Val Tyr Arg Gly Pro 130
135 140Met Val Asn Ala Pro His Pro Asp Ala Asp Ala Arg
Arg Pro Asp Pro145 150 155
160Ala Arg Ile Leu Ser Cys Tyr Arg Ala Ala Arg Arg Ala Val Glu Ser
165 170 175Leu Asp Arg Leu Gly
Arg Gly Glu Gly Ser Pro Ala Glu Thr Arg Val 180
185 190Trp Thr Ser His Glu Ala Leu Leu Leu Asp Tyr Glu
Leu Pro Leu Val 195 200 205Arg Arg
His Arg Ser Gly Arg Ser Tyr Leu Ala Ser Thr His Trp Pro 210
215 220Trp Val Gly Glu Arg Thr Arg Gln Pro Asp Gly
Ala His Val Arg Leu225 230 235
240Leu Ala Glu Val Asp Asn Pro Val Ala Cys Lys Val Gly Pro Thr Thr
245 250 255Thr Val Glu Gln
Val Leu Ala Leu Cys Thr Ala Leu Asp Pro Glu Arg 260
265 270Ser Pro Gly Arg Leu Ser Leu Val Ala Arg Phe
Gly Ala Ser Arg Ile 275 280 285Asp
Gly Leu Ala Pro Leu Val Arg Ala Val Arg Arg Ala Gly His Pro 290
295 300Val Leu Trp Leu Cys Asp Pro Met His Gly
Asn Gly Glu Arg Thr Ala305 310 315
320His Gly Leu Lys Thr Arg Arg Leu Ser Ala Val Met Ala Glu Ile
Ser 325 330 335Arg Phe Val
Asp Ile Val Ser Ala Glu Gly Gly Arg Ser Ala Gly Leu 340
345 350His Leu Glu Ala Ser Pro Asp Asp Ile Ala
Glu Cys Thr Gly Ala Gly 355 360
365Phe Thr Pro Ala Pro Gly Pro Ala Tyr Arg Thr Leu Cys Asp Pro Arg 370
375 380Leu Asn Leu Val Gln Ala Val Ala
Ala Thr Ala Tyr Trp Arg Leu Pro385 390
395 400Ala Leu Glu Ala Val Ala
4059660PRTStreptomyces species FH6421 9Met Ser Glu Ser Ala Arg Asn Ala
Arg Gly Leu Ala Ala Leu Leu Pro1 5 10
15Pro Pro Gly Thr Pro Phe Ala Val Leu His Arg Pro Gly Ala
Gly His 20 25 30Pro Gly Thr
Val Asp Val Val Ser Gly Pro Leu Arg Thr Ala Ala Thr 35
40 45Leu Ala Glu Leu Ser Leu Asp Asp Glu Ser Ala
Pro Ala Ser Gln Gly 50 55 60Pro Gly
Pro Ala His Arg Val Leu Ala Leu Val Pro His Arg Gln Ile65
70 75 80Ala Glu Arg Gly Phe Ala Ala
Pro Asp Asp Gly Thr Pro Leu Leu Ala 85 90
95Met Asp Ile Gly Thr Gln His Thr Val Pro Leu Glu Arg
Met Leu Ala 100 105 110Leu Leu
Pro Asp Arg Glu Leu His Val Glu Glu Thr Gly Phe Asp Leu 115
120 125Asp Asp Asp Arg Tyr Ala Ala Gly Val Asp
Ala Leu Thr Arg Gln Glu 130 135 140Ile
Gln Arg Gly Gln Gly Ala Asn Phe Val Leu Ala Arg Ser Leu His145
150 155 160Gly Arg Ile Arg Asp Phe
Asp Arg Thr Arg Ala Leu Ala Ala Leu Arg 165
170 175Arg Leu Leu Ile Ala Glu Ser Gly Ala Tyr Trp Thr
Tyr Leu Val Cys 180 185 190Thr
Gly Asp Arg Tyr Leu Ile Gly Ser Ser Pro Glu Gln His Val Arg 195
200 205Val Ala Gly Ser Arg Val Ser Met Asn
Pro Ile Ser Gly Thr Tyr Arg 210 215
220Tyr Pro Glu Gly Gly Arg Pro Asp Arg Glu Ser Leu Leu Arg Phe Leu225
230 235 240Ala Asp Pro Lys
Glu Ile His Glu Leu Tyr Met Val Val Asp Glu Glu 245
250 255Leu Lys Met Met Thr Glu Leu Cys Gly Ser
Arg Val Arg Val Ser Gly 260 265
270Pro Thr Leu Ala Trp Met Ser Arg Leu Ala His Thr Gln Tyr His Leu
275 280 285His Gly Glu Ser Pro Leu Pro
Leu Thr Asp Ile Leu Arg Gly Thr Leu 290 295
300Pro Ala Pro Thr Val Thr Gly Ser Pro Val Glu Asn Ala Cys Arg
Val305 310 315 320Ile Ala
Arg His Glu Pro Ala Gly Arg Gly Tyr Tyr Ser Gly Val Leu
325 330 335Ala Leu Ala Gly Gln Glu Gly
Gly Arg Arg Ala Leu Asp Ala Val Ile 340 345
350Val Leu Arg Thr Ala Asp Ile Thr Ala Asp Gly Ser Val Arg
Leu Thr 355 360 365Thr Gly Ala Thr
Val Val Arg Asp Ser Val Pro Arg Glu Glu Ala Ala 370
375 380Glu Thr Thr Ala Lys Ala Ala Gly Leu Leu Thr Ala
Leu Thr Arg Gly385 390 395
400Pro Ala Gly Arg Ser Ala Ala Pro Ala His Ala Ala Pro Asp Val Ser
405 410 415Leu Gly Ala Asp Pro
Ala Val Arg Ala Ala Leu Arg Ser Arg Asn Asp 420
425 430Gly Ile Ala Ala Phe Trp Leu Gly Gly Gly Ala Arg
Leu Pro Ala Pro 435 440 445Ser Pro
His Gly Pro Arg Val Ala Val Ile Asp Ala Glu Asp Arg Phe 450
455 460Thr Ser Met Leu Ala Gln Gln Leu Arg Ala Val
Gly Cys His Val Thr465 470 475
480Leu His Pro Trp Trp Ser Val Pro Glu Ala Ala Asp Asp Pro Gly Thr
