JP2001145490A - イコサペンタエン酸生合成遺伝子群をクローニングしたプラスミド及びイコサペンタエン酸を産生するラン藻 - Google Patents
イコサペンタエン酸生合成遺伝子群をクローニングしたプラスミド及びイコサペンタエン酸を産生するラン藻Info
- Publication number
- JP2001145490A JP2001145490A JP32916999A JP32916999A JP2001145490A JP 2001145490 A JP2001145490 A JP 2001145490A JP 32916999 A JP32916999 A JP 32916999A JP 32916999 A JP32916999 A JP 32916999A JP 2001145490 A JP2001145490 A JP 2001145490A
- Authority
- JP
- Japan
- Prior art keywords
- ala
- leu
- ser
- gly
- val
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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Abstract
(57)【要約】 （修正有）
【課題】 イコサペンタエン酸（EPA）などのn-3系列高
度不飽和脂肪酸をグラム陰性細菌で効率よく発現するプ
ラスミド及びこれを導入してn-3系列高度不飽和脂肪酸
を効率よく発現するラン藻の提供。 【解決手段】特定の６種類の塩基配列によってコードさ
れたイコサペンタエン酸生合成酵素群をコードする遺伝
子群を、広域宿主ベクターにクローニングして得られる
プラスミド及び該プラスミドを導入して得られるイコサ
ペンタエン酸を産生するラン藻。 【効果】 EPAなどのn-3系列高度不飽和脂肪酸は、医
薬、食品、飼料等に有用であり、物質生産に使用する生
物としてのラン藻は、価格、環境面からも有利である。
度不飽和脂肪酸をグラム陰性細菌で効率よく発現するプ
ラスミド及びこれを導入してn-3系列高度不飽和脂肪酸
を効率よく発現するラン藻の提供。 【解決手段】特定の６種類の塩基配列によってコードさ
れたイコサペンタエン酸生合成酵素群をコードする遺伝
子群を、広域宿主ベクターにクローニングして得られる
プラスミド及び該プラスミドを導入して得られるイコサ
ペンタエン酸を産生するラン藻。 【効果】 EPAなどのn-3系列高度不飽和脂肪酸は、医
薬、食品、飼料等に有用であり、物質生産に使用する生
物としてのラン藻は、価格、環境面からも有利である。
Description
【０００１】
【発明の属する技術分野】本発明は、イコサペンタエン
酸（以下EPAと称する）産生菌から得られたEPA生合成遺
伝子群を含有するプラスミド及び当該プラスミドを導入
して作製されるイコサペンタエン酸を産生する形質転換
体ラン藻に関する。
酸（以下EPAと称する）産生菌から得られたEPA生合成遺
伝子群を含有するプラスミド及び当該プラスミドを導入
して作製されるイコサペンタエン酸を産生する形質転換
体ラン藻に関する。
【０００２】
【従来の技術】イコサペンタエン酸・ドコサヘキサエン
酸に代表されるn-3系列高度不飽和脂肪酸は、細胞膜の
構成脂肪酸であり循環系・脳神経系の正常な機能の発現
に重要な役割を果たしていることが知られている。しか
し、人間はこれらを生合成することができないため主に
魚介類からの摂取に頼っている。また、慣習的に魚介類
を摂取しない民族も多く存在している。そのためn-3系
列高度不飽和脂肪酸は、生活習慣病の治療や予防のため
医薬品・健康食品として広く販売されている。さらに、
粉ミルク等の乳幼児向けの食品や養殖魚の稚魚の必須栄
養として飼料に添加するかたちでも用いられている。
酸に代表されるn-3系列高度不飽和脂肪酸は、細胞膜の
構成脂肪酸であり循環系・脳神経系の正常な機能の発現
に重要な役割を果たしていることが知られている。しか
し、人間はこれらを生合成することができないため主に
魚介類からの摂取に頼っている。また、慣習的に魚介類
を摂取しない民族も多く存在している。そのためn-3系
列高度不飽和脂肪酸は、生活習慣病の治療や予防のため
医薬品・健康食品として広く販売されている。さらに、
粉ミルク等の乳幼児向けの食品や養殖魚の稚魚の必須栄
養として飼料に添加するかたちでも用いられている。
【０００３】現在n-3系列高度不飽和脂肪酸の原料とし
ては、主としてマグロ・イワシなどの魚類が用いられて
いる。これらの魚類は自ら高度不飽和脂肪酸を生合成す
るのではなく餌として取り込んだものを蓄積している。
n-3系列高度不飽和脂肪酸を生合成できる生物として知
られているのは主に海洋細菌や海洋微細藻類等の海にお
ける食物連鎖の低次生産者となる海洋微生物であるが、
海洋ラン藻でこれを産生するものは知られていない。
ては、主としてマグロ・イワシなどの魚類が用いられて
いる。これらの魚類は自ら高度不飽和脂肪酸を生合成す
るのではなく餌として取り込んだものを蓄積している。
n-3系列高度不飽和脂肪酸を生合成できる生物として知
られているのは主に海洋細菌や海洋微細藻類等の海にお
ける食物連鎖の低次生産者となる海洋微生物であるが、
海洋ラン藻でこれを産生するものは知られていない。
【０００４】海洋ラン藻に遺伝子組換え技術を用いてn-
3系列高度不飽和脂肪酸の生合成遺伝群を導入した例と
して、海洋細菌由来のEPA生合成遺伝子群をシネココッ
カス(Synechoccus)sp.NKBG042902に導入した竹山らの報
告（H. Takeyama et al., Microbiolgy,143,2725-2731
(1997)）が有る。しかし、導入したプラスミドが約50kb
と巨大なものであったため複製の際問題が生じて安定な
EPA生産はみられなかった。
3系列高度不飽和脂肪酸の生合成遺伝群を導入した例と
して、海洋細菌由来のEPA生合成遺伝子群をシネココッ
カス(Synechoccus)sp.NKBG042902に導入した竹山らの報
告（H. Takeyama et al., Microbiolgy,143,2725-2731
(1997)）が有る。しかし、導入したプラスミドが約50kb
と巨大なものであったため複製の際問題が生じて安定な
EPA生産はみられなかった。
【０００５】
【発明が解決しようとする課題】海洋ラン藻は地球レベ
ルでの酸素の生産者として重要であり、その物質生産能
力を利用することにより二酸化炭素の削減にも寄与でき
る。また、ラン藻は光合成による独立栄養生物であるた
め、培養には安価な無機培地を用いることができ照明以
外には特殊な設備を要さない点で産業レベルに載せやす
いと考えられる。さらに、ラン藻は原核生物であるため
菌体全体を物質生産の原料として用いることにより廃棄
物の量を低く押さえることができるのも有利な点であ
る。このような性質を持ったラン藻に有用物質であるn-
3系列高度不飽和脂肪酸を生産させることは非常に意義
のあることと考えられる。本発明は、実用化のためにn-
3系列高度不飽和脂肪酸、とくにＥＰＡの産生能の向上
と導入するプラスミドの安定化を目的とする。
ルでの酸素の生産者として重要であり、その物質生産能
力を利用することにより二酸化炭素の削減にも寄与でき
る。また、ラン藻は光合成による独立栄養生物であるた
め、培養には安価な無機培地を用いることができ照明以
外には特殊な設備を要さない点で産業レベルに載せやす
いと考えられる。さらに、ラン藻は原核生物であるため
菌体全体を物質生産の原料として用いることにより廃棄
物の量を低く押さえることができるのも有利な点であ
る。このような性質を持ったラン藻に有用物質であるn-
3系列高度不飽和脂肪酸を生産させることは非常に意義
のあることと考えられる。本発明は、実用化のためにn-
3系列高度不飽和脂肪酸、とくにＥＰＡの産生能の向上
と導入するプラスミドの安定化を目的とする。
【０００６】
【課題を解決するための手段】本発明は、高度不飽和脂
肪酸を広範な細菌で発現するプラスミド及び該プラスミ
ドを導入し高度不飽和脂肪酸を安定に効率よく産生する
形質転換体ラン藻を提供する。すなわち、本発明は、
配列番号２，４，６，８，１０，及び１２で示される塩
基配列によってコードされたイコサペンタエン酸生合成
酵素群をコードする遺伝子群を、広域宿主ベクターにク
ローニングして得られるプラスミド及び３，５，７，
９，１１，及び１３で示される塩基配列で表されるイコ
サペンタエン酸生合成酵素群をコードする遺伝子群を、
広域宿主ベクターにクローニングして得られるプラスミ
ド、並びにこれらのプラスミドを導入して得られるイコ
サペンタエン酸を産生するラン藻に関する。
肪酸を広範な細菌で発現するプラスミド及び該プラスミ
ドを導入し高度不飽和脂肪酸を安定に効率よく産生する
形質転換体ラン藻を提供する。すなわち、本発明は、
配列番号２，４，６，８，１０，及び１２で示される塩
基配列によってコードされたイコサペンタエン酸生合成
酵素群をコードする遺伝子群を、広域宿主ベクターにク
ローニングして得られるプラスミド及び３，５，７，
９，１１，及び１３で示される塩基配列で表されるイコ
サペンタエン酸生合成酵素群をコードする遺伝子群を、
広域宿主ベクターにクローニングして得られるプラスミ
ド、並びにこれらのプラスミドを導入して得られるイコ
サペンタエン酸を産生するラン藻に関する。
【０００７】
【発明の実施の形態】本発明において用いられるＥＰＡ
生合成酵素群をコードする遺伝子群は、例えば実施例１
あるいは特開平８−２４２８６７号に記載の方法により
単離することができる。なお、本発明において、ＥＰＡ
生合成酵素群をコードする遺伝子群は、配列番号２，
４，６，８，１０，及び１２で示される塩基配列によっ
てコードされるものとストリンジェントな条件でハイブ
リダイズするものを包含する。
生合成酵素群をコードする遺伝子群は、例えば実施例１
あるいは特開平８−２４２８６７号に記載の方法により
単離することができる。なお、本発明において、ＥＰＡ
生合成酵素群をコードする遺伝子群は、配列番号２，
４，６，８，１０，及び１２で示される塩基配列によっ
てコードされるものとストリンジェントな条件でハイブ
リダイズするものを包含する。
【０００８】ＥＰＡ生合成酵素群をコードする遺伝子群
を他の生物に導入するためには、これらの遺伝子を運ん
で発現させる部品を備えたベクターが必要である。原核
生物に上述の遺伝子群を導入する際は、多くの場合には
そのままのプロモーター/ターミネーターが使用できる
ので、それぞれの微生物で複製のできる複製開始点を有
したベクターを用いることができる。一般的な広域宿主
ベクターが使用可能であり、例えばpJRD215（Davidson
et al.,Gene, 51,275-280(1987)）及びpBBR1MCSシリー
ズ（Kovach et al., Gene, 166,175-176(1995)）等を例
示することができる。これらのベクターへの上述の遺伝
子群のクローニングは、すべての遺伝子群を含むＤＮＡ
断片あるいはすべての遺伝子群を含むことになる複数の
ＤＮＡ断片を用いて、慣用の方法で行うことができる。
を他の生物に導入するためには、これらの遺伝子を運ん
で発現させる部品を備えたベクターが必要である。原核
生物に上述の遺伝子群を導入する際は、多くの場合には
そのままのプロモーター/ターミネーターが使用できる
ので、それぞれの微生物で複製のできる複製開始点を有
したベクターを用いることができる。一般的な広域宿主
ベクターが使用可能であり、例えばpJRD215（Davidson
et al.,Gene, 51,275-280(1987)）及びpBBR1MCSシリー
ズ（Kovach et al., Gene, 166,175-176(1995)）等を例
示することができる。これらのベクターへの上述の遺伝
子群のクローニングは、すべての遺伝子群を含むＤＮＡ
断片あるいはすべての遺伝子群を含むことになる複数の
ＤＮＡ断片を用いて、慣用の方法で行うことができる。
【０００９】一般的にベクターにクローニングした遺伝
子をそれぞれの原核生物に導入する方法としては、形質
転換法・接合法・エレクトロポーレーション法が挙げら
れる。細胞内への遺伝子の導入を確認する方法として
は、直接的には、その発現の結果である生産物、高度不
飽和脂肪酸を検出する。上述のように作製されたプラス
ミドを導入した生物から有機溶媒抽出を行うことにより
EPA等の高度不飽和脂肪酸を得ることができる。間接的
には、導入した遺伝子の一部をプローブやプライマーと
したハイブリダイゼーションやPCRにより遺伝子が導入
されたことを確認することが可能である。
子をそれぞれの原核生物に導入する方法としては、形質
転換法・接合法・エレクトロポーレーション法が挙げら
れる。細胞内への遺伝子の導入を確認する方法として
は、直接的には、その発現の結果である生産物、高度不
飽和脂肪酸を検出する。上述のように作製されたプラス
ミドを導入した生物から有機溶媒抽出を行うことにより
EPA等の高度不飽和脂肪酸を得ることができる。間接的
には、導入した遺伝子の一部をプローブやプライマーと
したハイブリダイゼーションやPCRにより遺伝子が導入
されたことを確認することが可能である。
【００１０】本発明の上記プラスミドをラン藻に導入す
る際にも、上記の方法を適用して行うことができる。用
いられるラン藻としては、特に制限はないが、例えばシ
ネココッカスsp.NKBG15041c、シネココッカスsp.NKBG04
2902等を挙げることができる。上記プラスミドを導入し
たイコサペンタエン酸を産生するラン藻として、平成11
年11月9日に工業技術院生命工学工業技術研究所に受託
番号FERM P-17634として寄託された、15041c/pJRDEPA-S
が挙げられる。
る際にも、上記の方法を適用して行うことができる。用
いられるラン藻としては、特に制限はないが、例えばシ
ネココッカスsp.NKBG15041c、シネココッカスsp.NKBG04
2902等を挙げることができる。上記プラスミドを導入し
たイコサペンタエン酸を産生するラン藻として、平成11
年11月9日に工業技術院生命工学工業技術研究所に受託
番号FERM P-17634として寄託された、15041c/pJRDEPA-S
が挙げられる。
【００１１】
【発明の効果】本発明では、高度不飽和脂肪酸の生合成
に必須でないORF及び遺伝子をコードしていていない部
分を可能な限り取り除いて短縮化することにより、他の
生物に導入した場合に細胞分裂に伴う脱落を防ぎ遺伝子
群の安定で効率良い発現が達成される。
に必須でないORF及び遺伝子をコードしていていない部
分を可能な限り取り除いて短縮化することにより、他の
生物に導入した場合に細胞分裂に伴う脱落を防ぎ遺伝子
群の安定で効率良い発現が達成される。
【００１２】
【実施例】次に、実施例及び参考例により本発明をさら
に具体的に説明する。
に具体的に説明する。
【００１３】実施例１ 小型化プラスミドの作製 特開平8-242867に記載のプラスミドpEPAに挿入されたEP
A生合成遺伝子群のうち、EPA生合成に必須であるORF3、
6、7、8および9のサブクローニングを行った。ORF5、
6、7、8および9については、クローニングベクターpBSI
IKS(+)（Stratagene社製）のXbaI-SpeI部位にXbaI-SpeI
断片（23,045-31,443）、XbaI部位にXbaI-XbaI断片（1
2,314-23,045）SpeI部位にSpeI-NheI断片（31,443-32,5
14）を順次サブクローニングを行いΔX4XbNh/pBS を作
製した。ΔX4XbNh/pBS をNotIで処理したものをT4DNAポ
リメラーゼにより平滑末端を作り、それをXhoIで処理し
て DNA断片Aを得た。 また、ORF3については、R/pSTV28
（HpaI断片7,951-9,129を宝酒造製ベクターpSTV28のSma
I部位に挿入したもの）をEcoRI及びPstIで処理して切り
出した断片をpBSIIKS(+)のEcoRI-PstI部位に挿入してR/
pBSを作製した。R/pBSをPstIで処理後T4DNAポリメラー
ゼにより平滑末端を作りXhoIリンカーを導入してからXh
oIでORF3を含む断片を切り出しDNA断片Bを得た。広域宿
主ベクターであるpJRD215（カナマイシン及びストレプ
トマイシン耐性)のXhoI-StuI部位に断片Aをパッカジン
ラムダDNAパケージングシステム（Promega社製）により
導入した後に、XhoI部位に断片BをDNA ライゲーション
キット （宝酒造製）を用いて導入しプラスミドを完成
させた。これをpJRDEPA-Sと命名した（図1）。
A生合成遺伝子群のうち、EPA生合成に必須であるORF3、
6、7、8および9のサブクローニングを行った。ORF5、
6、7、8および9については、クローニングベクターpBSI
IKS(+)（Stratagene社製）のXbaI-SpeI部位にXbaI-SpeI
断片（23,045-31,443）、XbaI部位にXbaI-XbaI断片（1
2,314-23,045）SpeI部位にSpeI-NheI断片（31,443-32,5
14）を順次サブクローニングを行いΔX4XbNh/pBS を作
製した。ΔX4XbNh/pBS をNotIで処理したものをT4DNAポ
リメラーゼにより平滑末端を作り、それをXhoIで処理し
て DNA断片Aを得た。 また、ORF3については、R/pSTV28
（HpaI断片7,951-9,129を宝酒造製ベクターpSTV28のSma
I部位に挿入したもの）をEcoRI及びPstIで処理して切り
出した断片をpBSIIKS(+)のEcoRI-PstI部位に挿入してR/
pBSを作製した。R/pBSをPstIで処理後T4DNAポリメラー
ゼにより平滑末端を作りXhoIリンカーを導入してからXh
oIでORF3を含む断片を切り出しDNA断片Bを得た。広域宿
主ベクターであるpJRD215（カナマイシン及びストレプ
トマイシン耐性)のXhoI-StuI部位に断片Aをパッカジン
ラムダDNAパケージングシステム（Promega社製）により
導入した後に、XhoI部位に断片BをDNA ライゲーション
キット （宝酒造製）を用いて導入しプラスミドを完成
させた。これをpJRDEPA-Sと命名した（図1）。
【００１４】参考例１ ｐJRDEPA-Sを導入した大腸菌で
のEPA生産 pJRDEPA-Sを用いて大腸菌K12/JM109を常法により形質転
換した。50μg/mlのカナマイシンを含むLB寒天培地（ト
リプトン1％、酵母エキス0.5％、NaCl 1％、寒天1.5
％）を用いて選別を行いJM109/pJRDEPA-Sのコロニーを
得た。このコロニーを50μg/mlのカナマイシンを含む2m
lLB液体培地に接種し25℃で24時間培養した。これを遠
心分離して菌体を集め培地を除いた後、10mMトリス塩酸
緩衝液 pH7.5を加えて懸濁し再度遠心分離を行って洗浄
した。洗浄菌体に少量の10mMトリス塩酸緩衝液 pH7.5を
加えて再懸濁し凍結乾燥をした。乾燥菌体に5％塩化水
素を含むメタノルーを1ml加え80℃1時間処理して脂肪酸
のメチルエステル化を行った。冷却後同量のn-ヘキサン
で3回抽出しn-ヘキサン層を減圧乾固して20μlのメタノ
ールに溶解し試料とした。この試料の一部をガスクロマ
トグラフィー（以下GLCと略す）により分析した。その
結果、標品のEPAメチルエステルと同様の保持時間にピ
ークが検出された。そのピークの面積比から算出される
総脂肪酸に対する割合は5.7％であった。また、このピ
ークはガスクロマトグラフィー質量スペクトル（以下GC
-MSと略す）分析により親イオン(M) m/z316、ベースピ
ークm/z79でありEPAメチルエステル標品と一致した。
のEPA生産 pJRDEPA-Sを用いて大腸菌K12/JM109を常法により形質転
換した。50μg/mlのカナマイシンを含むLB寒天培地（ト
リプトン1％、酵母エキス0.5％、NaCl 1％、寒天1.5
％）を用いて選別を行いJM109/pJRDEPA-Sのコロニーを
得た。このコロニーを50μg/mlのカナマイシンを含む2m
lLB液体培地に接種し25℃で24時間培養した。これを遠
心分離して菌体を集め培地を除いた後、10mMトリス塩酸
緩衝液 pH7.5を加えて懸濁し再度遠心分離を行って洗浄
した。洗浄菌体に少量の10mMトリス塩酸緩衝液 pH7.5を
加えて再懸濁し凍結乾燥をした。乾燥菌体に5％塩化水
素を含むメタノルーを1ml加え80℃1時間処理して脂肪酸
のメチルエステル化を行った。冷却後同量のn-ヘキサン
で3回抽出しn-ヘキサン層を減圧乾固して20μlのメタノ
ールに溶解し試料とした。この試料の一部をガスクロマ
トグラフィー（以下GLCと略す）により分析した。その
結果、標品のEPAメチルエステルと同様の保持時間にピ
ークが検出された。そのピークの面積比から算出される
総脂肪酸に対する割合は5.7％であった。また、このピ
ークはガスクロマトグラフィー質量スペクトル（以下GC
-MSと略す）分析により親イオン(M) m/z316、ベースピ
ークm/z79でありEPAメチルエステル標品と一致した。
【００１５】実施例２−１ ラン藻のトランスコンジュ
ゲーション ラン藻シネココッカス(Synechococcus) sp. NKBG15041c
（K.Sode et al., Appl. Microbiol. Biotechnol., 37,
369-373 (1992)）を3％NaClを含むBG11（表1）液体培地
（BG11M）1,000-1,500 Luxの光照射下23℃で 4-5日間
静置培養を行い（A550＜1）、室温で3,000rpm20分間遠
心して集め、BG11M液体培地に懸濁して3回洗った。この
ラン藻の濃度を分光光度計でA550＝1に合わせた。上記p
JRDEPA-Sでトランスコンジュゲーション用大腸菌S-17
（Simon et al., Bio/Technolgy,118,640-659(1983) ）
を形質転換し、50μg/mlのカナマイシンを含むLB寒天培
地に37℃一晩培養して生育したコロニーをBG11Mに懸濁
したものを分光光度計で濃度を測定しA650＝10に合わせ
た。以上のように調製したラン藻と大腸菌を等量ずつラ
ン藻A550＝1に対し大腸菌A650＝10になるように混合し
た。この菌液を遠心後元の10分の1になるように懸濁し
たものをBG11に15mMNaClを加えた1.2％寒天培地に50μl
ずつスポットし、照明下24〜48時間23℃で培養した。寒
天培地上にできた緑色のコロニーをメスで切り出して1m
lBG11Mに懸濁し75μg/mlカナマイシンを含むBG11M液体
培地に50分の1の濃度になるように加え照明下23℃で培
養した。
ゲーション ラン藻シネココッカス(Synechococcus) sp. NKBG15041c
（K.Sode et al., Appl. Microbiol. Biotechnol., 37,
369-373 (1992)）を3％NaClを含むBG11（表1）液体培地
（BG11M）1,000-1,500 Luxの光照射下23℃で 4-5日間
静置培養を行い（A550＜1）、室温で3,000rpm20分間遠
心して集め、BG11M液体培地に懸濁して3回洗った。この
ラン藻の濃度を分光光度計でA550＝1に合わせた。上記p
JRDEPA-Sでトランスコンジュゲーション用大腸菌S-17
（Simon et al., Bio/Technolgy,118,640-659(1983) ）
を形質転換し、50μg/mlのカナマイシンを含むLB寒天培
地に37℃一晩培養して生育したコロニーをBG11Mに懸濁
したものを分光光度計で濃度を測定しA650＝10に合わせ
た。以上のように調製したラン藻と大腸菌を等量ずつラ
ン藻A550＝1に対し大腸菌A650＝10になるように混合し
た。この菌液を遠心後元の10分の1になるように懸濁し
たものをBG11に15mMNaClを加えた1.2％寒天培地に50μl
ずつスポットし、照明下24〜48時間23℃で培養した。寒
天培地上にできた緑色のコロニーをメスで切り出して1m
lBG11Mに懸濁し75μg/mlカナマイシンを含むBG11M液体
培地に50分の1の濃度になるように加え照明下23℃で培
養した。
【００１６】表１ BG11の組成 ━━━━━━━━━━━━━━━━ NaNO3 1.5g MgSO4・7H2O 75mg CaCl2・2H2O 36mg クエン酸 6mg Na2EDTA 1mg Na2CO3 20mg 微量金属混合物 A5* 1ml K2HPO4 30mg クエン酸 鉄アンモニウム 6mg ビタミンB12 1mg 脱イオン蒸留水 -1ｌ pH 7.2-7.4 ━━━━━━━━━━━━━━━━ *微量金属混合物A5 ━━━━━━━━━━━━━━━ H3BO4 2.86g MnCl2・6H2O 1.81g ZnSO4・7H2O 222mg Na2MoO4・2H2O 390mg CuSO4・5H2O 79mg Co(NO3)2・7H2O 49.4mg 脱イオン蒸留水 -1ｌ ━━━━━━━━━━━━━━━
【００１７】実施例２−２ ラン藻のシングルコロニー
アイソレーション pJRDEPA-Sを有するラン藻をA730=3-4に生育させ、10-4-
10-5希釈したものを75μg/mlカナマイシンを含むBG11M
の寒天培地に塗布し23℃照明下約1ヶ月培養してシング
ルコロニーを形成させた。このコロニーを75μg/mlカナ
マイシンを含むBG11M液体培地に移して培養した。得ら
れた組換え体ラン藻を15041c/pJRDEPA-Sと命名した（受
託番号FERM P-17634）。
アイソレーション pJRDEPA-Sを有するラン藻をA730=3-4に生育させ、10-4-
10-5希釈したものを75μg/mlカナマイシンを含むBG11M
の寒天培地に塗布し23℃照明下約1ヶ月培養してシング
ルコロニーを形成させた。このコロニーを75μg/mlカナ
マイシンを含むBG11M液体培地に移して培養した。得ら
れた組換え体ラン藻を15041c/pJRDEPA-Sと命名した（受
託番号FERM P-17634）。
【００１８】実施例２−３ ラン藻の菌体脂質の調製及
び分析 pJRDEPA-S を有するラン藻及び非組換え体のラン藻を培
養しその菌体を実施例1-2と同様に遠心分離で集め洗浄
後凍結乾燥を行った。乾燥菌体に実施例1-2と同様の脂
肪酸のメチルエステル化を行いn-ヘキサン抽出し減圧乾
固後メタノールに溶解して粗試料とした。この粗試料を
シリカゲル薄層プレートにスポットしn-ヘキサン：エチ
ルエーテル（4：1、v/v）で展開してメチルエステルを
分離した。プリムリン発色により検出したメチルエステ
ル画分を掻き取り、メタノール：10％食塩（9：1、v/
v）1ml及び水1mlを加えたものから上述と同様にn-ヘキ
サン抽出を行い試料を調製した。この試料の一部をGLC
にかけて分析した。標品のEPAメチルエステルと保持時
間が一致するピーク及びその約1分前にもほぼ同じ高さ
のピークが検出された。2本ののピークはその面積比か
ら求めた総脂肪酸に対する割合はそれぞれ3.8％及び2.6
％であった。主な脂肪酸の総脂質に占める割合を表2に
示す。残りの試料を硝酸銀シリカゲル薄層プレートにス
ポットしn-ヘキサン：エチルエーテル（85：15、v/v）
で3回展開して2',7'-ジクロロフルオレセイン発色によ
り検出した多価不飽和脂肪酸画分を掻き取り上述と同様
な方法で GC-MS分析用試料を調製した。GC-MS分析によ
り、EPAメチルエステルと保持時間が一致するピークの
分子量は316でEPAと一致した。また、その前のピークは
分子量318で20：4であるが、アラキドン酸メチルエステ
ル（20：4（n-6））のガスクロマトグラフィーの保持時
間とは異なっていた。このピークは、標品の20：4（n-
3）メチルエステルの保持時間及びGC-MSのの解裂パター
ンと一致しており、20：4（n-3）メチルエステルと同定
された。
び分析 pJRDEPA-S を有するラン藻及び非組換え体のラン藻を培
養しその菌体を実施例1-2と同様に遠心分離で集め洗浄
後凍結乾燥を行った。乾燥菌体に実施例1-2と同様の脂
肪酸のメチルエステル化を行いn-ヘキサン抽出し減圧乾
固後メタノールに溶解して粗試料とした。この粗試料を
シリカゲル薄層プレートにスポットしn-ヘキサン：エチ
ルエーテル（4：1、v/v）で展開してメチルエステルを
分離した。プリムリン発色により検出したメチルエステ
ル画分を掻き取り、メタノール：10％食塩（9：1、v/
v）1ml及び水1mlを加えたものから上述と同様にn-ヘキ
サン抽出を行い試料を調製した。この試料の一部をGLC
にかけて分析した。標品のEPAメチルエステルと保持時
間が一致するピーク及びその約1分前にもほぼ同じ高さ
のピークが検出された。2本ののピークはその面積比か
ら求めた総脂肪酸に対する割合はそれぞれ3.8％及び2.6
％であった。主な脂肪酸の総脂質に占める割合を表2に
示す。残りの試料を硝酸銀シリカゲル薄層プレートにス
ポットしn-ヘキサン：エチルエーテル（85：15、v/v）
で3回展開して2',7'-ジクロロフルオレセイン発色によ
り検出した多価不飽和脂肪酸画分を掻き取り上述と同様
な方法で GC-MS分析用試料を調製した。GC-MS分析によ
り、EPAメチルエステルと保持時間が一致するピークの
分子量は316でEPAと一致した。また、その前のピークは
分子量318で20：4であるが、アラキドン酸メチルエステ
ル（20：4（n-6））のガスクロマトグラフィーの保持時
間とは異なっていた。このピークは、標品の20：4（n-
3）メチルエステルの保持時間及びGC-MSのの解裂パター
ンと一致しており、20：4（n-3）メチルエステルと同定
された。
【００１９】 表２ ｐJRDEPA-Sを導入したラン藻のおもな脂肪酸組成（総脂肪酸に対する％） ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ 脂肪酸 ｐJRDEPA-Sを導入したラン藻 非組換え体ラン藻 ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ 16:0 32.6 33.1 16:1 (n-7) 12.2 13.1 18:1 (n-9) 12.8 19.1 18:2 (n-6) 14.9 17.9 18:3 (n-3) 6.7 7.1 20:4 (n-3) 2.6 N.D. 20:5 (n-3) 3.8 N.D. ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ N.D. ：検出せず
【００２０】実施例3. プラスミドの比較 pJRDEPA-Sと同時にpJRDEPA（H. Takeyama et al., Micr
obiolgy,143,2725-2731(1997)）を実施例2-1と同様にシ
ネココッカスsp.NKBG15041cに導入して75μg/mlのカナ
マイシンを加えたBG11M液体培地1ｌで培養を行った。実
施例2-3と同様に脂肪酸メチルエステルを調製しGLCで分
析を行った。その結果を表3に示す。
obiolgy,143,2725-2731(1997)）を実施例2-1と同様にシ
ネココッカスsp.NKBG15041cに導入して75μg/mlのカナ
マイシンを加えたBG11M液体培地1ｌで培養を行った。実
施例2-3と同様に脂肪酸メチルエステルを調製しGLCで分
析を行った。その結果を表3に示す。
【００２１】 表３ NKBG15041CのEPA生産能と導入したプラスミド ━━━━━━━━━━━━━━━━━━━━━━━━━━━ プラスミド pJRDEPA-S pJRDEPA ━━━━━━━━━━━━━━━━━━━━━━━━━━━ EPA(%*) 1.79 0.03 サイズ(Kb) 31 47 ━━━━━━━━━━━━━━━━━━━━━━━━━━━ *総脂肪酸に対する％ このようにpJRDEPA-Sを導入したNKBG15041cでは、pJRDE
PAを導入したものに比較して飛躍的にEPA生産能が増加
した。
PAを導入したものに比較して飛躍的にEPA生産能が増加
した。
【００２２】参考例2 ラン藻の培養条件 pJRDEPA-S を有するラン藻を通常（23℃、1,000Lux、静
置）、低温（18℃、800Lux、振とう）及び弱照明（23
℃、40Lux、静置）の3条件で培養し、それぞれの脂肪酸
組成を実施例2-3を同様の方法でGLCにより分析した結果
を表3に示す。EPA及び20：4（n-3）の総脂肪酸に対する
割合は、通常に比べ生育を抑制した場合（低温及び弱照
明）の方が高く、それぞれ3.8％・2.6％に対し、6.0％
・5.7％及び5.2％・6.6％であった。
置）、低温（18℃、800Lux、振とう）及び弱照明（23
℃、40Lux、静置）の3条件で培養し、それぞれの脂肪酸
組成を実施例2-3を同様の方法でGLCにより分析した結果
を表3に示す。EPA及び20：4（n-3）の総脂肪酸に対する
割合は、通常に比べ生育を抑制した場合（低温及び弱照
明）の方が高く、それぞれ3.8％・2.6％に対し、6.0％
・5.7％及び5.2％・6.6％であった。
