阅读说明：本技术 吡唑霉素生物合成基因簇、重组菌及其应用 (Pyrazolomycin biosynthesis gene cluster, recombinant bacterium and application thereof ) 是由 陈文青 张梦 邓子新 张培潮 徐顾丹 周文婷 于 2019-08-01 设计创作，主要内容包括：本发明公开了一种吡唑霉素生物合成基因簇、重组菌及其应用,即是纯白色链霉菌NRRL 3601产生的治疗肿瘤的天然产物吡唑霉素的生物合成基因簇的克隆、测序、分析、功能研究及其应用。整个基因簇共包含24个基因。通过对上述生物合成基因的遗传操作可阻断或提高吡唑霉素的生物合成。本发明所提供的基因及其蛋白可用于该化合物的基因工程、蛋白表达、酶催化反应等,也可以用来寻找和发现可用于医药、工业或农业的化合物、基因、蛋白。(The invention discloses a pyrazolomycin biosynthesis gene cluster, a recombinant bacterium and application thereof, namely cloning, sequencing, analyzing, functional research and application of the biosynthesis gene cluster of a natural product pyrazolomycin for treating tumors, which is generated by pure streptomyces albus NRRL 3601. The whole gene cluster contains 24 genes. The biosynthesis of pyrazolomycin can be blocked or improved by genetic manipulation of the biosynthesis genes. The gene and the protein thereof provided by the invention can be used for genetic engineering, protein expression, enzyme catalytic reaction and the like of the compound, and can also be used for searching and discovering compounds, genes and proteins which can be used for medicine, industry or agriculture.)

1. A biosynthetic gene cluster of pyrazolycins characterized by: the nucleotide sequence of the gene cluster is shown as 798-27650 th site in SEQ ID NO.1, wherein the genes responsible for the biosynthesis of the pyrazolomycin comprise 24 genes, and specifically comprise:

prfA is located at bases 798-1988 in SEQ ID NO.1 and encodes an MFS transporter, and the amino acid sequence is SEQ ID NO. 2;

prfB is located at the 2035-3465 base of SEQ ID NO.1, encodes adenylosuccinate lyase, and has the amino acid sequence of SEQ ID NO. 3;

prfC is located at 3786-th 4460 th base in SEQ ID NO.1, encodes HAD family hydrolase, and has the amino acid sequence of SEQ ID NO. 4;

prfD is located at the 4640-th and 4996-th bases in SEQ ID NO.1, encodes isomerase and has an amino acid sequence of SEQ ID NO. 5;

prfE is located at 5056-6060 th base in SEQ ID NO.1 and codes beta-ribofuranosyl aminobenzene 5' -phosphate synthase, and the amino acid sequence is SEQ ID NO. 6;

prfF is located at base 6136-7167 in SEQ ID NO.1 and encodes SAICAR synthetase, and the amino acid sequence is SEQ ID NO. 7;

prfG is located at the 7321-8943 base in SEQ ID NO.1 and encodes fumarate reductase, and has the amino acid sequence of SEQ ID NO. 8;

The prfH is positioned at the base 9410-10399 in the SEQ ID NO.1 and codes for monooxygenase, and the amino acid sequence is SEQ ID NO. 9;

The prfI is positioned at the 10389-th base 11723 in the SEQ ID NO.1 and codes for monooxygenase, and the amino acid sequence is SEQ ID NO. 10;

The prfJ is positioned at the base 11789-12655 in the SEQ ID NO.1 and encodes a transporter, and the amino acid sequence is SEQ ID NO. 11;

prfK is located at the base 12701-13756 in SEQ ID NO.1, encodes oxidase, and has the amino acid sequence of SEQ ID NO. 12;

prfL is located at 13917-15158 th base in SEQ ID NO.1 and codes phosphoribosyl glycinamide synthetase, and the amino acid sequence is SEQ ID NO. 13;

The prfM is located at the 15220-16392 th base in SEQ ID NO.1 and encodes saccharopine dehydrogenase, and the amino acid sequence is SEQ ID NO. 14;

prfN is located at 16392-17705 base of SEQ ID NO.1, encodes L-lysine N ⁶ -monooxygenase, and has the amino acid sequence of SEQ ID NO. 15;

prfO is located at the 17724-th 19730 base of SEQ ID NO.1 and encodes methionine-tRNA ligase, and the amino acid sequence is SEQ ID NO. 16;

prfP is located at base 19774-20526 in SEQ ID NO.1 and encodes SAICAR synthetase, and the amino acid sequence is SEQ ID NO. 17;

prfQ is positioned at the 20516-21217 base in SEQ ID NO.1 and codes short-chain dehydrogenase, and the amino acid sequence is SEQ ID NO. 18;

prfR is located at the 21217-21999 base of SEQ ID NO.1, encodes hydrolase, and has the amino acid sequence of SEQ ID NO. 19;

the prfS is positioned at the 22004-22870 th base in the SEQ ID NO.1 and codes ATP phosphoribosyl transferase, and the amino acid sequence is SEQ ID NO. 20;

The prfT is positioned at the 22967-24175 th base in the SEQ ID NO.1 and codes the phthalic acid-4, 5-dioxygenase, and the amino acid sequence is SEQ ID NO. 21;

The prfU is positioned at the 24201 th 25364 th base in SEQ ID NO.1, encodes amidohydrolase, and has an amino acid sequence of SEQ ID NO. 22;

prfV is positioned at base 25403-26617 in SEQ ID NO.1, encodes MFS transporter, and has the amino acid sequence of SEQ ID NO. 23;

prfW is located at base 26674-27144 in SEQ ID NO.1 and encodes flavin reductase, and the amino acid sequence is SEQ ID NO. 24;

prfX is located at base 27141-27650 in SEQ ID NO.1 and encodes FMN reductase, and has the amino acid sequence SEQ ID NO. 25.

2. The biosynthetic gene cluster of pyrazolomycin of claim 1, wherein said gene prfE encodes a β -ribofuranosylaminobenzene 5' -phosphate synthase PrfE that catalyzes the assembly of the carbon nucleoside backbone and said gene prfN encodes an L-lysine N ⁶ -monooxygenase PrfN that catalyzes the initiation step of pyrazolomycin.

3. a recombinant bacterium comprising the biosynthetic gene cluster of pyrazolomycin according to claim 1 or 2.

4. Use of the recombinant bacterium of claim 3 for the synthesis of pyrazolomycin.

5. Use of the β -ribofuranosylaminobenzene 5' -phosphate synthase PrfE according to claim 2 for the synthesis of pyrazolomycin.

6. Use of the L-lysine N ⁶ -monooxygenase PrfN according to claim 2 for the synthesis of pyrazolomycin.

Technical Field

The invention belongs to the field of microbial genetic engineering, and particularly relates to a pyrazolomycin biosynthesis gene cluster, a recombinant bacterium and application thereof.

background

Pyrazolomycin was first isolated from a fermentation broth of pure Streptomyces albicans (Streptomyces candidus) by Lilly laboratories and its chemical structure of its alpha isomer, pyrazolomycin B, was determined by chromatography and X-ray crystallography (Buchanan, 1983). Pyrazolomycin, also known as antimycotic A23813, consists of a nitrogen-containing heterocyclic structure and a sugar moiety, is a C-nucleoside antibiotic like oxytetracycline (Showdomycin) and synomycin (Formycin), and is chemically stable (Suhadonik and Reischenbach, 1981). Pyrazoline has obvious biological activity of resisting virus and tumor, but the application of pyrazoline as an anticancer agent is greatly influenced by biotoxicity when the pyrazoline is used as the anticancer agent in clinical tests (Buchanan, 1983).

In 1976 Bernardo and Weigele synthesized pyrazolycins chemically, while FARKAS and SORM synthesized pyrazolycins chemically using ketoesters as precursors (Suhadolnik and Reischenbach, 1981). Buchanan et al have demonstrated that the C-3, C-4, C-5 and amide carbons of pyrazolomycin are C-1 to C-4 derived from glutamic acid (Buchanan, 1980). By isotope feeding experiments, ribose was shown to be a direct precursor to the ribofuranose residue of pyrazolomycin (Wang et al, 2019). Furthermore, the initial phase of the biosynthesis of pyrazolomycin has a certain similarity to the biosynthetic pathway of L-lysine (Ochi et al, 1979). However, the specific aspects of the biosynthesis mechanism are still unclear at present.

disclosure of Invention

aiming at the defects of the prior art, the invention is the research of the biosynthetic gene cluster of pyrazolomycin (Pyrazofurin), comprising the in vivo knockout verification of key genes, the in vitro function research and the application thereof. In order to analyze a biosynthesis mechanism of pyrazolomycin, the invention sequences the genome of pure Streptomyces albicans NRRL 3601(Streptomyces candidus NRRL 3601), finds a biosynthesis gene cluster of C-nucleoside antibiotic pyrazolomycin, and verifies the correctness of the biosynthesis gene cluster through sequence analysis, functional verification and biochemical experiments, thereby providing the pyrazolomycin biosynthesis gene cluster, the recombinant bacterium and the application thereof.

In order to achieve the above object, one aspect of the present invention provides a pyrazolomycin biosynthesis gene cluster, which is characterized in that: the nucleotide sequence of the gene cluster is shown as 798-27650 th site in SEQ ID NO.1, wherein the genes responsible for the biosynthesis of the pyrazolomycin comprise 24 genes, and specifically comprise:

prfA is located at bases 798-1988 in SEQ ID NO.1 and encodes an MFS transporter, and the amino acid sequence is SEQ ID NO. 2;

prfB is located at the 2035-3465 base of SEQ ID NO.1, encodes adenylosuccinate lyase, and has the amino acid sequence of SEQ ID NO. 3;

prfC is located at 3786-th 4460 th base in SEQ ID NO.1, encodes HAD family hydrolase, and has the amino acid sequence of SEQ ID NO. 4;

prfD is located at the 4640-th and 4996-th bases in SEQ ID NO.1, encodes isomerase and has an amino acid sequence of SEQ ID NO. 5;

prfE is located at 5056-6060 th base in SEQ ID NO.1 and codes beta-ribofuranosyl aminobenzene 5' -phosphate synthase, and the amino acid sequence is SEQ ID NO. 6;

prfF is located at base 6136-7167 in SEQ ID NO.1 and encodes SAICAR synthetase, and the amino acid sequence is SEQ ID NO. 7;

prfG is located at the 7321-8943 base in SEQ ID NO.1 and encodes fumarate reductase, and has the amino acid sequence of SEQ ID NO. 8;

the prfH is positioned at the base 9410-10399 in the SEQ ID NO.1 and codes for monooxygenase, and the amino acid sequence is SEQ ID NO. 9;

The prfI is positioned at the 10389-th base 11723 in the SEQ ID NO.1 and codes for monooxygenase, and the amino acid sequence is SEQ ID NO. 10;

The prfJ is positioned at the base 11789-12655 in the SEQ ID NO.1 and encodes a transporter, and the amino acid sequence is SEQ ID NO. 11;

prfK is located at the base 12701-13756 in SEQ ID NO.1, encodes oxidase, and has the amino acid sequence of SEQ ID NO. 12;

prfL is located at 13917-15158 th base in SEQ ID NO.1 and codes phosphoribosyl glycinamide synthetase, and the amino acid sequence is SEQ ID NO. 13;

The prfM is located at the 15220-16392 th base in SEQ ID NO.1 and encodes saccharopine dehydrogenase, and the amino acid sequence is SEQ ID NO. 14;

prfN is located at 16392-17705 base of SEQ ID NO.1, encodes L-lysine N ⁶ -monooxygenase, and has the amino acid sequence of SEQ ID NO. 15;

prfO is located at the 17724-th 19730 base of SEQ ID NO.1 and encodes methionine-tRNA ligase, and the amino acid sequence is SEQ ID NO. 16;

prfP is located at base 19774-20526 in SEQ ID NO.1 and encodes SAICAR synthetase, and the amino acid sequence is SEQ ID NO. 17;

prfQ is positioned at the 20516-21217 base in SEQ ID NO.1 and codes short-chain dehydrogenase, and the amino acid sequence is SEQ ID NO. 18;

prfR is located at the 21217-21999 base of SEQ ID NO.1, encodes hydrolase, and has the amino acid sequence of SEQ ID NO. 19;

The prfS is positioned at the 22004-22870 th base in the SEQ ID NO.1 and codes ATP phosphoribosyl transferase, and the amino acid sequence is SEQ ID NO. 20;

The prfT is positioned at the 22967-24175 th base in the SEQ ID NO.1 and codes the phthalic acid-4, 5-dioxygenase, and the amino acid sequence is SEQ ID NO. 21;

The prfU is positioned at the 24201 th 25364 th base in SEQ ID NO.1, encodes amidohydrolase, and has an amino acid sequence of SEQ ID NO. 22;

prfV is positioned at base 25403-26617 in SEQ ID NO.1, encodes MFS transporter, and has the amino acid sequence of SEQ ID NO. 23;

prfW is located at base 26674-27144 in SEQ ID NO.1 and encodes flavin reductase, and the amino acid sequence is SEQ ID NO. 24;

prfX is located at the 27141-27650 base in SEQ ID NO.1 and encodes FMN reductase with the amino acid sequence of SEQ ID NO. 25;

Preferably, the gene prfE encodes a beta-ribofuranosylaminobenzene 5' -phosphate synthase PrfE that catalyzes the assembly of the carbon-nucleoside skeleton, and the gene prfN encodes an L-lysine N ⁶ -monooxygenase PrfN that catalyzes the initial step of pyrazolomycin.

The invention also provides a recombinant bacterium containing the biosynthetic gene cluster of the pyrazolomycin.

the invention also provides an application of the recombinant bacterium in synthesis of pyrazolomycin.

The invention provides application of the beta-ribofuranosylaminobenzene 5' -phosphate synthase PrfE in synthesis of pyrazolomycin.

the invention also provides application of the L-lysine N ⁶ -monooxygenase PrfN in synthesis of pyrazolomycin.

The invention researches the biosynthesis pathway of the pyrazolomycin by adopting a method combining molecular biology, biochemistry and organic chemistry, analyzes the chemical structure of an intermediate-9 a compound accumulated by a mutant strain and verifies the function of related protein, and plays an important role in analyzing the assembly of a C-nucleoside skeleton of the pyrazolomycin and a post-heterocyclic modification mechanism. Meanwhile, the biosynthesis mechanism of the C-nucleoside antibiotics is researched, so that the biosynthesis of the natural product C-nucleoside antibiotics is enriched.

Applications of the pyrazolomycin biosynthesis gene cluster of the present invention include (but are not limited to):

(1) the invention also provides a way of producing a microorganism in which the pyrazolomycin biosynthesis gene cluster is interrupted, at least one of the genes comprising the nucleotide sequence of SEQ ID No. 1.

(2) Including intermediates accumulated during fermentation of a mutant in which the gene cluster is disrupted.

(3) Comprising the nucleotide sequence provided by the present invention or at least part of the nucleotide sequence may be modified or mutated. These include insertions, deletions, polymerase chain reaction, error-mediated polymerase chain reaction, reconnection of different sequences, directed evolution of different parts of a sequence or homologous sequences from other sources, or mutagenesis with chemical agents, etc.

(4) cloned genes comprising the nucleotide sequences provided by the invention or at least part of the nucleotide sequences can be expressed in an exogenous host by means of a suitable expression system to obtain the corresponding enzymes or other higher biological activity or yield. Such foreign hosts include Streptomyces, Pseudomonas, Escherichia, Bacillus, yeast, plants, and animals.

(5) The amino acid sequences provided by the invention can be used for separating the required protein and can be used for preparing antibodies.

(6) polypeptides comprising the amino acid sequences or at least partial sequences provided herein may have biological activity, even new biological activity, after removal or substitution of certain amino acids, or increased yield or optimized protein kinetics or other properties sought to be achieved.

(7) genes or gene clusters comprising the nucleotide sequences or at least part of the nucleotide sequences provided by the present invention can be expressed in heterologous hosts and their function in the metabolic chain of the host is understood by DNA chip technology.

(8) genes or gene clusters comprising the nucleotide sequences or at least part of the nucleotide sequences provided by the present invention can be genetically recombined to construct recombinant plasmids to obtain novel biosynthetic pathways, or can be inserted, substituted, deleted or inactivated to obtain novel biosynthetic pathways.

(9) contains the nucleotide sequence provided by the invention and encodes beta-ribofuranosylaminobenzene 5 '-phosphate synthase PrfE, which is characterized by catalyzing PRPP and P-nitrobenzoic acid (P-AB) to generate beta-ribofuranosylaminobenzene 5' -phosphate (beta-RFA-P), catalyzing compound 9a and PRPP to generate compound No.10 in the invention, and recombining with biosynthesis pathway or partial biosynthesis pathway of other natural products to obtain novel purine nucleoside compounds.

(11) the protein PrfN encoded by the amino acid sequence provided by the invention can catalyze L-Lys to form N ⁶ -OH Lys, which is the initial step of pyrazolomycin biosynthesis, and can be recombined with the biosynthesis pathway or partial biosynthesis pathway of other natural products to obtain novel purine nucleoside compounds.

(10) Comprises the biosynthesis of the m-type mycin B and the m-type mycin A related by the invention. The m-type mycin B is an alpha isomer of m-type mycin A, can be spontaneously formed by the m-type mycin A, and has low yield and no biological activity.

the invention has the following advantages and beneficial effects: the gene and protein information related to pyrazolomycin biosynthesis provided by the invention can help people to understand the biosynthesis mechanism of the nucleoside antibiotic pyrazolomycin, and provides materials and methods for further genetic modification. The gene and the protein thereof provided by the invention can also be used for searching and discovering compounds or genes and proteins which can be used for medicine, industry or agriculture.

drawings

FIG. 1: pyrazolomycin a chemical structure.

FIG. 2: pyrazolomycin B chemical structure.

FIG. 3: biosynthetic gene cluster of pyrazoles mycins.