485 490 495Val Leu Leu Leu
Gly Pro Gly Pro Gly Asp Pro Arg Asp Val Gly Asp 500
505 510Pro Arg Val Ala Arg Leu Arg Ser Leu Ala Gly
Arg Arg Leu Ala Arg 515 520 525Arg
Leu Pro Leu Ala Ala Val Cys Leu Gly His Gln Ala Val Cys Gly 530
535 540Val Leu Gly Leu Pro Leu Val Arg Leu Ala
Arg Pro Arg Gln Gly Ala545 550 555
560Arg Met Arg Val Gly Leu Trp Gly Arg Asp Arg His Val Gly Phe
Tyr 565 570 575Asn Ser Phe
Thr Ala Arg Ser Asp Thr Asp Arg Cys Pro Leu Pro Gly 580
585 590Arg Asp Ala Thr Ala Arg Val Trp Arg Arg
Asp Gly Gly Asp Val Val 595 600
605Ala Leu Asp Gly Pro Gly Leu Ala Thr Val Gln Phe His Ala Glu Ser 610
615 620Leu Leu Thr Glu Asp Gly Pro Asp
Ile Leu Arg Glu Leu Val Asp Arg625 630
635 640Ala Ala Arg Thr Glu Arg Arg Thr Glu Ala Leu Met
Ser Arg Arg Ala 645 650
655Lys Glu His Ala 66010199PRTStreptomyces species FH6421
10Val Asn Pro Pro Gly Thr Val Val Ala Asn Ala Ala Leu Asp Pro Arg1
5 10 15Glu Leu Arg Arg Thr Met
Gly His Phe Ala Thr Gly Val Thr Val Leu 20 25
30Thr Cys Arg Arg Gly Ala Arg Leu His Gly Ala Thr Val
Asn Ser Phe 35 40 45Thr Ser Val
Ser Leu Asp Pro Pro Leu Ala Leu Val Ala Leu Asp Arg 50
55 60Arg Thr Arg Ala Ala Ala Leu Leu Asp Asp Gly Pro
Phe Val Val Asn65 70 75
80Leu Leu Gly Glu His Gln Gln Asp Leu Ala Leu His Phe Ala Gly Gly
85 90 95Ser Pro Ala Asp Ser Val
Pro Trp Val Asp Gly Asp Gly Asp Arg Pro 100
105 110Arg Leu Ala Gly Thr Leu Gly His Leu Val Cys Arg
Pro Trp Arg Thr 115 120 125Tyr Asp
Gly Gly Asp His Thr Leu His Val Gly Arg Val Glu Glu Phe 130
135 140Ala Ala Gly Gly Gly Arg Pro Leu Leu Phe Tyr
Arg Gly Val Phe Pro145 150 155
160Arg Leu Met Pro Asp Gly Gly Gly Asp Pro Glu Gly Pro Glu Glu Val
165 170 175Trp Ser Leu Cys
Leu Asp Gly Pro Gly Pro Ala Thr Asp Gln Phe Val 180
185 190Thr Asp His Glu Thr Arg Lys
19511523PRTStreptomyces species FH6421 11Met Ala Pro Asp Asn Gly Gln Ser
Ala Ala Pro Gly Thr Ser Gly Ala1 5 10
15Ser Thr Gly Lys Ala Arg Val Thr Arg Pro Leu Thr Gly Asp
Glu Tyr 20 25 30Ile Glu Ser
Ile Arg Asp Gly Arg Glu Ile Trp Ala Tyr Gly Glu Lys 35
40 45Val Asp Asp Val Thr Lys His Pro Ala Phe Arg
Asn Thr Val Arg Met 50 55 60Thr Ala
Arg Leu Tyr Asp Ala Leu His Asp Pro Glu His His Asp Thr65
70 75 80Leu Thr Ala Pro Thr Asp Thr
Gly Ser Asp Gly Phe Thr His Lys Phe 85 90
95Tyr Arg Val Pro Arg Ser Val Gln Asp Leu Val Gly Asp
Arg Asp Ala 100 105 110Ile Ala
Asp Trp Ala Arg Leu Thr Tyr Gly Trp Met Gly Arg Ser Pro 115
120 125Asp Tyr Lys Ala Ser Phe Leu Val Thr Leu
Gly Ala Asn Pro Asp Tyr 130 135 140Tyr
Gly Asp Phe Ala Asp Asn Ala Arg Arg Trp Tyr Ala Thr Ala Gln145
150 155 160Glu Asn Val Leu Phe Trp
Asn His Ala Val Ile Asn Pro Pro Val Asp 165
170 175Arg His Arg Pro Ala Asp Glu Val Asp Asp Val Phe
Val His Val Glu 180 185 190Lys
Glu Cys Asp Asp Gly Leu Val Val Ser Gly Ala Lys Val Val Ala 195
200 205Thr Gly Ser Ala Leu Thr His Phe Asn
Phe Val Ala His Tyr Gly Leu 210 215
220Pro Val Lys Lys Lys Glu Phe Ala Leu Val Ala Thr Leu Pro Leu Ala225
230 235 240Ala Pro Gly Val
Lys Leu Ile Cys Arg Gln Ser Tyr Glu Leu Ala Ala 245
250 255Ser Arg Thr Gly Ser Pro Phe Asp Tyr Pro
Leu Ser Ser Arg Leu Asp 260 265
270Glu Asn Asp Thr Ile Phe Ile Leu Asp Lys Val Lys Ile Pro Trp Glu
275 280 285Asn Val Leu Ile Tyr Gly Asp
Thr Ala Arg Ala Gly Thr Phe Leu Gln 290 295
300Thr Ser Gly Phe Thr His Arg Leu Thr Phe His Gly Val Thr Arg
Leu305 310 315 320Ala Val
Lys Leu Asp Phe Leu Ala Gly Leu Leu Leu Lys Gly Val Glu
325 330 335Val Thr Gly Thr Lys Asp Phe
Arg Gly Ile Gln Thr Arg Val Gly Glu 340 345
350Val Leu Ala Trp Arg Asn Met Phe Trp Ala Leu Ser Asp Ala
Met Ala 355 360 365His Asn Pro Asp
Pro Trp His Asp Gly Ala Leu Leu Pro Asn Leu Asp 370