【００２３】 表４ おもな脂肪酸組成に及ぼす培養条件の影響（総脂肪酸に対する％） ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ 脂肪酸 低温 通常 弱照明 ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ 16:0 37.4 32.6 36.4 16:1 (n-7) 9.3 12.2 9.1 18:1 (n-9) 8.3 12.8 7.5 18:2 (n-6) 8.0 14.9 8.9 18:3 (n-3) 4.8 6.7 2.6 20:4 (n-3) 6.0 2.6 5.2 20:5 (n-3) 5.7 3.8 6.6 A730 4.28 4.48 1.51 ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ 低温：18℃、800Lux、振とう 通常：23℃、1,000lux、静置 弱照明：23℃、40Lux、静置
【００２４】参考例３ pJRDEPAに関しては、ラン藻を継代すると、ラン藻の分
裂にプラスミドの複製が間に合わなくなりプラスミドが
失われていったとの報告がある（H. Takeyama etal., M
icrobiolgy,143,2725-2731(1997)）。しかし、 pJRDEPA
-Sを有するラン藻ではそのような現象は観察されず、カ
ナマイシンの存在下で継代を繰り返すことができた。
裂にプラスミドの複製が間に合わなくなりプラスミドが
失われていったとの報告がある（H. Takeyama etal., M
icrobiolgy,143,2725-2731(1997)）。しかし、 pJRDEPA
-Sを有するラン藻ではそのような現象は観察されず、カ
ナマイシンの存在下で継代を繰り返すことができた。
【００２５】
【配列表】 <110> Sagami Chemical Reserach Center, Japan Bioindustry Association, Director-General of Agency of Industrial Science and Technology. <120> A plasmid carrying the eicosapentanoic acid synthesis gene claster and a transgenic cyanobacterium producing eicosapentanoic acid. <130> SO18226 <160> 13
【００２６】 <210> 1 <211> 37895 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 Position gatctcttac aaagaaacta tctcaatgtg aatttaacct taattccgtt taattacggc 60 ctgatagagc atcacccaat cagccataaa actgtaaagt gggtactcaa aggtggctgg 120 gcgattcttc tcaaatacaa agtgcccaac ccaagcaaat ccatatccga taacaggtaa 180 aagtagcaat aaaccccagc gctgagttag taatacataa gcgaataata ggatcactaa 240 actactgccg aaatagtgta atattcgaca gtttctatgc tgatgttgag ataaataaaa 300 agggtaaaat tcagcaaaag aacgatagcg cttactcatt actcacacct cggtaaaaaa 360 gcaactcgcc attaacttgg ccaatcgtca gttgttctat cgtctcaaag ttatgccgac 420 taaataactc tatatgtgca ttatgattag caaaaactcc gataccatca agatgaagtt 480 gttcatcaca ccaactcaaa actgcgtcga taagcttact gccatagccc ttgccttgct 540 ccacatttgc gatagcaata aactgtaaaa tgccacattg gccacttggt aagctctcta 600 taatctgatt ttctttgtta ataagtgcct gagttgaata ccaaccagta cttaacaaca 660 tctttaaacg ccaatgccaa aaacgcgctt cacctaaggg aacctgctga gtcactatgc 720 aggctacgcc tatcaatcta tccccaacga acataccaat aagtgcttgc tcctgttgcc 780 agagctcatt gagttcttct cgaatagccc cgcgaagctt ttgctcatac tgcgcttgat 840 caccactaaa aagtgtttcg ataaaaaagg gatcatcatg ataggcgtta tagagaatag 900 aggctgctat gcgtaaatct tctgccgtga gataaactgc acgacactct tccatggctt 960 gatcttccat tgttattgtc cttgaccttg atcacacaac accaatgtaa caagactgta 1020 tagaagtgca attaataatc aattcgtgca ttaagcaggt cagcatttct ttgctaaaca 1080 agctttattg gctttgacaa aactttgcct agactttaac gatagaaatc ataatgaaag 1140 agaaaagcta caacctagag gggaataatc aaacaactgc taagatctag ataatgtaat 1200 aaacaccgag tttatcgacc atacttagat agagtcatag caacgagaat agttatggat 1260 acaacgccgc aagatctatc acacctgttt ttacagctag gattagcaaa tgatcaaccc 1320 gcaattgaac agtttatcaa tgaccatcaa ttagcggaca atatattgct acatcaagca 1380 agcttttgga gcccatcgca aaagcacttc ttaattgagt catttaatga agatgcccag 1440 tggaccgaag tcatcgacca cttagacacc ttattaagaa aaaactaacc attacaacag 1500 caactttaaa ttttgccgta agccatctcc ccccacccca caacagcgtt gttgcttatg 1560 accactggag tacattcgtc tttagtcgtt ttaccatcac catgggtacg ttgagtgcga 1620 taaaaaagca cataaacttc tttatcggcc tgaatatagg cttcgttaaa atcagctgtt 1680 cccattaaag taaccacttg ctctttactc atgcctagag atatctttgt caaattgtca 1740 cggtttttat cttgagtttt ctcccaagca ccgtgattat cccagtcaga ttccccatca 1800 ccaacattga ccacacagcc cgttagccct aagcttgcaa tcccaaaaca tgctaaacct 1860 aataatttat ttttcatttt aacttcctgt tatgacatta tttttgctta gaagaaaagc 1920 aacttacatg ccaaaacaca agctgttgtt ttaaatgact ttatttatta ttagcctttt 1980 aggatatgcc tagagcaata ataattacca atgtttaagg aatttgacta actatgagtc 2040 cgattgagca agtgctaaca gctgctaaaa aaatcaatga acaaggtaga gaaccaacat 2100 tagcattgat taaaaccaaa cttggtaata gcatcccaat gcgcgagtta atccaaggtt 2160 tgcaacagtt taagtctatg agtgcagaag aaagacaagc aatacctagc agcttagcaa 2220 cagcaaaaga aactcaatat ggtcaatcaa gcttatctca atctgaacaa gctgatagga 2280 tcctccagct agaaaacgcc ctcaatgaat taagaaacga atttaatggg ctaaaaagtc 2340 aatttgataa cttacaacaa aacctgatga ataaagagcc tgacaccaaa tgcatgtaat 2400 tgaactacga tttgaatgtt ttgataacac cacgattact gcagcagaaa aagccattaa 2460 tggtttgctt gaagcttatc gagccaatgg ccaggttcta ggtcgtgaat ttgccgttgc 2520 atttaacgat ggtgagttta aagcacgcat gttaacccca gaaaaaagca gcttatctaa 2580 acgctttaat agtccttggg taaatagtgc actcgaagag ctaaccgaag ccaaattgct 2640 tgcgccacgt gaaaagtata ttggccaaga tattaattct gaagcatcta gccaagacac 2700 accaagttgg cagctacttt acacaagtta tgtgcacatg tgctcaccac taagaaatgg 2760 cgacaccttg cagcctattc cactgtatca aattccagca actgccaacg gcgatcataa 2820 acgaatgatc cgttggcaaa cagaatggca agcttgtgat gaattgcaaa tggccgcagc 2880 tactaaagct gaatttgccg cacttgaaga gctaaccagt catcagagtg atctatttag 2940 gcgtggttgg gacttacgtg gcagagtcga atacttgacg aaaattccga cctattacta 3000 tttataccgt gttggcggtg aaagcttagc agtagaaaag cagcgctctt gtcctaagtg 3060 tggcagtcaa gaatggctgc tcgataaacc attattggat atgttccatt ttcgctgtga 3120 cacctgccgc atcgtatcta atatctcttg ggaccattta taactcttcc gagtcttatc 3180 acactagagt ttagtcagca taaaaatggc gcttatattt caattaaaag aaatataagc 3240 gccattttca tcgatactat atatcagcag actattttcc gcgtaaatta gcccacatta 3300 atttcattct ttgccagatc cctggatgat ctagttgtgg catcgactct tcaataggtt 3360 taaccgcagg tgtaaccctt ggagtcaatt cgtttataaa ctcgtttaaa ctgtcactta 3420 atttaacgct ttgtacttca cctggaattt caatccatac gctgccatca ctattattaa 3480 ccgtcaacat tttatcttca tcatcaagaa taccaataaa ccaagtcggc tcttgcttaa 3540 gctttctctt catcattaaa tgaccaatga tgttttgttg taagtattca aaatcagttt 3600 gatcccacac ttggattagc tcaccttggc cccattgtga gtcaaaaaat agcggtgcag 3660 aaaaatgact gccaaaaaat ggattaattt ctgcagataa tgtcatttca agtgctgttt 3720 caacattagc aaattcacca ggttgttgac gtacaaccga ttgccaaaac actgcgccat 3780 cggagcccgc ttcggcgaca acacactcag acttttgtcc ttgcgcataa tatcttggct 3840 gttcaccaag cttatccatg taggcttgtt gatatttaga taaaaaaaga tctaaagcag 3900 gtaaagaaga cacttaagcc agttccaaaa tcagttataa taggggtcta ttttgacatg 3960 gaaaccgtat tgatgacaca acatcatgat ccctacagta acgcccccga actttctgaa 4020 ttaactttag gaaagtcgac cggttatcaa gagcagtatg atgcatcttt actacaagcg 4080 tgccgcgtaa attaaaccgt gatgctatcg gtctaaccaa tgagctacct tttcatggct 4140 gtgatatttg gactggctac gaactgtctt ggctaaatgc taaaggcaag ccaatgattg 4200 ctattgcaga ctttaaccta agttttgata gtaaaaatct gatcgagtct aagtcgttta 4260 agctgtattt aaacagctat aaccaaacac gatttgatag cgttcaagcg gttcaagaac 4320 gtttaactga agacttaagc gcctgtgccc aaggcacagt tacggtaaaa gtgattgaac 4380 ctaagcaatt taaccacctg agagtggttg atatgccagg tacctgcatt gacgatttag 4440 atattgaagt tgatgactat agctttaact ctgactatct caccgacagt gttgatgaca 4500 aagtcatggt tgctgaaacg ctaacgtcaa acttattgaa atcaaactgc ctaatcactt 4560 ctcagcctga ctggggtaca gtgatgatcc gttatcaagg gcctaagata gaccgtgaaa 4620 agctacttag atatctgatt tcatttagac agcacaatga atttcatgag cagtgtgttg 4680 agcgtatatt tgttgattta aagcactatt gccaatgtgc caaacttact gtctatgcac 4740 gttatacccg ccgtggtggt ttagatatca acccatatcg tagcgacttt gaaaaccctg 4800 cagaaaatca gcgcctagcg agacagtaat tgattgcagt acctacaaaa aacaatgcct 4860 ataagccaag cttatgggca tttttatatt atcaacttgt catcaaacct cagccgccaa 4920 gccttttagt tttatcgcta aattaagccg ctctctcagc caaatatttg caggattttg 4980 ctgtaattta tggctccaca ccatgaaata ctctatcggc tctaccgcaa aaggtaagtc 5040 aaatacctgt aagccaaaca gcttggcata ttcgtcagtg tgggcttttg acgcgatagc 5100 taacgcatca ctttttgagg caaccgacat catacttaat attgatgatt gctcgctgtg 5160 catttgcctt gccggtaaca cctgtttagt cagcaagtcg gcaacactta aattgtagcg 5220 gcgcatctta aaaataatat gcttttcatt aaagtattgc tcttgcgtca acccaccttg 5280 gatccttggg tgagcatttc gtgccacaca aactaattta tcctgcatta ctttttgact 5340 cttaaatgcc gcagattctg gcagccaaat atctaaggct aaatccacct tttctagttg 5400 taggtccatc tgcaactctt cttcaatgag cggcggctca cgaaatacaa tattaattgc 5460 agtgccctgt aacacttgct caatttgatc ttgcaagagt tgtattgccg actcgctggc 5520 atacacataa aaagttcgct cacttgaagt ggggtcaaat gcttcaaagc tagtcgcaac 5580 ttgctcaatt gttgacatag cgcccgcgag ctgttgataa agcgtcatcg cacttgcggt 5640 aggtttaact cccctaccca ctcgagtaaa caactcttct ccaacaatac tttttagcct 5700 cgaaatcgca ttactaaccg acgactgagt caaatccagc tcttctgccg cccggctaaa 5760 agatgaggtg cgatacaccg cagtaaaaac gcgaaataaa ttaagatcaa aagctttttg 5820 ctgcgacata aatcagctat ctccttatcc ttatccttat ccttataaaa agttagctcc 5880 agagcactct agctcaaaaa caactcagcg tattaagcca atattttggg aactcaatta 5940 atattcataa taaaagtatt cataatataa ataccaagtc ataatttagc cctaattatt 6000 aatcaattca agttacctat actggcctca attaagcaaa tgtctcatca gtctccctgc 6060 aactaaatgc aatattgaga cataaagctt tgaactgatt caatcttacg agggtaactt 6120 atgaaacaga ctctaatggc tatctcaatc atgtcgcttt tttcattcaa tgcgctagca 6180 gcgcaacatg aacatgacca catcactgtt gattacgaag ggaaagccgc aacagaacac 6240 accatagctc acaaccaagc tgtagctaaa acacttaact ttgccgacac gcgtgcattt 6300 gagcaatcgt ctaaaaatct agtcgccaag tttgataaag caactgccga tatattacgt 6360 gccgaatttg cttttattag cgatgaaatc cctgactcgg ttaacccgtc tctctaccgt 6420 caggctcagc ttaatatggt gcctaatggt ctgtataaag tgagcgatgg catttaccag 6480 gtccgcggta ccgacttatc taaccttaca cttatccgca gtgataacgg ttggatagca 6540 tacgatgttt tgttaaccaa agaagcagca aaagcctcac tacaatttgc gttaaagaat 6600 ctacctaaag atggcgattt acccgttgtt gcgatgattt actcccatag ccatgcggac 6660 cactttggcg gagctcgcgg tgttcaagag atgttccctg atgtcaaagt ctacggctca 6720 gataacatca ctaaagaaat tgtcgatgag aacgtacttg ccggtaacgc catgagccgc 6780 cgcgcagctt atcaatacgg cgcaacactg ggcaaacatg accacggtat tgttgatgct 6840 gcgctaggta aaggtctatc aaaaggtgaa atcacttacg tcgccccaga ctacacctta 6900 aacagtgaag gcaaatggga aacgctgacg attgatggtc tagagatggt gtttatggat 6960 gcctcgggca ccgaagctga gtcagaaatg atcacttata ttccctctaa aaaagcgctc 7020 tggacggcgg agcttaccta tcaaggtatg cacaacattt atacgctgcg cggcgctaaa 7080 gtacgtgatg cgctcaagtg gtcaaaagat atcaacgaaa tgatcaatgc ctttggtcaa 7140 gatgtcgaag tgctgtttgc ctcgcactct gcgccagtgt ggggtaacca agcgatcaac 7200 gatttcttac gcctacagcg tgataactac ggcctagtgc acaatcaaac cttgagactt 7260 gccaacgatg gtgtcggtat acaagatatt ggcgatgcga ttcaagacac gattccagag 7320 tctatctaca agacgtggca taccaatggt taccacggca cttatagcca taacgctaaa 7380 gcggtttata acaagtatct aggctacttc gatatgaacc cagccaacct taatccgctg 7440 ccaaccaagc aagaatctgc caagtttgtc gaatacatgg gcggcgcaga tgccgcaatt 7500 aagcgcgcta aagatgatta cgctcaaggt gaataccgct ttgttgcaac ggcattaaat 7560 aaggtggtga tggccgagcc agaaaatgac tccgctcgtc aattgctagc cgatacctat 7620 gagcaacttg gttatcaagc agaaggggct ggctggagaa acatttactt aactggcgca 7680 caagagctac gagtaggtat tcaagctggc gcgcctaaaa ccgcatcggc agatgtcatc 7740 agtgaaatgg acatgccgac tctatttgac ttcctcgcgg tgaagattga tagtcaacag 7800 gcggctaagc acggcttagt taagatgaat gttatcaccc ctgatactaa agatattctc 7860 tatattgagc taagcaacgg taacttaagc aacgcagtgg tcgacaaaga gcaagcagct 7920 gacgcaaacc ttatggttaa taaagctgac gttaaccgca tcttacttgg ccaagtaacc 7980 ctaaaagcgt tattagccag cggcgatgcc aagctcactg gtgataaaac ggcatttagt 8040 aaaatagccg atagcatggt cgagtttaca cctgacttcg aaatcgtacc aacgcctgtt 8100 aaatgaggca ttaatctcaa caagtgcaag ctagacataa aaatggggcg attagacgcc 8160 ccatttttta tgcaattttg aactagctag tcttagctga agctcgaaca acagctttaa 8220 aattcacttc ttctgctgca atacttattt gctgacactg accaatactc agtgcaaaac 8280 gataactatc atcaagatgg cccagtaaac aatgccaatt atcagcagcg ttcatttgct 8340 gttctttagc ctcaatcaaa cctaaaccag acttttgtgg ctcagcgtta ggcttattag 8400 aactcgactc tagtaaagca agaccaatat cttgttttaa caaaacctgt cgctgattaa 8460 gttgatgctc aaccttgtga tccgcaatag catcggaaat atcaacacaa tggctcaagc 8520 ttttaggtgc attaactcca agaaaagttt cgctcagtgc agagaagtca aacgcaaaag 8580 attttagcga taatgccagc ccaagtcctt tcgctttaat gtaagactcc ttgagcgccc 8640 acaaatcaaa aaagcggtct cgctgcaagg cctctggtaa cgctaacaag gctcgctttt 8700 ctgattcaga gaaataatga ctaagaatag agtggatatt ggtgctgtta cggcaacgct 8760 caatgtcgac gccaaactca atactagcag agtcagtttc ctccttgctt gcctgactgg 8820 cgcctttatt atcagcagtg caaatgccta ctaatagcca atctccacta tgactcacat 8880 taaagtggac cccggtttga gcaaattgcg catcactcaa tctaggctta cctttgtcgc 8940 catattcaaa gcgccattca ttggggcgta tttcactatg ttgtgacaat aaagcgcgca 9000 aatagcctct taccattaaa ccttgagttt tagcttcttg tttaatgtag cgattaacct 9060 taattaactc atcttcaggc agccatgact taaccaactc tgtagtctgg ttatcgcact 9120 cttgtattgt taacggacag aagtataagg aaatcaatcg agaagttagc aatttttcag 9180 gacactcttt aaagcaacaa acataacccc tatttttacc aatttaagat caaaactaaa 9240 gccaaaacta attgagaata gtgtcaaact agctttaaag gaaaaaaata taaaaagaac 9300 attatacttg tataaattat tttacacacc aaagccatga tcttcacaaa attagctccc 9360 tctccctaaa acaagattga ataaaaaaat aaaccttaac tttcatatag ataaaacaaa 9420 ccaatgggat aaagtatatt gaattcattt ttaaggaaaa attcaaattg aattcaagct 9480 cttcagtaaa agcatatttt gccgttagtg tgaaaaaaaa caaatttaaa aaccaacata 9540 gaacaaataa gcagacaata aaaccaaggc gcaacacaaa caacgcgctt acaattttca 9600 caaaaaagca acaagagtaa cgtttagtat ttggatatgg ttattgtaat tgagaatttt 9660 ataacaatta tattaaggga atgagtatgt ttttaaattc aaaactttcg cgctcagtca 9720 aacttgccat atccgcaggc ttaacagcct cgctagctat gcctgttttt gcagaagaaa 9780 ctgctgctga agaacaaata gaaagagtcg cagtgaccgg atcgcgaatc gctaaagcag 9840 agctaactca accagctcca gtcgtcagcc tttcagccga agaactgaca aaatttggta 9900 atcaagattt aggtagcgta ctagcagaat tacctgctat tggtgcaacc aacactatta 9960 ttggtaataa caatagcaac tcaagcgcag gtgttagctc agcagacttg cgtcgtctag 10020 gtgctaacag aaccttagta ttagtcaacg gtaagcgcta cgttgccggc caaccgggct 10080 cagctgaggt agatttgtca actataccaa ctagcatgat ctcgcgagtt gagattgtaa 10140 ccggcggtgc ttcagcaatt tatggttcgg acgctgtatc aggtgttatc aacgttatcc 10200 ttaaagaaga ctttgaaggc tttgagttta acgcacgtac tagcggttct actgaaagtg 10260 taggcactca agagcactct tttgacattt tgggtggtgc aaacgttgca gatggacgtg 10320 gtaatgtaac cttctacgca ggttatgaac gtacaaaaga agtcatggct accgacattc 10380 gccaattcga tgcttgggga acaattaaaa acgaagccga tggtggtgaa gatgatggta 10440 ttccagacag actacgtgta ccacgagttt attctgaaat gattaatgct accggtgtta 10500 tcaatgcatt tggtggtgga attggtcgct caacctttga cagtaacggc aatcctattg 10560 cacaacaaga acgtgatggg actaacagct ttgcatttgg ttcattccct aatggctgtg 10620 acacatgttt caacactgaa gcatacgaaa actatattcc aggggtagaa agaataaacg 10680 ttggctcatc attcaacttt gattttaccg ataacattca attttacact gacttcagat 10740 atgtaaagtc agatattcag caacaatttc agccttcatt ccgttttggt aacattaata 10800 tcaatgttga agataacgcc tttttgaatg acgacttgcg tcagcaaatg ctcgatgcgg 10860 gtcaaaccaa tgctagtttt gccaagtttt ttgatgaatt aggaaatcgc tcagcagaaa 10920 ataaacgcga acttttccgt tacgtaggtg gctttaaagg tggctttgat attagcgaaa 10980 ccatatttga ttacgacctt tactatgttt atggcgagac taataaccgt cgtaaaaccc 11040 ttaatgacct aattcctgat aactttgtcg cagctgtcga ctctgttatt gatcctgata 11100 ctggcttagc agcgtgtcgc tcacaagtag caagcgctca aggcgatgac tatacagatc 11160 ccgcgtctgt aaatggtagc gactgtgttg cttataaccc atttggcatg ggtcaagctt 11220 cagcagaagc ccgcgactgg gtttctgctg atgtgactcg tgaagacaaa ataactcaac 11280 aagtgattgg tggtactctc ggtaccgatt ctgaagaact atttgagctt caaggtggtg 11340 caatcgctat ggttgttggt tttgaatacc gtgaagaaac gtctggttca acaaccgatg 11400 aatttactaa agcaggtttc ttgacaagcg ctgcaacgcc agattcttat ggcgaatacg 11460 acgtgactga gtattttgtt gaggtgaaca tcccagtact aaaagaatta ccttttgcac 11520 atgagttgag ctttgacggt gcataccgta atgctgatta ctcacatgcc ggtaagactg 11580 aagcatggaa agctggtatg ttctactcac cattagagca acttgcatta cgtggtacgg 11640 taggtgaagc agtacgagca ccaaacattg cagaagcctt tagtccacgc tctcctggtt 