FIG. 4: high Performance Liquid Chromatography (HPLC) analysis of fermentation products of a synthetic gene cluster of pyrazolomycin is interrupted, wherein a Std-pyrazolomycin standard, an NRRL 3601-wild type, a delta prfD-mutant strain and a delta prfE-mutant strain are adopted.

FIG. 5: and (5) detecting PrfN in vitro biochemical reaction mass spectrum. NbtG, positive control; ForK, biochemical reaction of the ForK protein; PrfN, PrfN protein biochemical reaction; N.C, negative control.

FIG. 6: beta-ribofuranosylaminobenzene 5' -phosphate synthase-PrfE protein gel profiles.

FIG. 7 shows the liquid phase diagram of PrfE in vitro biochemical reaction, Complete-PrfE, PrfE full reaction, -9a, no substrate for reaction, -PRPP, no PRPP, -Mg ²⁺, no Mg ²⁺ for reaction, -N.C, no protein for reaction.

FIG. 8: the pathway of pyrazolomycin biosynthesis is presumed.

Detailed Description

The invention is explained in further detail below with reference to the figures and the specific examples:

The experimental procedures used in the following examples are conventional unless otherwise specified.

materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

1. Cloning and analyzing biosynthetic gene cluster of pyrazolomycin:

the method comprises the steps of firstly extracting the total DNA of pure streptomyces albus NRRL 3601, and carrying out genome-wide scanning sequencing on the total DNA by using a sequencing technology, carrying out BlastP analysis on a biosynthetic gene cluster (for) of the synechocystis using a bacterium producing the synechocystis-Procambrus crab ATCC 21070(S.kaniharaensis ATCC 21070) which codes methionine-tRNA ligase as an enzyme probe, wherein the gene cluster encodes PrfO (with 58% homology to ForJ) and a neighboring gene PrfN (L-lysine N ⁶ -monooxygenase with 62% homology to ForK), PrfP (SAICAR synthetase with 60% homology to CofB), which strongly indicates that the target gene cluster participates in the biosynthesis of PRF-A, thereby determining the approximate position of the biosynthetic gene cluster of the pyrazolomycin on the genome, further carrying out the sequence analysis on the biosynthetic gene cluster containing the pyrazolomycin, and carrying out the open reading frame DNA analysis of 27.1-kb, wherein the open reading frame DNA analysis includes 24 kb information.

Table 1: functional analysis of each gene and encoded protein in pyrazolomycin biosynthesis gene cluster

2. In vivo functional determination of genes involved in pyrazolomycin biosynthesis:

Key genes are sequentially knocked out one by one on a genome of pyrazolomycin producing bacteria NRRL 3601, detection and analysis are carried out on mutant strain fermentation liquor by a liquid phase mass spectrum combination instrument (HRLC-MS), and the two genes prfE and prfD are genes related to pyrazolomycin biosynthesis and are determined as shown in a legend 4. In addition, 13 genes including prfG, prfT, prfH, prfI may be involved in pyrazolomycin biosynthesis, and the remaining genes may be involved in transport regulation.

3. In vitro functional verification of N ⁶ hydroxylation of lysine encoded by prfN, a gene related to pyrazolomycin biosynthesis:

the prfN gene is amplified by PCR and cloned to an expression vector, heterologous overexpression and purification are carried out in escherichia coli, and relatively pure protein PrfN is obtained after SDS-PAGE analysis, bioinformatics analysis shows that PrfN is L-lysine N6-hydroxylase, the enzyme activity of the prfN is tested in vitro by using L-lysine as a substrate and NADPH as a cofactor, mass spectrometry detection results show that as shown in figure 5, the reaction is completely consistent with the fragmentation pattern of a positive control (NbtG reaction), a mixture without enzyme as a negative control cannot generate an expected MS peak, and experimental data prove that PrfN is lysine N ⁶ -monooxygenase for starting PRF-A biosynthesis.

4. In vitro functional validation of the β -ribofuranosyl aminophenyl 5' -phosphate synthase encoded by the gene prfE involved in pyrazolomycin biosynthesis:

the prfE gene is amplified by utilizing PCR and then cloned to an expression vector, heterologous overexpression and purification are carried out in escherichia coli, and relatively pure protein PrfE is obtained after SDS-PAGE analysis, bioinformatics analysis shows that PrfE is beta-ribofuranosylaminobenzene 5' -phosphate synthase, the invention takes a 9a compound and PRPP (5-phosphoribosyl 1-pyrophosphate) as substrates, and PrfE catalyzes in-vitro biochemical reaction under the condition that divalent metal ions Mg ²⁺ are used as cofactors, and as shown in figure 7, after liquid phase and mass spectrum detection, 10 compounds are formed through analysis.

5. Biosynthesis of pyrazolycins

Certain genes in the pyrazolomycin biosynthetic gene cluster are related in vivo to the biosynthesis of lysine and syndiomycin, with L-lysine and glutamate as starting compounds, lysine being hydroxylated first to form N ⁶ hydroxylated lysine while glutamate is dehydrogenated to form compound No.1, then both intermediates are subjected to the action of the two enzymes PrfO (methionine tRNA ligase) and PrfK (amino acid oxidase) to build the N-N bond (compound 4) while leaving compound 5. next, it is proposed that compound 4 is hydroxylated by PrfH/I in a sequential manner at the C-4 position to produce 6, which will be continuously dehydrated by spontaneous or unknown enzymatic reactions to produce 7. further studies of the PRF-a gene cluster speculate that PrfL (phosphoribosylglycinamide synthetase) is required for the next reaction, usually at the second step in the purine de novo biosynthetic pathway.

example 1 extraction of pure Streptomyces albus NRRL 3601 Total DNA of pyrazolomycin-producing bacteria:

30 mu.L of pure streptomyces albus NRRL 3601 spores are taken to be cultured in 50mL of TSB culture medium for 24h at the temperature of 30 ℃ and the rpm of 200 until the culture medium is in a turbid state. 50mL of pure Streptomyces albus NRRL 3601 bacterial liquid, centrifuging at 8,000rpm and 8 ℃ for 10min to remove supernatant, and collecting thalli. Dissolving the thalli in 25mL of 10.3% sucrose solution, shaking, uniformly mixing, washing the thalli, centrifuging at 6,000rpm at 4 ℃ for 10min, and removing a supernatant; dissolving the thalli in 25mL setbuffer, shaking and mixing uniformly, centrifuging at 6,000rpm at 4 ℃ for 10min to remove supernatant, and repeating twice; dissolving the thalli in 10mL of set buffer, shaking and uniformly mixing, adding 50 mu L of lysozyme solution (100mg/mL), and placing in a water bath kettle at 37 ℃ for warm bath for 30 min; then adding 200 mu L of proteinase K solution (50mg/mL), uniformly mixing, adding 600 mu L of 10% SDS, reversely mixing, placing in a 55 ℃ warm bath for 4h, reversely mixing every 15min in the period, adding 150 mu L of proteinase K solution every 30min until the mycelium is cracked and becomes transparent; then adding 4mL of 5M NaCl, reversing and uniformly mixing, and placing the bacterial liquid at room temperature to about 37 ℃; adding 10mL of chloroform, reversing and mixing uniformly to be milky white, 4,000rpm, 4 ℃, 15min, taking out supernatant, adding isopropanol with the volume of 0.6 time, and mixing uniformly; after mixing, flocculent DNA is separated out, the separated DNA is carefully selected out and washed twice by 75 percent ethanol, and the mixture is placed in a ventilated place for drying and is dissolved in proper amount of ultrapure water. The size of the DNA was determined by nucleic acid electrophoresis (40v, 12h) and the purity of the total DNA extracted was determined by measuring its concentration and OD value with Nanodrop 2000.

Example 2 fermentation conditions of pyrazolomycin-producing bacteria Streptomyces albus NRRL 3601 and detection conditions of product by High Performance Liquid Chromatography (HPLC).

inoculating spores of pure streptomyces albus NRRL 3601 into a seed culture medium, culturing for 48h at 30 ℃ and 220rmp, transferring into a fermentation shake flask according to the inoculum concentration of 2%, and culturing for 7 days at 30 ℃ and 220 rmp. Collecting fermentation liquid, adjusting pH to 3-4 with oxalic acid, centrifuging the fermentation liquid at 6,000rpm for 20min, removing supernatant, and detecting and analyzing by HPLC with 0.22 μm microporous filter membrane.

HPLC detection conditions: phase A was ultrapure water to which 0.15% trifluoroacetic acid (TFA) was added, and phase B was methanol. The initial 95% of phase A eluted with a gradient to 25% in 30min, phase A switched to 50% at 41min, phase A switched to 95% at 42min, and held for 60 min. The flow rate is 0.5mL/min, the detection wavelength is 254nm, and the column temperature is 30 ℃.

example 3.A. albidomyces-producing strain, S.albus NRRL 3601, and a method for heterologous expression conjugation transfer thereof.

the target plasmid to be conjugately transferred is firstly transformed into the competence of Escherichia coli E.coli ET12567/pUZ8002, after a transformant grows out, the positive monoclonal is inoculated into 5mL of LB culture solution, the culture is carried out overnight at 37 ℃, and the bacterial liquid is inoculated into 5mL of LB culture solution according to the proportion of 10% and cultured for 3-5h at 37 ℃. Taking host streptomycete spores, centrifuging at 4,500rpm for 3min, removing supernatant, washing twice with ultrapure water, and centrifuging at 4,500rpm for 3min to remove supernatant. Adding 700 μ L TES, mixing, thermally shocking at 45 deg.C for 10min, adding 750 μ L2 × spore pre-germination solution, and culturing at 30 deg.C for 3-5 h.

Centrifuging the cultured Escherichia coli at 4 deg.C and 4,000rpm for 3min, removing supernatant, adding 10mL LB, washing twice, centrifuging to remove supernatant, adding 1mL LB culture medium, and mixing Escherichia coli cells; centrifuging cultured streptomyces receptor spores at 4,500rpm for 3min, removing supernatant, washing with 1mL LB twice, mixing the treated escherichia coli and the treated spores uniformly, coating on a MYS culture dish, adding 1mL ultrapure water containing a proper amount of antibiotics for screening to cover after 24h, and culturing at 30 ℃ for one week until zygotes grow out.

Example 4 method of in-frame deletion on pyrazolomycin genome Using the pCRISPR-cas9 method

annealing the self-ligated gRNA enzyme to the Cas9 vector treated by XbaI/NcoI, performing enzyme digestion and recovery by StuI after verification is correct, amplifying the left and right arms (about 2.0kb) of the target gene by high fidelity enzyme, splicing the left arm, the right arm and the Cas9-gRNA in a Gibbson splicing mode, then performing transformation, culturing at 37 ℃ for 8h, and performing subsequent verification after a transformant grows out. The single clone which is verified to be correct is cultured in a culture medium at 37 ℃ overnight, transformed in E.coli ET12567/pUZ8002 competence after quality improvement, and then a subsequent test is carried out according to a method of heterologous expression conjugal transfer.

[ example 5 ] protein recombination, overexpression, isolation and purification containing genes encoding pyrazolomycin biosynthesis

Carrying out PCR amplification on a target gene, cloning the target gene onto an expression vector after DNA sequencing verification is correct, transforming an expression plasmid into Escherichia coli E.coli BL21(DE)/pLysE, selecting a positive monoclonal, carrying out overnight culture at 37 ℃ in 5mL LB culture medium, inoculating the positive monoclonal into 500mL LB at the temperature of 1 percent, carrying out culture until the thallus OD ₆₀₀ is 0.5-0.8 at the temperature of 37 ℃, adding IPTG (final concentration of 0.1-0.2mM) for induction, carrying out culture at 18 ℃ for 20h, and centrifuging at 6,000rmp for 15min to collect the thallus.

Adding a proper amount (20mL-30mL) of lysis buffer into the collected thalli, shaking for mixing, ultrasonically breaking escherichia coli cells by using an ultrasonic breaker, and centrifuging at 4 ℃ and 9,000rpm for 30min to obtain supernatant. Loading the supernatant into a gravity column filled with nickel at 4 ℃, eluting with Tris buffer containing imidazole at different concentrations (20mM-200mM), performing SDS-PAGE analysis on the eluted samples at different concentrations, and collecting purer protein samples.

Sequence listing

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tggccttagg tcagtgactc ctttactcgg ccggttacac ttggagacga cctggaagaa 480