375 380Tyr Gly Met Ala Tyr Arg Trp Phe Met Thr Val Gly
Tyr Pro Arg Val385 390 395
400Arg Glu Ile Ile Met Gln Asp Leu Ser Ser Gly Leu Ile Tyr Leu Thr
405 410 415Ser His Ala Lys Asp
Phe Asn Glu Pro Glu Leu Arg Pro His Leu Asp 420
425 430Arg Phe Met Arg Gly Ser Asn Gly Tyr Glu Ala Val
Glu Arg Ala Lys 435 440 445Leu Met
Lys Leu Ile Trp Asp Ser Val Gly Thr Glu Phe Ala Gly Arg 450
455 460His Glu Leu Tyr Glu Arg Asn Tyr Ser Gly Asn
His Glu Ser Val Arg465 470 475
480Ile Glu Leu Leu His Ala Gln Thr Ala Ser Gly Leu Val Asp Gln Tyr
485 490 495Arg Gly Phe Ala
Glu Gln Cys Met Ala Glu Tyr Asp Leu Asp Gly Trp 500
505 510Thr Ala Pro Asp Leu Val Pro Pro Asp Val Asp
515 52012222PRTStreptomyces species FH6421 12Met Glu
Gln Glu Arg Trp Asn Ser Val Asp Val Tyr Phe Ser Ser Leu1 5
10 15Leu Val Lys Glu Asp Glu Ala Leu
Ser Lys Ala Ala Gln Ala His Arg 20 25
30Glu Phe Asp Leu Pro Asp Leu Ala Val Ser Ala Pro Gln Gly Lys
Leu 35 40 45Leu His Leu Leu Ala
Arg Leu Arg Gln Ala Arg Arg Ile Leu Glu Ile 50 55
60Gly Thr Phe Gly Gly Tyr Ser Ser Ile Trp Leu Ala Arg Ala
Leu Pro65 70 75 80Pro
Asp Gly Arg Leu Val Thr Ile Glu Trp Glu Arg Ser Phe Ala Glu
85 90 95Ser Ala Ala Ser Arg Leu Ala
Glu Ala Gly Val Ala His Leu Val Glu 100 105
110Gln His Val Gly Arg Ala Leu Asp Ile Leu Pro Thr Leu Asp
Arg Pro 115 120 125Gly Thr Ala Pro
Phe Asp Met Val Phe Val Asp Ala Asn Lys Pro Asp 130
135 140Ile Pro Glu Tyr Phe Thr Trp Ala Leu Lys Leu Ser
Arg Pro Gly Ala145 150 155
160Val Val Val Val Asp Asn Val Val Leu Gly Gly Ala Val Thr Asp Pro
165 170 175Asp His Pro Asp Ala
Gly Val Gln Gly Val Arg Arg Phe His Glu Met 180
185 190Leu Ala Gly Arg Ser Asp Val Thr Ala Thr Ser Ile
Gln Thr Val Gly 195 200 205Thr Lys
Gly Tyr Asp Gly Phe Thr Leu Ala Leu Val Thr Gly 210
215 22013166PRTStreptomyces species FH6421 13Met Arg Glu
Asp Ser Ala Val Thr Thr Ala Ala Pro Pro Val His Leu1 5
10 15Val Pro Ala Met His His Leu Gly Val
Gln Thr Arg Asp Leu Asp Asn 20 25
30Ser Leu Ala Trp Tyr Lys Asp Phe Phe Gly Cys Ala Glu Thr Trp Thr
35 40 45Leu Thr Thr Phe Ser Asp Leu
Thr Arg Ser Arg Leu Pro Gly Ile Thr 50 55
60Arg Leu Thr Glu Ile Ser Val Ala Asp Val Arg Phe His Leu Phe Glu65
70 75 80Arg Ala Gly His
Asp Pro Ala Leu Pro Gly Gly Asn Lys Ala Gln Phe 85
90 95Gln His Val Cys Leu Ala Thr Gly Ser Pro
Glu Glu Leu Arg Ala Trp 100 105
110Arg Asp Arg Trp Ile Glu Leu Tyr Arg Ser Gly Arg Tyr Asp Phe Ala
115 120 125Thr Asp Glu Gln Pro Thr Asp
Ile Val Val Asp Ala Asp Gly Val His 130 135
140Ser Cys Tyr Leu Phe Asp Pro Asn Gly Leu Glu Phe Glu Phe Thr
Tyr145 150 155 160Val Pro
Gly Gly Ala Ala 16514268PRTStreptomyces species FH6421
14Met Ser Ala Gly Pro His Arg Thr Val Thr Glu Leu Pro Val Ala Glu1
5 10 15Gly Trp Asp Phe Gly Asp
Phe Pro Tyr Gly Leu Glu Pro Leu Thr Leu 20 25
30Pro Glu Pro Pro His Glu Pro Ala Ala Asp Val Pro Asp
Val Leu Cys 35 40 45Ala Glu Pro
Ala Pro Gly Gly Ala Arg Thr Ser Cys Pro Arg Thr Gly 50
55 60Pro Ala Pro Gly Leu Pro Glu Leu Ala His Gln Leu
Phe Trp Phe Arg65 70 75
80Trp Ile Thr Gly His Gln Leu Thr Phe Ala Ile Trp Gln Leu Leu Gly
85 90 95His Ala Leu His Gln Ala
His Ala Arg Pro Asp Pro Gly Pro Ser Leu 100
105 110Arg Ala Met Thr Asp Leu Thr Arg Ala Tyr Thr Ala
Met Leu Leu Tyr 115 120 125Thr Gly
Ser Cys Pro Lys Asp Val Tyr Ser Asp Val Ile Arg Pro Ser 130
135 140Met Phe Leu Gln His Arg Gly Phe Ser Gly Thr
Trp Ala Pro Asp Phe145 150 155
160Val Pro Val Arg Arg Leu Leu Arg Gly Arg Lys Thr Pro Trp His Glu
165 170 175Thr Pro Glu Gly
Gly Arg Leu Ala Asp Glu Val Arg Leu Tyr His Leu 180
185 190Val His Ser Gly Val Ala Ala Lys Leu Val Pro
Gly Gly Arg Ser Leu 195 200 205Leu
Gln Asp Thr Ala Pro Thr Ala Arg Pro His Asp Pro Arg Met Gln 210