11700 ttggccgcgt ttcagatcca tgtgatgcag ataacattaa tgacgatccg gatcgcgtgt 11760 caaactgtgc agcattgggg atccctccag gattccaagc taatgataac gtcagtgtag 11820 ataccttatc tggtggtaac ccagatctaa aacctgaaac atcaacatcc tttacaggtg 11880 gtcttgtttg gacaccaacg tttgctgaca atctatcatt cactgtcgat tattatgata 11940 ttcaaattga ggatgctatt ttgtcagtag ccacccagac tgtggctgat aactgtgttg 12000 actcaactgg cggacctgac accgacttct gtagtcaagt tgatcgtaat ccaacgacct 12060 atgatattga acttgttcgc tctggttatc taaatgccgc ggcattgaat accaaaggta 12120 ttgaatttca agctgcatac tcattagatc tagagtcttt caacgcgcct ggtgaactac 12180 gcttcaacct attggggaac caattacttg aactagaacg tcttgaattc caaaatcgtc 12240 ctgatgagat taatgatgaa aaaggcgaag taggtgatcc agagctgcag ttccgcctag 12300 gcatcgatta ccgtctagat gatctaagtg ttagctggaa cacgcgttat attgatagcg 12360 tagtaactta tgatgtctct gaaaatggtg gctctcctga agatttatat ccaggccaca 12420 taggctcaat gacaactcat gacttgagcg ctacatacta catcaatgag aacttcatga 12480 ttaacggtgg tgtacgtaac ctatttgacg cacttccacc tggatacact aacgatgcgc 12540 tatatgatct agttggtcgc cgtgcattcc taggtattaa ggtaatgatg taattaatta 12600 ttacgcctct aactaataaa aatgcaatct cttcgtagag attgcatttt tttatgaaat 12660 ccaatcttaa actggttctc cgagcatctt acgccttaaa aaccccgccc ctcaatgtaa 12720 cgccaaagtt aattgcttac acgcacttac acaaacgaac aatttcatta acacgagaca 12780 cagctcacgc tttttatttt acccttgatt ttactacata aaattgcgtt ttagcgcaca 12840 agtgttctcc caagctggtc gtatctgtaa ttattcagtc ccaggtgatt gtattgaccc 12900 ataagctcag gtagtctgct ctgccattag ctaaacaata ttgacaaaat ggcgataaaa 12960 tgtggcttag cgctaagttc accgtaagtt ttatcggcat taagtcccaa cagattatta 13020 acggaaaccc gctaaactga tggcaaaaat aaatagtgaa cacttggatg aagctactat 13080 tacttcgaat aagtgtacgc aaacagagac tgaggctcgg catagaaatg ccactacaac 13140 acctgagatg cgccgattca tacaagagtc ggatctcagt gttagccaac tgtctaaaat 13200 attaaatatc agtgaagcta ccgtacgtaa gtggcgcaag cgtgactctg tcgaaaactg 13260 tcctaatacc ccgcaccatc tcaataccac gctaacccct ttgcaagaat atgtggttgt 13320 gggcctgcgt tatcaattga aaatgccatt agacagattg ctcaaagcaa cccaagagtt 13380 tatcaatcca aacgtgtcgc gctcaggttt agcaagatgt ttgaagcgtt atggcgtttc 13440 acgggtgagt gatatccaaa gcccacacgt accaatgcgc tactttaatc aaattccagt 13500 cactcaaggc agcgatgtgc aaacctacac cctgcactat gaaacgctgg caaaaacctt 13560 agccttacct agtaccgatg gtgacaatgt ggtgcaagtg gtgtctctca ccattccacc 13620 aaagttaacc gaagaagcac ccagttcaat tttgctcggc attgatcctc atagcgactg 13680 gatctatctc gacatatacc aagatggcaa tacacaagcc acgaatagat atatggctta 13740 tgtgctaaaa cacgggccat tccatttacg aaagttactc gtgcgtaact atcacacctt 13800 tttacagcgc tttcctggag cgacgcaaaa tcgccgcccc tctaaagata tgcctgaaac 13860 aatcaacaag acgcctgaaa cacaggcacc cagtggagac tcataatgag ccagacctct 13920 aaacctacaa actcagcaac tgagcaagca caagactcac aagctgactc tcgtttaaat 13980 aaacgactaa aagatatgcc aattgctatt gttggcatgg cgagtatttt tgcaaactct 14040 cgctatttga ataagttttg ggacttaatc agcgaaaaaa ttgatgcgat tactgaatta 14100 ccatcaactc actggcagcc tgaagaatat tacgacgcag ataaaaccgc agcagacaaa 14160 agctactgta aacgtggtgg ctttttgcca gatgtagact tcaacccaat ggagtttggc 14220 ctgccgccaa acattttgga actgaccgat tcatcgcaac tattatcact catcgttgct 14280 aaagaagtgt tggctgatgc taacttacct gagaattacg accgcgataa aattggtatc 14340 accttaggtg tcggcggtgg tcaaaaaatt agccacagcc taacagcgcg tctgcaatac 14400 ccagtattga agaaagtatt cgccaatagc ggcattagtg acaccgacag cgaaatgctt 14460 atcaagaaat tccaagacca atatgtacac tgggaagaaa actcgttccc aggttcactt 14520 ggtaacgtta ttgcgggccg tatcgccaac cgcttcgatt ttggcggcat gaactgtgtg 14580 gttgatgctg cctgtgctgg atcacttgct gctatgcgta tggcgctaac agagctaact 14640 gaaggtcgct ctgaaatgat gatcaccggt ggtgtgtgta ctgataactc accctctatg 14700 tatatgagct tttcaaaaac gcccgccttt accactaacg aaaccattca gccatttgat 14760 atcgactcaa aaggcatgat gattggtgaa ggtattggca tggtggcgct aaagcgtctt 14820 gaagatgcag agcgcgatgg cgaccgcatt tactctgtaa ttaaaggtgt gggtgcatca 14880 tctgacggta agtttaaatc aatctatgcc cctcgcccat caggccaagc taaagcactt 14940 aaccgtgcct atgatgacgc aggttttgcg ccgcatacct taggtctaat tgaagctcac 15000 ggaacaggta ctgcagcagg tgacgcggca gagtttgccg gcctttgctc agtatttgct 15060 gaaggcaacg ataccaagca acacattgcg ctaggttcag ttaaatcaca aattggtcat 15120 actaaatcaa ctgcaggtac agcaggttta attaaagctg ctcttgcttt gcatcacaag 15180 gtactgccgc cgaccattaa cgttagtcag ccaagcccta aacttgatat cgaaaactca 15240 ccgttttatc taaacactga gactcgtcca tggttaccac gtgttgatgg tacgccgcgc 15300 cgcgcgggta ttagctcatt tggttttggt ggcactaact tccattttgt actagaagag 15360 tacaaccaag aacacagccg tactgatagc gaaaaagcta agtatcgtca acgccaagtg 15420 gcgcaaagct tccttgttag cgcaagcgat aaagcatcgc taattaacga gttaaacgta 15480 ctagcagcat ctgcaagcca agctgagttt atcctcaaag atgcagcagc aaactatggc 15540 gtacgtgagc ttgataaaaa tgcaccacgg atcggtttag ttgcaaacac agctgaagag 15600 ttagcaggcc taattaagca agcacttgcc aaactagcag ctagcgatga taacgcatgg 15660 cagctacctg gtggcactag ctaccgcgcc gctgcagtag aaggtaaagt tgccgcactg 15720 tttgctggcc aaggttcaca atatctcaat atgggccgtg accttacttg ttattaccca 15780 gagatgcgtc agcaatttgt aactgcagat aaagtatttg ccgcaaatga taaaacgccg 15840 ttatcgcaaa ctctgtatcc aaagcctgta tttaataaag atgaattaaa ggctcaagaa 15900 gccattttga ccaataccgc caatgcccaa agcgcaattg gtgcgatttc aatgggtcaa 15960 tacgatttgt ttactgcggc tggctttaat gccgacatgg ttgcaggcca tagctttggt 16020 gagctaagtg cactgtgtgc tgcaggtgtt atttcagctg atgactacta caagctggct 16080 tttgctcgtg gtgaggctat ggcaacaaaa gcaccggcta aagacggcgt tgaagcagat 16140 gcaggagcaa tgtttgcaat cataaccaag agtgctgcag accttgaaac cgttgaagcc 16200 accatcgcta aatttgatgg ggtgaaagtc gctaactata acgcgccaac gcaatcagta 16260 attgcaggcc caacagcaac taccgctgat gcggctaaag cgctaactga gcttggttac 16320 aaagcgatta acctgccagt atcaggtgca ttccacactg aacttgttgg tcacgctcaa 16380 gcgccatttg ctaaagcgat tgacgcagcc aaatttacta aaacaagccg agcactttac 16440 tcaaatgcaa ctggcggact ttatgaaagc actgctgcaa agattaaagc ctcgtttaag 16500 aaacatatgc ttcaatcagt gcgctttact agccagctag aagccatgta caacgacggc 16560 gcccgtgtat ttgttgaatt tggtccaaag aacatcttac aaaaattagt tcaaggcacg 16620 cttgtcaaca ctgaaaatga agtttgcact atctctatca accctaatcc taaagttgat 16680 agtgatctgc agcttaagca agcagcaatg cagctagcgg ttactggtgt ggtactcagt 16740 gaaattgacc cataccaagc cgatattgcc gcaccagcga aaaagtcgcc aatgagcatt 16800 tcgcttaatg ctgctaacca tatcagcaaa gcaactcgcg ctaagatggc caagtcttta 16860 gagacaggta tcgtcacctc gcaaatagaa catgttattg aagaaaaaat cgttgaagtt 16920 gagaaactgg ttgaagtcga aaagatcgtc gaaaaagtgg ttgaagtaga gaaagttgtt 16980 gaggttgaag ctcctgttaa ttcagtgcaa gccaatgcaa ttcaaacccg ttcagttgtc 17040 gctccagtaa tagagaacca agtcgtgtct aaaaacagta agccagcagt ccagagcatt 17100 agtggtgatg cactcagcaa cttttttgct gcacagcagc aaaccgcaca gttgcatcag 17160 cagttcttag ctattccgca gcaatatggt gagacgttca ctacgctgat gaccgagcaa 17220 gctaaactgg caagttctgg tgttgcaatt ccagagagtc tgcaacgctc aatggagcaa 17280 ttccaccaac tacaagcgca aacactacaa agccacaccc agttccttga gatgcaagcg 17340 ggtagcaaca ttgcagcgtt aaacctactc aatagcagcc aagcaactta cgctccagcc 17400 attcacaatg aagcgattca aagccaagtg gttcaaagcc aaactgcagt ccagccagta 17460 atttcaacac aagttaacca tgtgtcagag cagccaactc aagctccagc tccaaaagcg 17520 cagccagcac ctgtgacaac tccagttcaa actgctccgg cacaagttgt tcgtcaagcc 17580 gcaccagttc aagccgctat tgaaccgatt aatacaagtg ttgcgactac aacgccttca 17640 gccttcagcg ccgaaacagc cctgagcgca acaaaagtcc aagccactat gcttgaagtg 17700 gttgctgaga aaaccggtta cccaactgaa atgctagagc ttgaaatgga tatggaagcc 17760 gatttaggca tcgattctat caagcgtgta gaaattcttg gcacagtaca agatgagcta 17820 ccgggtctac ctgagcttag ccctgaagat ctagctgagt gtcgaacgct aggcgaaatc 17880 gttgactata tgggcagtaa actgccggct gaaggctcta tgaattctca gctgtctaca 17940 ggttccgcag ctgcgactcc tgcagcgaat ggtctttctg cggagaaagt tcaagcgact 18000 atgatgtctg tggttgccga aaagactggc tacccaactg aaatgctaga gcttgaaatg 18060 gatatggaag ccgatttagg catagattct atcaagcgcg ttgaaattct tggcacagta 18120 caagatgagc taccgggtct acctgagctt agccctgaag atctagctga gtgtcgtact 18180 ctaggcgaaa tcgttgacta tatgaactct aaactcgctg acggctctaa gctgccggct 18240 gaaggctcta tgaattctca gctgtctaca agtgccgcag ctgcgactcc tgcagcgaat 18300 ggtctctctg cggagaaagt tcaagcgact atgatgtctg tggttgccga aaagactggc 18360 tacccaactg aaatgctaga acttgaaatg gatatggaag ctgaccttgg catcgattca 18420 atcaagcgcg ttgaaattct tggcacagta caagatgagc taccgggttt acctgagcta 18480 aatccagaag atttggcaga gtgtcgtact cttggcgaaa tcgtgactta tatgaactct 18540 aaactcgctg acggctctaa gctgccagct gaaggctcta tgcactatca gctgtctaca 18600 agtaccgctg ctgcgactcc tgtagcgaat ggtctctctg cagaaaaagt tcaagcgacc 18660 atgatgtctg tagttgcaga taaaactggc tacccaactg aaatgcttga acttgaaatg 18720 gatatggaag ccgatttagg tatcgattct atcaagcgcg ttgaaattct tggcacagta 18780 caagatgagc taccgggttt acctgagcta aatccagaag atctagcaga gtgtcgcacc 18840 ctaggcgaaa tcgttgacta tatgggcagt aaactgccgg ctgaaggctc tgctaataca 18900 agtgccgctg cgtctcttaa tgttagtgcc gttgcggcgc ctcaagctgc tgcgactcct 18960 gtatcgaacg gtctctctgc agagaaagtg caaagcacta tgatgtcagt agttgcagaa 19020 aagaccggct acccaactga aatgctagaa cttggcatgg atatggaagc cgatttaggt 19080 atcgactcaa ttaaacgcgt tgagattctt ggcacagtac aagatgagct accgggtcta 19140 ccagagctta atcctgaaga tttagctgag tgccgtacgc tgggcgaaat cgttgactat 19200 atgaactcta agctggctga cggctctaag cttccagctg aaggctctgc taatacaagt 19260 gccactgctg cgactcctgc agtgaatggt ctttctgctg acaaggtaca ggcgactatg 19320 atgtctgtag ttgctgaaaa gaccggctac ccaactgaaa tgctagaact tggcatggat 19380 atggaagcag accttggtat tgattctatt aagcgcgttg aaattcttgg cacagtacaa 19440 gatgagctcc caggtttacc tgagcttaat cctgaagatc tcgctgagtg ccgcacgctt 19500 ggcgaaatcg ttagctatat gaactctcaa ctggctgatg gctctaaact ttctacaagt 19560 gcggctgaag gctctgctga tacaagtgct gcaaatgctg caaagccggc agcaatttcg 19620 gcagaaccaa gtgttgagct tcctcctcat agcgaggtag cgctaaaaaa gcttaatgcg 19680 gcgaacaagc tagaaaattg tttcgccgca gacgcaagtg ttgtgattaa cgatgatggt 19740 cacaacgcag gcgttttagc tgagaaactt attaaacaag gcctaaaagt agccgttgtg 19800 cgtttaccga aaggtcagcc tcaatcgcca ctttcaagcg atgttgctag ctttgagctt 19860 gcctcaagcc aagaatctga gcttgaagcc agtatcactg cagttatcgc gcagattgaa 19920 actcaggttg gcgctattgg tggctttatt cacttgcaac cagaagcgaa tacagaagag 19980 caaacggcag taaacctaga tgcgcaaagt tttactcacg ttagcaatgc gttcttgtgg 20040 gccaaattat tgcaaccaaa gctcgttgct ggagcagatg cgcgtcgctg ttttgtaaca 20100 gtaagccgta tcgacggtgg ctttggttac ctaaatactg acgccctaaa agatgctgag 20160 ctaaaccaag cagcattagc tggtttaact aaaaccttaa gccatgaatg gccacaagtg 20220 ttctgtcgcg cgctagatat tgcaacagat gttgatgcaa cccatcttgc tgatgcaatc 20280 accagtgaac tatttgatag ccaagctcag ctacctgaag tgggcttaag cttaattgat 20340 ggcaaagtta accgcgtaac tctagttgct gctgaagctg cagataaaac agcaaaagca 20400 gagcttaaca gcacagataa aatcttagtg actggtgggg caaaaggggt gacatttgaa 20460 tgtgcactgg cattagcatc tcgcagccag tctcacttta tcttagctgg gcgcagtgaa 20520 ttacaagctt taccaagctg ggctgagggt aagcaaacta gcgagctaaa atcagctgca 20580 atcgcacata ttatttctac tggtcaaaag ccaacgccta agcaagttga agccgctgtg 20640 tggccagtgc aaagcagcat tgaaattaat gccgccctag ccgcctttaa caaagttggc 20700 gcctcagctg aatacgtcag catggatgtt accgatagcg ccgcaatcac agcagcactt 20760 aatggtcgct caaatgagat caccggtctt attcatggcg caggtgtact agccgacaag 20820 catattcaag acaagactct tgctgaactt gctaaagttt atggcactaa agtcaacggc 20880 ctaaaagcgc tgctcgcggc acttgagcca agcaaaatta aattacttgc tatgttctca 20940 tctgcagcag gtttttacgg taatatcggc caaagcgatt acgcgatgtc gaacgatatt 21000 cttaacaagg cagcgctgca gttcaccgct cgcaacccac aagctaaagt catgagcttt 21060 aactggggtc cttgggatgg cggcatggtt aacccagcgc ttaaaaagat gtttaccgag 21120 cgtggtgtgt acgttattcc actaaaagca ggtgcagagc tatttgccac tcagctattg 21180 gctgaaactg gcgtgcagtt gctcattggt acgtcaatgc aaggtggcag cgacactaaa 21240 gcaactgaga ctgcttctgt aaaaaagctt aatgcgggtg aggtgctaag tgcatcgcat 21300 ccgcgtgctg gtgcacaaaa aacaccacta caagctgtca ctgcaacgcg tctgttaacc 21360 ccaagtgcca tggtcttcat tgaagatcac cgcattggcg gtaacagtgt gttgccaacg 21420 gtatgcgcca tcgactggat gcgtgaagcg gcaagcgaca tgcttggcgc tcaagttaag 21480 gtacttgatt acaagctatt aaaaggcatt gtatttgaga ctgatgagcc gcaagagtta 21540 acacttgagc taacgccaga cgattcagac gaagctacgc tacaagcatt aatcagctgt 21600 aatgggcgtc cgcaatacaa ggcgacgctt atcagtgata atgccgatat taagcaactt 21660 aacaagcagt ttgatttaag cgctaaggcg attaccacag caaaagagct ttatagcaac 21720 ggcaccttgt tccacggtcc gcgtctacaa gggatccaat ctgtagtgca gttcgatgat 21780 caaggcttaa ttgctaaagt cgctctgcct aaggttgaac ttagcgattg tggtgagttc 21840 ttgccgcaaa cccacatggg tggcagtcaa ccttttgctg aggacttgct attacaagct 21900 atgctggttt gggctcgcct taaaactggc tcggcaagtt tgccatcaag cattggtgag 21960 tttacctcat accaaccaat ggcctttggt gaaactggta ccatagagct tgaagtgatt 22020 aagcacaaca aacgctcact tgaagcgaat gttgcgctat atcgtgacaa cggcgagtta 22080 agtgccatgt ttaagtcagc taaaatcacc attagcaaaa gcttaaattc agcattttta 22140 cctgctgtct tagcaaacga cagtgaggcg aattagtgga acaaacgcct aaagctagtg 22200 cgatgccgct gcgcatcgca cttatcttac tgccaacacc gcagtttgaa gttaactctg 22260 tcgaccagtc agtattagcc agctatcaaa cactgcagcc tgagctaaat gccctgctta 22320 atagtgcgcc gacacctgaa atgctcagca tcactatctc agatgatagc gatgcaaaca 22380 gctttgagtc gcagctaaat gctgcgacca acgcaattaa caatggctat atcgtcaagc 22440 ttgctacggc aactcacgct ttgttaatgc tgcctgcatt aaaagcggcg caaatgcgga 22500 tccatcctca tgcgcagctt gccgctatgc agcaagctaa atcgacgcca atgagtcaag 22560 tatctggtga gctaaagctt ggcgctaatg cgctaagcct agctcagact aatgcgctgt 22620 ctcatgcttt aagccaagcc aagcgtaact taactgatgt cagcgtgaat gagtgttttg 22680 agaacctcaa aagtgaacag cagttcacag aggtttattc gcttattcag caacttgcta 22740 gccgcaccca tgtgagaaaa gaggttaatc aaggtgtgga acttggccct aaacaagcca 22800 aaagccacta ttggtttagc gaatttcacc aaaaccgtgt tgctgccatc aactttatta 22860 atggccaaca agcaaccagc tatgtgctta ctcaaggttc aggattgtta gctgcgaaat 22920 caatgctaaa ccagcaaaga ttaatgttta tcttgccggg taacagtcag caacaaataa 22980 ccgcatcaat aactcagtta atgcagcaat tagagcgttt gcaggtaact gaggttaatg 23040 agctttctct agaatgccaa ctagagctgc tcagcataat gtatgacaac ttagtcaacg 23100 cagacaaact cactactcgc gatagtaagc ccgcttatca ggctgtgatt caagcaagct 23160 ctgttagcgc tgcaaagcaa gagttaagcg cgcttaacga tgcactcaca gcgctgtttg 23220 ctgagcaaac aaacgccaca tcaacgaata aaggcttaat ccaatacaaa acaccggcgg 23280 gcagttactt aaccctaaca ccgcttggca gcaacaatga caacgcccaa gcgggtcttg 23340 cttttgtcta tccgggtgtg ggaacggttt acgccgatat gcttaatgag ctgcatcagt 23400 acttccctgc gctttacgcc aaacttgagc gtgaaggcga tttaaaggcg atgctacaag 23460 cagaagatat ctatcatctt gaccctaaac atgctgccca aatgagctta ggtgacttag 23520 ccattgctgg cgtggggagc agctacctgt taactcagct gctcaccgat gagtttaata 23580 ttaagcctaa ttttgcatta ggttactcaa tgggtgaagc atcaatgtgg gcaagcttag 23640 gcgtatggca aaacccgcat gcgctgatca gcaaaaccca aaccgacccg ctatttactt 23700 ctgctatttc cggcaaattg accgcggtta gacaagcttg gcagcttgat gataccgcag 23760 cggaaatcca gtggaatagc tttgtggtta gaagtgaagc agcgccgatt gaagccttgc 23820 taaaagatta cccacacgct tacctcgcga ttattcaagg ggatacctgc gtaatcgctg 23880 gctgtgaaat ccaatgtaaa gcgctacttg cagcactggg taaacgcggt attgcagcta 23940 atcgtgtaac ggcgatgcat acgcagcctg cgatgcaaga gcatcaaaat gtgatggatt 24000 tttatctgca accgttaaaa gcagagcttc ctagtgaaat aagctttatc agcgccgctg 24060 atttaactgc caagcaaacg