caggccggag cacgccgctc agtcgttcca cgactggggg tcaaggggtc gcaggttcaa 540

atcctgtcgt cccgacggtc gcagaaagcc cctctggcca ggtaaaatac ctggtcggag 600

gggctttctt atgtctccgg tctatcacgc tcgtgatctt ctctggacca ttccggggac 660

ctgagtgaca aagtgagtga caacggtttc tagctggccc tcgggaacag ccccacccgc 720

agttgccgaa gagcgtggcc gtcgagcacc gggcgcaccc gatcggatcg cggaaactcg 780

gcggccgtca gtggccgtca gtggctggct gtggagagtt cttttgctgg cgtcgccagc 840

agccggtagc ccgacgccag cagcacaagg cagcccagcg cgccgaaggc cccgcacgcc 900

gctgccaggt gcatgttccc gttgtgcagc accatgccca cgattgccgc gcccagcgag 960

ttgccggccg cgaacagcgt caccagccag gcgaacgcct ctgtcgcggt tcctgtcggg 1020

gccagctcgc cgatcagctt gaacaccgag accagtgttg gtgccaggaa gacgccggcc 1080

accaccatga tgccgaacat cggcactggc ggcggtacca gggttagcag ggcgtagccg 1140

aggagcaggc cgccggagct gaacagcagg cgcttgcgcg tctccatctt ccaggtcacc 1200

gcgccgtagg cgaggacgct gaccagcgag gccagcgagt gcaccgtcag caacatcggg 1260

gcgccgccga agatctcgtg ccgttcggcg taggaaacca cgagcacgtt gagcgtgccg 1320

atggccatgc cggcgcctgc cggggctacc agggcgatga tcagtcctgg cttgcgcagc 1380

gggcccagcc agtggcggtc gtgtgcggtg gggcgccagc ggcgggacgg ggcggccgtg 1440

gcgaacacca gcacgcctgc caggcagagg cctgcttgca gccacagtga ggtgactggc 1500

gtcgcgatcg ccaggcacag ggacaccagg agcgggccgg ccacgaagac catttcctgt 1560

gcggcggagt cgactgcgta aacgctgtcc agctgcttct tgtccgcgat gtccggccac 1620

agcacgcgca ggcacggctc tagtggtggc gtgaacaggc cggccagcac ggcgccggtc 1680

agcaccgtgc tgtgctgggc cggcgccacc gcgatcatga tgaaaccggc ggcggccagc 1740

agggcagtcg gcgccagtac cagggtctgg ccgacgcggt cgaccaggcg gcccagtgcc 1800

ggggccccga gtgcggacgc gatcgcgaag caggcggccg cggtgcccac gaaggcgtag 1860

tcggcgccgg cttcgcgcag tgacagcggg atcgacacgg cggccatcgc catcggcagc 1920

cggcccatcc agctgccgga caggacacgc aggaagtgcg gcattcgcag catctgcaag 1980

tagctcacgg acgaattccc ttgctgatga cccggtccgg gtcgtcgacg acgatcacaa 2040

caacggttcc ggagtgtagg tggccgcctc gggggcgctg gtgagcagct tggcgacccg 2100

tgcgcagatc tcggtgatct gctcggccgc taccccggtg aacgccgacc gatcggccag 2160

cagctcggtc aaggccgcct ggtccaacgg gaaccgctcg tccgaaccga ggttggtcag 2220

gagcgtgttg gtggtgccgc cctcgctcag ggcggtggcc gccgccaccg cgtgttcctt 2280

gatcagctcg tgcgccgtct ccctgccctg cccggcccgc accgccgcga ccagcatctt 2340

cgtggtggcc aggaacggca ggtagcgctc cagttcggct tcgactaccc gtggggccgc 2400

ggtgaagccg tcgagcaccg ccaaggtcgt ctggagaagt ccgtccaggg cgaagaacgc 2460

gttcggcagc gcgaccctgc gcaccaccga acaggacacg tcgccctcgt tccattggtc 2520

accggcgagc tcgccggcca tcgaggcgta accgcgcagt accaccatca gaccgctgat 2580

ccgctcgcag ttgcgcggat tcatcttgtg cggcatggcc gacgagccgg tctgcccctc 2640

cgcgaacccc tcaccggcca gctcgtgccc ggccatcagc cggatcgtct tcgccaggct 2700

ggtcggggct cctgccacct gcacgagcgc ggacacgacc tcgtagtcca gcgatcgggg 2760

gtagacctgg ccgacgctcg tgaaacggcg ttcgaagccg aggtgctcgg cgatccgcgt 2820

ctccagggcg aggaccttcc cggtgtcgcc gtcgaacagg tccagcatgt cctgcagggt 2880

gccgaccggt cccttgatgc cgcgcaacgg gtatcggttc agcaggtcgt cgacgcgctc 2940

gacggcgacc agcagctcgt tggccgcggt ggcgaaacgc ttgcccagcg ggagcacctg 3000

cgcggccacg ttgtgcgtgc ggcccgccat cgcggtggcg gcgtgctcgg tggcgagctc 3060

ggcgagtcgg gccagcacgc tgacgcaccg gtgcctgatc agccgcaggc tgtcgcgcac 3120

catcagctgc tcgatgttct cggtgaggtc gcgcgaggtc atgcccttgt gggcgtgctc 3180

gtgccccgcg agggcgttga actcctcgat ccgcgccttc acgtcgtgca gcgtcacccg 3240

ttcccgctgt ttgatcgact cgagatcgac ctggtccacc acggcggcgt agtcggcgat 3300

cacctgctca gggatgtcca cgcccagctc gcgttgggcg gtcatcaccg cgatccacag 3360

ccggcgctcg gtcgccaccc ggttgcgcgg tgcccagatg gccaccaggt cggatgaggc 3420

gtacctggca gcaaggacgt cgggcacgat ctctttggtg gtcatgtctt cctcggtagg 3480

tgggaccggt ggtggggctc ggcgccaccc ggagagtcct cagtggaagt ccgcggacgg 3540

ccgtgtgccg gagaactcgt ccgtccccgc gtcccgttgc cccctcggtg caccagcgag 3600

tcaaccagca ggctcgcgcc ggagacaacc atccgtccac agtggaccgc acgaagcggg 3660

tgcgggtagg gttgtcgtcg tcgtgtcctg ggtgcggtaa cgtcgaaatc tcgtgtgtcc 3720

gcgcgctgtg tggttggatt gtggccgaac gattcacatc gacctgaagt gaggctgcgg 3780

gcggcatgct tgaggcagtg gttttcgatc tcgacggcac actggtcgac acgccggccg 3840

ccatcggcgc gacgctggtc gaggtgctcg ccggagaagg gttcaccgtc gaccatgcgg 3900

cggtggccgc gaccatcggc aagccgctcg tacccagcgt ggccggcctg cttgagctcg 3960

ctgccgacga ccaggccgtg cggaccgtgg tcaccaggta ccagcggctg ttcgacgagc 4020

gagtgctcgg ccggggcacc gagctgctct acgccggagt cgccgacggc cttgccgcac 4080

tgcgcgggtc cgggctgtcc tgtgccatcg ccacctcgaa ggacctgcga gtggccaccg 4140

ctctgctggg cagttccggt atcgcggcgc acttcccggt ggtgatcacg cacgaccaag 4200

tcgcacaagg aaaaccgcac cccgaaatgg gtctgtgcgc agccgccgac ctcggcgtgg 4260

acgcgggtgc gtgcgcgtac gtgggagacg cggtggggga catggagatg gccgtcgcgg 4320

ccgggatgac cccgatcgga gtggcctacg gagtggcgag cgcggccgaa ctgaccgagc 4380

acggcgcggt acaggtctgc gcggacttcg cggaggtcgt caaagcggtg tcggccctcg 4440

ccgggcggcg cacgaactga ccgccgcgcc cgggagacga ccccgtcgat ggacaaccgc 4500

gagtggtgct gggattctct gctcgactcg tcctgacagt ccgagtcgat cgtcggctgt 4560

ccgccgtgtt tttcgcgtgt cccaaaggac aatcgcgtcg cggagccgat cagcgtccga 4620

atcgatggga gttcaattcg tgttcatcga ctgggagaag atgccgcgca cccggagccc 4680

acgtgcgaag gacagccggc ggatcgccag cgggcagaac atgtccttcg tgcgcatcga 4740

gatcgagccg accgcggagt tcaccggcga gacccactgg cacatgcacg agcagtgggt 4800

ggtcgtgctc gcgggagatg tccggatgac cgtcgccgac gaggaggtcc agttgaccac 4860

cggtgacgtg ctctacattc ccatcgacgc accgcacgcc ccgctggcgg tcggtcccgc 4920

gggcgcgacc tacctggaga tcttcgcgcc gccgaggatg gacctgctgc cggagagcat 4980

cgttccggcg gtctgacgac gaaacctgtg tgcccccggc acgatccgat cgtccggggg 5040

cacacaggtt ttctgctaca gccaggtcgc ggccaagccg ctgttgcgga ccgtggtgaa 5100

ccagtgggcc cggatcacgc cccgctgccg cagtccctcg gcccaggcgg ccgcggcctc 5160

ggtgtcccgg cagaacgcga aacaggtcgg tccgtggctg ctcatcgcga tggcgtccac 5220

actggactga ctgcggccct cgtccagcag atcgaccatc tcctgaccct gcaaggcgat 5280

ctgcgcgcgc ttgaagtagc cggtgaaggt cagctcgttg accacggagc agaacgtgtc 5340

gtagtcgcgc tcggccacgg cgggcgcgag ccccatgaac accaggtgcg aggtgcgcca 5400

cgactcggcc ggggggatcg gcatggtggt cgtgaaccac tcgagttcct cgggaccgcc 5460

gatgttccgg ccgttcacct gcaggatgag gatcggccag tccgggaagt ccatcttgat 5520

caccggagtg ggcggctgga cctcggtcgc gaaatggctg gggcgcaggt acttcgtcgg 5580

ctccgcggcg aagtcgggcg gattgaggtg gccgcagtcg accaggaacc ctccgtgttc 5640

ggccagcccg gtgctggcac cggaggtgcg gccgcggccg agagtggcgg agagccagcg 5700

gatgtccggg gtctggccgc tcagcagccc gtaggcgtag ccggccgcga tcagggtggt 5760

ggtcttggag ccgaacccgc tgtgcgcggg cagcgaggac tggacgtcca gggcgacgtc 5820

cgggccgtcc cagatccgct ggagcttggc cagtccgtcg accagcgcgt ggtgcgtctc 5880

ctcctcggcc ccgacgacgg tggtggcgcg gccgggggtc gcggtggcgg tgaccgtcgc 5940

gttcggtcgc tccgccccga tggaggcgat cccgttgcgg cggcgggatt cgccgtcgag 6000

gctgatcagg gtgaagctga gccgggcggg ggaggacacc cgcaccgcgg ttccctgcac 6060

cgtgctcgcg tcgttggtag tcgtcgatgc tgtcaaggcc gtgctcccgt cgtggctccg 6120

gtcgttggcg agtggtcagc gggcactgat cgacggcacg gcggccaggc cggtgcgctg 6180

cccgagttcc gaggagatcc cgcagacggc gatgaagttc tccaccgcgc gtgcgagccg 6240

ctgcgtccca gctgccgagg gcggggcggc ggtccacgcg gccgtcagcg cggcgtagat 6300

ctcggcgacg gcggcgacga cctcgggggc cagcggcttc ggcagcggcc actgcgctcg 6360

cggtcgtccc tcggcgacgt agccctgcac gacctcgcgc aggccggggt gcaggaagcg 6420

cagcagttcc ttgcagaccg ggatcccgtc caggtagaag cggttctcgt ccggggtgcc 6480

cgcgtggtcg atcatgatcg gctggcggtc ggcgtcgaac ccgaactccg ccttgccgtc 6540

caccagcgcc agaccgatgc tgccgcagtg ctgttccagc gtgcgcgcgg cggtcgtggt 6600

caggtcggcc accacctcga ggatctcagg ctcgaccgcg ccgacctcgg ctgcctccga 6660

ccggctgatg aaccggtcgg tctcctcgta cttggtggtg aactcgacca tcggctcgga 6720

cagccacggc gcgtcgcggt aagggggcac ggtgctctgg tcgagggtgc ctgcggcggc 6780

gcggcggtgc acagaggact cctgaggcag cgcgaggcgg tagatcacct ggagcgggat 6840

ggtcctgccc ggcccggttc gtcccttggt gcggtcggtg tccacgtcga gcaactggat 6900

ccggatggcg tcctccgcca cctgctccag gaagtgcgtg cggatgccgg cggcctcgaa 6960

cagctcgaac gaccgcaccg ccatcgcgca ggaagcggcg cccttgcctg ggatctcgtc 7020

gggcatgatg ccgaagtcga acacggagta gcggtcggag aagacgaaga caccctcacc 7080

cggccggccg tcgctcggcg ggaccagtac ttcaagatcc ttgcttgacc agcgtttcac 7140

gtgtttcctt ccgtgggcgt gcagcatcaa cacgacgccg atcccggcgt cgcgcgcgat 7200

ggacgatctc tttctccccc aggggaagcc ccggcgcggg aacctatacg cgagctggtt 7260

cgctgccaag acaagccatt cgtgccgggc acgccgtgcg ccgtcaagtg aggaaattcc 7320

tcagtcggtg gcgaacgccc caccgcagcg ctcgaccagc gccccgccca ccgcgtagcc 7380

ggcggtcagc gcgggaccca gcgtgccgcc cgatcccggg tagccgggcc cgaacacgcc 7440

cgcggagatg ttgccgcacg cgtacagacc cggaaccgcc gtcccggtcg ccgtgagcac 7500

ctcaccggac gtgctggtga ccgggccgcc cttggagccg agcgcgcccg gcacgacccg 7560

cacggcgtgg aacggcgggg tgacgagttc accgaggcac ggattgccgg ggttgcgcgg 7620

gtcgccgtag tagcggtcgt gcgagctctc gccgcggtgg aagtccgggt cgacgccggc 7680

acgggcgtgg gcggagaagc ggtcgacggt cttcgtgagc tccgccgggt cgacgccgat 7740

cgcctcggcc agcgcggcgg gtgtggcggc actggtgagc caggagggaa ccggatcgcg 7800

cggtgcggcg ggaccgacgt tgtagctggt gcggaacttc tcgtcgaaca cgagccacac 7860

cggcaggtgg gccggctggt gcaggtcggc gtcgaagttg aacaggaccc tggtgatgtc 7920

gtggtagttc accgcctcgt tcacgaaccg gcggccgtgg cggtcgacca tgatcgagcc 7980

cggcaggcag cgctcaccga ccaggtggcg ggtcagcggc gcgccgtcgt actcctcggg 8040

cacgccctgc agtgcgggca cccaccaggc ctggttcatg cgttccaccg cggcgccggc 8100

ggacatcgcc atgcgcagac cgtcgccggt gttcaccggc ggactgaccg ggacgaccgg 8160

gatgtcgccg aggaaagccc cggaggccgg atcccattcg aagccgccgt tggccagaac 8220

gacggctctg gcgctgaacc ggcggccgtc ctcacaccgc acgcccgtca cggcggcggc 8280

gtcgttgtcg tcgcgcagca aggacgtggc acgcgcccgc aggacgaacc tgacaccgag 8340

gtcgtcgcac gccgcgacca gcgcggccac cagtgccgcg cccacggtca gcacgccgtc 8400

agcaagtcgc tgggccagca gggaagcgtc cgggccggtg aaacgggcac ggtgacgttc 8460

gtcgtaggtc agcggaggga actgcgggcc cctgcgcaca cgctcgagca ggccgggacg 8520

cgacgccgtg gggaacggct cgttgtcgag gcaacggccg acgtcgacgg cgccgggcca 8580

ggtggagtgg tagtccgggc ggtcgatcgg gaagaaccgg gcgccggtgt gcgccagcag 8640

ccagtcgacc atctccggcg cggtgcggac gtagtgctcc aggcgctcgc gcgggaccgt 8700

gccttccacg acccgctcga ggtaccgcag cgcgtcctct gcggagtcgg gcagaccggc 8760

ctgcttcatc acggagctgt tcggcaccca caacatgccg ccggacaccg cggtggtgcc 8820

accgagcacg tccgagcgtt cgagcacgac cacggacagg cctgcggccg ccgccctggc 8880

ggcggcgagc aggccggcgc ctcccgagcc cacgacgacg acatcgaact tttccaggtc 8940

cacccaatta ccttccccga aaccggacga cggggacagt atttgtggtt gttgtggatg 9000

ggcaagtgag agaacagttt tacgtctcac tcggacggag catcggaatt cacgctgggt 9060

gacgaccggc cgggtgtcgg agggggcctg ttgttgccga gtcggcagta atatcggggt 9120

gttgttccgg catgtggacg tgatgtggcg tcgaagtggt cgtacgcgtg tgtactgcgg 9180

cgattggctc gatcgccgcc tcggccggcc gtgacggtgg cctgcgtctg gctgggacgt 9240

taaatccgcg tacgcgaacg tcggcgaccg gcaacttctt cacggaatga ttgatttggt 9300

tgcgaacgcg ccaccgtgcc ggtttactgg cggtcggccg ctggggagcg gtgcgagggg 9360

ttccagcgag ctgtgtctcg aaaccagcaa ctgtgtgggg ggttggcgca tgcgctccgg 9420

cgcttattcg tcatcagcaa cgatcgtcgg ctatcacttg tccggaatgg acctgacggg 9480

tccgtcctcc gagctcgtgg cgaccgcgcg cacggtggaa gaggccggtg ccgactacct 9540

ggtgctgcct gaccgcacgg cagcgaggcc ggacggcgcg agccctccgg ccgcgctgat 9600

cgccgccgcc ttcctggccg cgcacaccag cgagatcggg atcgtcgtca ccgcggcgac 9660

cggctaccac gagccgtaca gcctggcgcg tcagatcgcc tcgctggacc acatcagcgc 9720

cggtcgcgtc ggctggttcg ccgacagcgc caccgatgcc gcctccgacg ccaaccaccg 9780

gcgtgagggc cacgacccgg cagccgcccc ggagaccagg gcgctcgagt tcgtgccgct 9840

ggtccgcgac ctgtgggaca gctgggagga cgacgcgttc gtgcacgaga aggacaccgg 9900

caggttcgtc gacccggcga agatccacgt cgtcgaccac gtcggcacgg cgctcgccgt 9960