215 220Ala Leu Val Tyr Asp Asn Tyr Phe Leu Thr
Leu Arg Ala Asp Val Pro225 230 235
240Thr Ala Glu Val Val Glu Gln Leu Arg Arg Arg Leu Ala Ala Val
Arg 245 250 255Leu Asp Val
Ser Val Asn Gly Leu Tyr Pro Gly Leu 260
26515299PRTStreptomyces species FH6421 15Val Arg Tyr Gly Val Val Val Leu
Pro Glu Arg Arg Trp Ala Gln Ala1 5 10
15Arg Glu Gln Trp Val Arg Ala Glu Glu Phe Gly Phe Asp His
Ala Trp 20 25 30Thr Tyr Asp
Gln Leu Met Trp Arg Trp Leu Arg Asp Glu Pro Trp Phe 35
40 45Gly Ala Val Pro Thr Leu Ala Ala Ala Ala Glu
Ala Thr Ser Thr Leu 50 55 60Thr Val
Gly Thr Met Val Ala Thr Pro Thr Tyr Arg His Pro Val Thr65
70 75 80Leu Ala Lys Glu Val Met Thr
Leu Glu Asp Ile Ala Gly Gly Arg Phe 85 90
95Val Cys Gly Leu Gly Ala Gly Ala Gly Gly Leu Asp Asp
Arg Val Val 100 105 110Asp Pro
Ala Ala Tyr Ser Pro Arg Gln Arg Ala Asp Arg Phe Thr Glu 115
120 125Phe Val Asp Leu Leu Asp Lys Leu Leu Ser
Arg Arg Ser Thr Thr His 130 135 140Thr
Gly Thr Tyr Tyr Asp Val Arg Glu Val Pro Val His Pro Gly Cys145
150 155 160Leu Ala Thr Pro Arg Val
Pro Phe Ala Ile Ala Ala Thr Gly Pro Arg 165
170 175Gly Met Arg Leu Ala Ala Arg His Ala Asp Met Trp
Ile Thr Ala Gly 180 185 190Arg
Pro Gly Asp Phe Asp Ala Leu Pro Tyr Glu Glu Thr Leu Pro Val 195
200 205Ile Lys Glu Gln Leu Ala Arg Leu Asp
Glu Ala Cys Glu Arg Thr Gly 210 215
220Arg Asp Pro Ala Thr Leu Arg Arg Leu Leu Leu Thr Gly Ala Met Val225
230 235 240Gly Gly Thr Leu
Asp Ser Val Glu Ala Tyr Arg Asp Ala Ala Gly Arg 245
250 255Phe Gly Glu Leu Gly Ile Thr Asp Phe Val
Val His Trp Pro Arg Pro 260 265
270Ser Phe Pro Tyr Gln Gly Arg Val Glu Val Leu Glu Gln Ile Ala Arg
275 280 285Asp Val Leu Thr Val Arg Gly
Gly Glu Arg Pro 290 29516291PRTStreptomyces species
FH6421 16Val Ile Ala Tyr Glu Ile Val Asp Met Phe Thr Gly Thr Pro Phe Gln1
5 10 15Gly Cys Ala Leu
Gly Val Val Pro Asp Ala Thr Ala Leu Asp Asp Asp 20
25 30Gly Met Arg Ala Val Ala Arg Glu Ile Gly Leu
Thr Glu Thr Ala Phe 35 40 45Val
Leu Pro Pro Glu Ser Pro Asp Ala Thr His Arg Val Arg Val Phe 50
55 60Thr Pro Glu Arg Glu Ser Pro Tyr Gly Gly
His Ser Ala Ile Gly Thr65 70 75
80Ala Thr Thr Leu Val Arg Leu Gly Arg Leu Arg Ala Gly Glu Leu
Val 85 90 95Gln Glu Cys
Gly Gly Arg Leu Met Thr Val Arg Ala Ser Ala Arg Arg 100
105 110Ala Thr Leu Gly Val Arg Gly Glu Pro Val
Pro Pro Gly Ala Trp Asp 115 120
125Pro Val Pro Leu Leu Glu Ala Cys Gly Leu Thr Glu Asp Asp Leu Val 130
135 140Ala Gly Pro Arg Val Thr Gly Phe
Gly Pro Ala Phe His Val Leu Pro145 150
155 160Val Gly Pro Glu Ala Val Ala Arg Ala Ala His Asp
Pro Ala His Pro 165 170
175Val Trp Ser Thr Cys Pro Asp Ala Val Val Val Ala Tyr Asp Arg Arg
180 185 190Gly His Leu Ala Asp Val
Arg Val Phe Ala Pro Gly Tyr Gly Met Pro 195 200
205Glu Asp Pro Ala Cys Ala Ser Ala Ala Leu Ala Leu Gly Ala
Trp Leu 210 215 220Thr Gly Ala Gly Leu
Val Pro Ala Thr Asp Gly Thr Arg Leu Tyr Arg225 230
235 240Val Arg Gln Gly His Gly Leu Gly Arg Pro
Ala Arg Leu Asp Cys Ala 245 250
255Val Thr Val Arg Asp Gly Arg Ala Val Ala Ala Glu Val Thr Gly Glu
260 265 270Val Ala Ala Thr Ala
Ala Gly Arg Met His Leu Pro Arg Thr Ala Ala 275
280 285Val Ala Arg 29017597PRTStreptomyces species
FH6421 17Val Leu Phe Arg Pro Glu Leu Arg Gly Thr Arg Gly Ala Val Ala Ser1
5 10 15Thr His Trp Leu
Ala Ser Ala Ala Gly Phe Arg Met Tyr Asp Lys Gly 20
25 30Gly Asn Ala Phe Asp Ala Ala Val Ala Ala Ala
Phe Val Ile Gln Val 35 40 45Val
Glu Pro His Leu Asn Gly Pro Gly Gly Asp Val Pro Val Leu Val 50
55 60His Arg Ala Gly Ser Gly Arg Val Asp Val
Val Cys Gly Gln Gly Pro65 70 75
80Met Pro Arg Ala Ala Thr Ile Glu Arg Phe Glu Gln Leu Gly Leu
Ser 85 90 95Val Val Pro
Gly Ser Gly Leu Leu Pro Ala Val Val Pro Gly Ala Phe 100