gtgagtgagc aagcacttag cagccaagtc gttgctcagt 24120 ctattgccga caccttctgc caaaccttgg actttaccgc gctagtacat cacgcccaac 24180 atcaaggcgc taagctgttt gttgaaattg gcgcggatag acaaaactgc accttgatag 24240 acaagattgt taaacaagat ggtgccagca gtgtacaaca tcaaccttgt tgcacagtgc 24300 ctatgaacgc aaaaggtagc caagatatta ccagcgtgat taaagcgctt ggccaattaa 24360 ttagccatca ggtgccatta tcggtgcaac catttattga tggactcaag cgcgagctaa 24420 cactttgcca attgaccagc caacagctgg cagcacatgc aaatgttgac agcaagtttg 24480 agtctaacca agaccattta cttcaagggg aagtctaatg tcattaccag acaatgcttc 24540 taaccacctt tctgccaacc agaaaggcgc atctcaggca agtaaaacca gtaagcaaag 24600 caaaatcgcc attgtcggtt tagccactct gtatccagac gctaaaaccc cgcaagaatt 24660 ttggcagaat ttgctggata aacgcgactc tcgcagcacc ttaactaacg aaaaactcgg 24720 cgctaacagc caagattatc aaggtgtgca aggccaatct gaccgttttt attgtaataa 24780 aggcggctac attgagaact tcagctttaa tgctgcaggc tacaaattgc cggagcaaag 24840 cttaaatggc ttggacgaca gcttcctttg ggcgctcgat actagccgta acgcactaat 24900 tgatgctggt attgatatca acggcgctga tttaagccgc gcaggtgtag tcatgggcgc 24960 gctgtcgttc ccaactaccc gctcaaacga tctgtttttg ccaatttatc acagcgccgt 25020 tgaaaaagcc ctgcaagata aactaggcgt aaaggcattt aagctaagcc caactaatgc 25080 tcataccgct cgcgcggcaa atgagagcag cctaaatgca gccaatggtg ccattgccca 25140 taacagctca aaagtggtgg ccgatgcact tggccttggc ggcgcacaac taagcctaga 25200 tgctgcctgt gctagttcgg tttactcatt aaagcttgcc tgcgattacc taagcactgg 25260 caaagccgat atcatgctag caggcgcagt atctggcgcg gatcctttct ttattaatat 25320 gggattctca atcttccacg cctacccaga ccatggtatc tcagtaccgt ttgatgccag 25380 cagtaaaggt ttgtttgctg gcgaaggcgc tggcgtatta gtgcttaaac gtcttgaaga 25440 tgccgagcgc gacaatgaca aaatctatgc ggttgttagc ggcgtaggtc tatcaaacga 25500 cggtaaaggc cagtttgtat taagccctaa tccaaaaggt caggtgaagg cctttgaacg 25560 tgcttatgct gccagtgaca ttgagccaaa agacattgaa gtgattgagt gccacgcaac 25620 aggcacaccg cttggcgata aaattgagct cacttcaatg gaaaccttct ttgaagacaa 25680 gctgcaaggc accgatgcac cgttaattgg ctcagctaag tctaacttag gccacctatt 25740 aactgcagcg ggcatgccgg ggatcatgaa gatgatcttc gccatgaaag aaggttacct 25800 gccgccaagt atcaatatta gtgatgctat cgcttcgccg aaaaaactct tcggtaaacc 25860 aaccctgcct agcatggttc aaggctggcc agataagcca tcgaataatc attttggtgt 25920 aagaacccgt cacgcaggcg tatcggtatt tggctttggt ggctgtaacg cccatctgtt 25980 gcttgagtca tacaacggca aaggaacagt aaaggcagaa gccactcaag taccgcgtca 26040 agctgagccg ctaaaagtgg ttggccttgc ctcgcacttt gggcctctta gcagcattaa 26100 tgcactcaac aatgctgtga cccaagatgg gaatggcttt atcgaactgc cgaaaaagcg 26160 ctggaaaggc cttgaaaagc acagtgaact gttagctgaa tttggcttag catctgcgcc 26220 aaaaggtgct tatgttgata acttcgagct ggacttttta cgctttaaac tgccgccaaa 26280 cgaagatgac cgtttgatct cacagcagct aatgctaatg cgagtaacag acgaagccat 26340 tcgtgatgcc aagcttgagc cggggcaaaa agtagctgta ttagtggcaa tggaaactga 26400 gcttgaactg catcagttcc gcggccgggt taacttgcat actcaattag cgcaaagtct 26460 tgccgccatg ggcgtgagtt tatcaacgga tgaataccaa gcgcttgaag ccatcgccat 26520 ggacagcgtg cttgatgctg ccaagctcaa tcagtacacc agctttattg gtaatattat 26580 ggcgtcacgc gtggcgtcac tatgggactt taatggccca gccttcacta tttcagcagc 26640 agagcaatct gtgagccgct gtatcgatgt ggcgcaaaac ctcatcatgg aggataacct 26700 agatgcggtg gtgattgcag cggtcgatct ctctggtagc tttgagcaag tcattcttaa 26760 aaatgccatt gcacctgtag ccattgagcc aaacctcgaa gcaagcctta atccaacatc 26820 agcaagctgg aatgtcggtg aaggtgctgg cgcggtcgtg cttgttaaaa atgaagctac 26880 atcgggctgc tcatacggcc aaattgatgc acttggcttt gctaaaactg ccgaaacagc 26940 gttggctacc gacaagctac tgagccaaac tgccacagac tttaataagg ttaaagtgat 27000 tgaaactatg gcagcgcctg ctagccaaat tcaattagcg ccaatagtta gctctcaagt 27060 gactcacact gctgcagagc agcgtgttgg tcactgcttt gctgcagcgg gtatggcaag 27120 cctattacac ggcttactta acttaaatac tgtagcccaa accaataaag ccaattgcgc 27180 gcttatcaac aatatcagtg aaaaccaatt atcacagctg ttgattagcc aaacagcgag 27240 cgaacaacaa gcattaaccg cgcgtttaag caatgagctt aaatccgatg ctaaacacca 27300 actggttaag caagtcacct taggtggccg tgatatctac cagcatattg ttgatacacc 27360 gcttgcaagc cttgaaagca ttactcagaa attggcgcaa gcgacagcat cgacagtggt 27420 caaccaagtt aaacctatta aggccgctgg ctcagtcgaa atggctaact cattcgaaac 27480 ggaaagctca gcagagccac aaataacaat tgcagcacaa cagactgcaa acattggcgt 27540 caccgctcag gcaaccaaac gtgaattagg taccccacca atgacaacaa ataccattgc 27600 taatacagca aataatttag acaagactct tgagactgtt gctggcaata ctgttgctag 27660 caaggttggc tctggcgaca tagtcaattt tcaacagaac caacaattgg ctcaacaagc 27720 tcacctcgcc tttcttgaaa gccgcagtgc gggtatgaag gtggctgatg ctttattgaa 27780 gcaacagcta gctcaagtaa caggccaaac tatcgataat caggccctcg atactcaagc 27840 cgtcgatact caaacaagcg agaatgtagc gattgccgca gaatcaccag ttcaagttac 27900 aacacctgtt caagttacaa cacctgttca aatcagtgtt gtggagttaa aaccagatca 27960 cgctaatgtg ccaccataca cgccgccagt gcctgcatta aagccgtgta tctggaacta 28020 tgccgattta gttgagtacg cagaaggcga tatcgccaag gtatttggca gtgattatgc 28080 cattatcgac agctactcgc gccgcgtacg tctaccgacc actgactacc tgttggtatc 28140 gcgcgtgacc aaacttgatg cgaccatcaa tcaatttaag ccatgctcaa tgaccactga 28200 gtacgacatc cctgttgatg cgccgtactt agtagacgga caaatccctt gggcggtagc 28260 agtagaatca ggccaatgtg acttgatgct tattagctat ctcggtatcg actttgagaa 28320 caaaggcgag cgggtttatc gactactcga ttgtaccctc accttcctag gcgacttgcc 28380 acgtggcgga gataccctac gttacgacat taagatcaat aactatgctc gcaacggcga 28440 caccctgctg ttcttcttct cgtatgagtg ttttgttggc gacaagatga tcctcaagat 28500 ggatggcggc tgcgctggct tcttcactga tgaagagctt gccgacggta aaggcgtgat 28560 tcgcacagaa gaagagatta aagctcgcag cctagtgcaa aagcaacgct ttaatccgtt 28620 actagattgt cctaaaaccc aatttagtta tggtgatatt cataagctat taactgctga 28680 tattgagggt tgttttggcc caagccacag tggcgtccac cagccgtcac tttgtttcgc 28740 atctgaaaaa ttcttgatga ttgaacaagt cagcaaggtt gatcgcactg gcggtacttg 28800 gggacttggc ttaattgagg gtcataagca gcttgaagca gaccactggt acttcccatg 28860 tcatttcaag ggcgaccaag tgatggctgg ctcgctaatg gctgaaggtt gtggccagtt 28920 attgcagttc tatatgctgc accttggtat gcatacccaa actaaaaatg gtcgtttcca 28980 acctcttgaa aacgcctcac agcaagtacg ctgtcgcggt caagtgctgc cacaatcagg 29040 cgtgctaact taccgtatgg aagtgactga aatcggtttc agtccacgcc catatgctaa 29100 agctaacatc gatatcttgc ttaatggcaa agcggtagtg gatttccaaa acctaggggt 29160 gatgataaaa gaggaagatg agtgtactcg ttatccactt ttgactgaat caacaacggc 29220 tagcactgca caagtaaacg ctcaaacaag tgcgaaaaag gtatacaagc cagcatcagt 29280 caatgcgcca ttaatggcac aaattcctga tctgactaaa gagccaaaca agggcgttat 29340 tccgatttcc catgttgaag caccaattac gccagactac ccgaaccgtg tacctgatac 29400 agtgccattc acgccgtatc acatgtttga gtttgctaca ggcaatatcg aaaactgttt 29460 cgggccagag ttctcaatct atcgcggcat gatcccacca cgtacaccat gcggtgactt 29520 acaagtgacc acacgtgtga ttgaagttaa cggtaagcgt ggcgacttta aaaagccatc 29580 atcgtgtatc gctgaatatg aagtgcctgc agatgcgtgg tatttcgata aaaacagcca 29640 cggcgcagtg atgccatatt caattttaat ggagatctca ctgcaaccta acggctttat 29700 ctcaggttac atgggcacaa ccctaggctt ccctggcctt gagctgttct tccgtaactt 29760 agacggtagc ggtgagttac tacgtgaagt agatttacgt ggtaaaacca tccgtaacga 29820 ctcacgttta ttatcaacag tgatggccgg cactaacatc atccaaagct ttagcttcga 29880 gctaagcact gacggtgagc ctttctatcg cggcactgcg gtatttggct attttaaagg 29940 tgacgcactt aaagatcagc taggcctaga taacggtaaa gtcactcagc catggcatgt 30000 agctaacggc gttgctgcaa gcactaaggt gaacctgctt gataagagct gccgtcactt 30060 taatgcgcca gctaaccagc cacactatcg tctagccggt ggtcagctga actttatcga 30120 cagtgttgaa attgttgata atggcggcac cgaaggttta ggttacttgt atgccgagcg 30180 caccattgac ccaagtgatt ggttcttcca gttccacttc caccaagatc cggttatgcc 30240 aggctcctta ggtgttgaag caattattga aaccatgcaa gcttacgcta ttagtaaaga 30300 cttgggcgca gatttcaaaa atcctaagtt tggtcagatt ttatcgaaca tcaagtggaa 30360 gtatcgcggt caaatcaatc cgctgaacaa gcagatgtct atggatgtca gcattacttc 30420 aatcaaagat gaagacggta agaaagtcat cacaggtaat gccagcttga gtaaagatgg 30480 tctgcgcata tacgaggtct tcgatatagc tatcagcatc gaagaatctg tataaatcgg 30540 agtgactgtc tggctatttt actcaatttc tgtgtcaaaa gtgctcacct atattcatag 30600 gctgcgcgct tttttctgga aattgagcaa aagtatctgc gtcctaactc gatttataag 30660 aatggtttaa ttgaaaagaa caacagctaa gagccgcaag ctcaatataa ataattaagg 30720 gtcttacaaa taatgaatcc tacagcaact aacgaaatgc tttctccgtg gccatgggct 30780 gtgacagagt caaatatcag ttttgacgtg caagtgatgg aacaacaact taaagatttt 30840 agccgggcat gttacgtggt caatcatgcc gaccacggct ttggtattgc gcaaactgcc 30900 gatatcgtga ctgaacaagc ggcaaacagc acagatttac ctgttagtgc ttttactcct 30960 gcattaggta ccgaaagcct aggcgacaat aatttccgcc gcgttcacgg cgttaaatac 31020 gcttattacg caggcgctat ggcaaacggt atttcatctg aagagctagt gattgcccta 31080 ggtcaagctg gcattttgtg ttcgtttgga gcagccggtc ttattccaag tcgcgttgaa 31140 gcggcaatta accgtattca agcagcgctg ccaaatggcc cttatatgtt taaccttatc 31200 catagtccta gcgagccagc attagagcgt ggcagcgtag agctattttt aaagcataag 31260 gtacgcaccg ttgaagcatc agctttctta ggtctaacac cacaaatcgt ctattaccgt 31320 gcagcaggat tgagccgaga cgcacaaggt aaagttgtgg ttggtaacaa ggttatcgct 31380 aaagtaagtc gcaccgaagt ggctgaaaag tttatgatgc cagcgcccgc aaaaatgcta 31440 caaaaactag ttgatgacgg ttcaattacc gctgagcaaa tggagctggc gcaacttgta 31500 cctatggctg acgacatcac tgcagaggcc gattcaggtg gccatactga taaccgtcca 31560 ttagtaacat tgctgccaac cattttagcg ctgaaagaag aaattcaagc taaataccaa 31620 tacgacactc ctattcgtgt cggttgtggt ggcggtgtgg gtacgcctga tgcagcgctg 31680 gcaacgttta acatgggcgc ggcgtatatt gttaccggct ctatcaacca agcttgtgtt 31740 gaagcgggcg caagtgatca cactcgtaaa ttacttgcca ccactgaaat ggccgatgtg 31800 actatggcac cagctgcaga tatgttcgag atgggcgtaa aactgcaggt ggttaagcgc 31860 ggcacgctat tcccaatgcg cgctaacaag ctatatgaga tctacacccg ttacgattca 31920 atcgaagcga tcccattaga cgagcgtgaa aagcttgaga aacaagtatt ccgctcaagc 31980 ctagatgaaa tatgggcagg tacagtggcg cactttaacg agcgcgaccc taagcaaatc 32040 gaacgcgcag agggtaaccc taagcgtaaa atggcattga ttttccgttg gtacttaggt 32100 ctttctagtc gctggtcaaa ctcaggcgaa gtgggtcgtg aaatggatta tcaaatttgg 32160 gctggccctg ctctcggtgc atttaaccaa tgggcaaaag gcagttactt agataactat 32220 caagaccgaa atgccgtcga tttggcaaag cacttaatgt acggcgcggc ttacttaaat 32280 cgtattaact cgctaacggc tcaaggcgtt aaagtgccag cacagttact tcgctggaag 32340 ccaaaccaaa gaatggccta atacacttac aaagcaccag tctaaaaagc cactaatctt 32400 gattagtggc tttttttatt gtggtcaata tgaggctatt tagcctgtaa gcctgaaaat 32460 atcagcactc tgactttaca agcaaattat aattaaggca gggctctact catttatact 32520 gctagcaaac aagcaagttg cccagtaaaa caacaaggta cctgatttat atcgtcataa 32580 aagttggcta gagattcgtt attgatcttt actgattaga gtcgctctgt ttggaaaaag 32640 gtttctcgtt atcatcaaaa tacactctca aacctttaat caattacaac ttaggctttc 32700 tgcgggcatt tttatcttat ttgccacagc tgtatttgcc tttaggtttt gggtgcaact 32760 accattaatt gaggcctcat tagttaaatt atctgagcaa gagctcacct ctttaaatta 32820 cgcttttcag caaatgagaa agccactaca aaccattaat tacgactatg cggtgtggga 32880 cagaacctac agctatatga aatcaaactc agcgagcgct aaaaggtact atgaaaaaca 32940 tgagtaccca gatgatacgt tcaagagttt aaaagtcgac ggagtattta tattcaaccg 33000 tacaaatcag ccagttttta gtaaaggttt taatcataga aatgatatac cgctggtctt 33060 tgaattaact gactttaaac aacatccaca aaacatcgca ttatctccac aaaccaaaca 33120 ggcacaccca ccggcaagta agccgttaga ctcccctgat gatgtgcctt ctacccatgg 33180 ggttatcgcc acacgatacg gtccagcaat ttatagctct accagcattt taaaatctga 33240 tcgtagcggc tcccaacttg gttatttagt cttcattagg ttaattgatg aatggttcat 33300 cgctgagcta tcgcaataca ctgccgcagg tgttgaaatc gctatggctg atgccgcaga 33360 cgcacaatta gcgagattag gcgcaaacac taagcttaat aaagtaaccg ctacatccga 33420 acggttaata actaatgtcg atggtaagcc tctgttgaag ttagtgcttt accataccaa 33480 taaccaaccg ccgccgatgc tagattacag tataataatt ctattagttg agatgtcatt 33540 tttactgatc ctcgcttatt tcctttactc ctacttctta gtcaggccag ttagaaagct 33600 ggcttcagat attaaaaaaa tggataaaag tcgtgaaatt aaaaagctaa ggtatcacta 33660 ccctattact gagctagtca aagttgcgac tcacttcaac gccctaatgg ggacgattca 33720 ggaacaaact aaacagctta atgaacaagt ttttattgat aaattaacca atattcccaa 33780 tcgtcgcgct tttgagcagc gacttgaaac ctattgccaa ctgctagccc ggcaacaaat 33840 tggctttact ctcatcattg ccgatgtgga tcattttaaa gagtacaacg atactcttgg 33900 gcaccttgct ggggatgaag cattaataaa agtggcacaa acactatcgc aacagtttta 33960 ccgtgcagaa gatatttgtg cccgttttgg tggtgaagaa tttattatgt tatttcgaga 34020 catacctgat gagcccttgc agagaaagct cgatgcgatg ctgcactctt ttgcagagct 34080 caacctacct catccaaact catcaaccgc taattacgtt actgtgagcc ttggggtttg 34140 cacagttgtt gctgttgatg attttgaatt taaaagtgag tcgcatatta ttggcagtca 34200 ggctgcatta atcgcagata aggcgcttta tcatgctaaa gcctgtggtc gtaaccagtt 34260 gtcaaaaact actattactg ttgatgagat tgagcaatta gaagcaaata aaatcggtca 34320 tcaagcctaa actcgttcga gtactttccc ctaagtcaga gctatttgcc acttcaagat 34380 gtggctacaa ggcttactct ttcaaaacct gcatcaatag aacacagcaa aatacaataa 34440 tttaagtcaa tttagcctat taaacagagt taatgacagc tcatggtcgc aacttattag 34500 ctatttctag caatataaaa acttatccat tagtagtaac caataaaaaa actaatatat 34560 aaaactattt aatcattatt ttacagatga ttagctacca cccaccttaa gctggctata 34620 ttcgcactag taaaaataaa cattagatcg ggttcagatc aatttacgag tctcgtataa 34680 aatgtacaat aattcactta atttaatact gcatattttt acaagtagag agcggtgatg 34740 aaacaaaata cgaaaggctt tacattaatt gaattagtca tcgtgattat tattctcggt 34800 atacttgctg ctgtggcact gccgaaattc atcaatgttc aagatgacgc taggatctct 34860 gcgatgagcg gtcagttttc atcatttgaa agtgccgtaa aactatacca tagcggttgg 34920 ttagccaaag gctacaacac tgcggttgaa aagctctcag gctttggcca aggtaatgtt 34980 gcatcaagtg acacaggttt tccgtactca acatcaggca cgagtactga tgtgcataaa 35040 gcttgtggtg aactatggca tggcattacc gatacagact tcacaattgg tgcggttagt 35100 gatggcgatc taatgactgc agatgtcgat attgcttaca cctatcgtgg tgatatgtgt 35160 atctatcgcg atctgtattt tattcagcgc tcattaccta ctaaggtgat gaactacaaa 35220 tttaaaactg gtgaaataga aattattgat gctttctaca accctgacgg ctcaactggt 35280 caattaccat aaatttggcg cttatctaag ttgtacttgc tctgaccgac acaaataatg 35340 tcgtttctca gcatatatca aaatacacag caaaaatttg gggttagcta tatagctaac 35400 cccaaatcat atctaacttt acactgcatc taattccaaa cagtatccag ccaaaagcct 35460 aaactattgt tgactcagcg ctaaaatatg cgatgcaaca aacaagtctt ggatcgcaat 35520 acctgagcta tcaaaaatgg tcacctcatc agcactttga cgtcctgttg cggactcgtt 35580 tatcacctga ccaatctcaa ttatcggcgt atttctgcta tgttgaaact caccaataac 35640 aatagattga gaagcaaagt cgcaaaacaa gcgagcatga ctatataggt cagttggcaa 35700 ctcttgctta cccactttat cagcgcccat tgcagaaata tgcgttcctg cttgtaccca 35760 ctgcgcttca aataaaggcg cttgagctgt ggttgctgtg ataataatat ctgcttgttc 35820 acaagcagct tgtgcatcac aagcttcggc attaatgcct ttttctaata aacgcttaac 35880 caagttttca gttttgctag cactacggcc aactaccaat accttagtta atgaacgaac 35940 cttgctcact gctagcactt catattcagc ctgatgaccg gtaccaaaaa cagttaatac 36000 cgtagcatct tctctcgcga ggtaactcac tgctactgca tcggcagcac cagtgcggta 36060 agcattaacg gtagtggcag caatcaccgn ctgcaacata ccggttaatg gatcgagtaa 36120 aaatacgtta gtgccgtggc atggtaaacc atgtttatgg ttatcaggcc aatagctgcc 36180 tgttttccag ccgacaaggt ttggcgttga agccgacttt aatgagaaca tttcattaag 36240 gttcgcgccc tgtgcattaa ctaccgggaa caaggttgct ttatcatcta cggcagcgac 36300 aaacgcttct ttaacagcga tataagccag ctcatgggag atgagctttg atgtttgcgc 36360 ttcagttaaa tagatcatat taccacccct gcactcgatt ccagatctca tagccaccat 36420 tatcaccatc agtatcaaat acatggtact