gcgcgggccg ttgaacgtca tccgcccgcc gcaggggcac cccgtcgtct tcgcgcgggc 10020

cgccgacacc tcggtcgcgc accacgccga cgtgctggtt tccgatgtcc cgcaacaggg 10080

tcacatcctc gcggtcgagc ccttcgtcgc ggcggacgaa gtcgccgcgc gcgaactcca 10140

cgcagcggcg ggaagccccg ccggcggcga cgtcctcgtc ggtacccccg agcaggtggc 10200

cgcccagctg cgagaccgcc acgtcctggt gcggttcacg actcgggcac agctggtcgc 10260

gttcacgaca cacgtactgc cgttgctgga caagagcgct cccaccgggg cgacgctgcg 10320

cgagcggctc gggctgcccg cagtggccaa ccgattcgcg ccgacccagg acaaggagct 10380

catcggcaat gcccgctgac cgcgaactgc acctcggcct gatgttctgg gccaccggca 10440

cgcaccaggc cggctggcgg cacccggacg ccagcgccga cgcggcctac gacatcgaac 10500

tcatccaaga ggtgtgccgc acggccgagc gcgccaagtt cgacttcgcg ttcctcggcg 10560

accggctcgc ctgcgacccc gcgttgcagc acagcaaccc ggcgcagatc tcccggctgg 10620

agccgttcgc cacggccgcc gcgatcgccg ccgccaccac gcacatcggg gtcgtcgtca 10680

cggcgaaccc gacgtactcg gacccgttcg gcgtggccag gaacctcgcc tcgctggacc 10740

acatcagcgg gggccgcgcg gcgtggaacc tggtcaccgg cgcggacgcc gcggcggcgt 10800

tgaactacag ccgcgaccag cactgggaca cccagaagcg gtacgactgg gccgaggaga 10860

cgctggagat cgcccgcgcg ctgtgggact ccggcgacca gccgatcaac caccgcagcg 10920

agtacttctc gatcgacggg ccgctcggcc tgccccggcc gccgcagggc cacgtcgtcc 10980

tgctcaacgc gggcacctcg gaccaggcgc gggaactggg cgcgcggcag gccgacatgg 11040

tgttcgcggg cccgcagccg acgctggcca agcgcaagga gtactacgcc gacatcaagg 11100

cccgcgcggc gaagtacggc cgcgaggacc acgtcacggt gatgcccggc ctcaccccga 11160

tcgtcgcgcc cacgaccgag gaagccgtgg cgctgcacga ccagctgaac tcgctgatcg 11220

tgctcgaccc cgaggtcgag gtcggtgagg tcgtgcgcgc gggtgggatc ggtgagggct 11280

tcaagcgcaa cctggtctcg gcctcggagg cgttcggcgt gaacctgcgg ggcaacgacc 11340

tgaacgccgt ggtgcccgcg gaaacgctgg cacgcgtttc cgaggacggg cgcaggcggc 11400

tcgccgagat cacccggctc acccggcgca ccgccgacgg cccggagcgc atcacctacg 11460

cagacctcgt gcacgccaca ccgcagcagc tgtcgcacgt ggtcgtcggc aacccggagg 11520

agatcgcgga cgagatccag ctctggttag aggagcggat ggcggacggc ttcaacatct 11580

acccggccta tgtgcccggc tcggtcaccg cgttcaccga actcgtggtg cccgtgctgc 11640

agcggcgggg gttgttccgc aaggactacc ggggtcgcac cctgcgcgac catctcggct 11700

tggcggtacc tgcggtgcgt tgacaccgtg agaagtagac gccgcgtcgt ccatccacca 11760

cacggccggt tccggccgtg gttcgaccat gcgttcactg ggggaagagt tgtctgtgca 11820

gttgcttcgt gaggaaagga cttcctggtc catccgggag aaatcgggtt gtgtcaccgt 11880

gccgatctcg ttgctggggg aggcggactc tcctcggcag gcaggtgtca acgcggagca 11940

cgtgcgggtg ctcacggggg tggagaacct gccgccgatc ctggtgcacc gccacacgat 12000

gcgcgtcatc gacggcatgc accgcgtgca ggccgcgctc gcccgcgggg aggacaccat 12060

cgaggtggtg ttcttcgacg gcgacgaggc cgcggcgttc gtgctcgccg tcgagttgaa 12120

cggcggacac ggcctgccgc tcacgctcgc cgaccgcagg acggccgctg cacgcatcgt 12180

gcgggcctat cccgagtggt cggaccggcg catcgccgcg gcggccggca tctcgcccaa 12240

aaccgccggc gcggtgcggg cgcaactgtc aagtgaggaa attcctcaga tgcgcgaacg 12300

gctgggactc gacgggcgag tgaggcgggt ggcaccgcgt accgtgccgg ctcataccac 12360

cgagccggag cgggtgcagg ccaggccggc gcgggcaaaa caaccgcagc ctgagaggca 12420

gctcgacttc ggcgcggtca tctacgcgct gcggcgcgat ccctcgttgc ggttcaacga 12480

gaacgggcgg acgttgctcc ggatgctgga catgcaccac ctccccgcgg cggggtggag 12540

cggcatcatc ggcgcggtgc ccgcgcactg cgcgtccgtg gtggcggagc tggcgcgcga 12600

gtgcgccgag gcgtggttgg tcctcgcgga cgaactcgac gagaagtcgg actagtggta 12660

cgtcttccgg cgcccctgcc gggcgccgga agacgcggtt tcaggcgaac ctggtgagct 12720

ggaacggttc ggccagcggg tgcgggtcac cgagcacctc gcgcgccgcg atctcgccga 12780

gcagcggcgc cagggtcagg ccgctgtgcg tggcgatcac gtagacgcca ttcctgcccg 12840

gcaccgcgcc gacgatggac atcccgtctc gcggcatcgg ccgcacgctc acgaacgcct 12900

tctccacctc ggcgtccgcc gcaccgggca ggacctcggg cagccgggcg agcgccatct 12960

ccgccagcgc cgccacatcg ctcgactcgg tgagcgtgtg gtcgaggtcg tggcagtgca 13020

gcaccagccg gttgccggtg tgcggccgga tgctcagccc tggcgcgtgg atgatccggt 13080

tgggcggcgc ggacaccggc gtggtgcgca ccagcaggcc tcgggtcagc cgctccgggg 13140

cgccgggtgg caacagcggc agcccgggca gcagagtggc gttgccccgg cccgcgcagc 13200

acagcaccgc gtccgcgcgc accgcgtcca gtgactcgac ctgttccccg atccgcacgt 13260

ccgccccggc cgccctggcc ctgtccagca ccgcgggcag cagcacgtcg ccgagcacgt 13320

gcatgtcgcg cgggtagaag tagacgggtc cggtgacgtt ctccagcgcc agtcccggtt 13380

ccaggtcgcg aatgacctcc tcgggcgtca gttcctccac cggatagccg agttcgcggt 13440

agccgtccgc gatggcccgc tgcgcggccg cgctcgcgct gtcgcggccc cagtgcagat 13500

tcccctgccg gaaaagccat tgccgtggtc ccgcgacgtc ggcggccagc cgctcctgcg 13560

ccgccagcgc ctcgatgctc agctcgaagt acgcgctgtt cttctcgtcg atcgcgttgg 13620

cccagccgaa actgtgcgcg gacagggtgt ccagtggttc gtcacgcgac atcacggtca 13680

cctgtgcgcc cgcctcggcg agccgcagtg cggcgcacgc gccgatcgcg cccgcaccga 13740

tcaccaggac cttcatgctc tctcctccgc tgaacgtgtg gcgcaacgat ggtgggtccg 13800

gtgcccgccg cgcaacgtgt cctttatgga cggatttttg atggcttccg ggacgcgtgg 13860

ttgactcgcg gtcgtcgtca gcaagcagaa gccgggcaaa aggtgtggag gatcgcttga 13920

gcagccgcag cgtcctgttc gtcaacctga ggcgcatcaa gcgcgagggc ttcgagtcac 13980

tggtggccgc ccgtcgtctc ggctaccgcg tggtgctgct gggccgggcg ctgccggagt 14040

tcgctcagcc gctggtggac gagttccacc aggtcgacac ctacgacacg gacctcgctc 14100

ttaagacggc gcgggagatc gccgctgcca acgacctcgc cggtgtcgtg aacttcaccg 14160

agatcgacgt ccagctcgtc gcggcgatcg ccgccgagct cgggctgccc ggcatgccgc 14220

cggaggccgc ggtgctggcg cgcaacaagc tcgcgatgaa gcaggcgctc gccggaatcg 14280

acggcgtgct gccgaagttc gcgcgcgtca gtgatctcga cgacctgtgc gccgccgtcg 14340

cggacatcgg tttcccctgt gttgtcaaac ccaccggtgc ctccgggtcc aaggggatct 14400

tcgagctgca cggtgaaagc gatctcgaac cggcgatggc cgagttgcag cgcatcgcgg 14460

atccctcgtt cgacgcggtg ttccgccagt tcggtgcgga gttcatcgtc gaggagtacc 14520

tgaccggaga cgaactgtcg gtcgagggct tcgtcgcgga cggcaccgtg caccagctgc 14580

tgctgaccga caaggtcacc accgtcccgt tccacctcga ggtttcgcac cggctgccga 14640

gcgtgctgcc cgccgccgcc caggaccaga tcctggcgtg cagcgagaag atcgtgcgcg 14700

cgctcgggtt cgacaactgc gccttccacc tggaggccaa gtgggacggc gagcgcatgc 14760

ggttcatcga ggtggccgcg cggcccgccg gtgactacat cgcctcgcac ctggtgaagg 14820

ccgcgaccgg catcgacttc ttcgccaacg tcatccgggt cgcgacggga gagccgttgc 14880

agctcgtgcc cgaccgggac ctgcaggcgg ggctgcggtt cgtgttcgcc gagagcgccg 14940

gcacgttcga cggcctcgac ggtgtcacgg aactgttcga ggaaccgggc tacgaccacg 15000

tgttcaccga agtgccgatc ggcgcccagg tcgcgttgcc gcccgcgaac ttcggctcgc 15060

agcgcgttgc cgcggtctgt gcccgcgcgc atgaccgcgc cgggctcgac gcgctgctcg 15120

acaccgccgg agaggcgatc aaagtccgga tcggctagtc acgaccacag ccgggggaga 15180

gggggaaccg tgcggctgac gttgctggga tgtggccgga tggggcgcgg tgcgggctac 15240

gcgctcgccc gcgaccagcg caccacgtcg atcacggtcg tcgaccacga tcgggagcgc 15300

gcggaggagc tcgcgcgctg gctctccgga tacgcggcgt gccctgtgaa gaccggggac 15360

tcggacgcga tcgcgggcag tgacgcggtc gcggcggcgc tgccgtggtc gggaacgcgc 15420

tcggtgatcg aactcgccgc ggcggcgggc gttccggtcg cgagcatcac ccgcccaccc 15480

ggcgacgaat cgtccgatgt ggacgcaatg gcgaacgccg ccggtgtccc ggtgctgctg 15540

ccggtcggcc tcgagccggg actgaccgag ctggccgccg cggacgtcgc gcgcaggctg 15600

accgctctca ccggaggtgt gcgcacgctg gaggtgttct gcggaggcgt cccggccact 15660

ccacgcgcgc cctgggggta caccgcgttc ttcggcggtg agctcgcgaa ccacctgccg 15720

atcgcgcaac gctcctcgat cgccgtcgag cacggcgaga tcgtcgacca cccgcgcttc 15780

tccggggtcg aggaacggca cgtggccggc gtcggcctgc tggaggccta ccacgacggc 15840

atggtgccct ggctggccga ccacccggcg ctgcgcgacg ccgactgcac gcagaaaacc 15900

ctgcgctggc ccggattcgc cgacgccgtc acgggtctgg cgaacctcgg gctgctggac 15960

gagtcaccgg tcgcggtcga cggtgtcgcc gtggccccga aacgccttgt cgaaaacgtg 16020

ctgtcgccgc ggttgcgcgc gcagcccgac gacgaggacg tcgtggtgct ggacgtgtcc 16080

gcgaccggcc tgccggacgc ggacggcagc accctcagca tcaggtcgct gctggtggat 16140

cgcgccgacc gggagaccgg gctggccgcg atgacccgca cgaccggatt caccctggcc 16200

gcggcggtca cgctgctggc cgaccgcacg gtcgggggcg cgggctggct gcgcccgcac 16260

ctggcgttgt cggagcggca cttcgcccgc atgaagaccg atctggccgc tctcggcgtg 16320

cggtggaccg ccgacggaga ggccctgcac ggtcaccggg cggggacggt cgcatcgaaa 16380

ggcactgact gatgagcacc ggacacgacg tcctggacgt ggtggggatc ggtatcggac 16440

cggcgaacct gagtctcgcg gcactgctgg ccgccgagag cagcgagctg aacgtggcgt 16500

tcctcgaccg caagccgcac ttcggctggc attccggcct gatgctgccc gacgcgcaga 16560

tgcaggtgca ctacctcaag gacctggtga cgccggtcga cccgacgaac ccgttctcgt 16620

tcacggcgtt tctcgtggcg accaagcgct tctaccggct gctcgtcacc ggccgcagca 16680

aggtgccgcg ccgcgagttc gagcagtact gccggtgggc ggccgagcgg atcccgaacc 16740

tgcgcttcgg cgtcgaggtg cgcgaggtga cgtggaacgg cgagctgttc gtcgtgcaca 16800

ccgaccaggg cgtcctgcac gcgcgcaacg tggtcagcgg taccggtctg gtgccgcggc 16860

tgccgtcgtt cgccaccggg cacgatccac aggaggtgtt ccacgcttcg acgctgctcg 16920

acgtgccgag aacgttcgcg ggccgccggg tcgcggtggt cggcggcggg cagagcggtg 16980

ccgaggtcgt gcaccacctg ctgacctcac cggaccggcc ggcgtcgatc gtgtggggca 17040

cctcgcgttc gaacctgctg ccgctggacg acagcccgtt cgtcgacgag ctgttcgtgc 17100

cgaactacag ccgctacttc cacggtctgc cggccaagcg ccggatggaa ctggtggacg 17160

agcagaagat ggccagcgac ggcgtctcgg tgagcctgct ggaggcgata taccaccggc 17220

tctacgacct ggagatgctg gagggcgagg aacgtccgtg ccggatgctg ctcggccatt 17280

cgctgaccac tgtggacaag acgccgaccg gcctgctcac gcggtggatg ccgggcgccg 17340

gcgctgaagt cggtcacgag gtcgacgtcg tcatctgcgc caccggttac cggcacgggt 17400

tgccgaggcc gctgcacggc ctgccgcggc acctgcgcga ggcgatgtgc ggcgacgacg 17460

gcgagatcac cgtgcggccg gacttcagcc ttgactggga aggcccgcag gaccggcgga 17520

tgtacgtgca gaacgccgcc aggcacagct tcggcgtcgc cgacccgaac ctgagcctgc 17580

tcgcctggcg cagcgccacc atcgcgaaca gcgtgctcgg gtacgagcgt tacgacgtcg 17640

gcgaggtggg agcggtgctc gactggacct cctcagcaga ttccgacgtc gcgctgacgt 17700

tctgaccgaa caggaaccgt ctgatgatcg tcagtgagat cccccgcgag ctcgccgacc 17760

tggcgcgggc ggacttcgtg cgcagtgccg ccccggcgtg ggccgaggcg gcgggagtgc 17820

cgttcaacgt gcgccgggtg acgttgcggc cgggcgagac gactgccgag cacaaccacc 17880

acgacctgga ggtctgggtg atgctcgacg gtgtcggcga ggtcggctgg gacggccatc 17940

agcgcgtgct gaccgcgggc gacagcgtct acctgccacc tctcgcgccg cacaccctgc 18000

gcaacctctc cagcgaccgg cccttgtcgt tcttctcgat gtggtgggag aacctgtccg 18060

cgctcgctgc ggtgcacgcg gaacgccggg aacaggctgt ggaacggcag ggccgccccg 18120

tgctgctgct gccgtcgttc cccacgccga acggtgaact gcacctcggg cacctgtccg 18180

gaccgttcct caacgctgac gcgtgccggc gtgcgttgct ggcggcgggc gaacgcgcgc 18240

acctgctgct cggcacggtc ggccatcaga gccaggtgtc cgcggccgcg gaggcggaag 18300

gcctgtcgtt ccacgagctc gccgaacgca acaccgacgc gatcatcgaa ggtctccagg 18360

cggcgggcat cgactgggac gtgttcgtgc ggccgtcgga gcccgcctac ccggcgatgg 18420

cgacctcggt cttcgagtcg ttgcgcgaca ggggagtcct cgtccgccgc actgaaccga 18480

cgaactactg cgaaccgtgc gggcggttcc tgctcgaagc gttcgtcgcc ggccactgcc 18540

cgcactgcgg gtcgaaccag acggccggca tcgaatgcga gttgtgcgcg ttgccctacg 18600

acgaccgcga tctcgtcgat ccgtcgtgcg ccacgtgtgg tgccgccgcg acgcaacggc 18660

cactgacccg gtacttcatg ccgttggaac cactgcggga cgagctcagc ggctacctgc 18720

gcggtgcggc gatgcacggc aggctgcgcg cctacaccga gcgcgtgctc gcgaagacgt 18780

tgcccgacct gccggtgagc atcccggccg agcacggcat cccgatccac gtcgaggacg 18840

cgtcggggcc cgccgagcag cggatgtact cggcgttcga gctggccgca cggttcctca 18900

ccgcgctcga cggattcgcg gacggctggg aggcgtacgc gcggcaggag aacccccgca 18960

ccgtgctgtt cttcggcttc gacaacgcgt tcctgcgagc gttcgcgttt ccggcggtgc 19020

tgggcgcgtt caccgatgcg ctgccgctgc cagaagcgct ggtgtgcaac gacttctacc 19080

tcctcgacgg ggagaagttc tccaccggcc gcaagcacgc ggtgtgggcg cggcaggccg 19140

tcaccccggc caacgccgac cagttgcggc tctacctcgc ggccacctcg ccggacgtgc 19200

gccggcgcga cttcaccacg cggggctacg cggagttcgt gacggccgag ttgatcggca 19260

ggtggcagcg ccggctcgac gacgtcggcg ggcgggtggc cgaacacttc ggcggcctca 19320