105 110Gly Ala Trp Leu Arg Val Leu Ala Glu Tyr
Gly Thr Leu Arg Leu Glu 115 120
125Asp Val Leu Glu Pro Ala Ile Gly Tyr Ala Glu Arg Gly Tyr Pro Leu 130
135 140Leu Pro Lys Ala Ala Ala Met Ile
Glu Ala Leu Gln Glu Leu Phe Arg145 150
155 160Asp Glu Trp Thr Glu Ser Ala Arg Thr Tyr Leu Val
Gly Gly Ala Ala 165 170
175Pro Arg Pro Gly Gln Arg Met Thr Asn Pro Asp Leu Ala Arg Thr Tyr
180 185 190Arg Arg Val Leu Asp Glu
Ala Arg Ala Ala Gly Ala Asp Arg Asp Lys 195 200
205Gln Ile Asp Ala Ala Leu Arg Ala Phe Tyr Glu Gly Phe Val
Ala Glu 210 215 220Ala Ile Asp Gly Tyr
Leu Ala Lys Ala Glu Glu Ile Asp Ala Thr Gly225 230
235 240Arg Arg His Arg Gly Leu Leu Thr Gly Ala
Asp Leu Ala Gly Trp Arg 245 250
255Ala Thr Val Glu Pro Ser Leu Ser Phe Asp His Arg Gly Leu Thr Val
260 265 270His Lys Ala Gly Pro
Trp Ser Gln Gly Pro Val Phe Leu Gln Gln Leu 275
280 285Ala Leu Leu Arg Glu Phe Asp Leu Ala Gly Met Gly
Pro His Ser Ala 290 295 300Glu Phe Val
His Thr Val Thr Glu Ala Ala Lys Leu Ala Phe Ala Asp305
310 315 320Arg Glu Ala Trp Tyr Gly Asp
Pro Ala His Ala Glu Val Pro Val Gly 325
330 335Asp Leu Leu Asp Pro Ala Tyr Thr Ala Ala Arg Arg
Glu Leu Ile Gly 340 345 350Ser
Glu Ala Ser Thr Glu Leu Arg Pro Gly Ser Pro Gly Gly Arg Thr 355
360 365Pro Val Leu Pro Pro Val His Asp Glu
Ser Ala Gly Pro Ala Gly Pro 370 375
380Ser Trp Leu Gly Glu Leu Glu Glu Gly Ile Pro Ala Val Val Arg Ser385
390 395 400Thr Ala Ala Arg
Gly Asp Thr Cys Cys Val Thr Ala Thr Asp Ala His 405
410 415Gly Asn Met Val Val Ala Thr Pro Ser Gly
Gly Trp Leu Lys Ser Ser 420 425
430Pro Val Val Pro Gly Leu Gly Phe Pro Leu Gly Thr Arg Gly Gln Met
435 440 445Ala Thr Leu Thr Arg Gly His
Ala Asn Ala Leu Ala Pro Gly Lys Arg 450 455
460Pro Arg Thr Thr Leu Ser Pro Thr Leu Val Leu Arg Glu Gly Arg
Pro465 470 475 480Ala Leu
Ala Phe Gly Thr Pro Gly Gly Asp Gln Gln Asp Gln Trp Thr
485 490 495Leu Gln Phe Phe Leu Arg His
Thr Glu His Gly Met Gly Leu Gln Glu 500 505
510Ala Val Glu Ala Arg Thr Phe His Thr Asp His Val Pro Thr
Ser Phe 515 520 525Thr Pro Arg Arg
Phe Ala Pro Gly Thr Val Thr Val Glu Ser Gly Met 530
535 540Pro Glu Glu Thr Ile Gln Glu Leu Arg Arg Arg Gly
His Gln Val Arg545 550 555
560Thr Val Ala Asp Tyr Ser Leu Ser Lys Val Cys Val Thr Gly Leu Ala
565 570 575Ser Asp Asp Met Val
Ile Ala Ala Ala Ser Pro Arg Gly Ala Gln Ala 580
585 590Tyr Ala Val Ala Asp
59518769PRTStreptomyces species FH6421 18Met Glu Gly Glu Arg Met Val Pro
His Pro Ser Leu Asp Pro Gly Asp1 5 10
15His Ile Val Leu Gly Glu Ala Arg Gln Asn Asn Leu Lys Gly
Val Ser 20 25 30Leu Arg Ile
Pro Lys Gly Arg Leu Thr Val Phe Thr Gly Val Ser Gly 35
40 45Ser Gly Lys Ser Ser Leu Val Phe Gly Thr Ile
Ala Val Glu Ser Gln 50 55 60Arg Gln
Met Asn Glu Thr Tyr Pro Ala Phe Ile Arg Asn Arg Leu Pro65
70 75 80Lys Phe Glu Arg Pro Asp Ala
Glu Val Ile Glu Asn Leu Ser Thr Ala 85 90
95Ile Val Ile Asp Gln Arg Pro Val Gly Gly Asn Ala Arg
Ser Thr Val 100 105 110Gly Thr
Met Thr Glu Ile His Ala Met Leu Arg Val Leu Phe Ser Arg 115
120 125His Gly Arg Pro Ser Ala Gly Pro Ser His
Met Tyr Ser Phe Asn Asp 130 135 140Pro
Arg Gly Met Cys Pro Glu Cys Glu Gly Leu Gly Ser Arg Val Arg145
150 155 160Leu Asp Leu Asn Arg Leu
Leu Asp Glu Asp Lys Ser Leu Asn Glu Gly 165
170 175Ala Ile Arg Phe Gln Pro Phe Ala Val Gly Thr Phe
Pro Trp Gln Leu 180 185 190Tyr
Ala Glu Ser Gly Leu Phe Asp Pro Asp Leu Pro Leu Arg Glu Phe 195
200 205Ser Ala Asp Asp Arg Glu Leu Leu Leu
His Gly Ser Gly Phe Lys Val 210 215
220Asp Arg Ala Gly Arg His Gly Val Tyr Lys Asn Glu Tyr Glu Gly Ile225
230 235 240Val Leu Arg Phe
Thr Arg Arg Tyr Leu Lys Ala Gly Leu Asp Thr Leu 245
250 255Lys Pro Lys Glu Arg Ala Ala Val Gln Glu
Val Val Thr Glu Gly Pro 260 265