gagcgtgcat tgaagctgtt gcacaggcgt 36480 ggttcggcaa aatatgtaga cgactaccta ccgggaactg cgctaaatca ataacgccgc 36540 catcaactgc ttcaataatg ccgtgctctt gattaacagt tataacctgt agacctgata 36600 acacgtgacc gctgtcgtca cacactaaac cataaccaca atcttttggc tgctctgcag 36660 tacctctatc acccgaaaga gccatccaac ccgcatcaat gaaaatccag tttttatcag 36720 gattatgacc aataacactg gtcactaccg ttgcggcaat atcagttaac tgacacacgt 36780 ttagccctgc catgactaaa tcgaagaagg tgtacacacc cgctctaacc tcggtgatcc 36840 catcaaggtt ttgatagctt tgcgctgttg gtgttgaacc aatactaacg atgtcacatt 36900 gcatacccgc tgcgcgaatg cgtcagcagc ttgtacagcc gctgcaactt cattttgcgc 36960 cgcatcaatt aattgctgtt tttcaaaaca ttgatatgac tcaccagcgt gagtgagtac 37020 gccgtgaaaa ctcgctgcgc cagacgttag tatctgagca atttcaatca acttatcggc 37080 ttccggtgga ataccaccac gatggccatc acaatcaatt tcaattaatg ctggtatttg 37140 gcagtcataa gaaccacaga aatgatttag ctgatgcgct tgctcaacac tatcaagtaa 37200 aactcttgca ttaatacctt ggtccaacat tttagcaata cgcggcaact taccatcggc 37260 aatacctact gcataaataa tgtctgtgta acctttagat gctaaggcct cggcctcttt 37320 taccgttgat acagtgactg gtgagttttt agtgggtaat aaaaactcgg ctgcttcaag 37380 tgatcttaac gttttaaaat gcggtcttag gtttgcacct aatccttcaa ttttttggcg 37440 tagttgactg aggttattaa taaatactgg cttatttaca tataaaaacg gtgtatcaat 37500 tgcttgatac tgactttgct gagtcgtgga aagtatttga gtagatggca tctttaatat 37560 cctagttcat caatcaatct aacaagtttg atgcctagcc acagtggctt gtattcatga 37620 tgctttggaa aatgcttata ttcaaagtat ttgaaagaca tcaaacttct tgtttaatgc 37680 tcagtatcca ccagcacgca tttattttat attaactatt atcaagatat agattaggtt 37740 caaaccaaat gattagtact gaagatctac gttttatcag cgtaatcgcc agtcatcgca 37800 ccttagctga tgccgctaga acactaaata tcacgccacc atcagtgaca ttaaggttgc 37860 agcatattga aaagaaacta tcgattagcc tgatc 37895 <210> ２ <211> 831 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 atg gta aga ggc tat ttg cgc gct tta ttg tca caa cat agt gaa ata 48 Met Val Arg Gly Tyr Leu Arg Ala Leu Leu Ser Gln His Ser Glu Ile 1 5 10 15 cgc ccc aat gaa tgg cgc ttt gaa tat ggc gac aaa ggt aag cct aga 96 Arg Pro Asn Glu Trp Arg Phe Glu Tyr Gly Asp Lys Gly Lys Pro Arg 20 25 30 ttg agt gat gcg caa ttt gct caa acc ggg gtc cac ttt aat gtg agt 144 Leu Ser Asp Ala Gln Phe Ala Gln Thr Gly Val His Phe Asn Val Ser 35 40 45 cat agt gga gat tgg cta tta gta ggc att tgc act gct gat aat aaa 192 His Ser Gly Asp Trp Leu Leu Val Gly Ile Cys Thr Ala Asp Asn Lys 50 55 60 ggc gcc agt cag gca agc aag gag gaa act gac tct gct agt att gag 240 Gly Ala Ser Gln Ala Ser Lys Glu Glu Thr Asp Ser Ala Ser Ile Glu 65 70 75 80 ttt ggc gtc gac att gag cgt tgc cgt aac agc acc aat atc cac tct 288 Phe Gly Val Asp Ile Glu Arg Cys Arg Asn Ser Thr Asn Ile His Ser 85 90 95 att ctt agt cat tat ttc tct gaa tca gaa aag cga gcc ttg tta gcg 336 Ile Leu Ser His Tyr Phe Ser Glu Ser Glu Lys Arg Ala Leu Leu Ala 100 105 110 tta cca gag gcc ttg cag cga gac cgc ttt ttt gat ttg tgg gcg ctc 384 Leu Pro Glu Ala Leu Gln Arg Asp Arg Phe Phe Asp Leu Trp Ala Leu 115 120 125 aag gag tct tac att aaa gcg aaa gga ctt ggg ctg gca tta tcg cta 432 Lys Glu Ser Tyr Ile Lys Ala Lys Gly Leu Gly Leu Ala Leu Ser Leu 130 135 140 aaa tct ttt gcg ttt gac ttc tct gca ctg agc gaa act ttt ctt gga 480 Lys Ser Phe Ala Phe Asp Phe Ser Ala Leu Ser Glu Thr Phe Leu Gly 145 150 155 160 gtt aat gca cct aaa agc ttg agc cat tgt gtt gat att tcc gat gct 528 Val Asn Ala Pro Lys Ser Leu Ser His Cys Val Asp Ile Ser Asp Ala 165 170 175 att gcg gat cac aag gtt gag cat caa ctt aat cag cga cag gtt ttg 576 Ile Ala Asp His Lys Val Glu His Gln Leu Asn Gln Arg Gln Val Leu 180 185 190 tta aaa caa gat att ggt ctt gct tta cta gag tcg agt tct aat aag 624 Leu Lys Gln Asp Ile Gly Leu Ala Leu Leu Glu Ser Ser Ser Asn Lys 195 200 205 cct aac gct gag cca caa aag tct ggt tta ggt ttg att gag gct aaa 672 Pro Asn Ala Glu Pro Gln Lys Ser Gly Leu Gly Leu Ile Glu Ala Lys 210 215 220 gaa cag caa atg aac gct gct gat aat tgg cat tgt tta ctg ggc cat 720 Glu Gln Gln Met Asn Ala Ala Asp Asn Trp His Cys Leu Leu Gly His 225 230 235 240 ctt gat gat agt tat cgt ttt gca ctg agt att ggt cag tgt cag caa 768 Leu Asp Asp Ser Tyr Arg Phe Ala Leu Ser Ile Gly Gln Cys Gln Gln 245 250 255 ata agt att gca gca gaa gaa gtg aat ttt aaa gct gtt gtt cga gct 816 Ile Ser Ile Ala Ala Glu Glu Val Asn Phe Lys Ala Val Val Arg Ala 260 265 270 tca gct aag act agc 831 Ser Ala Lys Thr Ser 275 <210> 3 <211> 277 <212> PRT <400> 1 Met Val Arg Gly Tyr Leu Arg Ala Leu Leu Ser Gln His Ser Glu Ile 1 5 10 15 Arg Pro Asn Glu Trp Arg Phe Glu Tyr Gly Asp Lys Gly Lys Pro Arg 20 25 30 Leu Ser Asp Ala Gln Phe Ala Gln Thr Gly Val His Phe Asn Val Ser 35 40 45 His Ser Gly Asp Trp Leu Leu Val Gly Ile Cys Thr Ala Asp Asn Lys 50 55 60 Gly Ala Ser Gln Ala Ser Lys Glu Glu Thr Asp Ser Ala Ser Ile Glu 65 70 75 80 Phe Gly Val Asp Ile Glu Arg Cys Arg Asn Ser Thr Asn Ile His Ser 85 90 95 Ile Leu Ser His Tyr Phe Ser Glu Ser Glu Lys Arg Ala Leu Leu Ala 100 105 110 Leu Pro Glu Ala Leu Gln Arg Asp Arg Phe Phe Asp Leu Trp Ala Leu 115 120 125 Lys Glu Ser Tyr Ile Lys Ala Lys Gly Leu Gly Leu Ala Leu Ser Leu 130 135 140 Lys Ser Phe Ala Phe Asp Phe Ser Ala Leu Ser Glu Thr Phe Leu Gly 145 150 155 160 Val Asn Ala Pro Lys Ser Leu Ser His Cys Val Asp Ile Ser Asp Ala 165 170 175 Ile Ala Asp His Lys Val Glu His Gln Leu Asn Gln Arg Gln Val Leu 180 185 190 Leu Lys Gln Asp Ile Gly Leu Ala Leu Leu Glu Ser Ser Ser Asn Lys 195 200 205 Pro Asn Ala Glu Pro Gln Lys Ser Gly Leu Gly Leu Ile Glu Ala Lys 210 215 220 Glu Gln Gln Met Asn Ala Ala Asp Asn Trp His Cys Leu Leu Gly His 225 230 235 240 Leu Asp Asp Ser Tyr Arg Phe Ala Leu Ser Ile Gly Gln Cys Gln Gln 245 250 255 Ile Ser Ile Ala Ala Glu Glu Val Asn Phe Lys Ala Val Val Arg Ala 260 265 270 Ser Ala Lys Thr Ser 275 <210> 4 <211> 864 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 atg gca aaa ata aat agt gaa cac ttg gat gaa gct act att act tcg 48 Met Ala Lys Ile Asn Ser Glu His Leu Asp Glu Ala Thr Ile Thr Ser 1 5 10 15 aat aag tgt acg caa aca gag act gag gct cgg cat aga aat gcc act 96 Asn Lys Cys Thr Gln Thr Glu Thr Glu Ala Arg His Arg Asn Ala Thr 20 25 30 aca aca cct gag atg cgc cga ttc ata caa gag tcg gat ctc agt gtt 144 Thr Thr Pro Glu Met Arg Arg Phe Ile Gln Glu Ser Asp Leu Ser Val 35 40 45 agc caa ctg tct aaa ata tta aat atc agt gaa gct acc gta cgt aag 192 Ser Gln Leu Ser Lys Ile Leu Asn Ile Ser Glu Ala Thr Val Arg Lys 50 55 60 tgg cgc aag cgt gac tct gtc gaa aac tgt cct aat acc ccg cac cat 240 Trp Arg Lys Arg Asp Ser Val Glu Asn Cys Pro Asn Thr Pro His His 65 70 75 80 ctc aat acc acg cta acc cct ttg caa gaa tat gtg gtt gtg ggc ctg 288 Leu Asn Thr Thr Leu Thr Pro Leu Gln Glu Tyr Val Val Val Gly Leu 85 90 95 cgt tat caa ttg aaa atg cca tta gac aga ttg ctc aaa gca acc caa 336 Arg Tyr Gln Leu Lys Met Pro Leu Asp Arg Leu Leu Lys Ala Thr Gln 100 105 110 gag ttt atc aat cca aac gtg tcg cgc tca ggt tta gca aga tgt ttg 384 Glu Phe Ile Asn Pro Asn Val Ser Arg Ser Gly Leu Ala Arg Cys Leu 115 120 125 aag cgt tat ggc gtt tca cgg gtg agt gat atc caa agc cca cac gta 432 Lys Arg Tyr Gly Val Ser Arg Val Ser Asp Ile Gln Ser Pro His Val 130 135 140 cca atg cgc tac ttt aat caa att cca gtc act caa ggc agc gat gtg 480 Pro Met Arg Tyr Phe Asn Gln Ile Pro Val Thr Gln Gly Ser Asp Val 145 150 155 160 caa acc tac acc ctg cac tat gaa acg ctg gca aaa acc tta gcc tta 528 Gln Thr Tyr Thr Leu His Tyr Glu Thr Leu Ala Lys Thr Leu Ala Leu 165 170 175 cct agt acc gat ggt gac aat gtg gtg caa gtg gtg tct ctc acc att 576 Pro Ser Thr Asp Gly Asp Asn Val Val Gln Val Val Ser Leu Thr Ile 180 185 190 cca cca aag tta acc gaa gaa gca ccc agt tca att ttg ctc ggc att 624 Pro Pro Lys Leu Thr Glu Glu Ala Pro Ser Ser Ile Leu Leu Gly Ile 195 200 205 gat cct cat agc gac tgg atc tat ctc gac ata tac caa gat ggc aat 672 Asp Pro His Ser Asp Trp Ile Tyr Leu Asp Ile Tyr Gln Asp Gly Asn 210 215 220 aca caa gcc acg aat aga tat atg gct tat gtg cta aaa cac ggg cca 720 Thr Gln Ala Thr Asn Arg Tyr Met Ala Tyr Val Leu Lys His Gly Pro 225 230 235 240 ttc cat tta cga aag tta ctc gtg cgt aac tat cac acc ttt tta cag 768 Phe His Leu Arg Lys Leu Leu Val Arg Asn Tyr His Thr Phe Leu Gln 245 250 255 cgc ttt cct gga gcg acg caa aat cgc cgc ccc tct aaa gat atg cct 816 Arg Phe Pro Gly Ala Thr Gln Asn Arg Arg Pro Ser Lys Asp Met Pro 260 265 270 gaa aca atc aac aag acg cct gaa aca cag gca ccc agt gga gac tca 864 Glu Thr Ile Asn Lys Thr Pro Glu Thr Gln Ala Pro Ser Gly Asp Ser 275 280 285 <210> 5 <211> 288 <212> PRT <400> 1 Met Ala Lys Ile Asn Ser Glu His Leu Asp Glu Ala Thr Ile Thr Ser 1 5 10 15 Asn Lys Cys Thr Gln Thr Glu Thr Glu Ala Arg His Arg Asn Ala Thr 20 25 30 Thr Thr Pro Glu Met Arg Arg Phe Ile Gln Glu Ser Asp Leu Ser Val 35 40 45 Ser Gln Leu Ser Lys Ile Leu Asn Ile Ser Glu Ala Thr Val Arg Lys 50 55 60 Trp Arg Lys Arg Asp Ser Val Glu Asn Cys Pro Asn Thr Pro His His 65 70 75 80 Leu Asn Thr Thr Leu Thr Pro Leu Gln Glu Tyr Val Val Val Gly Leu 85 90 95 Arg Tyr Gln Leu Lys Met Pro Leu Asp Arg Leu Leu Lys Ala Thr Gln 100 105 110 Glu Phe Ile Asn Pro Asn Val Ser Arg Ser Gly Leu Ala Arg Cys Leu 115 120 125 Lys Arg Tyr Gly Val Ser Arg Val Ser Asp Ile Gln Ser Pro His Val 130 135 140 Pro Met Arg Tyr Phe Asn Gln Ile Pro Val Thr Gln Gly Ser Asp Val 145 150 155 160 Gln Thr Tyr Thr Leu His Tyr Glu Thr Leu Ala Lys Thr Leu Ala Leu 165 170 175 Pro Ser Thr Asp Gly Asp Asn Val Val Gln Val Val Ser Leu Thr Ile 180 185 190 Pro Pro Lys Leu Thr Glu Glu Ala Pro Ser Ser Ile Leu Leu Gly Ile 195 200 205 Asp Pro His Ser Asp Trp Ile Tyr Leu Asp Ile Tyr Gln Asp Gly Asn 210 215 220 Thr Gln Ala Thr Asn Arg Tyr Met Ala Tyr Val Leu Lys His Gly Pro 225 230 235 240 Phe His Leu Arg Lys Leu Leu Val Arg Asn Tyr His Thr Phe Leu Gln 245 250 255 Arg Phe Pro Gly Ala Thr Gln Asn Arg Arg Pro Ser Lys Asp Met Pro 260 265 270 Glu Thr Ile Asn Lys Thr Pro Glu Thr Gln Ala Pro Ser Gly Asp Ser 275 280 285 <210> 6 <211> 8268 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 atg agc cag acc tct aaa cct aca aac tca gca act gag caa gca caa 48 Met Ser Gln Thr Ser Lys Pro Thr Asn Ser Ala Thr Glu Gln Ala Gln 1 5 10 15 gac tca caa gct gac tct cgt tta aat aaa cga cta aaa gat atg cca 96 Asp Ser Gln Ala Asp Ser Arg Leu Asn Lys Arg Leu Lys Asp Met Pro 20 25 30 att gct att gtt ggc atg gcg agt att ttt gca aac tct cgc tat ttg 144 Ile Ala Ile Val Gly Met Ala Ser Ile Phe Ala Asn Ser Arg Tyr Leu 35 40 45 aat aag ttt tgg gac tta atc agc gaa aaa att gat gcg att act gaa 192 Asn Lys Phe Trp Asp Leu Ile Ser Glu Lys Ile Asp Ala Ile Thr Glu 50 55 60 tta cca tca act cac tgg cag cct gaa gaa tat tac gac gca gat aaa 240 Leu Pro Ser Thr His Trp Gln Pro Glu Glu Tyr Tyr Asp Ala Asp Lys 65 70 75 80 acc gca gca gac aaa agc tac tgt aaa cgt ggt ggc ttt ttg cca gat 288 Thr Ala Ala Asp Lys Ser Tyr Cys Lys Arg Gly Gly Phe Leu Pro Asp 85 90 95 gta gac ttc aac cca atg gag ttt ggc ctg ccg cca aac att ttg gaa 336 Val Asp Phe Asn Pro Met Glu Phe Gly Leu Pro Pro Asn Ile Leu Glu 100 105 110 ctg acc gat tca tcg caa cta tta tca ctc atc gtt gct aaa gaa gtg 384 Leu Thr Asp Ser Ser Gln Leu Leu Ser Leu Ile Val Ala Lys Glu Val 115 120 125 ttg gct gat gct aac tta cct gag aat tac gac cgc gat aaa att ggt 432 Leu Ala Asp Ala Asn Leu Pro Glu Asn Tyr Asp Arg Asp Lys Ile Gly 130 135 140 atc acc tta ggt gtc ggc ggt ggt caa aaa att agc cac agc cta aca 480 Ile Thr Leu Gly Val Gly Gly Gly Gln Lys Ile Ser His Ser Leu Thr 145 150 155 160 gcg cgt ctg caa tac cca gta ttg aag aaa gta ttc gcc aat agc ggc 528 Ala Arg Leu Gln Tyr Pro Val Leu Lys Lys Val Phe Ala Asn Ser Gly 165 170 175 att agt gac acc gac agc gaa atg ctt atc aag aaa ttc caa gac caa 576 Ile Ser Asp Thr Asp Ser Glu Met Leu Ile Lys Lys Phe Gln Asp Gln 180 185 190 tat gta cac tgg gaa gaa aac tcg ttc cca ggt tca ctt ggt aac gtt 624 Tyr Val His Trp Glu Glu Asn Ser Phe Pro Gly Ser Leu Gly Asn Val 195 200 205 att gcg ggc cgt atc gcc aac cgc ttc gat ttt ggc ggc atg aac tgt 672 Ile Ala Gly Arg Ile Ala Asn Arg Phe Asp Phe Gly Gly Met Asn Cys 210 215 220 gtg gtt gat gct gcc tgt gct gga tca ctt gct gct atg cgt atg gcg 720 Val Val Asp Ala Ala Cys Ala Gly Ser Leu Ala Ala Met Arg Met Ala 225 230 235 240 cta aca gag cta act gaa ggt cgc tct gaa atg atg atc acc ggt ggt 768 Leu Thr Glu Leu Thr Glu Gly Arg Ser Glu Met Met Ile Thr Gly Gly 245 250 255 gtg tgt act gat aac tca ccc tct atg tat atg agc ttt tca aaa acg 816 Val Cys Thr Asp Asn Ser Pro Ser Met Tyr Met Ser Phe Ser Lys Thr 260 265 270 ccc gcc ttt acc act aac gaa acc att cag cca ttt gat atc gac tca 864 Pro Ala Phe Thr Thr Asn Glu Thr Ile Gln Pro Phe Asp Ile Asp Ser 275 280 285 aaa ggc atg atg att ggt gaa ggt att ggc atg gtg gcg cta aag cgt 912 Lys Gly Met Met Ile Gly Glu Gly Ile Gly Met Val Ala Leu Lys Arg 290 295 300 ctt gaa gat gca gag cgc gat ggc gac cgc att tac tct gta att aaa 960 Leu Glu Asp Ala Glu Arg Asp Gly Asp Arg Ile Tyr Ser Val Ile Lys 305 310 315 320 ggt gtg ggt gca tca tct gac ggt aag ttt aaa tca atc tat gcc cct 1008 Gly Val Gly Ala Ser Ser Asp Gly Lys Phe Lys Ser Ile Tyr Ala Pro 325 330 335 cgc cca tca ggc caa gct aaa gca ctt aac cgt gcc tat gat gac gca 1056 Arg Pro Ser Gly Gln Ala Lys Ala Leu Asn Arg Ala Tyr Asp Asp Ala 340 345 350 ggt ttt gcg ccg cat acc tta ggt cta att gaa gct cac gga aca ggt 1104 Gly Phe Ala Pro His Thr Leu Gly Leu Ile Glu Ala His Gly Thr Gly 355 360 365 act gca gca ggt gac gcg gca gag ttt gcc ggc ctt tgc tca gta ttt 1152 Thr Ala Ala Gly Asp Ala Ala Glu Phe Ala Gly Leu Cys Ser Val Phe 370 375 380 gct gaa ggc aac gat acc aag caa cac att gcg cta ggt tca gtt aaa 1200 Ala Glu Gly Asn Asp Thr Lys Gln His Ile Ala Leu Gly Ser Val Lys 385 390 395 400 tca caa att ggt cat act aaa tca act gca ggt aca gca ggt tta att 1248 Ser Gln Ile Gly His Thr Lys Ser Thr Ala Gly Thr Ala Gly Leu Ile 405 410 415 aaa gct gct ctt gct ttg cat cac aag gta ctg ccg ccg acc att aac 1296 Lys Ala Ala Leu Ala Leu His His Lys Val Leu Pro Pro Thr Ile Asn 420 425 430 gtt agt cag cca agc cct aaa ctt gat atc gaa aac tca ccg ttt tat 1344 Val Ser Gln Pro Ser Pro Lys Leu Asp Ile Glu Asn Ser Pro Phe Tyr 435 440 445 cta aac act gag act cgt cca tgg tta cca cgt gtt gat ggt acg ccg 1392 Leu Asn Thr Glu Thr Arg Pro Trp Leu Pro Arg Val Asp Gly Thr Pro 450 455 460 cgc cgc gcg ggt att agc tca ttt ggt ttt ggt ggc act aac ttc cat 1440 Arg Arg Ala Gly Ile Ser Ser Phe Gly Phe Gly Gly Thr Asn Phe His 465 470 475 480 ttt gta cta gaa gag tac aac caa gaa cac agc cgt act gat agc gaa 1488 Phe Val Leu Glu Glu Tyr Asn Gln Glu His Ser Arg Thr Asp Ser Glu 485 490 495 aaa gct aag tat cgt caa cgc caa gtg gcg caa agc ttc ctt gtt agc 1536 Lys Ala Lys Tyr Arg Gln Arg Gln Val Ala Gln Ser Phe Leu Val Ser 500 505 510 gca agc gat aaa gca tcg cta att aac gag tta aac gta cta gca gca 1584 Ala Ser Asp Lys Ala Ser Leu Ile Asn Glu Leu Asn Val Leu Ala Ala 515 520 525 tct gca agc caa gct gag ttt atc ctc aaa gat gca gca gca aac tat 1632 Ser Ala Ser Gln Ala Glu Phe Ile Leu Lys Asp Ala Ala Ala Asn Tyr 530 535 540 ggc gta cgt gag ctt gat aaa aat gca cca cgg atc ggt tta gtt gca 1680 Gly Val Arg Glu Leu Asp Lys Asn Ala Pro Arg Ile Gly Leu Val Ala 545 550 555 560 aac aca gct gaa gag