ccccggaagc gggaggctgg cacgccgagg cggaacgctt ctacggtcag atcaaggagt 19380

tcgcgtcgtg cgcgacgctg gactacctgc ccggccgctt caagccgcgt gcggtggtgg 19440

cggcggcctg cgcgttcatc cggcaggccg aggacttcgc cgaggtgagc gcggacgcca 19500

cccccggctc cgggatcgcc aggacgtgcg cggcgctgga gctgatggcg ttgcgcacgc 19560

tggccatggc cgtctggccg ctggctcccg agttcggccg gcgggtcgcc gccgcgctcg 19620

gagaggacac catcgcgctg gagccgacgc cgcgctgggt gcgaccggat acggagatca 19680

agttcgccac cgatcacttc agccccgatg aggtcgtcgc gggacgctga gcgtgatccg 19740

gaccgccgaa cagcacctgg gaggcagggg agcatgcgcg acaccgacac cgacaccgac 19800

accgacaccg atgccgacgt cgccgggccg gcgagtgtgg tggacgagcg ggagccggac 19860

atcaccggcc gcagcaagaa actgtggctg gtgcccggcg accggtgcgt cgtggaactg 19920

gttcccagcc tgcgcagctt cacccgcgcc cgggacgagc tcgtcgacga gaccggtccg 19980

ttgcggctgg acttctacga gaaagccgcg gcacgactgt ccgatgcggg tgtcgagtgc 20040

gcgttccgcg cccgtctcgg gccggtcacc tacctcgccg actaccggcc ggcgccaccg 20100

ttcgaggtga tcgtcaagaa cgtggcgacg ggttcgacga cacgcaaata ccccggcctc 20160

ttccccgacg gtgagccgtt gccccgcccg gtggtcaagt tcgactaccg cgtcgacccc 20220

gaggaccagc cgatcggtga ggactacctg cgggtgctcg acctgccggt cgacctgatg 20280

cgcgagcagg cgctgctggt caacgacgtg ttgcgggact ggctgaaccc gctgcggctg 20340

ctggacttct gcgtgatctt cggtttctcc tccgccggtg agatcagcct gatctcggag 20400

gtgtcgcagg actgcatgcg gttgcggcac ccggacggct cgccgctgga caaggacctg 20460

ttccgcggcg gtgcgtccgc cggcgagctc gtcgagcagt ggaagagggc gttcgatggg 20520

ctctgagtcc cctgtcgcac tggtcaccgg ttcgaaccgg ggcagcggac gtgccatcgc 20580

cgcgcggttg cacgagcgcg gctatcgggt gttctcgttg aaccgcaccg tgaccggcga 20640

ggactggctg ggtgagcggg agtgcgacct cgcgagtcgt gagtcgctgg cggccggggt 20700

ggcggcggtg ctggagcggg ccggccggct ggacgtggtg atcgccaacg cggtcgagcg 20760

cgtgctggac ccgatcgaga agatgtccga acaggactgg gaccggcagc tggagatcaa 20820

cctgtcgtcg gtgttccggt tggtgaagca ggtgctgccg gcgttgcgcc gctcctgcgg 20880

actgttcctc gtgatgggca gccacgcggg gtcgcgttac ttcgagggcg ggagcgcgta 20940

cagcgccacc aaggccgcgc tgaaggcgtt ggtggagacg ttgttgctgg aggagcggaa 21000

caacggcgtg cgggcgtgtc tgctggctcc cggcgcgatc tcgaacctgg acggtgacga 21060

cgcgcccacc aaggtgagcg tcgagtccgt cgggacgtgc gtggcgagca tcgtcgcgga 21120

ctggccggcc gatctcgtgg tgggggagct ggagttccgc ccggcgctgc tgcccgagtc 21180

cccggtgacc ggcatcgacc gtctgctgca cgtctgatgt tcttcgtcgt caccgacctc 21240

gacggcacgt tgctcaccag cgaccggctg gtcaccgagc gggcgaagag cgcgctgacg 21300

aaggtgcgct cctccggtgg tgtcgtcgtg ctgatcaccg cgaggccgtt gcgggatgtg 21360

accgagatcg gccaggaggt cggtgcgaac ttcctggtgt gctcgggcgg cgcggtgctg 21420

tacgacccgg tgcgcggcga gctggtgcgc gcgaccacgc tgggctcccg ccgggcgacg 21480

agcctgatct cgttgctgcg ccagacgttt cccggaatcc ggctcggtgt cgaccacctg 21540

gcccgctgcg acctcgaccc cggcttccac gtgggcgcgc cgggcgtcgc ggaccggcac 21600

gcgacggagc cgttgcggca ggtcgccgaa ccagcggtga agctgatcgc gcagtccgac 21660

gagatgccgg tcgaccggct cgcgcgggcg atcagcgcgc tggtcggggc ggcgtgctcg 21720

gtggcggtgc cgtgcagcta cttcgttgac gtgctgcccg ccggggtgca caaggccgtc 21780

ggactcggcg agctgcacga gtgccacggc cgcgtgctgc cgtcgatcgc ctttggtgac 21840

atgccgtcgg atctgccgat gttgaggtgg gcggacgtct cggtcgccac ggcgaacgcg 21900

caccccgcgg tgctggaggc gtgcgaccac gtcacggcgc accacgacgc cgacggtgtc 21960

gctgtctatc tggaaagcct gatcagcact gggggatgag gaaatggtca ctctcgtgct 22020

gcccacgggg tcgttgcacg agccgactct gcgcctgttc gacgcggcgg gcctgaccgt 22080

ccaccggcag gcgtcgcggg cgctgcgggc acacgtcgac ttccccggca tcgagcgggt 22140

ggtgttcggc aaacctcgcg agattccggg tctggtcgcg gacggcgtcg tcgacctcgg 22200

gctgaccggc acggactgga tcgaggagag cggcgccaag gtcgaggtgg tgcacgagtt 22260

ccagtactcg aagaccacca gcgccggttg gcgcgtcgtg ctcgccgtac cggtcgacca 22320

tcccgcgcgg accgcggcgg atctgcccgc gggcgtacgc gtcgcgtcgg agtaccccgc 22380

catcgcacgg cgctacttcg aggaggaact gggccaggag gcgcgcatcg tgcactcgca 22440

cggttcgacc gaggcgaagg tgccggatct cgcggacgct gtgctggaga tcgtggagac 22500

cggggacacg ttgcggcaca acgacctacg cgagctggtg gtcgtgcggc gatgcgcggt 22560

gcagctggtg gtcgccacgc aggcggaccc cgtcgtgcgc gctacggcgg agaaggtcgc 22620

gttgctgctc aagggcgcgc gggcagccac ggagcacgcg ctgctgaccg tggtggcgcc 22680

cgctgactgc tgggaccggg tgcggcagga actgccgcgg tactggtggc tgctcggcgg 22740

agacccggaa accgtggtgg cacaagcgac ctacgagctt cggggcgtcg cggaagcgat 22800

cacgcggctc acacaggccg gtgcggtgca ggtcgtggag acgccgatgc gaaagctggt 22860

gacggcgtga caggtctccg gagttcctgc cgttcaacgg tcctcgatgt catgtttgcg 22920

cagaatccac tgtggtgaga gaggttttcc aatggccacc aaatccatga acgagactct 22980

gacccgagtc ggcaggggca cgcccatggg cgagctgctg cgcgagtact ggatgccggt 23040

gatgcggtcc gagcgcgtcg tcgcgggtgg tgaacccatc gctgtcgagc tgctcggcga 23100

ccggtacgtg atcttccgcg gcgacgacgg caccgtcgcc tgcttcgacg aggcgtgccc 23160

gcaccgcggc gcttcgctcg cgttggcgcg caacgaggac tgtgcgctgc gctgcatcta 23220

ccacggctgg aagttcacga ccgacggccg gaccgtcgag acgccctccg aaccggagga 23280

cggagggcgg ttcgcgtcca aggtgaagct caaccaccac cccgtcgtcg acgcgggcgg 23340

cgtgatctgg gtgtgggccg gcggcaccgg tcgtgagccg agcccgttcc cgcgcttcgc 23400

cttcaccgac ctgccctcct cgcacgtgtt cggcatcgtg gcgatcctcg actgcaactg 23460

ggtgcaggga ctggaagcgg acatcgactc cgcgcacgtg tcgctgttgc acgagaccga 23520

ggcggcgaag ggtccgctga aggatctgct cgacgaccgc gcgccacgcg acgagatcga 23580

gaagacctcc tggggcatcc gctacgcggc gatccgccga ctgtccactg gggacagcct 23640

ggtgcgggtg aaaccgatgg tcatgccgtg gtacaccatc gtgcccgagc ttcccaacgg 23700

tgaccgcctg tggcacgcgt gggtgccgat caacgaccac cagaccatct tctggtacct 23760

ctggtacaac gaggaccagc cggtcgaccc gaactacttc gccgaccagt tcggcctgga 23820

cctcgagaac atgaacaagg acaacttccg cgagggttac tcgcggaaga acatgtgggg 23880

ccaggaccgc gccgcgatgc gcgcgggcac gtcgttctcc ggcatcagcg gcctcgccgc 23940

ccaggacatc gccgtgcagg agagcatggg cgcgatcgtg gaccgttcga tcgagaaccc 24000

cggcaagagc gacaccggtc tcgtccgggc acgccacttc ctgctggacg cgatcaagga 24060

caaggcggag ggaagggtgc cggccgggct cggcgacgag gtcgactacc gcaagatctc 24120

gtcggcgaac gtcgtggtga gcgacgggga cgactggcgt cagctggtgg tgtagcacgg 24180

tgccggtgcg ggtgacggtg tcagcgcagt tcggccgcga ctgccgggcg tggcgtcatc 24240

ctgcccggca tcgcgcgcac cgtcccggcg gtcaccacga cggtgatcgc cgggatgtcg 24300

ccgacgcgca tggcctccgc caccgtgtcc gcctgggaac cagcgggcgc cgcgagaccc 24360

agcagatccg cgggcgcgcc ccggcgcacc cgtccttccg gtaggtgcca cacgtcggca 24420

accgtgcccg tcgcgagcga ccacgccacc tcgggttcga gcccgccgag cgcggtcagc 24480

tccagcaccg tcttcagcac cgcgagcggc atcacaccga acccggacgg cgtgtccgag 24540

ccgaccacca cgcggtgcag ctcgtcgcgg gacgccgcat gctccacgat gcgcagcgcc 24600

gccttgaggt tgcccgcctg caccacctgc aacgccatgt ccgtctcggc gaacaacgcc 24660

tcgacctccg cgaacggcaa cgcagtcggc ccgccgttcg cgtgcccgca cacgtcgggc 24720

cgcaacgcga gcaacgcctc gcctcccagc gaggccccac ccgcggcact cgccccgccc 24780

gcgtggcaca tcacggtcag cccggcctcc cgcgcccagc gcacgtgctc gacgccgtcg 24840

accggatcgg tgaacgcgcc gaacccgtac ttcgcgagcc gcaccccggt cgccgcgagc 24900

tcccggaagt cctcttcgga cagcgtaggc tcgatcagca ccgcgcccgc gcgcacccgc 24960

atgccgccgg gccggtactc gctgaagcac cgctgcgcgg cgacggcaag cgctttcacc 25020

ccggccctgt cgcgtggccg tcccggcacg tgcacctcgc ccgccgacac cgtcgtcgtg 25080

atgccgccct ggacgtaccc ggcgaggtaa tcgaccgcgc gctgccgcgg gctgtagtcc 25140

ccgaacgtca cgtgcccgtg tgagtcgatc agtcctggcg cgatcgtgcc gccgcgcagg 25200

tcgatcacct cgtcggcgcc gtcgatctcg tgcgtcaggc tgctcgcctt ccccgtcgcg 25260

tggatcatgc cgtcccggca gaccacggtg tcggcgtcga gcagcggtac cgcgagatct 25320

ccggagaaaa cggttgccgc gccggtcagc acgagcgttg ccatgcgtcc tccacatcgg 25380

actcgagtgc gacaaagccg cgtcagtcgg ccgcgaccgg ttgacccgcg ggcacgaaca 25440

accggtagcc ggcaccgagc actgccagac agaacaacgc accggcaccc gcgtgcgccg 25500

ccgccaggtg cagttcgccg cggtccagca ccgggcccac caccgcggcg ccgagcgagg 25560

ttccggccgc gaagagcgtg accagccagg cgaacgcctc ggtgaccgtg ccgcgcggcg 25620

cgaggtcgct gatcaacccg aacaacgcgg ccagcatcgg cgccaggaag aacccggtga 25680

gcagcatcag gccggccatc ggcagaggcg tcggcacggt cgcgagcatc ccgtatccca 25740

ccagcagccc ggccgcgaag gacacgatcc gcgtcctcgg ctggatcgtc caccggatca 25800

tgccgtagat cagagcgccg gccaacgccg cgaaagcgtt cagcgccagc aacatcggcg 25860

cgccgccgaa cacctcgaac cgctcggcgt agctcaccac cagcacgttg agcgtgccga 25920

tcgccacacc ggtgccgacc ggggcgagca gcaggatcgt caggccgcgg tgccgcagcg 25980

ggccgagcca gtgccggtgc tccgtcttct ccggcgccca cctcctggcc ggtgcggcgg 26040

tggcgaacgt cgccacgccc gccagcgtga gcagcgcctg cgcccacagc gcgctgaccg 26100

gagcgaacac cgcgaggcac agcgacacca cgagcgggcc gaagacgaac accagctcct 26160

gtgcggccga gtcgaccgcg taggccttct ccagctcgtc ctcggcgacg aggcccggcc 26220

agagcacgcg caggcacggt tccagcggag gtgtgagcgc gcccgccagc accgcaccga 26280

tcagcaccac cgcgtggttg gcgggtgcga gtgcgatcgt gacgaatccg atcgccgcga 26340

gcagcccggt cggcacgagc acccgcacct ggcccacccg gtcgacgatg cggccgatca 26400

cgggggagcc gatcgcggac gagagggcga aggcgccgga caccagtccg atgaacacgt 26460

agtcggcacc cgcgtcgcgc agcgccagcg ggatcgcgac ggccgccatg gccatcgcga 26520

gcctgccgac ccagctgccg aacagcacgc ggaacacgtg ctgcctgcgc aggacggacc 26580

agtacgtcac gcgagcctcc cggacgtcag tgagcacgtg ccaggcccgg tctcgcccgt 26640

gacgtctccc cggacggact cgccgcacac cgctcaggcg actgacgtcc agcgtccctg 26700

atgccgcaca agcggcggaa cgtcctgccc ggacgtgatc tcgaccggtg cgccgacgag 26760

cgccacgtgg tcgccgaccg cgatgcgctc aacggcgcgg cacctcatcc ggaccggcgg 26820

gtcgagcagg tgcggcacgc cgtcgtcgtc cggcttccac gtggtgcccg gtccgaaacg 26880

atcagcgccg ctgcgggcga acaggtcggc caggccgcgc tgctcggtgc cgagcaggtg 26940

caccgcgaag agcggcgccg cgctcagcac cggccacgcg gacgccttcg taccgatcca 27000

cgccaccacc agcggcgggt gcatgctcgc cgagcagaag gagctgaccg tgacgccgat 27060

cggcccgtcc ggcccggctg ccgtgatgat cgcgacggcc tgcgggtggt gccgcagcgc 27120

gcaccgcagc tcttcggcga tcatgacgcc accaacgcgt tcagcgcgtc cgcgtcgagg 27180

aggtccacga gtgcggctgt gagagcgtcg tcatcgacac cgagggcgga ctcgttgagg 27240

aagatgccgc gaccgggcac ctgacctccc agctcgacga gcagcggccg gaggtgcacc 27300

tccaccgcga gcgcgtgccg gtcggaggcc gcgatctgca acggaagcac cgccttgccg 27360

cgcaaccagt tcggtggcgc ctggtccagc accgccttca acaacccggt gtaggtcgct 27420

ttgtaggtcg gagtcgccac gacgagcacg tccgactgtg cgagctcgtc gagcaccgcc 27480

cgcatccgcg tggagtcgaa cacctcgtgg gtgatttccg ccgcgtcgac ggtcggtccg 27540

acgacgggat ccttgagcac ttgacgagcc gcctcgcgca acgcgagcgc cgcccgcagc 27600

gtacgggaag ccgggcgggg atttcctacc agcacaccga tcgcagtcat gtgctgaacc 27660

ttgcgcgatg gcggcgggcg cgtcgacttc gagtcctgat tggacggtgc cccgggcgtg 27720

atctgagcca tggtcgccga gtgcggcctc ggttgtgttg cttcctccgt cctttcaggt 27780

acgttccagc cgtttggacg agcgccgtca ccgggctgcg gggagcggta cggctggatg 27840

gatccggtgc tcgtgccaaa agggcgatcc ggtcaacgaa ccgtgtgccg cgcatcgacg 27900

cgcggccgga cagcggtgcc ggtagcgcgc gcgtcggttc aggtctcacc tcggctcgat 27960

gcgggcgcag cccgatcttg ttgcaggcca atggatatgt gggggcatgc cgacatggca 28020

gctgggggaa cgaacgtggc cgtcgccgtg cctgcgggtg ctggcgagcc cacctgcttc 28080

atcaggcgaa gtgtggccga gatgttcggc gacgcggtgg ccgcgcaccc ggaccgggtc 28140

gcggtggccg gggacgaccg ctcactcacc taccgcgagc tggccgcgag cgtcaagacg 28200

gcggcatcgc ggttgaacgc ggccggggtg gcgccagacg actgcgtcgg cgtcttcgtc 28260

gaaccgtccg cagagctcgt cgtggccgta tggtccgtgg tgtgcgcggg tggcggttat 28320

gtgccgttgt cgcctgacta cccggacgaa cgcctccgct acatgctgca cgacgcccgg 28380

cctcgggtgg tgctcaccca gcggagcttg cgcgcccggc tggccgcgat cgcgccgccg 28440

ggcatcgagg tcgtctgcgt cgaggagatc acgcggacga cgggaccggt tcccgcgcgg 28500

cagtgggcgg ctccgcggcc cgagcacctc gcctacgtca tctacacctc cggcagcacc 28560

ggtgtgccga agggggtgat ggtcgagcac ggcgcggtgg cgaaccagct cggctggctg 28620

cacgacgtgc acgggctggg ccccggttcg gtggtgttgc agaagacgcc gctgagcttc 28680