270Cys Glu Ala Cys Gly Gly Ala Arg Leu Gly Pro Ala Ala Leu Ala Ser
275 280 285Arg Ile Ala Gly Glu Asn Ile
Ala Asp Tyr Ser Ala Leu Glu Val Thr 290 295
300Asp Leu Ile Gly Arg Leu Glu Arg Asn Asp Ala Pro Pro Val Lys
Pro305 310 315 320Val Val
Gln Ala Ala Leu Ala Ala Leu Arg Arg Ile Glu Ala Val Gly
325 330 335Leu Gly Tyr Leu Ser Leu Asp
Arg Gln Thr Ala Thr Leu Ser Gly Gly 340 345
350Glu Ala Gln Arg Leu Lys Thr Val Arg His Leu Gly Ser Ser
Leu Thr 355 360 365Gly Leu Thr Tyr
Ile Phe Asp Glu Pro Ser Val Gly Leu His Pro Arg 370
375 380Asp Val Arg Arg Leu Asn Glu Leu Leu Leu Ala Leu
Arg Asp Lys Gly385 390 395
400Asn Thr Val Leu Val Val Glu His Asp Arg Asp Val Ile Ala Ile Ala
405 410 415Asp His Val Val Asp
Met Gly Pro Gly Ala Gly Ser Gln Gly Gly Glu 420
425 430Val Val Tyr Glu Gly Ser Pro Thr Gly Leu Arg Gly
Ser Asp Ser Pro 435 440 445Thr Gly
Arg Gly Leu Arg Ser Val Pro Gly Leu Lys Arg Arg Leu Arg 450
455 460Ala Pro Asp Gly Arg Leu Thr Val Arg Gly Ala
Arg Leu His Asn Leu465 470 475
480Lys Asp Val Thr Val Asp Val Pro Thr Gly Val Leu Val Ala Leu Ser
485 490 495Gly Val Ala Gly
Ser Gly Lys Ser Ser Leu Ala Arg Glu Leu Ala Ala 500
505 510Arg His Pro Glu Glu Thr Val Val Val Asp Gln
Ser Ser Ile Gly Ile 515 520 525Ser
Ser Arg Ser Thr Pro Ala Thr Tyr Thr Asp Ile Met Asp Thr Val 530
535 540Arg Arg Leu Phe Ala Arg Ala Ser Gly Thr
Asp Pro Gly Leu Phe Ser545 550 555
560Phe Asn Ser Ala Gly Ala Cys Pro Glu Cys Gln Gly Arg Gly Val
Ile 565 570 575Glu Thr Asp
Leu Ala Phe Met Asp Pro Val Thr Thr Val Cys Glu Arg 580
585 590Cys Glu Gly Arg Arg Phe Asn Asp Glu Ala
Leu Ser His Thr Leu Ser 595 600
605Gly Arg Asn Ile Ala Asp Val Leu Ala Met Thr Ala Glu Glu Ala Ile 610
615 620Gly Phe Phe Ala Glu Asp Ser Val
Arg Arg Lys Leu Ala Leu Leu Thr625 630
635 640Glu Val Gly Leu Gly Tyr Leu Thr Leu Gly Arg Ser
Leu Ser Thr Leu 645 650
655Ser Gly Gly Glu Arg Gln Arg Leu Lys Leu Ala His Arg Leu His Ala
660 665 670Ser Gly Ser Val Tyr Ile
Phe Asp Glu Pro Ser Thr Gly Leu His Met 675 680
685Thr Asp Val Gly Lys Leu Leu Thr Leu Phe Asp Arg Leu Val
Asp Gly 690 695 700Gly Asn Thr Val Val
Val Ile Glu His Asp Leu Asp Val Leu Lys Tyr705 710
715 720Ala Asp Trp Ile Ile Asp Leu Gly Pro Glu
Ala Gly Arg His Gly Gly 725 730
735Arg Val Val Phe Glu Gly Thr Pro Ala Asp Leu Ala Arg Val Arg Glu
740 745 750Ser His Thr Gly Arg
Cys Leu Ala Glu Asp Leu Ala Ala His Gly His 755
760 765Leu1967PRTStreptomyces species FH 6421 19Met Pro
Leu Ile His Val Thr Leu Leu Ser Gly Arg Gly Glu Glu Glu1 5
10 15Ile Ala Ala Leu Gly Arg Ala Val
Thr Glu Ala Val His Thr Thr Leu 20 25
30Gly Thr Pro Arg Glu Ala Ile Arg Val Thr Val Asp Ala Cys Pro
Pro 35 40 45Glu His Trp Phe Val
Gly Gly Val Ser Met Ala Glu Lys Lys Ala Ala 50 55
60Arg Gly Gly6520400PRTStreptomyces species FH6421 20Leu His
Val His Pro Ser Leu Ser Glu Pro Gln Arg Gly Val Met Arg1 5
10 15Ile Ala Val Ile Gly Ala Gly Pro
Gly Gly Leu Tyr Thr Ala Cys Leu 20 25
30Val Lys Arg Leu Arg Pro His Asp Val Val Glu Val Trp Glu Ala
Asn 35 40 45Ala Ala His Asp Thr
Phe Gly Phe Gly Val Val Phe Ser Asp Gly Ala 50 55
60Leu Gly Gly Ile Glu Ala Ala Asp Pro Ala Leu Phe Glu Ala
Val Glu65 70 75 80Ala
Glu Phe Ala Arg Trp Thr Arg Ile Asp Val Cys Tyr Arg Gly Arg
85 90 95Val Gln Arg Asn Glu Gly Tyr
Gly Phe Ala Ala Ile Gly Arg His Thr 100 105
110Leu Leu Arg Leu Leu Gln Glu Arg Cys Ala Asp Leu Gly Val
Arg Leu 115 120 125His Phe Gly Ser
Pro Ala Pro Glu Val Ser Arg Leu Arg Ala Ser His 130
135 140Asp Leu Val Val Ala Ala Asp Gly Val Gly Ser Arg
Thr Arg Ala Arg145 150 155
160Tyr Glu Ala Ala Phe Gly Thr Glu Arg Glu Gln Ala Gly Ser Arg Tyr
165 170 175Met Trp Leu Gly Thr