tta gca ggc cta att aag caa gca ctt gcc aaa 1728 Asn Thr Ala Glu Glu Leu Ala Gly Leu Ile Lys Gln Ala Leu Ala Lys 565 570 575 cta gca gct agc gat gat aac gca tgg cag cta cct ggt ggc act agc 1776 Leu Ala Ala Ser Asp Asp Asn Ala Trp Gln Leu Pro Gly Gly Thr Ser 580 585 590 tac cgc gcc gct gca gta gaa ggt aaa gtt gcc gca ctg ttt gct ggc 1824 Tyr Arg Ala Ala Ala Val Glu Gly Lys Val Ala Ala Leu Phe Ala Gly 595 600 605 caa ggt tca caa tat ctc aat atg ggc cgt gac ctt act tgt tat tac 1872 Gln Gly Ser Gln Tyr Leu Asn Met Gly Arg Asp Leu Thr Cys Tyr Tyr 610 615 620 cca gag atg cgt cag caa ttt gta act gca gat aaa gta ttt gcc gca 1920 Pro Glu Met Arg Gln Gln Phe Val Thr Ala Asp Lys Val Phe Ala Ala 625 630 635 640 aat gat aaa acg ccg tta tcg caa act ctg tat cca aag cct gta ttt 1968 Asn Asp Lys Thr Pro Leu Ser Gln Thr Leu Tyr Pro Lys Pro Val Phe 645 650 655 aat aaa gat gaa tta aag gct caa gaa gcc att ttg acc aat acc gcc 2016 Asn Lys Asp Glu Leu Lys Ala Gln Glu Ala Ile Leu Thr Asn Thr Ala 660 665 670 aat gcc caa agc gca att ggt gcg att tca atg ggt caa tac gat ttg 2064 Asn Ala Gln Ser Ala Ile Gly Ala Ile Ser Met Gly Gln Tyr Asp Leu 675 680 685 ttt act gcg gct ggc ttt aat gcc gac atg gtt gca ggc cat agc ttt 2112 Phe Thr Ala Ala Gly Phe Asn Ala Asp Met Val Ala Gly His Ser Phe 690 695 700 ggt gag cta agt gca ctg tgt gct gca ggt gtt att tca gct gat gac 2160 Gly Glu Leu Ser Ala Leu Cys Ala Ala Gly Val Ile Ser Ala Asp Asp 705 710 715 720 tac tac aag ctg gct ttt gct cgt ggt gag gct atg gca aca aaa gca 2208 Tyr Tyr Lys Leu Ala Phe Ala Arg Gly Glu Ala Met Ala Thr Lys Ala 725 730 735 ccg gct aaa gac ggc gtt gaa gca gat gca gga gca atg ttt gca atc 2256 Pro Ala Lys Asp Gly Val Glu Ala Asp Ala Gly Ala Met Phe Ala Ile 740 745 750 ata acc aag agt gct gca gac ctt gaa acc gtt gaa gcc acc atc gct 2304 Ile Thr Lys Ser Ala Ala Asp Leu Glu Thr Val Glu Ala Thr Ile Ala 755 760 765 aaa ttt gat ggg gtg aaa gtc gct aac tat aac gcg cca acg caa tca 2352 Lys Phe Asp Gly Val Lys Val Ala Asn Tyr Asn Ala Pro Thr Gln Ser 770 775 780 gta att gca ggc cca aca gca act acc gct gat gcg gct aaa gcg cta 2400 Val Ile Ala Gly Pro Thr Ala Thr Thr Ala Asp Ala Ala Lys Ala Leu 785 790 795 800 act gag ctt ggt tac aaa gcg att aac ctg cca gta tca ggt gca ttc 2448 Thr Glu Leu Gly Tyr Lys Ala Ile Asn Leu Pro Val Ser Gly Ala Phe 805 810 815 cac act gaa ctt gtt ggt cac gct caa gcg cca ttt gct aaa gcg att 2496 His Thr Glu Leu Val Gly His Ala Gln Ala Pro Phe Ala Lys Ala Ile 820 825 830 gac gca gcc aaa ttt act aaa aca agc cga gca ctt tac tca aat gca 2544 Asp Ala Ala Lys Phe Thr Lys Thr Ser Arg Ala Leu Tyr Ser Asn Ala 835 840 845 act ggc gga ctt tat gaa agc act gct gca aag att aaa gcc tcg ttt 2592 Thr Gly Gly Leu Tyr Glu Ser Thr Ala Ala Lys Ile Lys Ala Ser Phe 850 855 860 aag aaa cat atg ctt caa tca gtg cgc ttt act agc cag cta gaa gcc 2640 Lys Lys His Met Leu Gln Ser Val Arg Phe Thr Ser Gln Leu Glu Ala 865 870 875 880 atg tac aac gac ggc gcc cgt gta ttt gtt gaa ttt ggt cca aag aac 2688 Met Tyr Asn Asp Gly Ala Arg Val Phe Val Glu Phe Gly Pro Lys Asn 885 890 895 atc tta caa aaa tta gtt caa ggc acg ctt gtc aac act gaa aat gaa 2736 Ile Leu Gln Lys Leu Val Gln Gly Thr Leu Val Asn Thr Glu Asn Glu 900 905 910 gtt tgc act atc tct atc aac cct aat cct aaa gtt gat agt gat ctg 2784 Val Cys Thr Ile Ser Ile Asn Pro Asn Pro Lys Val Asp Ser Asp Leu 915 920 925 cag ctt aag caa gca gca atg cag cta gcg gtt act ggt gtg gta ctc 2832 Gln Leu Lys Gln Ala Ala Met Gln Leu Ala Val Thr Gly Val Val Leu 930 935 940 agt gaa att gac cca tac caa gcc gat att gcc gca cca gcg aaa aag 2880 Ser Glu Ile Asp Pro Tyr Gln Ala Asp Ile Ala Ala Pro Ala Lys Lys 945 950 955 960 tcg cca atg agc att tcg ctt aat gct gct aac cat atc agc aaa gca 2928 Ser Pro Met Ser Ile Ser Leu Asn Ala Ala Asn His Ile Ser Lys Ala 965 970 975 act cgc gct aag atg gcc aag tct tta gag aca ggt atc gtc acc tcg 2976 Thr Arg Ala Lys Met Ala Lys Ser Leu Glu Thr Gly Ile Val Thr Ser 980 985 990 caa ata gaa cat gtt att gaa gaa aaa atc gtt gaa gtt gag aaa ctg 3024 Gln Ile Glu His Val Ile Glu Glu Lys Ile Val Glu Val Glu Lys Leu 995 1000 1005 gtt gaa gtc gaa aag atc gtc gaa aaa gtg gtt gaa gta gag aaa gtt 3072 Val Glu Val Glu Lys Ile Val Glu Lys Val Val Glu Val Glu Lys Val 1010 1015 1020 gtt gag gtt gaa gct cct gtt aat tca gtg caa gcc aat gca att caa 3120 Val Glu Val Glu Ala Pro Val Asn Ser Val Gln Ala Asn Ala Ile Gln 1025 1030 1035 1040 acc cgt tca gtt gtc gct cca gta ata gag aac caa gtc gtg tct aaa 3168 Thr Arg Ser Val Val Ala Pro Val Ile Glu Asn Gln Val Val Ser Lys 1045 1050 1055 aac agt aag cca gca gtc cag agc att agt ggt gat gca ctc agc aac 3216 Asn Ser Lys Pro Ala Val Gln Ser Ile Ser Gly Asp Ala Leu Ser Asn 1060 1065 1070 ttt ttt gct gca cag cag caa acc gca cag ttg cat cag cag ttc tta 3264 Phe Phe Ala Ala Gln Gln Gln Thr Ala Gln Leu His Gln Gln Phe Leu 1075 1080 1085 gct att ccg cag caa tat ggt gag acg ttc act acg ctg atg acc gag 3312 Ala Ile Pro Gln Gln Tyr Gly Glu Thr Phe Thr Thr Leu Met Thr Glu 1090 1095 1100 caa gct aaa ctg gca agt tct ggt gtt gca att cca gag agt ctg caa 3360 Gln Ala Lys Leu Ala Ser Ser Gly Val Ala Ile Pro Glu Ser Leu Gln 1105 1110 1115 1120 cgc tca atg gag caa ttc cac caa cta caa gcg caa aca cta caa agc 3408 Arg Ser Met Glu Gln Phe His Gln Leu Gln Ala Gln Thr Leu Gln Ser 1125 1130 1135 cac acc cag ttc ctt gag atg caa gcg ggt agc aac att gca gcg tta 3456 His Thr Gln Phe Leu Glu Met Gln Ala Gly Ser Asn Ile Ala Ala Leu 1140 1145 1150 aac cta ctc aat agc agc caa gca act tac gct cca gcc att cac aat 3504 Asn Leu Leu Asn Ser Ser Gln Ala Thr Tyr Ala Pro Ala Ile His Asn 1155 1160 1165 gaa gcg att caa agc caa gtg gtt caa agc caa act gca gtc cag cca 3552 Glu Ala Ile Gln Ser Gln Val Val Gln Ser Gln Thr Ala Val Gln Pro 1170 1175 1180 gta att tca aca caa gtt aac cat gtg tca gag cag cca act caa gct 3600 Val Ile Ser Thr Gln Val Asn His Val Ser Glu Gln Pro Thr Gln Ala 1185 1190 1195 1200 cca gct cca aaa gcg cag cca gca cct gtg aca act cca gtt caa act 3648 Pro Ala Pro Lys Ala Gln Pro Ala Pro Val Thr Thr Pro Val Gln Thr 1205 1210 1215 gct ccg gca caa gtt gtt cgt caa gcc gca cca gtt caa gcc gct att 3696 Ala Pro Ala Gln Val Val Arg Gln Ala Ala Pro Val Gln Ala Ala Ile 1220 1225 1230 gaa ccg att aat aca agt gtt gcg act aca acg cct tca gcc ttc agc 3744 Glu Pro Ile Asn Thr Ser Val Ala Thr Thr Thr Pro Ser Ala Phe Ser 1235 1240 1245 gcc gaa aca gcc ctg agc gca aca aaa gtc caa gcc act atg ctt gaa 3792 Ala Glu Thr Ala Leu Ser Ala Thr Lys Val Gln Ala Thr Met Leu Glu 1250 1255 1260 gtg gtt gct gag aaa acc ggt tac cca act gaa atg cta gag ctt gaa 3840 Val Val Ala Glu Lys Thr Gly Tyr Pro Thr Glu Met Leu Glu Leu Glu 1265 1270 1275 1280 atg gat atg gaa gcc gat tta ggc atc gat tct atc aag cgt gta gaa 3888 Met Asp Met Glu Ala Asp Leu Gly Ile Asp Ser Ile Lys Arg Val Glu 1285 1290 1295 att ctt ggc aca gta caa gat gag cta ccg ggt cta cct gag ctt agc 3936 Ile Leu Gly Thr Val Gln Asp Glu Leu Pro Gly Leu Pro Glu Leu Ser 1300 1305 1310 cct gaa gat cta gct gag tgt cga acg cta ggc gaa atc gtt gac tat 3984 Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly Glu Ile Val Asp Tyr 1315 1320 1325 atg ggc agt aaa ctg ccg gct gaa ggc tct atg aat tct cag ctg tct 4032 Met Gly Ser Lys Leu Pro Ala Glu Gly Ser Met Asn Ser Gln Leu Ser 1330 1335 1340 aca ggt tcc gca gct gcg act cct gca gcg aat ggt ctt tct gcg gag 4080 Thr Gly Ser Ala Ala Ala Thr Pro Ala Ala Asn Gly Leu Ser Ala Glu 1345 1350 1355 1360 aaa gtt caa gcg act atg atg tct gtg gtt gcc gaa aag act ggc tac 4128 Lys Val Gln Ala Thr Met Met Ser Val Val Ala Glu Lys Thr Gly Tyr 1365 1370 1375 cca act gaa atg cta gag ctt gaa atg gat atg gaa gcc gat tta ggc 4176 Pro Thr Glu Met Leu Glu Leu Glu Met Asp Met Glu Ala Asp Leu Gly 1380 1385 1390 ata gat tct atc aag cgc gtt gaa att ctt ggc aca gta caa gat gag 4224 Ile Asp Ser Ile Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu 1395 1400 1405 cta ccg ggt cta cct gag ctt agc cct gaa gat cta gct gag tgt cgt 4272 Leu Pro Gly Leu Pro Glu Leu Ser Pro Glu Asp Leu Ala Glu Cys Arg 1410 1415 1420 act cta ggc gaa atc gtt gac tat atg aac tct aaa ctc gct gac ggc 4320 Thr Leu Gly Glu Ile Val Asp Tyr Met Asn Ser Lys Leu Ala Asp Gly 1425 1430 1435 1440 tct aag ctg ccg gct gaa ggc tct atg aat tct cag ctg tct aca agt 4368 Ser Lys Leu Pro Ala Glu Gly Ser Met Asn Ser Gln Leu Ser Thr Ser 1445 1450 1455 gcc gca gct gcg act cct gca gcg aat ggt ctc tct gcg gag aaa gtt 4416 Ala Ala Ala Ala Thr Pro Ala Ala Asn Gly Leu Ser Ala Glu Lys Val 1460 1465 1470 caa gcg act atg atg tct gtg gtt gcc gaa aag act ggc tac cca act 4464 Gln Ala Thr Met Met Ser Val Val Ala Glu Lys Thr Gly Tyr Pro Thr 1475 1480 1485 gaa atg cta gaa ctt gaa atg gat atg gaa gct gac ctt ggc atc gat 4512 Glu Met Leu Glu Leu Glu Met Asp Met Glu Ala Asp Leu Gly Ile Asp 1490 1495 1500 tca atc aag cgc gtt gaa att ctt ggc aca gta caa gat gag cta ccg 4560 Ser Ile Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu Leu Pro 1505 1510 1515 1520 ggt tta cct gag cta aat cca gaa gat ttg gca gag tgt cgt act ctt 4608 Gly Leu Pro Glu Leu Asn Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu 1525 1530 1535 ggc gaa atc gtg act tat atg aac tct aaa ctc gct gac ggc tct aag 4656 Gly Glu Ile Val Thr Tyr Met Asn Ser Lys Leu Ala Asp Gly Ser Lys 1540 1545 1550 ctg cca gct gaa ggc tct atg cac tat cag ctg tct aca agt acc gct 4704 Leu Pro Ala Glu Gly Ser Met His Tyr Gln Leu Ser Thr Ser Thr Ala 1555 1560 1565 gct gcg act cct gta gcg aat ggt ctc tct gca gaa aaa gtt caa gcg 4752 Ala Ala Thr Pro Val Ala Asn Gly Leu Ser Ala Glu Lys Val Gln Ala 1570 1575 1580 acc atg atg tct gta gtt gca gat aaa act ggc tac cca act gaa atg 4800 Thr Met Met Ser Val Val Ala Asp Lys Thr Gly Tyr Pro Thr Glu Met 1585 1590 1595 1600 ctt gaa ctt gaa atg gat atg gaa gcc gat tta ggt atc gat tct atc 4848 Leu Glu Leu Glu Met Asp Met Glu Ala Asp Leu Gly Ile Asp Ser Ile 1605 1610 1615 aag cgc gtt gaa att ctt ggc aca gta caa gat gag cta ccg ggt tta 4896 Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu Leu Pro Gly Leu 1620 1625 1630 cct gag cta aat cca gaa gat cta gca gag tgt cgc acc cta ggc gaa 4944 Pro Glu Leu Asn Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly Glu 1635 1640 1645 atc gtt gac tat atg ggc agt aaa ctg ccg gct gaa ggc tct gct aat 4992 Ile Val Asp Tyr Met Gly Ser Lys Leu Pro Ala Glu Gly Ser Ala Asn 1650 1655 1660 aca agt gcc gct gcg tct ctt aat gtt agt gcc gtt gcg gcg cct caa 5040 Thr Ser Ala Ala Ala Ser Leu Asn Val Ser Ala Val Ala Ala Pro Gln 1665 1670 1675 1680 gct gct gcg act cct gta tcg aac ggt ctc tct gca gag aaa gtg caa 5088 Ala Ala Ala Thr Pro Val Ser Asn Gly Leu Ser Ala Glu Lys Val Gln 1685 1690 1695 agc act atg atg tca gta gtt gca gaa aag acc ggc tac cca act gaa 5136 Ser Thr Met Met Ser Val Val Ala Glu Lys Thr Gly Tyr Pro Thr Glu 1700 1705 1710 atg cta gaa ctt ggc atg gat atg gaa gcc gat tta ggt atc gac tca 5184 Met Leu Glu Leu Gly Met Asp Met Glu Ala Asp Leu Gly Ile Asp Ser 1715 1720 1725 att aaa cgc gtt gag att ctt ggc aca gta caa gat gag cta ccg ggt 5232 Ile Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu Leu Pro Gly 1730 1735 1740 cta cca gag ctt aat cct gaa gat tta gct gag tgc cgt acg ctg ggc 5280 Leu Pro Glu Leu Asn Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly 1745 1750 1755 1760 gaa atc gtt gac tat atg aac tct aag ctg gct gac ggc tct aag ctt 5328 Glu Ile Val Asp Tyr Met Asn Ser Lys Leu Ala Asp Gly Ser Lys Leu 1765 1770 1775 cca gct gaa ggc tct gct aat aca agt gcc act gct gcg act cct gca 5376 Pro Ala Glu Gly Ser Ala Asn Thr Ser Ala Thr Ala Ala Thr Pro Ala 1780 1785 1790 gtg aat ggt ctt tct gct gac aag gta cag gcg act atg atg tct gta 5424 Val Asn Gly Leu Ser Ala Asp Lys Val Gln Ala Thr Met Met Ser Val 1795 1800 1805 gtt gct gaa aag acc ggc tac cca act gaa atg cta gaa ctt ggc atg 5472 Val Ala Glu Lys Thr Gly Tyr Pro Thr Glu Met Leu Glu Leu Gly Met 1810 1815 1820 gat atg gaa gca gac ctt ggt att gat tct att aag cgc gtt gaa att 5520 Asp Met Glu Ala Asp Leu Gly Ile Asp Ser Ile Lys Arg Val Glu Ile 1825 1830 1835 1840 ctt ggc aca gta caa gat gag ctc cca ggt tta cct gag ctt aat cct 5568 Leu Gly Thr Val Gln Asp Glu Leu Pro Gly Leu Pro Glu Leu Asn Pro 1845 1850 1855 gaa gat ctc gct gag tgc cgc acg ctt ggc gaa atc gtt agc tat atg 5616 Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly Glu Ile Val Ser Tyr Met 1860 1865 1870 aac tct caa ctg gct gat ggc tct aaa ctt tct aca agt gcg gct gaa 5664 Asn Ser Gln Leu Ala Asp Gly Ser Lys Leu Ser Thr Ser Ala Ala Glu 1875 1880 1885 ggc tct gct gat aca agt gct gca aat gct gca aag ccg gca gca att 5712 Gly Ser Ala Asp Thr Ser Ala Ala Asn Ala Ala Lys Pro Ala Ala Ile 1890 1895 1900 tcg gca gaa cca agt gtt gag ctt cct cct cat agc gag gta gcg cta 5760 Ser Ala Glu Pro Ser Val Glu Leu Pro Pro His Ser Glu Val Ala Leu 1905 1910 1915 1920 aaa aag ctt aat gcg gcg aac aag cta gaa aat tgt ttc gcc gca gac 5808 Lys Lys Leu Asn Ala Ala Asn Lys Leu Glu Asn Cys Phe Ala Ala Asp 1925 1930 1935 gca agt gtt gtg att aac gat gat ggt cac aac gca ggc gtt tta gct 5856 Ala Ser Val Val Ile Asn Asp Asp Gly His Asn Ala Gly Val Leu Ala 1940 1945 1950 gag aaa ctt att aaa caa ggc cta aaa gta gcc gtt gtg cgt tta ccg 5904 Glu Lys Leu Ile Lys Gln Gly Leu Lys Val Ala Val Val Arg Leu Pro 1955 1960 1965 aaa ggt cag cct caa tcg cca ctt tca agc gat gtt gct agc ttt gag 5952 Lys Gly Gln Pro Gln Ser Pro Leu Ser Ser Asp Val Ala Ser Phe Glu 1970 1975 1980 ctt gcc tca agc caa gaa tct gag ctt gaa gcc agt atc act gca gtt 6000 Leu Ala Ser Ser Gln Glu Ser Glu Leu Glu Ala Ser Ile Thr Ala Val 1985 1990 1995 2000 atc gcg cag att gaa act cag gtt ggc gct att ggt ggc ttt att cac 6048 Ile Ala Gln Ile Glu Thr Gln Val Gly Ala Ile Gly Gly Phe Ile His 2005 2010 2015 ttg caa cca gaa gcg aat aca gaa gag caa acg gca gta aac cta gat 6096 Leu Gln Pro Glu Ala Asn Thr Glu Glu Gln Thr Ala Val Asn Leu Asp 2020 2025 2030 gcg caa agt ttt act cac gtt agc aat gcg ttc ttg tgg gcc aaa tta 6144 Ala Gln Ser Phe Thr His Val Ser Asn Ala Phe Leu Trp Ala Lys Leu 2035 2040 2045 ttg caa cca aag ctc gtt gct gga gca gat gcg cgt cgc tgt ttt gta 6192 Leu Gln Pro Lys Leu Val Ala Gly Ala Asp Ala Arg Arg Cys Phe Val 2050 2055 2060 aca gta agc cgt atc gac ggt ggc ttt ggt tac cta aat act gac gcc 6240 Thr Val Ser Arg Ile Asp Gly Gly Phe Gly Tyr Leu Asn Thr Asp Ala 2065 2070 2075 2080 cta aaa gat gct gag cta aac caa gca gca tta gct ggt tta act aaa 6288 Leu Lys Asp Ala Glu Leu Asn Gln Ala Ala Leu Ala Gly Leu Thr Lys 2085 2090 2095 acc tta agc cat gaa tgg cca caa gtg ttc tgt cgc gcg cta gat att 6336 Thr Leu Ser His Glu Trp Pro Gln Val Phe Cys Arg Ala Leu Asp Ile 2100 2105 2110 gca aca gat gtt gat gca acc cat ctt gct gat gca atc acc agt gaa 6384 Ala Thr Asp Val Asp Ala Thr His Leu Ala Asp Ala Ile Thr Ser Glu 2115 2120 2125 cta ttt gat agc caa gct cag cta cct gaa gtg ggc tta agc tta att 6432 Leu Phe Asp Ser Gln Ala Gln Leu Pro Glu Val Gly Leu Ser Leu Ile 2130 2135 2140 gat ggc aaa gtt aac cgc gta act cta gtt gct gct gaa gct gca gat 6480 Asp Gly Lys Val Asn Arg Val Thr Leu Val Ala Ala Glu Ala