gacgcggcgc agtgggaggt gctggccccc gcttgtggtg ccaccgtggt cgcgggcacc 28740

cccggcctgc accgggaccc ggaccggctc gtcgacacga tcgtgctgca ccgggtcacg 28800

acgttgcagt gcgtgccgac gttgctgcgg gccctcgtcg acggcggcag ggcgaccgag 28860

tgcacgtcgt tgcgacagat ctgcagcggg ggcgagacgc tcacccgctc actggccgac 28920

gagtgcctgc gggaggtgcc gtgggccgag ctcgtgaacc tgtacgggcc gaccgagtgc 28980

accatcaacg cctccgccca cgtcgtgagc cgggaggagc tcctcgacgg cccgccggcg 29040

gttccgctcg ggattcccgc gcacggcacg tacttccgcg tgctcgacga gcgctgcgca 29100

ccggtgtccc ccggcgagat cggcgagctg ttcatcggcg gccgccagct ggcccgcggc 29160

tacctcggca ggccggacct gacggcccag cggttcgtgc ccgatccgtt cacgccggag 29220

cccgccaggc tgtttcgttc cggtgacctg gtccgggccg acgcggacgg caccgtccac 29280

ttcgtcggcc gcgccgacaa ccaggtgaag ctgcgcgggt tccgggtgga gctcgacgag 29340

atcaggctcg cgatcgaggc gcacgactgg atcagaaccg ccgccgtgct cgtcaagccc 29400

gacgcccgca ccgggttcga gaatctggtg gcctgcatcg aactcgaccc gcgccaggcg 29460

ttcctgatgg accagggcaa ccacggcgcg caccaccagt ccaaagcgtc gcggctgcag 29520

gtgcgggctc agctgtccaa cccggggctg cgcccgcagg ccgacctggc cggcctgccg 29580

cgggtggccc tgcccgggcg ggagccgacc gccgcgcagc gccgcgaggt gttcgcccgc 29640

aagacgtacc ggttctacga cggcggtgaa gtcacggcgg ccgacctgct cgacgtgctc 29700

aaccggcggc cggcaggcag cgggtccctg cgtcccgagg acgtggacgc cggcacgctc 29760

ggcgaggtcc ttcgctggtt cgggcagttc cacagcggag agcgattgct gcccaagcac 29820

ggctacgccg cgcccggtgc gctctacgcc gtccagctct acctcgaggt ccgcggtgtg 29880

gccgggctcg cggccggtca ctactactac cacccggtcg atcacgaact cgtgctcgtc 29940

cggcccctgg cgcaagccgg tcccgcccgg ctcctggtgc actgcgtcgg gcggcgctcg 30000

gtcgtcgaaa cggtgtaccg caacaacgtc 30030

<210> 2

<211> 396

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 2

Val Ser Tyr Leu Gln Met Leu Arg Met Pro His Phe Leu Arg Val Leu

1 5 10 15

Ser Gly Ser Trp Met Gly Arg Leu Pro Met Ala Met Ala Ala Val Ser

20 25 30

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

35 40 45

Thr Ala Ala Ala Cys Phe Ala Ile Ala Ser Ala Leu Gly Ala Pro Ala

50 55 60

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

65 70 75 80

Thr Ala Leu Leu Ala Ala Ala Gly Phe Ile Met Ile Ala Val Ala Pro

85 90 95

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

100 105 110

Thr Pro Pro Leu Glu Pro Cys Leu Arg Val Leu Trp Pro Asp Ile Ala

115 120 125

Asp Lys Lys Gln Leu Asp Ser Val Tyr Ala Val Asp Ser Ala Ala Gln

130 135 140

Glu Met Val Phe Val Ala Gly Pro Leu Leu Val Ser Leu Cys Leu Ala

145 150 155 160

Ile Ala Thr Pro Val Thr Ser Leu Trp Leu Gln Ala Gly Leu Cys Leu

165 170 175

Ala Gly Val Leu Val Phe Ala Thr Ala Ala Pro Ser Arg Arg Trp Arg

180 185 190

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

195 200 205

Leu Ile Ile Ala Leu Val Ala Pro Ala Gly Ala Gly Met Ala Ile Gly

210 215 220

Thr Leu Asn Val Leu Val Val Ser Tyr Ala Glu Arg His Glu Ile Phe

225 230 235 240

Gly Gly Ala Pro Met Leu Leu Thr Val His Ser Leu Ala Ser Leu Val

245 250 255

Ser Val Leu Ala Tyr Gly Ala Val Thr Trp Lys Met Glu Thr Arg Lys

260 265 270

Arg Leu Leu Phe Ser Ser Gly Gly Leu Leu Leu Gly Tyr Ala Leu Leu

275 280 285

Thr Leu Val Pro Pro Pro Val Pro Met Phe Gly Ile Met Val Val Ala

290 295 300

Gly Val Phe Leu Ala Pro Thr Leu Val Ser Val Phe Lys Leu Ile Gly

305 310 315 320

Glu Leu Ala Pro Thr Gly Thr Ala Thr Glu Ala Phe Ala Trp Leu Val

325 330 335

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

340 345 350

Val Leu His Asn Gly Asn Met His Leu Ala Ala Ala Cys Gly Ala Phe

355 360 365

Gly Ala Leu Gly Cys Leu Val Leu Leu Ala Ser Gly Tyr Arg Leu Leu

370 375 380

Ala Thr Pro Ala Lys Glu Leu Ser Thr Ala Ser His

385 390 395

<210> 3

<211> 476

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 3

Met Thr Thr Lys Glu Ile Val Pro Asp Val Leu Ala Ala Arg Tyr Ala

1 5 10 15

Ser Ser Asp Leu Val Ala Ile Trp Ala Pro Arg Asn Arg Val Ala Thr

20 25 30

Glu Arg Arg Leu Trp Ile Ala Val Met Thr Ala Gln Arg Glu Leu Gly

35 40 45

Val Asp Ile Pro Glu Gln Val Ile Ala Asp Tyr Ala Ala Val Val Asp

50 55 60

Gln Val Asp Leu Glu Ser Ile Lys Gln Arg Glu Arg Val Thr Leu His

65 70 75 80

Asp Val Lys Ala Arg Ile Glu Glu Phe Asn Ala Leu Ala Gly His Glu

85 90 95

His Ala His Lys Gly Met Thr Ser Arg Asp Leu Thr Glu Asn Ile Glu

100 105 110

Gln Leu Met Val Arg Asp Ser Leu Arg Leu Ile Arg His Arg Cys Val

115 120 125

Ser Val Leu Ala Arg Leu Ala Glu Leu Ala Thr Glu His Ala Ala Thr

130 135 140

Ala Met Ala Gly Arg Thr His Asn Val Ala Ala Gln Val Leu Pro Leu

145 150 155 160

Gly Lys Arg Phe Ala Thr Ala Ala Asn Glu Leu Leu Val Ala Val Glu

165 170 175

Arg Val Asp Asp Leu Leu Asn Arg Tyr Pro Leu Arg Gly Ile Lys Gly

180 185 190

Pro Val Gly Thr Leu Gln Asp Met Leu Asp Leu Phe Asp Gly Asp Thr

195 200 205

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

210 215 220

Glu Arg Arg Phe Thr Ser Val Gly Gln Val Tyr Pro Arg Ser Leu Asp

225 230 235 240

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

245 250 255

Leu Ala Lys Thr Ile Arg Leu Met Ala Gly His Glu Leu Ala Gly Glu

260 265 270

Gly Phe Ala Glu Gly Gln Thr Gly Ser Ser Ala Met Pro His Lys Met

275 280 285

Asn Pro Arg Asn Cys Glu Arg Ile Ser Gly Leu Met Val Val Leu Arg

290 295 300

Gly Tyr Ala Ser Met Ala Gly Glu Leu Ala Gly Asp Gln Trp Asn Glu

305 310 315 320

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

325 330 335

Phe Phe Ala Leu Asp Gly Leu Leu Gln Thr Thr Leu Ala Val Leu Asp

340 345 350

Gly Phe Thr Ala Ala Pro Arg Val Val Glu Ala Glu Leu Glu Arg Tyr

355 360 365

Leu Pro Phe Leu Ala Thr Thr Lys Met Leu Val Ala Ala Val Arg Ala

370 375 380

Gly Gln Gly Arg Glu Thr Ala His Glu Leu Ile Lys Glu His Ala Val

385 390 395 400

Ala Ala Ala Thr Ala Leu Ser Glu Gly Gly Thr Thr Asn Thr Leu Leu

405 410 415

Thr Asn Leu Gly Ser Asp Glu Arg Phe Pro Leu Asp Gln Ala Ala Leu

420 425 430

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

435 440 445

Gln Ile Thr Glu Ile Cys Ala Arg Val Ala Lys Leu Leu Thr Ser Ala

450 455 460

Pro Glu Ala Ala Thr Tyr Thr Pro Glu Pro Leu Leu

465 470 475

<210> 4

<211> 224

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 4

Met Leu Glu Ala Val Val Phe Asp Leu Asp Gly Thr Leu Val Asp Thr

1 5 10 15

Pro Ala Ala Ile Gly Ala Thr Leu Val Glu Val Leu Ala Gly Glu Gly

20 25 30

Phe Thr Val Asp His Ala Ala Val Ala Ala Thr Ile Gly Lys Pro Leu

35 40 45

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

50 55 60

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

65 70 75 80

Leu Gly Arg Gly Thr Glu Leu Leu Tyr Ala Gly Val Ala Asp Gly Leu

85 90 95

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

100 105 110

Asp Leu Arg Val Ala Thr Ala Leu Leu Gly Ser Ser Gly Ile Ala Ala

115 120 125

His Phe Pro Val Val Ile Thr His Asp Gln Val Ala Gln Gly Lys Pro

130 135 140

His Pro Glu Met Gly Leu Cys Ala Ala Ala Asp Leu Gly Val Asp Ala

145 150 155 160

Gly Ala Cys Ala Tyr Val Gly Asp Ala Val Gly Asp Met Glu Met Ala

165 170 175

Val Ala Ala Gly Met Thr Pro Ile Gly Val Ala Tyr Gly Val Ala Ser

180 185 190

Ala Ala Glu Leu Thr Glu His Gly Ala Val Gln Val Cys Ala Asp Phe

195 200 205

Ala Glu Val Val Lys Ala Val Ser Ala Leu Ala Gly Arg Arg Thr Asn

210 215 220

<210> 5

<211> 118

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 5

Val Phe Ile Asp Trp Glu Lys Met Pro Arg Thr Arg Ser Pro Arg Ala

1 5 10 15

Lys Asp Ser Arg Arg Ile Ala Ser Gly Gln Asn Met Ser Phe Val Arg

20 25 30

Ile Glu Ile Glu Pro Thr Ala Glu Phe Thr Gly Glu Thr His Trp His

35 40 45

Met His Glu Gln Trp Val Val Val Leu Ala Gly Asp Val Arg Met Thr

50 55 60

Val Ala Asp Glu Glu Val Gln Leu Thr Thr Gly Asp Val Leu Tyr Ile

65 70 75 80

Pro Ile Asp Ala Pro His Ala Pro Leu Ala Val Gly Pro Ala Gly Ala

85 90 95

Thr Tyr Leu Glu Ile Phe Ala Pro Pro Arg Met Asp Leu Leu Pro Glu

100 105 110

Ser Ile Val Pro Ala Val

115

<210> 6

<211> 334

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 6

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

1 5 10 15

Thr Leu Ile Ser Leu Asp Gly Glu Ser Arg Arg Arg Asn Gly Ile Ala

20 25 30

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

35 40 45

Pro Gly Arg Ala Thr Thr Val Val Gly Ala Glu Glu Glu Thr His His

50 55 60

Ala Leu Val Asp Gly Leu Ala Lys Leu Gln Arg Ile Trp Asp Gly Pro

65 70 75 80

Asp Val Ala Leu Asp Val Gln Ser Ser Leu Pro Ala His Ser Gly Phe

85 90 95

Gly Ser Lys Thr Thr Thr Leu Ile Ala Ala Gly Tyr Ala Tyr Gly Leu

100 105 110

Leu Ser Gly Gln Thr Pro Asp Ile Arg Trp Leu Ser Ala Thr Leu Gly

115 120 125

Arg Gly Arg Thr Ser Gly Ala Ser Thr Gly Leu Ala Glu His Gly Gly

130 135 140

Phe Leu Val Asp Cys Gly His Leu Asn Pro Pro Asp Phe Ala Ala Glu

145 150 155 160

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

165 170 175

Pro Thr Pro Val Ile Lys Met Asp Phe Pro Asp Trp Pro Ile Leu Ile

180 185 190

Leu Gln Val Asn Gly Arg Asn Ile Gly Gly Pro Glu Glu Leu Glu Trp

195 200 205

Phe Thr Thr Thr Met Pro Ile Pro Pro Ala Glu Ser Trp Arg Thr Ser

210 215 220

His Leu Val Phe Met Gly Leu Ala Pro Ala Val Ala Glu Arg Asp Tyr

225 230 235 240

Asp Thr Phe Cys Ser Val Val Asn Glu Leu Thr Phe Thr Gly Tyr Phe

245 250 255

Lys Arg Ala Gln Ile Ala Leu Gln Gly Gln Glu Met Val Asp Leu Leu

260 265 270

Asp Glu Gly Arg Ser Gln Ser Ser Val Asp Ala Ile Ala Met Ser Ser

275 280 285

His Gly Pro Thr Cys Phe Ala Phe Cys Arg Asp Thr Glu Ala Ala Ala

290 295 300

Ala Trp Ala Glu Gly Leu Arg Gln Arg Gly Val Ile Arg Ala His Trp

305 310 315 320

Phe Thr Thr Val Arg Asn Ser Gly Leu Ala Ala Thr Trp Leu

325 330

<210> 7

<211> 343

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 7

Met Leu His Ala His Gly Arg Lys His Val Lys Arg Trp Ser Ser Lys

1 5 10 15

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

20 25 30

Val Phe Val Phe Ser Asp Arg Tyr Ser Val Phe Asp Phe Gly Ile Met

35 40 45

Pro Asp Glu Ile Pro Gly Lys Gly Ala Ala Ser Cys Ala Met Ala Val

50 55 60

Arg Ser Phe Glu Leu Phe Glu Ala Ala Gly Ile Arg Thr His Phe Leu

65 70 75 80

Glu Gln Val Ala Glu Asp Ala Ile Arg Ile Gln Leu Leu Asp Val Asp

85 90 95

Thr Asp Arg Thr Lys Gly Arg Thr Gly Pro Gly Arg Thr Ile Pro Leu

100 105 110

Gln Val Ile Tyr Arg Leu Ala Leu Pro Gln Glu Ser Ser Val His Arg

115 120 125

Arg Ala Ala Ala Gly Thr Leu Asp Gln Ser Thr Val Pro Pro Tyr Arg

130 135 140

Asp Ala Pro Trp Leu Ser Glu Pro Met Val Glu Phe Thr Thr Lys Tyr

145 150 155 160

Glu Glu Thr Asp Arg Phe Ile Ser Arg Ser Glu Ala Ala Glu Val Gly

165 170 175

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

180 185 190

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

195 200 205

Val Asp Gly Lys Ala Glu Phe Gly Phe Asp Ala Asp Arg Gln Pro Ile

210 215 220

Met Ile Asp His Ala Gly Thr Pro Asp Glu Asn Arg Phe Tyr Leu Asp

225 230 235 240

Gly Ile Pro Val Cys Lys Glu Leu Leu Arg Phe Leu His Pro Gly Leu

245 250 255

Arg Glu Val Val Gln Gly Tyr Val Ala Glu Gly Arg Pro Arg Ala Gln

260 265 270

Trp Pro Leu Pro Lys Pro Leu Ala Pro Glu Val Val Ala Ala Val Ala

275 280 285

Glu Ile Tyr Ala Ala Leu Thr Ala Ala Trp Thr Ala Ala Pro Pro Ser

290 295 300

Ala Ala Gly Thr Gln Arg Leu Ala Arg Ala Val Glu Asn Phe Ile Ala

305 310 315 320

Val Cys Gly Ile Ser Ser Glu Leu Gly Gln Arg Thr Gly Leu Ala Ala

325 330 335

Val Pro Ser Ile Ser Ala Arg

340

<210> 8

<211> 540

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 8

Val Asp Leu Glu Lys Phe Asp Val Val Val Val Gly Ser Gly Gly Ala

1 5 10 15

Gly Leu Leu Ala Ala Ala Arg Ala Ala Ala Ala Gly Leu Ser Val Val

20 25 30