Asp Arg Pro Phe Asp Ala Leu Thr Phe Ala Val 180
185 190Val Glu Thr Asp His Gly Pro Val Gln Ala His Ala
Tyr Pro Tyr Ala 195 200 205Pro Gly
Arg Ser Thr Phe Ile Val Glu Ile Gly Asp Ala Val Trp Arg 210
215 220Ala Ala Gly Phe Arg Ala Ala Glu Arg Arg Ser
Asp Asp Pro Val Gly225 230 235
240Asp Arg Glu Ser Ile Gly Arg Val Arg Glu Tyr Phe Thr Gly Leu Leu
245 250 255Asp Gly His Gly
Leu Trp Gly Asn Arg Ser Tyr Trp Gly Arg Phe Ala 260
265 270Ala Val Arg Asn Arg Asp Trp Ser His Gly Asn
Leu Val Leu Leu Gly 275 280 285Asp
Ser Ala His Thr Thr His Phe Ser Ile Gly Ser Gly Thr Lys Leu 290
295 300Ala Met Glu Asp Gly Leu Ser Leu Ala Thr
Ala Leu His Asp His Gly305 310 315
320Thr Val Pro Glu Ala Leu Ala Ala Tyr Glu Ala Glu Arg Arg Pro
Ala 325 330 335Val Glu Arg
Met Gln Arg Thr Ala Leu Thr Ser Leu Glu Trp Phe Glu 340
345 350Asn Ile Asp Arg Cys Val Gly Leu Ala Pro
Glu Ala Phe Asn Ala His 355 360
365Leu Leu Thr Arg Ser Gly Arg Leu Thr Arg Asp Asp Leu Pro Ala Val 370
375 380Asp Pro Glu His Val Gly Arg Ile
Arg Arg Arg Phe Ala Asp Ala Ala385 390
395 40021804PRTStreptomyces species FH6421 21Met Pro Asp
Val Leu Thr Pro Thr Pro Leu Pro Ala Ala Asp Leu Ala1 5
10 15Gly Leu Phe Arg Ala Leu Asp Pro Pro
Pro Phe Ala Leu Val Arg Arg 20 25
30Ala Ala Pro Asp Gly Thr Ser Thr Gly Pro Phe Asp Val Phe Ile Gly
35 40 45Thr Met Asp Thr Val Arg Arg
Val Thr Asp Leu Pro Ser Gly Pro Ala 50 55
60Val Pro Gly Gly Gly Pro His Thr Leu Ala Leu Leu Pro Tyr Arg Cys65
70 75 80Leu Ala Glu Arg
Gly Leu Asp Cys His Asp Asp Gly Thr Pro Leu Arg 85
90 95Val Leu Arg Ile Arg Arg Arg His Thr Ala
Asp His Ala Ala Leu Thr 100 105
110Ala Ala Leu Ala Ala Val Arg Pro Ala Gly Asp Leu Leu Gly Glu Gly
115 120 125Ala Gly Phe Asp Gly Ser Asp
Glu Asp Tyr Ala Asp Leu Val Arg Asp 130 135
140Leu Met Ala Asp Glu Val Ala Arg Thr Gly Leu His Val Leu Ile
Arg145 150 155 160Arg Asp
Phe Thr Ala Arg Leu Pro Gly His Gly Pro Val Val Val Gly
165 170 175Glu Leu Phe Arg Arg Leu Leu
Ala Val Glu His Gly Ala Tyr Trp Thr 180 185
190Phe Ala Val Tyr Thr Gly Gly Pro Asp Gly Ala Ala Leu Ala
Gly Ala 195 200 205Ser Pro Gln Gly
His Val Thr Leu Arg Asn Gly Arg Val Val Met Arg 210
215 220Pro Met Cys Gly Thr Leu Arg Leu Pro Pro Gly Gly
Arg Pro Ser Ala225 230 235
240Ala Asp Leu Val Ala Phe Leu Arg Asp Gly Lys Glu Ser Glu Glu Leu
245 250 255Gly Ala Val Val Asp
Ala Glu Leu Ala Met Leu Cys Arg Ile Ser Glu 260
265 270Gly Asp Val Arg Leu Glu Gly Pro Arg Leu Arg Pro
Met Ala Arg Val 275 280 285Leu His
Thr Glu Cys Arg Ile Ser Ala Thr Ala Ala Leu Pro Ala Arg 290
295 300His Thr Leu Ala Gly Ser Leu Phe Ala Ala Thr
Ala Val Gly Arg Pro305 310 315
320Phe Ala Asp Ala Cys Arg Val Ile Thr Arg Arg Glu Pro Thr Gly Arg
325 330 335Gly Tyr Tyr Gly
Gly Leu Ile Ala Leu Leu Gly His Asp Asp Ala Gly 340
345 350Asn Glu Glu Leu Asp Thr Ala Val Leu Ile Arg
Thr Phe Glu Val Ser 355 360 365Gly
Gln Gly Arg Leu Lys Leu Ser Val Gly Ala Thr Leu Gly Pro Arg 370
375 380Ser Val Ala Ala Asp Glu Thr Ala Glu Thr
Arg Ala Lys Ala Ser Ala385 390 395
400Leu Val Ser Ala Leu Ala Ser Gly Gly Pro Thr Ala Glu Gly Gly
Ala 405 410 415Gly Arg His
Ala Arg Ala Gly Leu Gly Arg Gly Pro Glu Ala Ala Gly 420
425 430Gly Pro Ala Thr Gly Glu Arg Ser Gly Val
Pro Gly Asp Arg Thr Arg 435 440
445His Gln Gln Ala Ala Gly Arg Gln Pro Thr Ser Pro Ala Asp Pro Ala 450
455 460Trp Arg Pro Ser Val Thr Ala Glu
Gly Gly Ala Gly Asp His Ala Arg465 470
475 480Ala Gly Leu Gly Arg Gly Pro Glu Ala Ala Gly Gly
Pro Ala Thr Gly 485 490
495Glu Gly Gly Gly Val Pro Gly Asp Arg Thr Arg His Gln Gln Ala Ala
500 505 510Gly Arg Gln Pro Thr Ser
Pro Ala Asp Pro Ala Trp Arg Pro Ser Val 515 520
525Thr Ala Glu Gly Gly Ala Gly Gly His Ala Arg Ala Gly Leu