Ala Asp 2145 2150 2155 2160 aaa aca gca aaa gca gag ctt aac agc aca gat aaa atc tta gtg act 6528 Lys Thr Ala Lys Ala Glu Leu Asn Ser Thr Asp Lys Ile Leu Val Thr 2165 2170 2175 ggt ggg gca aaa ggg gtg aca ttt gaa tgt gca ctg gca tta gca tct 6576 Gly Gly Ala Lys Gly Val Thr Phe Glu Cys Ala Leu Ala Leu Ala Ser 2180 2185 2190 cgc agc cag tct cac ttt atc tta gct ggg cgc agt gaa tta caa gct 6624 Arg Ser Gln Ser His Phe Ile Leu Ala Gly Arg Ser Glu Leu Gln Ala 2195 2200 2205 tta cca agc tgg gct gag ggt aag caa act agc gag cta aaa tca gct 6672 Leu Pro Ser Trp Ala Glu Gly Lys Gln Thr Ser Glu Leu Lys Ser Ala 2210 2215 2220 gca atc gca cat att att tct act ggt caa aag cca acg cct aag caa 6720 Ala Ile Ala His Ile Ile Ser Thr Gly Gln Lys Pro Thr Pro Lys Gln 2225 2230 2235 2240 gtt gaa gcc gct gtg tgg cca gtg caa agc agc att gaa att aat gcc 6768 Val Glu Ala Ala Val Trp Pro Val Gln Ser Ser Ile Glu Ile Asn Ala 2245 2250 2255 gcc cta gcc gcc ttt aac aaa gtt ggc gcc tca gct gaa tac gtc agc 6816 Ala Leu Ala Ala Phe Asn Lys Val Gly Ala Ser Ala Glu Tyr Val Ser 2260 2265 2270 atg gat gtt acc gat agc gcc gca atc aca gca gca ctt aat ggt cgc 6864 Met Asp Val Thr Asp Ser Ala Ala Ile Thr Ala Ala Leu Asn Gly Arg 2275 2280 2285 tca aat gag atc acc ggt ctt att cat ggc gca ggt gta cta gcc gac 6912 Ser Asn Glu Ile Thr Gly Leu Ile His Gly Ala Gly Val Leu Ala Asp 2290 2295 2300 aag cat att caa gac aag act ctt gct gaa ctt gct aaa gtt tat ggc 6960 Lys His Ile Gln Asp Lys Thr Leu Ala Glu Leu Ala Lys Val Tyr Gly 2305 2310 2315 2320 act aaa gtc aac ggc cta aaa gcg ctg ctc gcg gca ctt gag cca agc 7008 Thr Lys Val Asn Gly Leu Lys Ala Leu Leu Ala Ala Leu Glu Pro Ser 2325 2330 2335 aaa att aaa tta ctt gct atg ttc tca tct gca gca ggt ttt tac ggt 7056 Lys Ile Lys Leu Leu Ala Met Phe Ser Ser Ala Ala Gly Phe Tyr Gly 2340 2345 2350 aat atc ggc caa agc gat tac gcg atg tcg aac gat att ctt aac aag 7104 Asn Ile Gly Gln Ser Asp Tyr Ala Met Ser Asn Asp Ile Leu Asn Lys 2355 2360 2365 gca gcg ctg cag ttc acc gct cgc aac cca caa gct aaa gtc atg agc 7152 Ala Ala Leu Gln Phe Thr Ala Arg Asn Pro Gln Ala Lys Val Met Ser 2370 2375 2380 ttt aac tgg ggt cct tgg gat ggc ggc atg gtt aac cca gcg ctt aaa 7200 Phe Asn Trp Gly Pro Trp Asp Gly Gly Met Val Asn Pro Ala Leu Lys 2385 2390 2395 2400 aag atg ttt acc gag cgt ggt gtg tac gtt att cca cta aaa gca ggt 7248 Lys Met Phe Thr Glu Arg Gly Val Tyr Val Ile Pro Leu Lys Ala Gly 2405 2410 2415 gca gag cta ttt gcc act cag cta ttg gct gaa act ggc gtg cag ttg 7296 Ala Glu Leu Phe Ala Thr Gln Leu Leu Ala Glu Thr Gly Val Gln Leu 2420 2425 2430 ctc att ggt acg tca atg caa ggt ggc agc gac act aaa gca act gag 7344 Leu Ile Gly Thr Ser Met Gln Gly Gly Ser Asp Thr Lys Ala Thr Glu 2435 2440 2445 act gct tct gta aaa aag ctt aat gcg ggt gag gtg cta agt gca tcg 7392 Thr Ala Ser Val Lys Lys Leu Asn Ala Gly Glu Val Leu Ser Ala Ser 2450 2455 2460 cat ccg cgt gct ggt gca caa aaa aca cca cta caa gct gtc act gca 7440 His Pro Arg Ala Gly Ala Gln Lys Thr Pro Leu Gln Ala Val Thr Ala 2465 2470 2475 2480 acg cgt ctg tta acc cca agt gcc atg gtc ttc att gaa gat cac cgc 7488 Thr Arg Leu Leu Thr Pro Ser Ala Met Val Phe Ile Glu Asp His Arg 2485 2490 2495 att ggc ggt aac agt gtg ttg cca acg gta tgc gcc atc gac tgg atg 7536 Ile Gly Gly Asn Ser Val Leu Pro Thr Val Cys Ala Ile Asp Trp Met 2500 2505 2510 cgt gaa gcg gca agc gac atg ctt ggc gct caa gtt aag gta ctt gat 7584 Arg Glu Ala Ala Ser Asp Met Leu Gly Ala Gln Val Lys Val Leu Asp 2515 2520 2525 tac aag cta tta aaa ggc att gta ttt gag act gat gag ccg caa gag 7632 Tyr Lys Leu Leu Lys Gly Ile Val Phe Glu Thr Asp Glu Pro Gln Glu 2530 2535 2540 tta aca ctt gag cta acg cca gac gat tca gac gaa gct acg cta caa 7680 Leu Thr Leu Glu Leu Thr Pro Asp Asp Ser Asp Glu Ala Thr Leu Gln 2545 2550 2555 2560 gca tta atc agc tgt aat ggg cgt ccg caa tac aag gcg acg ctt atc 7728 Ala Leu Ile Ser Cys Asn Gly Arg Pro Gln Tyr Lys Ala Thr Leu Ile 2565 2570 2575 agt gat aat gcc gat att aag caa ctt aac aag cag ttt gat tta agc 7776 Ser Asp Asn Ala Asp Ile Lys Gln Leu Asn Lys Gln Phe Asp Leu Ser 2580 2585 2590 gct aag gcg att acc aca gca aaa gag ctt tat agc aac ggc acc ttg 7824 Ala Lys Ala Ile Thr Thr Ala Lys Glu Leu Tyr Ser Asn Gly Thr Leu 2595 2600 2605 ttc cac ggt ccg cgt cta caa ggg atc caa tct gta gtg cag ttc gat 7872 Phe His Gly Pro Arg Leu Gln Gly Ile Gln Ser Val Val Gln Phe Asp 2610 2615 2620 gat caa ggc tta att gct aaa gtc gct ctg cct aag gtt gaa ctt agc 7920 Asp Gln Gly Leu Ile Ala Lys Val Ala Leu Pro Lys Val Glu Leu Ser 2625 2630 2635 2640 gat tgt ggt gag ttc ttg ccg caa acc cac atg ggt ggc agt caa cct 7968 Asp Cys Gly Glu Phe Leu Pro Gln Thr His Met Gly Gly Ser Gln Pro 2645 2650 2655 ttt gct gag gac ttg cta tta caa gct atg ctg gtt tgg gct cgc ctt 8016 Phe Ala Glu Asp Leu Leu Leu Gln Ala Met Leu Val Trp Ala Arg Leu 2660 2665 2670 aaa act ggc tcg gca agt ttg cca tca agc att ggt gag ttt acc tca 8064 Lys Thr Gly Ser Ala Ser Leu Pro Ser Ser Ile Gly Glu Phe Thr Ser 2675 2680 2685 tac caa cca atg gcc ttt ggt gaa act ggt acc ata gag ctt gaa gtg 8112 Tyr Gln Pro Met Ala Phe Gly Glu Thr Gly Thr Ile Glu Leu Glu Val 2690 2695 2700 att aag cac aac aaa cgc tca ctt gaa gcg aat gtt gcg cta tat cgt 8160 Ile Lys His Asn Lys Arg Ser Leu Glu Ala Asn Val Ala Leu Tyr Arg 2705 2710 2715 2720 gac aac ggc gag tta agt gcc atg ttt aag tca gct aaa atc acc att 8208 Asp Asn Gly Glu Leu Ser Ala Met Phe Lys Ser Ala Lys Ile Thr Ile 2725 2730 2735 agc aaa agc tta aat tca gca ttt tta cct gct gtc tta gca aac gac 8256 Ser Lys Ser Leu Asn Ser Ala Phe Leu Pro Ala Val Leu Ala Asn Asp 2740 2745 2750 agt gag gcg aat 8268 Ser Glu Ala Asn 2755 <210> 7 <211> 2756 <212> PRT <400> 1 Met Ser Gln Thr Ser Lys Pro Thr Asn Ser Ala Thr Glu Gln Ala Gln 1 5 10 15 Asp Ser Gln Ala Asp Ser Arg Leu Asn Lys Arg Leu Lys Asp Met Pro 20 25 30 Ile Ala Ile Val Gly Met Ala Ser Ile Phe Ala Asn Ser Arg Tyr Leu 35 40 45 Asn Lys Phe Trp Asp Leu Ile Ser Glu Lys Ile Asp Ala Ile Thr Glu 50 55 60 Leu Pro Ser Thr His Trp Gln Pro Glu Glu Tyr Tyr Asp Ala Asp Lys 65 70 75 80 Thr Ala Ala Asp Lys Ser Tyr Cys Lys Arg Gly Gly Phe Leu Pro Asp 85 90 95 Val Asp Phe Asn Pro Met Glu Phe Gly Leu Pro Pro Asn Ile Leu Glu 100 105 110 Leu Thr Asp Ser Ser Gln Leu Leu Ser Leu Ile Val Ala Lys Glu Val 115 120 125 Leu Ala Asp Ala Asn Leu Pro Glu Asn Tyr Asp Arg Asp Lys Ile Gly 130 135 140 Ile Thr Leu Gly Val Gly Gly Gly Gln Lys Ile Ser His Ser Leu Thr 145 150 155 160 Ala Arg Leu Gln Tyr Pro Val Leu Lys Lys Val Phe Ala Asn Ser Gly 165 170 175 Ile Ser Asp Thr Asp Ser Glu Met Leu Ile Lys Lys Phe Gln Asp Gln 180 185 190 Tyr Val His Trp Glu Glu Asn Ser Phe Pro Gly Ser Leu Gly Asn Val 195 200 205 Ile Ala Gly Arg Ile Ala Asn Arg Phe Asp Phe Gly Gly Met Asn Cys 210 215 220 Val Val Asp Ala Ala Cys Ala Gly Ser Leu Ala Ala Met Arg Met Ala 225 230 235 240 Leu Thr Glu Leu Thr Glu Gly Arg Ser Glu Met Met Ile Thr Gly Gly 245 250 255 Val Cys Thr Asp Asn Ser Pro Ser Met Tyr Met Ser Phe Ser Lys Thr 260 265 270 Pro Ala Phe Thr Thr Asn Glu Thr Ile Gln Pro Phe Asp Ile Asp Ser 275 280 285 Lys Gly Met Met Ile Gly Glu Gly Ile Gly Met Val Ala Leu Lys Arg 290 295 300 Leu Glu Asp Ala Glu Arg Asp Gly Asp Arg Ile Tyr Ser Val Ile Lys 305 310 315 320 Gly Val Gly Ala Ser Ser Asp Gly Lys Phe Lys Ser Ile Tyr Ala Pro 325 330 335 Arg Pro Ser Gly Gln Ala Lys Ala Leu Asn Arg Ala Tyr Asp Asp Ala 340 345 350 Gly Phe Ala Pro His Thr Leu Gly Leu Ile Glu Ala His Gly Thr Gly 355 360 365 Thr Ala Ala Gly Asp Ala Ala Glu Phe Ala Gly Leu Cys Ser Val Phe 370 375 380 Ala Glu Gly Asn Asp Thr Lys Gln His Ile Ala Leu Gly Ser Val Lys 385 390 395 400 Ser Gln Ile Gly His Thr Lys Ser Thr Ala Gly Thr Ala Gly Leu Ile 405 410 415 Lys Ala Ala Leu Ala Leu His His Lys Val Leu Pro Pro Thr Ile Asn 420 425 430 Val Ser Gln Pro Ser Pro Lys Leu Asp Ile Glu Asn Ser Pro Phe Tyr 435 440 445 Leu Asn Thr Glu Thr Arg Pro Trp Leu Pro Arg Val Asp Gly Thr Pro 450 455 460 Arg Arg Ala Gly Ile Ser Ser Phe Gly Phe Gly Gly Thr Asn Phe His 465 470 475 480 Phe Val Leu Glu Glu Tyr Asn Gln Glu His Ser Arg Thr Asp Ser Glu 485 490 495 Lys Ala Lys Tyr Arg Gln Arg Gln Val Ala Gln Ser Phe Leu Val Ser 500 505 510 Ala Ser Asp Lys Ala Ser Leu Ile Asn Glu Leu Asn Val Leu Ala Ala 515 520 525 Ser Ala Ser Gln Ala Glu Phe Ile Leu Lys Asp Ala Ala Ala Asn Tyr 530 535 540 Gly Val Arg Glu Leu Asp Lys Asn Ala Pro Arg Ile Gly Leu Val Ala 545 550 555 560 Asn Thr Ala Glu Glu Leu Ala Gly Leu Ile Lys Gln Ala Leu Ala Lys 565 570 575 Leu Ala Ala Ser Asp Asp Asn Ala Trp Gln Leu Pro Gly Gly Thr Ser 580 585 590 Tyr Arg Ala Ala Ala Val Glu Gly Lys Val Ala Ala Leu Phe Ala Gly 595 600 605 Gln Gly Ser Gln Tyr Leu Asn Met Gly Arg Asp Leu Thr Cys Tyr Tyr 610 615 620 Pro Glu Met Arg Gln Gln Phe Val Thr Ala Asp Lys Val Phe Ala Ala 625 630 635 640 Asn Asp Lys Thr Pro Leu Ser Gln Thr Leu Tyr Pro Lys Pro Val Phe 645 650 655 Asn Lys Asp Glu Leu Lys Ala Gln Glu Ala Ile Leu Thr Asn Thr Ala 660 665 670 Asn Ala Gln Ser Ala Ile Gly Ala Ile Ser Met Gly Gln Tyr Asp Leu 675 680 685 Phe Thr Ala Ala Gly Phe Asn Ala Asp Met Val Ala Gly His Ser Phe 690 695 700 Gly Glu Leu Ser Ala Leu Cys Ala Ala Gly Val Ile Ser Ala Asp Asp 705 710 715 720 Tyr Tyr Lys Leu Ala Phe Ala Arg Gly Glu Ala Met Ala Thr Lys Ala 725 730 735 Pro Ala Lys Asp Gly Val Glu Ala Asp Ala Gly Ala Met Phe Ala Ile 740 745 750 Ile Thr Lys Ser Ala Ala Asp Leu Glu Thr Val Glu Ala Thr Ile Ala 755 760 765 Lys Phe Asp Gly Val Lys Val Ala Asn Tyr Asn Ala Pro Thr Gln Ser 770 775 780 Val Ile Ala Gly Pro Thr Ala Thr Thr Ala Asp Ala Ala Lys Ala Leu 785 790 795 800 Thr Glu Leu Gly Tyr Lys Ala Ile Asn Leu Pro Val Ser Gly Ala Phe 805 810 815 His Thr Glu Leu Val Gly His Ala Gln Ala Pro Phe Ala Lys Ala Ile 820 825 830 Asp Ala Ala Lys Phe Thr Lys Thr Ser Arg Ala Leu Tyr Ser Asn Ala 835 840 845 Thr Gly Gly Leu Tyr Glu Ser Thr Ala Ala Lys Ile Lys Ala Ser Phe 850 855 860 Lys Lys His Met Leu Gln Ser Val Arg Phe Thr Ser Gln Leu Glu Ala 865 870 875 880 Met Tyr Asn Asp Gly Ala Arg Val Phe Val Glu Phe Gly Pro Lys Asn 885 890 895 Ile Leu Gln Lys Leu Val Gln Gly Thr Leu Val Asn Thr Glu Asn Glu 900 905 910 Val Cys Thr Ile Ser Ile Asn Pro Asn Pro Lys Val Asp Ser Asp Leu 915 920 925 Gln Leu Lys Gln Ala Ala Met Gln Leu Ala Val Thr Gly Val Val Leu 930 935 940 Ser Glu Ile Asp Pro Tyr Gln Ala Asp Ile Ala Ala Pro Ala Lys Lys 945 950 955 960 Ser Pro Met Ser Ile Ser Leu Asn Ala Ala Asn His Ile Ser Lys Ala 965 970 975 Thr Arg Ala Lys Met Ala Lys Ser Leu Glu Thr Gly Ile Val Thr Ser 980 985 990 Gln Ile Glu His Val Ile Glu Glu Lys Ile Val Glu Val Glu Lys Leu 995 1000 1005 Val Glu Val Glu Lys Ile Val Glu Lys Val Val Glu Val Glu Lys Val 1010 1015 1020 Val Glu Val Glu Ala Pro Val Asn Ser Val Gln Ala Asn Ala Ile Gln 1025 1030 1035 1040 Thr Arg Ser Val Val Ala Pro Val Ile Glu Asn Gln Val Val Ser Lys 1045 1050 1055 Asn Ser Lys Pro Ala Val Gln Ser Ile Ser Gly Asp Ala Leu Ser Asn 1060 1065 1070 Phe Phe Ala Ala Gln Gln Gln Thr Ala Gln Leu His Gln Gln Phe Leu 1075 1080 1085 Ala Ile Pro Gln Gln Tyr Gly Glu Thr Phe Thr Thr Leu Met Thr Glu 1090 1095 1100 Gln Ala Lys Leu Ala Ser Ser Gly Val Ala Ile Pro Glu Ser Leu Gln 1105 1110 1115 1120 Arg Ser Met Glu Gln Phe His Gln Leu Gln Ala Gln Thr Leu Gln Ser 1125 1130 1135 His Thr Gln Phe Leu Glu Met Gln Ala Gly Ser Asn Ile Ala Ala Leu 1140 1145 1150 Asn Leu Leu Asn Ser Ser Gln Ala Thr Tyr Ala Pro Ala Ile His Asn 1155 1160 1165 Glu Ala Ile Gln Ser Gln Val Val Gln Ser Gln Thr Ala Val Gln Pro 1170 1175 1180 Val Ile Ser Thr Gln Val Asn His Val Ser Glu Gln Pro Thr Gln Ala 1185 1190 1195 1200 Pro Ala Pro Lys Ala Gln Pro Ala Pro Val Thr Thr Pro Val Gln Thr 1205 1210 1215 Ala Pro Ala Gln Val Val Arg Gln Ala Ala Pro Val Gln Ala Ala Ile 1220 1225 1230 Glu Pro Ile Asn Thr Ser Val Ala Thr Thr Thr Pro Ser Ala Phe Ser 1235 1240 1245 Ala Glu Thr Ala Leu Ser Ala Thr Lys Val Gln Ala Thr Met Leu Glu 1250 1255 1260 Val Val Ala Glu Lys Thr Gly Tyr Pro Thr Glu Met Leu Glu Leu Glu 1265 1270 1275 1280 Met Asp Met Glu Ala Asp Leu Gly Ile Asp Ser Ile Lys Arg Val Glu 1285 1290 1295 Ile Leu Gly Thr Val Gln Asp Glu Leu Pro Gly Leu Pro Glu Leu Ser 1300 1305 1310 Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly Glu Ile Val Asp Tyr 1315 1320 1325 Met Gly Ser Lys Leu Pro Ala Glu Gly Ser Met Asn Ser Gln Leu Ser 1330 1335 1340 Thr Gly Ser Ala Ala Ala Thr Pro Ala Ala Asn Gly Leu Ser Ala Glu 1345 1350 1355 1360 Lys Val Gln Ala Thr Met Met Ser Val Val Ala Glu Lys Thr Gly Tyr 1365 1370 1375 Pro Thr Glu Met Leu Glu Leu Glu Met Asp Met Glu Ala Asp Leu Gly 1380 1385 1390 Ile Asp Ser Ile Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu 1395 1400 1405 Leu Pro Gly Leu Pro Glu Leu Ser Pro Glu Asp Leu Ala Glu Cys Arg 1410 1415 1420 Thr Leu Gly Glu Ile Val Asp Tyr Met Asn Ser Lys Leu Ala Asp Gly 1425 1430 1435 1440 Ser Lys Leu Pro Ala Glu Gly Ser Met Asn Ser Gln Leu Ser Thr Ser 1445 1450 1455 Ala Ala Ala Ala Thr Pro Ala Ala Asn Gly Leu Ser Ala Glu Lys Val 1460 1465 1470 Gln Ala Thr Met Met Ser Val Val Ala Glu Lys Thr Gly Tyr Pro Thr 1475 1480 1485 Glu Met Leu Glu Leu Glu Met Asp Met Glu Ala Asp Leu Gly Ile Asp 1490 1495 1500 Ser Ile Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu Leu Pro 1505 1510 1515 1520 Gly Leu Pro Glu Leu Asn Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu 1525 1530 1535 Gly Glu Ile Val Thr Tyr Met Asn Ser Lys Leu Ala Asp Gly Ser Lys 1540 1545 1550 Leu Pro Ala Glu Gly Ser Met His Tyr Gln Leu Ser Thr Ser Thr Ala 1555 1560 1565 Ala Ala Thr Pro Val Ala Asn Gly Leu Ser Ala Glu Lys Val Gln Ala 1570 1575 1580 Thr Met Met Ser Val Val Ala Asp Lys Thr Gly Tyr Pro Thr Glu Met 1585 1590 1595 1600 Leu Glu Leu Glu Met Asp Met Glu Ala Asp Leu Gly Ile Asp Ser Ile 1605 1610 1615 Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu Leu Pro Gly Leu 1620 1625 1630 Pro Glu Leu Asn Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly Glu 1635 1640 1645 Ile Val Asp Tyr Met Gly Ser Lys Leu Pro Ala Glu Gly Ser Ala Asn 1650 1655 1660 Thr Ser Ala Ala Ala Ser Leu Asn Val Ser Ala Val Ala Ala Pro Gln 1665 1670 1675 1680 Ala Ala Ala Thr Pro Val Ser Asn Gly Leu Ser Ala Glu Lys Val Gln 1685 1690 1695 Ser Thr Met Met Ser Val Val Ala Glu Lys Thr Gly Tyr Pro Thr Glu 1700 1705 1710 Met Leu Glu Leu Gly Met Asp Met Glu Ala Asp Leu Gly Ile Asp Ser 1715 1720 1725 Ile Lys Arg Val Glu Ile Leu Gly Thr Val Gln Asp Glu Leu Pro Gly 1730 1735 1740 Leu Pro Glu Leu Asn Pro Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly 1745 1750 1755 1760 Glu Ile Val Asp Tyr Met Asn Ser Lys Leu Ala Asp Gly Ser Lys Leu 1765 1770 1775 Pro Ala Glu Gly Ser Ala Asn Thr Ser Ala Thr Ala Ala Thr Pro Ala 1780 1785 1790 Val Asn Gly Leu Ser Ala Asp Lys Val Gln Ala Thr Met Met Ser Val 1795 1800 1805 Val Ala Glu Lys Thr Gly Tyr Pro Thr Glu Met Leu Glu Leu Gly Met 1810 1815 1820 Asp Met Glu Ala Asp Leu Gly Ile Asp Ser Ile Lys Arg Val Glu Ile 1825 1830 1835 1840 Leu Gly Thr Val Gln Asp Glu Leu Pro Gly Leu Pro Glu Leu Asn Pro 1845 1850 1855 Glu Asp Leu Ala Glu Cys Arg Thr Leu Gly Glu Ile Val Ser Tyr Met 1860 1865 1870 Asn Ser Gln Leu Ala Asp