Val Leu Glu Arg Ser Asp Val Leu Gly Gly Thr Thr Ala Val Ser Gly

35 40 45

Gly Met Leu Trp Val Pro Asn Ser Ser Val Met Lys Gln Ala Gly Leu

50 55 60

Pro Asp Ser Ala Glu Asp Ala Leu Arg Tyr Leu Glu Arg Val Val Glu

65 70 75 80

Gly Thr Val Pro Arg Glu Arg Leu Glu His Tyr Val Arg Thr Ala Pro

85 90 95

Glu Met Val Asp Trp Leu Leu Ala His Thr Gly Ala Arg Phe Phe Pro

100 105 110

Ile Asp Arg Pro Asp Tyr His Ser Thr Trp Pro Gly Ala Val Asp Val

115 120 125

Gly Arg Cys Leu Asp Asn Glu Pro Phe Pro Thr Ala Ser Arg Pro Gly

130 135 140

Leu Leu Glu Arg Val Arg Arg Gly Pro Gln Phe Pro Pro Leu Thr Tyr

145 150 155 160

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

165 170 175

Ala Gln Arg Leu Ala Asp Gly Val Leu Thr Val Gly Ala Ala Leu Val

180 185 190

Ala Ala Leu Val Ala Ala Cys Asp Asp Leu Gly Val Arg Phe Val Leu

195 200 205

Arg Ala Arg Ala Thr Ser Leu Leu Arg Asp Asp Asn Asp Ala Ala Ala

210 215 220

Val Thr Gly Val Arg Cys Glu Asp Gly Arg Arg Phe Ser Ala Arg Ala

225 230 235 240

Val Val Leu Ala Asn Gly Gly Phe Glu Trp Asp Pro Ala Ser Gly Ala

245 250 255

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

260 265 270

Gly Asp Gly Leu Arg Met Ala Met Ser Ala Gly Ala Ala Val Glu Arg

275 280 285

Met Asn Gln Ala Trp Trp Val Pro Ala Leu Gln Gly Val Pro Glu Glu

290 295 300

Tyr Asp Gly Ala Pro Leu Thr Arg His Leu Val Gly Glu Arg Cys Leu

305 310 315 320

Pro Gly Ser Ile Met Val Asp Arg His Gly Arg Arg Phe Val Asn Glu

325 330 335

Ala Val Asn Tyr His Asp Ile Thr Arg Val Leu Phe Asn Phe Asp Ala

340 345 350

Asp Leu His Gln Pro Ala His Leu Pro Val Trp Leu Val Phe Asp Glu

355 360 365

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

370 375 380

Val Pro Ser Trp Leu Thr Ser Ala Ala Thr Pro Ala Ala Leu Ala Glu

385 390 395 400

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

405 410 415

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

420 425 430

Ser Ser His Asp Arg Tyr Tyr Gly Asp Pro Arg Asn Pro Gly Asn Pro

435 440 445

Cys Leu Gly Glu Leu Val Thr Pro Pro Phe His Ala Val Arg Val Val

450 455 460

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

465 470 475 480

Glu Val Leu Thr Ala Thr Gly Thr Ala Val Pro Gly Leu Tyr Ala Cys

485 490 495

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

500 505 510

Gly Thr Leu Gly Pro Ala Leu Thr Ala Gly Tyr Ala Val Gly Gly Ala

515 520 525

Leu Val Glu Arg Cys Gly Gly Ala Phe Ala Thr Asp

530 535 540

<210> 9

<211> 329

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 9

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

1 5 10 15

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

20 25 30

Ala Arg Thr Val Glu Glu Ala Gly Ala Asp Tyr Leu Val Leu Pro Asp

35 40 45

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

50 55 60

Ala Ala Ala Phe Leu Ala Ala His Thr Ser Glu Ile Gly Ile Val Val

65 70 75 80

Thr Ala Ala Thr Gly Tyr His Glu Pro Tyr Ser Leu Ala Arg Gln Ile

85 90 95

Ala Ser Leu Asp His Ile Ser Ala Gly Arg Val Gly Trp Phe Ala Asp

100 105 110

Ser Ala Thr Asp Ala Ala Ser Asp Ala Asn His Arg Arg Glu Gly His

115 120 125

Asp Pro Ala Ala Ala Pro Glu Thr Arg Ala Leu Glu Phe Val Pro Leu

130 135 140

Val Arg Asp Leu Trp Asp Ser Trp Glu Asp Asp Ala Phe Val His Glu

145 150 155 160

Lys Asp Thr Gly Arg Phe Val Asp Pro Ala Lys Ile His Val Val Asp

165 170 175

His Val Gly Thr Ala Leu Ala Val Arg Gly Pro Leu Asn Val Ile Arg

180 185 190

Pro Pro Gln Gly His Pro Val Val Phe Ala Arg Ala Ala Asp Thr Ser

195 200 205

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

210 215 220

His Ile Leu Ala Val Glu Pro Phe Val Ala Ala Asp Glu Val Ala Ala

225 230 235 240

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

245 250 255

Val Gly Thr Pro Glu Gln Val Ala Ala Gln Leu Arg Asp Arg His Val

260 265 270

Leu Val Arg Phe Thr Thr Arg Ala Gln Leu Val Ala Phe Thr Thr His

275 280 285

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

290 295 300

Glu Arg Leu Gly Leu Pro Ala Val Ala Asn Arg Phe Ala Pro Thr Gln

305 310 315 320

Asp Lys Glu Leu Ile Gly Asn Ala Arg

325

<210> 10

<211> 444

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 10

Met Pro Ala Asp Arg Glu Leu His Leu Gly Leu Met Phe Trp Ala Thr

1 5 10 15

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

20 25 30

Ala Tyr Asp Ile Glu Leu Ile Gln Glu Val Cys Arg Thr Ala Glu Arg

35 40 45

Ala Lys Phe Asp Phe Ala Phe Leu Gly Asp Arg Leu Ala Cys Asp Pro

50 55 60

Ala Leu Gln His Ser Asn Pro Ala Gln Ile Ser Arg Leu Glu Pro Phe

65 70 75 80

Ala Thr Ala Ala Ala Ile Ala Ala Ala Thr Thr His Ile Gly Val Val

85 90 95

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

100 105 110

Leu Ala Ser Leu Asp His Ile Ser Gly Gly Arg Ala Ala Trp Asn Leu

115 120 125

Val Thr Gly Ala Asp Ala Ala Ala Ala Leu Asn Tyr Ser Arg Asp Gln

130 135 140

His Trp Asp Thr Gln Lys Arg Tyr Asp Trp Ala Glu Glu Thr Leu Glu

145 150 155 160

Ile Ala Arg Ala Leu Trp Asp Ser Gly Asp Gln Pro Ile Asn His Arg

165 170 175

Ser Glu Tyr Phe Ser Ile Asp Gly Pro Leu Gly Leu Pro Arg Pro Pro

180 185 190

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

195 200 205

Glu Leu Gly Ala Arg Gln Ala Asp Met Val Phe Ala Gly Pro Gln Pro

210 215 220

Thr Leu Ala Lys Arg Lys Glu Tyr Tyr Ala Asp Ile Lys Ala Arg Ala

225 230 235 240

Ala Lys Tyr Gly Arg Glu Asp His Val Thr Val Met Pro Gly Leu Thr

245 250 255

Pro Ile Val Ala Pro Thr Thr Glu Glu Ala Val Ala Leu His Asp Gln

260 265 270

Leu Asn Ser Leu Ile Val Leu Asp Pro Glu Val Glu Val Gly Glu Val

275 280 285

Val Arg Ala Gly Gly Ile Gly Glu Gly Phe Lys Arg Asn Leu Val Ser

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

Glu Arg Ile Thr Tyr Ala Asp Leu Val His Ala Thr Pro Gln Gln Leu

355 360 365

Ser His Val Val Val Gly Asn Pro Glu Glu Ile Ala Asp Glu Ile Gln

370 375 380

Leu Trp Leu Glu Glu Arg Met Ala Asp Gly Phe Asn Ile Tyr Pro Ala

385 390 395 400

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

405 410 415

Leu Gln Arg Arg Gly Leu Phe Arg Lys Asp Tyr Arg Gly Arg Thr Leu

420 425 430

Arg Asp His Leu Gly Leu Ala Val Pro Ala Val Arg

435 440

<210> 11

<211> 288

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 11

Met Arg Ser Leu Gly Glu Glu Leu Ser Val Gln Leu Leu Arg Glu Glu

1 5 10 15

Arg Thr Ser Trp Ser Ile Arg Glu Lys Ser Gly Cys Val Thr Val Pro

20 25 30

Ile Ser Leu Leu Gly Glu Ala Asp Ser Pro Arg Gln Ala Gly Val Asn

35 40 45

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

50 55 60

Leu Val His Arg His Thr Met Arg Val Ile Asp Gly Met His Arg Val

65 70 75 80

Gln Ala Ala Leu Ala Arg Gly Glu Asp Thr Ile Glu Val Val Phe Phe

85 90 95

Asp Gly Asp Glu Ala Ala Ala Phe Val Leu Ala Val Glu Leu Asn Gly

100 105 110

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

115 120 125

Arg Ile Val Arg Ala Tyr Pro Glu Trp Ser Asp Arg Arg Ile Ala Ala

130 135 140

Ala Ala Gly Ile Ser Pro Lys Thr Ala Gly Ala Val Arg Ala Gln Leu

145 150 155 160

Ser Ser Glu Glu Ile Pro Gln Met Arg Glu Arg Leu Gly Leu Asp Gly

165 170 175

Arg Val Arg Arg Val Ala Pro Arg Thr Val Pro Ala His Thr Thr Glu

180 185 190

Pro Glu Arg Val Gln Ala Arg Pro Ala Arg Ala Lys Gln Pro Gln Pro

195 200 205

Glu Arg Gln Leu Asp Phe Gly Ala Val Ile Tyr Ala Leu Arg Arg Asp

210 215 220

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

225 230 235 240

Asp Met His His Leu Pro Ala Ala Gly Trp Ser Gly Ile Ile Gly Ala

245 250 255

Val Pro Ala His Cys Ala Ser Val Val Ala Glu Leu Ala Arg Glu Cys

260 265 270

Ala Glu Ala Trp Leu Val Leu Ala Asp Glu Leu Asp Glu Lys Ser Asp

275 280 285

<210> 12

<211> 351

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 12

Met Lys Val Leu Val Ile Gly Ala Gly Ala Ile Gly Ala Cys Ala Ala

1 5 10 15

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

20 25 30

Glu Pro Leu Asp Thr Leu Ser Ala His Ser Phe Gly Trp Ala Asn Ala

35 40 45

Ile Asp Glu Lys Asn Ser Ala Tyr Phe Glu Leu Ser Ile Glu Ala Leu

50 55 60

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

65 70 75 80

Leu Phe Arg Gln Gly Asn Leu His Trp Gly Arg Asp Ser Ala Ser Ala

85 90 95

Ala Ala Gln Arg Ala Ile Ala Asp Gly Tyr Arg Glu Leu Gly Tyr Pro

100 105 110

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

115 120 125

Leu Ala Leu Glu Asn Val Thr Gly Pro Val Tyr Phe Tyr Pro Arg Asp

130 135 140

Met His Val Leu Gly Asp Val Leu Leu Pro Ala Val Leu Asp Arg Ala

145 150 155 160

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

165 170 175

Asp Ala Val Arg Ala Asp Ala Val Leu Cys Cys Ala Gly Arg Gly Asn

180 185 190

Ala Thr Leu Leu Pro Gly Leu Pro Leu Leu Pro Pro Gly Ala Pro Glu

195 200 205

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

210 215 220

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

225 230 235 240

Gly Asn Arg Leu Val Leu His Cys His Asp Leu Asp His Thr Leu Thr

245 250 255

Glu Ser Ser Asp Val Ala Ala Leu Ala Glu Met Ala Leu Ala Arg Leu

260 265 270

Pro Glu Val Leu Pro Gly Ala Ala Asp Ala Glu Val Glu Lys Ala Phe

275 280 285

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

290 295 300

Val Pro Gly Arg Asn Gly Val Tyr Val Ile Ala Thr His Ser Gly Leu

305 310 315 320

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

325 330 335

Asp Pro His Pro Leu Ala Glu Pro Phe Gln Leu Thr Arg Phe Ala

340 345 350

<210> 13

<211> 413

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 13

Leu Ser Ser Arg Ser Val Leu Phe Val Asn Leu Arg Arg Ile Lys Arg

1 5 10 15

Glu Gly Phe Glu Ser Leu Val Ala Ala Arg Arg Leu Gly Tyr Arg Val

20 25 30

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

35 40 45

Glu Phe His Gln Val Asp Thr Tyr Asp Thr Asp Leu Ala Leu Lys Thr

50 55 60

Ala Arg Glu Ile Ala Ala Ala Asn Asp Leu Ala Gly Val Val Asn Phe

65 70 75 80

Thr Glu Ile Asp Val Gln Leu Val Ala Ala Ile Ala Ala Glu Leu Gly

85 90 95

Leu Pro Gly Met Pro Pro Glu Ala Ala Val Leu Ala Arg Asn Lys Leu

100 105 110

Ala Met Lys Gln Ala Leu Ala Gly Ile Asp Gly Val Leu Pro Lys Phe

115 120 125

Ala Arg Val Ser Asp Leu Asp Asp Leu Cys Ala Ala Val Ala Asp Ile

130 135 140

Gly Phe Pro Cys Val Val Lys Pro Thr Gly Ala Ser Gly Ser Lys Gly

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Val Glu Gly Phe Val Ala Asp Gly Thr Val His Gln Leu Leu Leu Thr

210 215 220

Asp Lys Val Thr Thr Val Pro Phe His Leu Glu Val Ser His Arg Leu

225 230 235 240

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

245 250 255

Glu Lys Ile Val Arg Ala Leu Gly Phe Asp Asn Cys Ala Phe His Leu

260 265 270

Glu Ala Lys Trp Asp Gly Glu Arg Met Arg Phe Ile Glu Val Ala Ala

275 280 285

Arg Pro Ala Gly Asp Tyr Ile Ala Ser His Leu Val Lys Ala Ala Thr

290 295 300

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

305 310 315 320

Leu Gln Leu Val Pro Asp Arg Asp Leu Gln Ala Gly Leu Arg Phe Val

325 330 335

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

340 345 350

Leu Phe Glu Glu Pro Gly Tyr Asp His Val Phe Thr Glu Val Pro Ile

355 360 365

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

370 375 380

Ala Ala Val Cys Ala Arg Ala His Asp Arg Ala Gly Leu Asp Ala Leu

385 390 395 400

Leu Asp Thr Ala Gly Glu Ala Ile Lys Val Arg Ile Gly

405 410

<210> 14

<211> 390

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 14

Met Gly Arg Gly Ala Gly Tyr Ala Leu Ala Arg Asp Gln Arg Thr Thr

1 5 10 15

Ser Ile Thr Val Val Asp His Asp Arg Glu Arg Ala Glu Glu Leu Ala

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Met Ala Asn Ala Ala Gly Val Pro Val Leu Leu Pro Val Gly Leu Glu