Gly Arg 530 535 540Gly Pro Glu Ala Ala
Gly Gly Pro Ala Thr Gly Glu Gly Gly Gly Val545 550
555 560Pro Gly Asp Arg Thr Arg His Gln Gln Ala
Ala Gly Arg Arg Pro Thr 565 570
575Ser Ser Ala Asp Pro Ala Trp Cys Pro Ser Val Thr Ala Glu Leu Asp
580 585 590Arg Arg Arg Ala Arg
Leu Ser Ala Tyr Trp Gln Arg Pro Arg Arg Pro 595
600 605Gly Ser Arg Pro Ala Pro Arg Pro Pro Val Leu Leu
Val Asp Thr Gly 610 615 620Gly Glu Glu
Thr Ala Pro Leu Ala Ala Met Leu Arg Gly Leu Gly Arg625
630 635 640Thr Val Asp Val Arg Pro Ala
Tyr Pro Ala Ala Ala Ala Pro Arg Thr 645
650 655Val Ala Pro Gly Thr Thr Val Val Leu Gly Pro Gly
Pro Gly Asp Pro 660 665 670Leu
Ala His Gly Asp Asp Arg Ile Thr Ala Leu Arg Ala Met Thr Ser 675
680 685Ala Leu Leu Ser Ser Gly Ala Pro Thr
Phe Gly Val Gly Leu Gly Phe 690 695
700His Leu Leu Leu Ala Val Leu Gly Leu Ala Gly Ala Ala Arg Ala Trp705
710 715 720Asp Gly Ala Thr
Gly Gln Arg Glu Ile Glu Val Phe Gly Arg Arg Ala 725
730 735Thr Val Gly Tyr Gly Gly Thr His Thr Val
Val Ala Gly Pro His Thr 740 745
750Asp Thr Leu Ala Arg Arg Leu Ser Leu Thr Leu Cys Tyr Gly Pro Ala
755 760 765His Gly Glu Leu Val Ala Met
Arg Gly Pro Arg Thr Gly Gly Val Ala 770 775
780Phe Leu Pro Ala Ser Val Leu Ser Val Glu Gly Ala Glu Leu Leu
Asp785 790 795 800Leu Leu
Leu Pro22481PRTStreptomyces species FH6421 22Met Ala Glu Val Thr Gln Thr
Asp Val Arg Arg Glu Phe Asp Arg Gly1 5 10
15Met Pro Gly Leu Arg Val Pro Gly Arg Val Val Val Ile
Gly Ala Gly 20 25 30Leu Ser
Gly Leu Ala Val Ala Tyr Glu Leu Ala Arg Arg Gly Thr Asp 35
40 45Val Thr Val Leu Glu Ala Ser Asn Arg Pro
Gly Gly Arg Ala Tyr Thr 50 55 60Leu
Arg Glu Pro Phe Thr Asp Gly Leu Tyr Ala Glu Ala Gly Ala Met65
70 75 80Thr Leu Thr Pro His Cys
His Tyr Ala Met His Tyr Leu Arg Glu Leu 85
90 95Gly Val Glu Leu Glu Thr Ala Asp Leu Val Gly Ser
Gln Phe Ser Tyr 100 105 110Phe
Val Gly Asn Arg Phe Phe Gly Pro Asp Ala Asp Ser Leu Asp Arg 115
120 125Ala Gly Leu Pro Leu Ala Pro His Glu
Lys Gly Leu Ser Val Thr Asp 130 135
140Met Ile Asp Arg Tyr Val Arg Arg Ala Tyr Glu Ala Leu Glu Pro Asp145
150 155 160Ile Thr Ala Ala
Asp Trp Ala Pro Thr Pro Leu Leu Glu Pro Tyr Asp 165
170 175Arg Arg Ser Val Tyr Glu Val Leu Ser Gly
Arg Gly Ala Ser Pro Ala 180 185
190Ala Ile Asp Leu Val Glu Pro His Phe Leu Glu Met Arg Gly Gly Asp
195 200 205Leu Lys Thr Ala Ser Ala Leu
Ser Trp Leu Arg His Glu Ser Ser Pro 210 215
220His Ser Leu Ala Asn Ala Asp Pro Arg Trp Ser Lys Val Lys Gly
Gly225 230 235 240Thr Asp
Arg Phe Pro Arg Ala Phe Ala Glu Arg Leu Lys Asp Arg Ile
245 250 255Arg Tyr Arg Ala Pro Val Val
Arg Val Ala Gln Asp Asp Glu Gly Ala 260 265
270Arg Val Thr Phe Leu Asp Gly Thr Arg Met Arg Ser Val Asp
Ala Asp 275 280 285Arg Val Val Val
Thr Val Pro Phe Ser Ala Ile Arg His Ile Asp Phe 290
295 300Thr Asp Ala Gly Leu Ser Asp Ala Lys Gln Ala Val
Met Arg Arg Val305 310 315
320Lys Tyr Ser Ser Ile Val Arg Val Tyr Leu Gln Met Arg Arg Arg Phe
325 330 335Trp Ala Gln Asp Asn
Ala Ser Phe Ser Thr Asp Leu Pro Val Arg Trp 340
345 350Val Arg Asp Ala Thr Pro Arg Leu Pro Gly Pro Arg
Lys Ile Leu Glu 355 360 365Cys Leu
Ile Thr Gly Trp Arg Ala Arg Ala Leu Ala Val Leu Ser Pro 370
375 380Glu Glu Arg Ile Arg Phe Ala Leu Glu His Val
Glu Ser Met Leu Pro385 390 395
400Gly Ala Arg Glu His Phe Glu Thr Gly Thr Ser Val Val Trp Asp Gln
405 410 415Gln Pro Tyr Ile
Glu Gly Ala Tyr Ile Leu Pro Glu Met Gly His Ser 420
425 430Ser Leu Met Pro Ala Met Arg Arg Pro Glu Gly
Arg Ile His Phe Ala 435 440 445Gly
Asp His Thr Ser Phe Glu Pro Asn Gly Gly Ser Met Thr Leu Ala 450
455 460Leu Glu Ser Ala Ala Arg Thr Val Leu Glu
Leu Gly Gly Thr Ala Asn465 470 475
480Gly
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