100 105 110

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

115 120 125

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

130 135 140

Pro Ala Thr Pro Arg Ala Pro Trp Gly Tyr Thr Ala Phe Phe Gly Gly

145 150 155 160

Glu Leu Ala Asn His Leu Pro Ile Ala Gln Arg Ser Ser Ile Ala Val

165 170 175

Glu His Gly Glu Ile Val Asp His Pro Arg Phe Ser Gly Val Glu Glu

180 185 190

Arg His Val Ala Gly Val Gly Leu Leu Glu Ala Tyr His Asp Gly Met

195 200 205

Val Pro Trp Leu Ala Asp His Pro Ala Leu Arg Asp Ala Asp Cys Thr

210 215 220

Gln Lys Thr Leu Arg Trp Pro Gly Phe Ala Asp Ala Val Thr Gly Leu

225 230 235 240

Ala Asn Leu Gly Leu Leu Asp Glu Ser Pro Val Ala Val Asp Gly Val

245 250 255

Ala Val Ala Pro Lys Arg Leu Val Glu Asn Val Leu Ser Pro Arg Leu

260 265 270

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

275 280 285

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

290 295 300

Leu Val Asp Arg Ala Asp Arg Glu Thr Gly Leu Ala Ala Met Thr Arg

305 310 315 320

Thr Thr Gly Phe Thr Leu Ala Ala Ala Val Thr Leu Leu Ala Asp Arg

325 330 335

Thr Val Gly Gly Ala Gly Trp Leu Arg Pro His Leu Ala Leu Ser Glu

340 345 350

Arg His Phe Ala Arg Met Lys Thr Asp Leu Ala Ala Leu Gly Val Arg

355 360 365

Trp Thr Ala Asp Gly Glu Ala Leu His Gly His Arg Ala Gly Thr Val

370 375 380

Ala Ser Lys Gly Thr Asp

385 390

<210> 15

<211> 437

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 15

Met Ser Thr Gly His Asp Val Leu Asp Val Val Gly Ile Gly Ile Gly

1 5 10 15

Pro Ala Asn Leu Ser Leu Ala Ala Leu Leu Ala Ala Glu Ser Ser Glu

20 25 30

Leu Asn Val Ala Phe Leu Asp Arg Lys Pro His Phe Gly Trp His Ser

35 40 45

Gly Leu Met Leu Pro Asp Ala Gln Met Gln Val His Tyr Leu Lys Asp

50 55 60

Leu Val Thr Pro Val Asp Pro Thr Asn Pro Phe Ser Phe Thr Ala Phe

65 70 75 80

Leu Val Ala Thr Lys Arg Phe Tyr Arg Leu Leu Val Thr Gly Arg Ser

85 90 95

Lys Val Pro Arg Arg Glu Phe Glu Gln Tyr Cys Arg Trp Ala Ala Glu

100 105 110

Arg Ile Pro Asn Leu Arg Phe Gly Val Glu Val Arg Glu Val Thr Trp

115 120 125

Asn Gly Glu Leu Phe Val Val His Thr Asp Gln Gly Val Leu His Ala

130 135 140

Arg Asn Val Val Ser Gly Thr Gly Leu Val Pro Arg Leu Pro Ser Phe

145 150 155 160

Ala Thr Gly His Asp Pro Gln Glu Val Phe His Ala Ser Thr Leu Leu

165 170 175

Asp Val Pro Arg Thr Phe Ala Gly Arg Arg Val Ala Val Val Gly Gly

180 185 190

Gly Gln Ser Gly Ala Glu Val Val His His Leu Leu Thr Ser Pro Asp

195 200 205

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

210 215 220

Leu Asp Asp Ser Pro Phe Val Asp Glu Leu Phe Val Pro Asn Tyr Ser

225 230 235 240

Arg Tyr Phe His Gly Leu Pro Ala Lys Arg Arg Met Glu Leu Val Asp

245 250 255

Glu Gln Lys Met Ala Ser Asp Gly Val Ser Val Ser Leu Leu Glu Ala

260 265 270

Ile Tyr His Arg Leu Tyr Asp Leu Glu Met Leu Glu Gly Glu Glu Arg

275 280 285

Pro Cys Arg Met Leu Leu Gly His Ser Leu Thr Thr Val Asp Lys Thr

290 295 300

Pro Thr Gly Leu Leu Thr Arg Trp Met Pro Gly Ala Gly Ala Glu Val

305 310 315 320

Gly His Glu Val Asp Val Val Ile Cys Ala Thr Gly Tyr Arg His Gly

325 330 335

Leu Pro Arg Pro Leu His Gly Leu Pro Arg His Leu Arg Glu Ala Met

340 345 350

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

355 360 365

Trp Glu Gly Pro Gln Asp Arg Arg Met Tyr Val Gln Asn Ala Ala Arg

370 375 380

His Ser Phe Gly Val Ala Asp Pro Asn Leu Ser Leu Leu Ala Trp Arg

385 390 395 400

Ser Ala Thr Ile Ala Asn Ser Val Leu Gly Tyr Glu Arg Tyr Asp Val

405 410 415

Gly Glu Val Gly Ala Val Leu Asp Trp Thr Ser Ser Ala Asp Ser Asp

420 425 430

Val Ala Leu Thr Phe

435

<210> 16

<211> 668

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 16

Met Ile Val Ser Glu Ile Pro Arg Glu Leu Ala Asp Leu Ala Arg Ala

1 5 10 15

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

20 25 30

Pro Phe Asn Val Arg Arg Val Thr Leu Arg Pro Gly Glu Thr Thr Ala

35 40 45

Glu His Asn His His Asp Leu Glu Val Trp Val Met Leu Asp Gly Val

50 55 60

Gly Glu Val Gly Trp Asp Gly His Gln Arg Val Leu Thr Ala Gly Asp

65 70 75 80

Ser Val Tyr Leu Pro Pro Leu Ala Pro His Thr Leu Arg Asn Leu Ser

85 90 95

Ser Asp Arg Pro Leu Ser Phe Phe Ser Met Trp Trp Glu Asn Leu Ser

100 105 110

Ala Leu Ala Ala Val His Ala Glu Arg Arg Glu Gln Ala Val Glu Arg

115 120 125

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

130 135 140

Glu Leu His Leu Gly His Leu Ser Gly Pro Phe Leu Asn Ala Asp Ala

145 150 155 160

Cys Arg Arg Ala Leu Leu Ala Ala Gly Glu Arg Ala His Leu Leu Leu

165 170 175

Gly Thr Val Gly His Gln Ser Gln Val Ser Ala Ala Ala Glu Ala Glu

180 185 190

Gly Leu Ser Phe His Glu Leu Ala Glu Arg Asn Thr Asp Ala Ile Ile

195 200 205

Glu Gly Leu Gln Ala Ala Gly Ile Asp Trp Asp Val Phe Val Arg Pro

210 215 220

Ser Glu Pro Ala Tyr Pro Ala Met Ala Thr Ser Val Phe Glu Ser Leu

225 230 235 240

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

245 250 255

Glu Pro Cys Gly Arg Phe Leu Leu Glu Ala Phe Val Ala Gly His Cys

260 265 270

Pro His Cys Gly Ser Asn Gln Thr Ala Gly Ile Glu Cys Glu Leu Cys

275 280 285

Ala Leu Pro Tyr Asp Asp Arg Asp Leu Val Asp Pro Ser Cys Ala Thr

290 295 300

Cys Gly Ala Ala Ala Thr Gln Arg Pro Leu Thr Arg Tyr Phe Met Pro

305 310 315 320

Leu Glu Pro Leu Arg Asp Glu Leu Ser Gly Tyr Leu Arg Gly Ala Ala

325 330 335

Met His Gly Arg Leu Arg Ala Tyr Thr Glu Arg Val Leu Ala Lys Thr

340 345 350

Leu Pro Asp Leu Pro Val Ser Ile Pro Ala Glu His Gly Ile Pro Ile

355 360 365

His Val Glu Asp Ala Ser Gly Pro Ala Glu Gln Arg Met Tyr Ser Ala

370 375 380

Phe Glu Leu Ala Ala Arg Phe Leu Thr Ala Leu Asp Gly Phe Ala Asp

385 390 395 400

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

405 410 415

Phe Gly Phe Asp Asn Ala Phe Leu Arg Ala Phe Ala Phe Pro Ala Val

420 425 430

Leu Gly Ala Phe Thr Asp Ala Leu Pro Leu Pro Glu Ala Leu Val Cys

435 440 445

Asn Asp Phe Tyr Leu Leu Asp Gly Glu Lys Phe Ser Thr Gly Arg Lys

450 455 460

His Ala Val Trp Ala Arg Gln Ala Val Thr Pro Ala Asn Ala Asp Gln

465 470 475 480

Leu Arg Leu Tyr Leu Ala Ala Thr Ser Pro Asp Val Arg Arg Arg Asp

485 490 495

Phe Thr Thr Arg Gly Tyr Ala Glu Phe Val Thr Ala Glu Leu Ile Gly

500 505 510

Arg Trp Gln Arg Arg Leu Asp Asp Val Gly Gly Arg Val Ala Glu His

515 520 525

Phe Gly Gly Leu Thr Pro Glu Ala Gly Gly Trp His Ala Glu Ala Glu

530 535 540

Arg Phe Tyr Gly Gln Ile Lys Glu Phe Ala Ser Cys Ala Thr Leu Asp

545 550 555 560

Tyr Leu Pro Gly Arg Phe Lys Pro Arg Ala Val Val Ala Ala Ala Cys

565 570 575

Ala Phe Ile Arg Gln Ala Glu Asp Phe Ala Glu Val Ser Ala Asp Ala

580 585 590

Thr Pro Gly Ser Gly Ile Ala Arg Thr Cys Ala Ala Leu Glu Leu Met

595 600 605

Ala Leu Arg Thr Leu Ala Met Ala Val Trp Pro Leu Ala Pro Glu Phe

610 615 620

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

625 630 635 640

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

645 650 655

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

660 665

<210> 17

<211> 250

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 17

Met Arg Asp Thr Asp Thr Asp Thr Asp Thr Asp Thr Asp Ala Asp Val

1 5 10 15

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

20 25 30

Arg Ser Lys Lys Leu Trp Leu Val Pro Gly Asp Arg Cys Val Val Glu

35 40 45

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

50 55 60

Asp Glu Thr Gly Pro Leu Arg Leu Asp Phe Tyr Glu Lys Ala Ala Ala

65 70 75 80

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

85 90 95

Pro Val Thr Tyr Leu Ala Asp Tyr Arg Pro Ala Pro Pro Phe Glu Val

100 105 110

Ile Val Lys Asn Val Ala Thr Gly Ser Thr Thr Arg Lys Tyr Pro Gly

115 120 125

Leu Phe Pro Asp Gly Glu Pro Leu Pro Arg Pro Val Val Lys Phe Asp

130 135 140

Tyr Arg Val Asp Pro Glu Asp Gln Pro Ile Gly Glu Asp Tyr Leu Arg

145 150 155 160

Val Leu Asp Leu Pro Val Asp Leu Met Arg Glu Gln Ala Leu Leu Val

165 170 175

Asn Asp Val Leu Arg Asp Trp Leu Asn Pro Leu Arg Leu Leu Asp Phe

180 185 190

Cys Val Ile Phe Gly Phe Ser Ser Ala Gly Glu Ile Ser Leu Ile Ser

195 200 205

Glu Val Ser Gln Asp Cys Met Arg Leu Arg His Pro Asp Gly Ser Pro

210 215 220

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

225 230 235 240

Glu Gln Trp Lys Arg Ala Phe Asp Gly Leu

245 250

<210> 18

<211> 233

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 18

Met Gly Ser Glu Ser Pro Val Ala Leu Val Thr Gly Ser Asn Arg Gly

1 5 10 15

Ser Gly Arg Ala Ile Ala Ala Arg Leu His Glu Arg Gly Tyr Arg Val

20 25 30

Phe Ser Leu Asn Arg Thr Val Thr Gly Glu Asp Trp Leu Gly Glu Arg

35 40 45

Glu Cys Asp Leu Ala Ser Arg Glu Ser Leu Ala Ala Gly Val Ala Ala

50 55 60

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

65 70 75 80

Glu Arg Val Leu Asp Pro Ile Glu Lys Met Ser Glu Gln Asp Trp Asp

85 90 95

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

100 105 110

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

115 120 125

Ser His Ala Gly Ser Arg Tyr Phe Glu Gly Gly Ser Ala Tyr Ser Ala

130 135 140

Thr Lys Ala Ala Leu Lys Ala Leu Val Glu Thr Leu Leu Leu Glu Glu

145 150 155 160

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

165 170 175

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

180 185 190

Gly Thr Cys Val Ala Ser Ile Val Ala Asp Trp Pro Ala Asp Leu Val

195 200 205

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

210 215 220

Thr Gly Ile Asp Arg Leu Leu His Val

225 230

<210> 19

<211> 260

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 19

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

1 5 10 15

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

20 25 30

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

35 40 45

Thr Glu Ile Gly Gln Glu Val Gly Ala Asn Phe Leu Val Cys Ser Gly

50 55 60

Gly Ala Val Leu Tyr Asp Pro Val Arg Gly Glu Leu Val Arg Ala Thr

65 70 75 80

Thr Leu Gly Ser Arg Arg Ala Thr Ser Leu Ile Ser Leu Leu Arg Gln

85 90 95

Thr Phe Pro Gly Ile Arg Leu Gly Val Asp His Leu Ala Arg Cys Asp

100 105 110

Leu Asp Pro Gly Phe His Val Gly Ala Pro Gly Val Ala Asp Arg His

115 120 125

Ala Thr Glu Pro Leu Arg Gln Val Ala Glu Pro Ala Val Lys Leu Ile

130 135 140

Ala Gln Ser Asp Glu Met Pro Val Asp Arg Leu Ala Arg Ala Ile Ser

145 150 155 160

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

165 170 175

Val Asp Val Leu Pro Ala Gly Val His Lys Ala Val Gly Leu Gly Glu

180 185 190

Leu His Glu Cys His Gly Arg Val Leu Pro Ser Ile Ala Phe Gly Asp

195 200 205

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

210 215 220

Thr Ala Asn Ala His Pro Ala Val Leu Glu Ala Cys Asp His Val Thr

225 230 235 240

Ala His His Asp Ala Asp Gly Val Ala Val Tyr Leu Glu Ser Leu Ile

245 250 255

Ser Thr Gly Gly

260

<210> 20

<211> 288

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 20

Met Val Thr Leu Val Leu Pro Thr Gly Ser Leu His Glu Pro Thr Leu

1 5 10 15

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

20 25 30

Ala Leu Arg Ala His Val Asp Phe Pro Gly Ile Glu Arg Val Val Phe

35 40 45

Gly Lys Pro Arg Glu Ile Pro Gly Leu Val Ala Asp Gly Val Val Asp

50 55 60

Leu Gly Leu Thr Gly Thr Asp Trp Ile Glu Glu Ser Gly Ala Lys Val

65 70 75 80

Glu Val Val His Glu Phe Gln Tyr Ser Lys Thr Thr Ser Ala Gly Trp

85 90 95

Arg Val Val Leu Ala Val Pro Val Asp His Pro Ala Arg Thr Ala Ala

100 105 110

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

115 120 125

Arg Arg Tyr Phe Glu Glu Glu Leu Gly Gln Glu Ala Arg Ile Val His

130 135 140

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

145 150 155 160

Leu Glu Ile Val Glu Thr Gly Asp Thr Leu Arg His Asn Asp Leu Arg

165 170 175

Glu Leu Val Val Val Arg Arg Cys Ala Val Gln Leu Val Val Ala Thr

180 185 190

Gln Ala Asp Pro Val Val Arg Ala Thr Ala Glu Lys Val Ala Leu Leu

195 200 205

Leu Lys Gly Ala Arg Ala Ala Thr Glu His Ala Leu Leu Thr Val Val

210 215 220

Ala Pro Ala Asp Cys Trp Asp Arg Val Arg Gln Glu Leu Pro Arg Tyr

225 230 235 240

Trp Trp Leu Leu Gly Gly Asp Pro Glu Thr Val Val Ala Gln Ala Thr

245 250 255

Tyr Glu Leu Arg Gly Val Ala Glu Ala Ile Thr Arg Leu Thr Gln Ala

260 265 270

Gly Ala Val Gln Val Val Glu Thr Pro Met Arg Lys Leu Val Thr Ala

275 280 285

<210> 21

<211> 402

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 21

Met Asn Glu Thr Leu Thr Arg Val Gly Arg Gly Thr Pro Met Gly Glu

1 5 10 15

Leu Leu Arg Glu Tyr Trp Met Pro Val Met Arg Ser Glu Arg Val Val

20 25 30

Ala Gly Gly Glu Pro Ile Ala Val Glu Leu Leu Gly Asp Arg Tyr Val

35 40 45

Ile Phe Arg Gly Asp Asp Gly Thr Val Ala Cys Phe Asp Glu Ala Cys

50 55 60

Pro His Arg Gly Ala Ser Leu Ala Leu Ala Arg Asn Glu Asp Cys Ala

65 70 75 80

Leu Arg Cys Ile Tyr His Gly Trp Lys Phe Thr Thr Asp Gly Arg Thr

85 90 95

Val Glu Thr Pro Ser Glu Pro Glu Asp Gly Gly Arg Phe Ala Ser Lys

100 105 110

Val Lys Leu Asn His His Pro Val Val Asp Ala Gly Gly Val Ile Trp

115 120 125

Val Trp Ala Gly Gly Thr Gly Arg Glu Pro Ser Pro Phe Pro Arg Phe

130 135 140

Ala Phe Thr Asp Leu Pro Ser Ser His Val Phe Gly Ile Val Ala Ile

145 150 155 160

Leu Asp Cys Asn Trp Val Gln Gly Leu Glu Ala Asp Ile Asp Ser Ala

165 170 175

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

180 185 190

Asp Leu Leu Asp Asp Arg Ala Pro Arg Asp Glu Ile Glu Lys Thr Ser

195 200 205

Trp Gly Ile Arg Tyr Ala Ala Ile Arg Arg Leu Ser Thr Gly Asp Ser

210 215 220

Leu Val Arg Val Lys Pro Met Val Met Pro Trp Tyr Thr Ile Val Pro

225 230 235 240

Glu Leu Pro Asn Gly Asp Arg Leu Trp His Ala Trp Val Pro Ile Asn

245 250 255

Asp His Gln Thr Ile Phe Trp Tyr Leu Trp Tyr Asn Glu Asp Gln Pro

260 265 270

Val Asp Pro Asn Tyr Phe Ala Asp Gln Phe Gly Leu Asp Leu Glu Asn

275 280 285

Met Asn Lys Asp Asn Phe Arg Glu Gly Tyr Ser Arg Lys Asn Met Trp

290 295 300

Gly Gln Asp Arg Ala Ala Met Arg Ala Gly Thr Ser Phe Ser Gly Ile

305 310 315 320

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

325 330 335

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

340 345 350

Val Arg Ala Arg His Phe Leu Leu Asp Ala Ile Lys Asp Lys Ala Glu

355 360 365

Gly Arg Val Pro Ala Gly Leu Gly Asp Glu Val Asp Tyr Arg Lys Ile

370 375 380

Ser Ser Ala Asn Val Val Val Ser Asp Gly Asp Asp Trp Arg Gln Leu

385 390 395 400

Val Val

<210> 22

<211> 387

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 22

Met Ala Thr Leu Val Leu Thr Gly Ala Ala Thr Val Phe Ser Gly Asp

1 5 10 15

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

20 25 30

Met Ile His Ala Thr Gly Lys Ala Ser Ser Leu Thr His Glu Ile Asp

35 40 45

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

50 55 60

Leu Ile Asp Ser His Gly His Val Thr Phe Gly Asp Tyr Ser Pro Arg

65 70 75 80

Gln Arg Ala Val Asp Tyr Leu Ala Gly Tyr Val Gln Gly Gly Ile Thr

85 90 95

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

100 105 110

Arg Ala Gly Val Lys Ala Leu Ala Val Ala Ala Gln Arg Cys Phe Ser

115 120 125

Glu Tyr Arg Pro Gly Gly Met Arg Val Arg Ala Gly Ala Val Leu Ile

130 135 140

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

145 150 155 160

Val Arg Leu Ala Lys Tyr Gly Phe Gly Ala Phe Thr Asp Pro Val Asp

165 170 175

Gly Val Glu His Val Arg Trp Ala Arg Glu Ala Gly Leu Thr Val Met

180 185 190

Cys His Ala Gly Gly Ala Ser Ala Ala Gly Gly Ala Ser Leu Gly Gly

195 200 205

Glu Ala Leu Leu Ala Leu Arg Pro Asp Val Cys Gly His Ala Asn Gly

210 215 220

Gly Pro Thr Ala Leu Pro Phe Ala Glu Val Glu Ala Leu Phe Ala Glu

225 230 235 240

Thr Asp Met Ala Leu Gln Val Val Gln Ala Gly Asn Leu Lys Ala Ala

245 250 255

Leu Arg Ile Val Glu His Ala Ala Ser Arg Asp Glu Leu His Arg Val

260 265 270

Val Val Gly Ser Asp Thr Pro Ser Gly Phe Gly Val Met Pro Leu Ala

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

Leu Ala Ala Pro Ala Gly Ser Gln Ala Asp Thr Val Ala Glu Ala Met

340 345 350

Arg Val Gly Asp Ile Pro Ala Ile Thr Val Val Val Thr Ala Gly Thr

355 360 365

Val Arg Ala Met Pro Gly Arg Met Thr Pro Arg Pro Ala Val Ala Ala

370 375 380

Glu Leu Arg

385

<210> 23

<211> 404

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 23

Val Leu Thr Asp Val Arg Glu Ala Arg Val Thr Tyr Trp Ser Val Leu

1 5 10 15

Arg Arg Gln His Val Phe Arg Val Leu Phe Gly Ser Trp Val Gly Arg

20 25 30

Leu Ala Met Ala Met Ala Ala Val Ala Ile Pro Leu Ala Leu Arg Asp

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Gly Phe Val Thr Ile Ala Leu Ala Pro Ala Asn His Ala Val Val Leu

100 105 110

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

115 120 125

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

130 135 140

Ala Tyr Ala Val Asp Ser Ala Ala Gln Glu Leu Val Phe Val Phe Gly

145 150 155 160

Pro Leu Val Val Ser Leu Cys Leu Ala Val Phe Ala Pro Val Ser Ala

165 170 175

Leu Trp Ala Gln Ala Leu Leu Thr Leu Ala Gly Val Ala Thr Phe Ala

180 185 190

Thr Ala Ala Pro Ala Arg Arg Trp Ala Pro Glu Lys Thr Glu His Arg

195 200 205

His Trp Leu Gly Pro Leu Arg His Arg Gly Leu Thr Ile Leu Leu Leu

210 215 220

Ala Pro Val Gly Thr Gly Val Ala Ile Gly Thr Leu Asn Val Leu Val

225 230 235 240

Val Ser Tyr Ala Glu Arg Phe Glu Val Phe Gly Gly Ala Pro Met Leu

245 250 255

Leu Ala Leu Asn Ala Phe Ala Ala Leu Ala Gly Ala Leu Ile Tyr Gly

260 265 270

Met Ile Arg Trp Thr Ile Gln Pro Arg Thr Arg Ile Val Ser Phe Ala

275 280 285

Ala Gly Leu Leu Val Gly Tyr Gly Met Leu Ala Thr Val Pro Thr Pro

290 295 300

Leu Pro Met Ala Gly Leu Met Leu Leu Thr Gly Phe Phe Leu Ala Pro

305 310 315 320

Met Leu Ala Ala Leu Phe Gly Leu Ile Ser Asp Leu Ala Pro Arg Gly

325 330 335

Thr Val Thr Glu Ala Phe Ala Trp Leu Val Thr Leu Phe Ala Ala Gly

340 345 350

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

355 360 365

Leu His Leu Ala Ala Ala His Ala Gly Ala Gly Ala Leu Phe Cys Leu

370 375 380

Ala Val Leu Gly Ala Gly Tyr Arg Leu Phe Val Pro Ala Gly Gln Pro

385 390 395 400

Val Ala Ala Asp

<210> 24

<211> 156

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 24

Met Ile Ala Glu Glu Leu Arg Cys Ala Leu Arg His His Pro Gln Ala

1 5 10 15

Val Ala Ile Ile Thr Ala Ala Gly Pro Asp Gly Pro Ile Gly Val Thr

20 25 30

Val Ser Ser Phe Cys Ser Ala Ser Met His Pro Pro Leu Val Val Ala

35 40 45

Trp Ile Gly Thr Lys Ala Ser Ala Trp Pro Val Leu Ser Ala Ala Pro

50 55 60

Leu Phe Ala Val His Leu Leu Gly Thr Glu Gln Arg Gly Leu Ala Asp

65 70 75 80

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

85 90 95

Lys Pro Asp Asp Asp Gly Val Pro His Leu Leu Asp Pro Pro Val Arg

100 105 110

Met Arg Cys Arg Ala Val Glu Arg Ile Ala Val Gly Asp His Val Ala

115 120 125

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

130 135 140

Leu Val Arg His Gln Gly Arg Trp Thr Ser Val Ala

145 150 155

<210> 25

<211> 169

<212> PRT

<213> Streptomyces albidus (Streptomyces candidus)

<400> 25

Met Thr Ala Ile Gly Val Leu Val Gly Asn Pro Arg Pro Ala Ser Arg

1 5 10 15

Thr Leu Arg Ala Ala Leu Ala Leu Arg Glu Ala Ala Arg Gln Val Leu

20 25 30

Lys Asp Pro Val Val Gly Pro Thr Val Asp Ala Ala Glu Ile Thr His

35 40 45

Glu Val Phe Asp Ser Thr Arg Met Arg Ala Val Leu Asp Glu Leu Ala

50 55 60

Gln Ser Asp Val Leu Val Val Ala Thr Pro Thr Tyr Lys Ala Thr Tyr

65 70 75 80

Thr Gly Leu Leu Lys Ala Val Leu Asp Gln Ala Pro Pro Asn Trp Leu

85 90 95

Arg Gly Lys Ala Val Leu Pro Leu Gln Ile Ala Ala Ser Asp Arg His

100 105 110

Ala Leu Ala Val Glu Val His Leu Arg Pro Leu Leu Val Glu Leu Gly

115 120 125

Gly Gln Val Pro Gly Arg Gly Ile Phe Leu Asn Glu Ser Ala Leu Gly

130 135 140

Val Asp Asp Asp Ala Leu Thr Ala Ala Leu Val Asp Leu Leu Asp Ala

145 150 155 160

Asp Ala Leu Asn Ala Leu Val Ala Ser

165