Gly Ser Lys Leu Ser Thr Ser Ala Ala Glu 1875 1880 1885 Gly Ser Ala Asp Thr Ser Ala Ala Asn Ala Ala Lys Pro Ala Ala Ile 1890 1895 1900 Ser Ala Glu Pro Ser Val Glu Leu Pro Pro His Ser Glu Val Ala Leu 1905 1910 1915 1920 Lys Lys Leu Asn Ala Ala Asn Lys Leu Glu Asn Cys Phe Ala Ala Asp 1925 1930 1935 Ala Ser Val Val Ile Asn Asp Asp Gly His Asn Ala Gly Val Leu Ala 1940 1945 1950 Glu Lys Leu Ile Lys Gln Gly Leu Lys Val Ala Val Val Arg Leu Pro 1955 1960 1965 Lys Gly Gln Pro Gln Ser Pro Leu Ser Ser Asp Val Ala Ser Phe Glu 1970 1975 1980 Leu Ala Ser Ser Gln Glu Ser Glu Leu Glu Ala Ser Ile Thr Ala Val 1985 1990 1995 2000 Ile Ala Gln Ile Glu Thr Gln Val Gly Ala Ile Gly Gly Phe Ile His 2005 2010 2015 Leu Gln Pro Glu Ala Asn Thr Glu Glu Gln Thr Ala Val Asn Leu Asp 2020 2025 2030 Ala Gln Ser Phe Thr His Val Ser Asn Ala Phe Leu Trp Ala Lys Leu 2035 2040 2045 Leu Gln Pro Lys Leu Val Ala Gly Ala Asp Ala Arg Arg Cys Phe Val 2050 2055 2060 Thr Val Ser Arg Ile Asp Gly Gly Phe Gly Tyr Leu Asn Thr Asp Ala 2065 2070 2075 2080 Leu Lys Asp Ala Glu Leu Asn Gln Ala Ala Leu Ala Gly Leu Thr Lys 2085 2090 2095 Thr Leu Ser His Glu Trp Pro Gln Val Phe Cys Arg Ala Leu Asp Ile 2100 2105 2110 Ala Thr Asp Val Asp Ala Thr His Leu Ala Asp Ala Ile Thr Ser Glu 2115 2120 2125 Leu Phe Asp Ser Gln Ala Gln Leu Pro Glu Val Gly Leu Ser Leu Ile 2130 2135 2140 Asp Gly Lys Val Asn Arg Val Thr Leu Val Ala Ala Glu Ala Ala Asp 2145 2150 2155 2160 Lys Thr Ala Lys Ala Glu Leu Asn Ser Thr Asp Lys Ile Leu Val Thr 2165 2170 2175 Gly Gly Ala Lys Gly Val Thr Phe Glu Cys Ala Leu Ala Leu Ala Ser 2180 2185 2190 Arg Ser Gln Ser His Phe Ile Leu Ala Gly Arg Ser Glu Leu Gln Ala 2195 2200 2205 Leu Pro Ser Trp Ala Glu Gly Lys Gln Thr Ser Glu Leu Lys Ser Ala 2210 2215 2220 Ala Ile Ala His Ile Ile Ser Thr Gly Gln Lys Pro Thr Pro Lys Gln 2225 2230 2235 2240 Val Glu Ala Ala Val Trp Pro Val Gln Ser Ser Ile Glu Ile Asn Ala 2245 2250 2255 Ala Leu Ala Ala Phe Asn Lys Val Gly Ala Ser Ala Glu Tyr Val Ser 2260 2265 2270 Met Asp Val Thr Asp Ser Ala Ala Ile Thr Ala Ala Leu Asn Gly Arg 2275 2280 2285 Ser Asn Glu Ile Thr Gly Leu Ile His Gly Ala Gly Val Leu Ala Asp 2290 2295 2300 Lys His Ile Gln Asp Lys Thr Leu Ala Glu Leu Ala Lys Val Tyr Gly 2305 2310 2315 2320 Thr Lys Val Asn Gly Leu Lys Ala Leu Leu Ala Ala Leu Glu Pro Ser 2325 2330 2335 Lys Ile Lys Leu Leu Ala Met Phe Ser Ser Ala Ala Gly Phe Tyr Gly 2340 2345 2350 Asn Ile Gly Gln Ser Asp Tyr Ala Met Ser Asn Asp Ile Leu Asn Lys 2355 2360 2365 Ala Ala Leu Gln Phe Thr Ala Arg Asn Pro Gln Ala Lys Val Met Ser 2370 2375 2380 Phe Asn Trp Gly Pro Trp Asp Gly Gly Met Val Asn Pro Ala Leu Lys 2385 2390 2395 2400 Lys Met Phe Thr Glu Arg Gly Val Tyr Val Ile Pro Leu Lys Ala Gly 2405 2410 2415 Ala Glu Leu Phe Ala Thr Gln Leu Leu Ala Glu Thr Gly Val Gln Leu 2420 2425 2430 Leu Ile Gly Thr Ser Met Gln Gly Gly Ser Asp Thr Lys Ala Thr Glu 2435 2440 2445 Thr Ala Ser Val Lys Lys Leu Asn Ala Gly Glu Val Leu Ser Ala Ser 2450 2455 2460 His Pro Arg Ala Gly Ala Gln Lys Thr Pro Leu Gln Ala Val Thr Ala 2465 2470 2475 2480 Thr Arg Leu Leu Thr Pro Ser Ala Met Val Phe Ile Glu Asp His Arg 2485 2490 2495 Ile Gly Gly Asn Ser Val Leu Pro Thr Val Cys Ala Ile Asp Trp Met 2500 2505 2510 Arg Glu Ala Ala Ser Asp Met Leu Gly Ala Gln Val Lys Val Leu Asp 2515 2520 2525 Tyr Lys Leu Leu Lys Gly Ile Val Phe Glu Thr Asp Glu Pro Gln Glu 2530 2535 2540 Leu Thr Leu Glu Leu Thr Pro Asp Asp Ser Asp Glu Ala Thr Leu Gln 2545 2550 2555 2560 Ala Leu Ile Ser Cys Asn Gly Arg Pro Gln Tyr Lys Ala Thr Leu Ile 2565 2570 2575 Ser Asp Asn Ala Asp Ile Lys Gln Leu Asn Lys Gln Phe Asp Leu Ser 2580 2585 2590 Ala Lys Ala Ile Thr Thr Ala Lys Glu Leu Tyr Ser Asn Gly Thr Leu 2595 2600 2605 Phe His Gly Pro Arg Leu Gln Gly Ile Gln Ser Val Val Gln Phe Asp 2610 2615 2620 Asp Gln Gly Leu Ile Ala Lys Val Ala Leu Pro Lys Val Glu Leu Ser 2625 2630 2635 2640 Asp Cys Gly Glu Phe Leu Pro Gln Thr His Met Gly Gly Ser Gln Pro 2645 2650 2655 Phe Ala Glu Asp Leu Leu Leu Gln Ala Met Leu Val Trp Ala Arg Leu 2660 2665 2670 Lys Thr Gly Ser Ala Ser Leu Pro Ser Ser Ile Gly Glu Phe Thr Ser 2675 2680 2685 Tyr Gln Pro Met Ala Phe Gly Glu Thr Gly Thr Ile Glu Leu Glu Val 2690 2695 2700 Ile Lys His Asn Lys Arg Ser Leu Glu Ala Asn Val Ala Leu Tyr Arg 2705 2710 2715 2720 Asp Asn Gly Glu Leu Ser Ala Met Phe Lys Ser Ala Lys Ile Thr Ile 2725 2730 2735 Ser Lys Ser Leu Asn Ser Ala Phe Leu Pro Ala Val Leu Ala Asn Asp 2740 2745 2750 Ser Glu Ala Asn 2755 <210> 8 <211> 2340 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 gtg gaa caa acg cct aaa gct agt gcg atg ccg ctg cgc atc gca ctt 48 Val Glu Gln Thr Pro Lys Ala Ser Ala Met Pro Leu Arg Ile Ala Leu 1 5 10 15 atc tta ctg cca aca ccg cag ttt gaa gtt aac tct gtc gac cag tca 96 Ile Leu Leu Pro Thr Pro Gln Phe Glu Val Asn Ser Val Asp Gln Ser 20 25 30 gta tta gcc agc tat caa aca ctg cag cct gag cta aat gcc ctg ctt 144 Val Leu Ala Ser Tyr Gln Thr Leu Gln Pro Glu Leu Asn Ala Leu Leu 35 40 45 aat agt gcg ccg aca cct gaa atg ctc agc atc act atc tca gat gat 192 Asn Ser Ala Pro Thr Pro Glu Met Leu Ser Ile Thr Ile Ser Asp Asp 50 55 60 agc gat gca aac agc ttt gag tcg cag cta aat gct gcg acc aac gca 240 Ser Asp Ala Asn Ser Phe Glu Ser Gln Leu Asn Ala Ala Thr Asn Ala 65 70 75 80 att aac aat ggc tat atc gtc aag ctt gct acg gca act cac gct ttg 288 Ile Asn Asn Gly Tyr Ile Val Lys Leu Ala Thr Ala Thr His Ala Leu 85 90 95 tta atg ctg cct gca tta aaa gcg gcg caa atg cgg atc cat cct cat 336 Leu Met Leu Pro Ala Leu Lys Ala Ala Gln Met Arg Ile His Pro His 100 105 110 gcg cag ctt gcc gct atg cag caa gct aaa tcg acg cca atg agt caa 384 Ala Gln Leu Ala Ala Met Gln Gln Ala Lys Ser Thr Pro Met Ser Gln 115 120 125 gta tct ggt gag cta aag ctt ggc gct aat gcg cta agc cta gct cag 432 Val Ser Gly Glu Leu Lys Leu Gly Ala Asn Ala Leu Ser Leu Ala Gln 130 135 140 act aat gcg ctg tct cat gct tta agc caa gcc aag cgt aac tta act 480 Thr Asn Ala Leu Ser His Ala Leu Ser Gln Ala Lys Arg Asn Leu Thr 145 150 155 160 gat gtc agc gtg aat gag tgt ttt gag aac ctc aaa agt gaa cag cag 528 Asp Val Ser Val Asn Glu Cys Phe Glu Asn Leu Lys Ser Glu Gln Gln 165 170 175 ttc aca gag gtt tat tcg ctt att cag caa ctt gct agc cgc acc cat 576 Phe Thr Glu Val Tyr Ser Leu Ile Gln Gln Leu Ala Ser Arg Thr His 180 185 190 gtg aga aaa gag gtt aat caa ggt gtg gaa ctt ggc cct aaa caa gcc 624 Val Arg Lys Glu Val Asn Gln Gly Val Glu Leu Gly Pro Lys Gln Ala 195 200 205 aaa agc cac tat tgg ttt agc gaa ttt cac caa aac cgt gtt gct gcc 672 Lys Ser His Tyr Trp Phe Ser Glu Phe His Gln Asn Arg Val Ala Ala 210 215 220 atc aac ttt att aat ggc caa caa gca acc agc tat gtg ctt act caa 720 Ile Asn Phe Ile Asn Gly Gln Gln Ala Thr Ser Tyr Val Leu Thr Gln 225 230 235 240 ggt tca gga ttg tta gct gcg aaa tca atg cta aac cag caa aga tta 768 Gly Ser Gly Leu Leu Ala Ala Lys Ser Met Leu Asn Gln Gln Arg Leu 245 250 255 atg ttt atc ttg ccg ggt aac agt cag caa caa ata acc gca tca ata 816 Met Phe Ile Leu Pro Gly Asn Ser Gln Gln Gln Ile Thr Ala Ser Ile 260 265 270 act cag tta atg cag caa tta gag cgt ttg cag gta act gag gtt aat 864 Thr Gln Leu Met Gln Gln Leu Glu Arg Leu Gln Val Thr Glu Val Asn 275 280 285 gag ctt tct cta gaa tgc caa cta gag ctg ctc agc ata atg tat gac 912 Glu Leu Ser Leu Glu Cys Gln Leu Glu Leu Leu Ser Ile Met Tyr Asp 290 295 300 aac tta gtc aac gca gac aaa ctc act act cgc gat agt aag ccc gct 960 Asn Leu Val Asn Ala Asp Lys Leu Thr Thr Arg Asp Ser Lys Pro Ala 305 310 315 320 tat cag gct gtg att caa gca agc tct gtt agc gct gca aag caa gag 1008 Tyr Gln Ala Val Ile Gln Ala Ser Ser Val Ser Ala Ala Lys Gln Glu 325 330 335 tta agc gcg ctt aac gat gca ctc aca gcg ctg ttt gct gag caa aca 1056 Leu Ser Ala Leu Asn Asp Ala Leu Thr Ala Leu Phe Ala Glu Gln Thr 340 345 350 aac gcc aca tca acg aat aaa ggc tta atc caa tac aaa aca ccg gcg 1104 Asn Ala Thr Ser Thr Asn Lys Gly Leu Ile Gln Tyr Lys Thr Pro Ala 355 360 365 ggc agt tac tta acc cta aca ccg ctt ggc agc aac aat gac aac gcc 1152 Gly Ser Tyr Leu Thr Leu Thr Pro Leu Gly Ser Asn Asn Asp Asn Ala 370 375 380 caa gcg ggt ctt gct ttt gtc tat ccg ggt gtg gga acg gtt tac gcc 1200 Gln Ala Gly Leu Ala Phe Val Tyr Pro Gly Val Gly Thr Val Tyr Ala 385 390 395 400 gat atg ctt aat gag ctg cat cag tac ttc cct gcg ctt tac gcc aaa 1248 Asp Met Leu Asn Glu Leu His Gln Tyr Phe Pro Ala Leu Tyr Ala Lys 405 410 415 ctt gag cgt gaa ggc gat tta aag gcg atg cta caa gca gaa gat atc 1296 Leu Glu Arg Glu Gly Asp Leu Lys Ala Met Leu Gln Ala Glu Asp Ile 420 425 430 tat cat ctt gac cct aaa cat gct gcc caa atg agc tta ggt gac tta 1344 Tyr His Leu Asp Pro Lys His Ala Ala Gln Met Ser Leu Gly Asp Leu 435 440 445 gcc att gct ggc gtg ggg agc agc tac ctg tta act cag ctg ctc acc 1392 Ala Ile Ala Gly Val Gly Ser Ser Tyr Leu Leu Thr Gln Leu Leu Thr 450 455 460 gat gag ttt aat att aag cct aat ttt gca tta ggt tac tca atg ggt 1440 Asp Glu Phe Asn Ile Lys Pro Asn Phe Ala Leu Gly Tyr Ser Met Gly 465 470 475 480 gaa gca tca atg tgg gca agc tta ggc gta tgg caa aac ccg cat gcg 1488 Glu Ala Ser Met Trp Ala Ser Leu Gly Val Trp Gln Asn Pro His Ala 485 490 495 ctg atc agc aaa acc caa acc gac ccg cta ttt act tct gct att tcc 1536 Leu Ile Ser Lys Thr Gln Thr Asp Pro Leu Phe Thr Ser Ala Ile Ser 500 505 510 ggc aaa ttg acc gcg gtt aga caa gct tgg cag ctt gat gat acc gca 1584 Gly Lys Leu Thr Ala Val Arg Gln Ala Trp Gln Leu Asp Asp Thr Ala 515 520 525 gcg gaa atc cag tgg aat agc ttt gtg gtt aga agt gaa gca gcg ccg 1632 Ala Glu Ile Gln Trp Asn Ser Phe Val Val Arg Ser Glu Ala Ala Pro 530 535 540 att gaa gcc ttg cta aaa gat tac cca cac gct tac ctc gcg att att 1680 Ile Glu Ala Leu Leu Lys Asp Tyr Pro His Ala Tyr Leu Ala Ile Ile 545 550 555 560 caa ggg gat acc tgc gta atc gct ggc tgt gaa atc caa tgt aaa gcg 1728 Gln Gly Asp Thr Cys Val Ile Ala Gly Cys Glu Ile Gln Cys Lys Ala 565 570 575 cta ctt gca gca ctg ggt aaa cgc ggt att gca gct aat cgt gta acg 1776 Leu Leu Ala Ala Leu Gly Lys Arg Gly Ile Ala Ala Asn Arg Val Thr 580 585 590 gcg atg cat acg cag cct gcg atg caa gag cat caa aat gtg atg gat 1824 Ala Met His Thr Gln Pro Ala Met Gln Glu His Gln Asn Val Met Asp 595 600 605 ttt tat ctg caa ccg tta aaa gca gag ctt cct agt gaa ata agc ttt 1872 Phe Tyr Leu Gln Pro Leu Lys Ala Glu Leu Pro Ser Glu Ile Ser Phe 610 615 620 atc agc gcc gct gat tta act gcc aag caa acg gtg agt gag caa gca 1920 Ile Ser Ala Ala Asp Leu Thr Ala Lys Gln Thr Val Ser Glu Gln Ala 625 630 635 640 ctt agc agc caa gtc gtt gct cag tct att gcc gac acc ttc tgc caa 1968 Leu Ser Ser Gln Val Val Ala Gln Ser Ile Ala Asp Thr Phe Cys Gln 645 650 655 acc ttg gac ttt acc gcg cta gta cat cac gcc caa cat caa ggc gct 2016 Thr Leu Asp Phe Thr Ala Leu Val His His Ala Gln His Gln Gly Ala 660 665 670 aag ctg ttt gtt gaa att ggc gcg gat aga caa aac tgc acc ttg ata 2064 Lys Leu Phe Val Glu Ile Gly Ala Asp Arg Gln Asn Cys Thr Leu Ile 675 680 685 gac aag att gtt aaa caa gat ggt gcc agc agt gta caa cat caa cct 2112 Asp Lys Ile Val Lys Gln Asp Gly Ala Ser Ser Val Gln His Gln Pro 690 695 700 tgt tgc aca gtg cct atg aac gca aaa ggt agc caa gat att acc agc 2160 Cys Cys Thr Val Pro Met Asn Ala Lys Gly Ser Gln Asp Ile Thr Ser 705 710 715 720 gtg att aaa gcg ctt ggc caa tta att agc cat cag gtg cca tta tcg 2208 al Ile Lys Ala Leu Gly Gln Leu Ile Ser His Gln Val Pro Leu Ser 725 730 735 gtg caa cca ttt att gat gga ctc aag cgc gag cta aca ctt tgc caa 2256 Val Gln Pro Phe Ile Asp Gly Leu Lys Arg Glu Leu Thr Leu Cys Gln 740 745 750 ttg acc agc caa cag ctg gca gca cat gca aat gtt gac agc aag ttt 2304 Leu Thr Ser Gln Gln Leu Ala Ala His Ala Asn Val Asp Ser Lys Phe 755 760 765 gag tct aac caa gac cat tta ctt caa ggg gaa gtc 2340 Glu Ser Asn Gln Asp His Leu Leu Gln Gly Glu Val 770 775 780 <210> 9 <211> 780 <212> PRT <400> 1 Val Glu Gln Thr Pro Lys Ala Ser Ala Met Pro Leu Arg Ile Ala Leu 1 5 10 15 Ile Leu Leu Pro Thr Pro Gln Phe Glu Val Asn Ser Val Asp Gln Ser 20 25 30 Val Leu Ala Ser Tyr Gln Thr Leu Gln Pro Glu Leu Asn Ala Leu Leu 35 40 45 Asn Ser Ala Pro Thr Pro Glu Met Leu Ser Ile Thr Ile Ser Asp Asp 50 55 60 Ser Asp Ala Asn Ser Phe Glu Ser Gln Leu Asn Ala Ala Thr Asn Ala 65 70 75 80 Ile Asn Asn Gly Tyr Ile Val Lys Leu Ala Thr Ala Thr His Ala Leu 85 90 95 Leu Met Leu Pro Ala Leu Lys Ala Ala Gln Met Arg Ile His Pro His 100 105 110 Ala Gln Leu Ala Ala Met Gln Gln Ala Lys Ser Thr Pro Met Ser Gln 115 120 125 Val Ser Gly Glu Leu Lys Leu Gly Ala Asn Ala Leu Ser Leu Ala Gln 130 135 140 Thr Asn Ala Leu Ser His Ala Leu Ser Gln Ala Lys Arg Asn Leu Thr 145 150 155 160 Asp Val Ser Val Asn Glu Cys Phe Glu Asn Leu Lys Ser Glu Gln Gln 165 170 175 Phe Thr Glu Val Tyr Ser Leu Ile Gln Gln Leu Ala Ser Arg Thr His 180 185 190 Val Arg Lys Glu Val Asn Gln Gly Val Glu Leu Gly Pro Lys Gln Ala 195 200 205 Lys Ser His Tyr Trp Phe Ser Glu Phe His Gln Asn Arg Val Ala Ala 210 215 220 Ile Asn Phe Ile Asn Gly Gln Gln Ala Thr Ser Tyr Val Leu Thr Gln 225 230 235 240 Gly Ser Gly Leu Leu Ala Ala Lys Ser Met Leu Asn Gln Gln Arg Leu 245 250 255 Met Phe Ile Leu Pro Gly Asn Ser Gln Gln Gln Ile Thr Ala Ser Ile 260 265 270 Thr Gln Leu Met Gln Gln Leu Glu Arg Leu Gln Val Thr Glu Val Asn 275 280 285 Glu Leu Ser Leu Glu Cys Gln Leu Glu Leu Leu Ser Ile Met Tyr Asp 290 295 300 Asn Leu Val Asn Ala Asp Lys Leu Thr Thr Arg Asp Ser Lys Pro Ala 305 310 315 320 Tyr Gln Ala Val Ile Gln Ala Ser Ser Val Ser Ala Ala Lys Gln Glu 325 330 335 Leu Ser Ala Leu Asn Asp Ala Leu Thr Ala Leu Phe Ala Glu Gln Thr 340 345 350 Asn Ala Thr Ser Thr Asn Lys Gly Leu Ile Gln Tyr Lys Thr Pro Ala 355 360 365 Gly Ser Tyr Leu Thr Leu Thr Pro Leu Gly Ser Asn Asn Asp Asn Ala 370 375 380 Gln Ala Gly Leu Ala Phe Val Tyr Pro Gly Val Gly Thr Val Tyr Ala 385 390 395 400 Asp Met Leu Asn Glu Leu His Gln Tyr Phe Pro Ala Leu Tyr Ala Lys 405 410 415 Leu Glu Arg Glu Gly Asp Leu Lys Ala Met Leu Gln Ala Glu Asp Ile 420 425 430 Tyr His Leu Asp Pro Lys His Ala Ala Gln Met Ser Leu Gly Asp Leu 435 440 445 Ala Ile Ala Gly Val Gly Ser Ser Tyr Leu Leu Thr Gln Leu Leu Thr 450 455 460 Asp Glu Phe Asn Ile Lys Pro Asn Phe Ala Leu Gly Tyr Ser Met Gly 465 470 475 480 Glu Ala Ser Met Trp Ala Ser Leu Gly Val Trp Gln Asn Pro His Ala 485 490 495 Leu Ile Ser Lys Thr Gln Thr Asp Pro Leu Phe Thr Ser Ala Ile Ser 500 505 510 Gly Lys Leu Thr Ala Val Arg Gln Ala Trp Gln Leu Asp Asp Thr Ala 515 520 525 Ala Glu Ile Gln Trp Asn Ser Phe Val Val Arg Ser Glu Ala Ala Pro 530 535 540 Ile Glu Ala Leu Leu Lys Asp Tyr Pro His Ala Tyr Leu Ala Ile Ile 545 550 555 560 Gln Gly Asp Thr Cys Val Ile Ala Gly Cys Glu Ile Gln Cys Lys Ala 565 570 575 Leu Leu Ala Ala Leu Gly Lys Arg Gly Ile Ala Ala Asn Arg Val Thr 580 585 590 Ala Met His Thr Gln Pro Ala Met Gln Glu His Gln Asn Val Met Asp 595 600 605 Phe Tyr Leu Gln Pro Leu Lys Ala Glu Leu Pro Ser Glu Ile Ser Phe 610 615 620 Ile Ser Ala Ala Asp Leu Thr Ala Lys Gln Thr Val Ser Glu Gln Ala 625 630 635 640 Leu Ser Ser Gln Val Val Ala Gln Ser Ile Ala Asp Thr Phe Cys Gln 645 650 655 Thr Leu Asp Phe Thr Ala Leu Val His His Ala Gln His Gln Gly Ala 660 665 670 Lys Leu Phe Val Glu Ile Gly Ala Asp Arg Gln Asn Cys Thr Leu Ile 675 680 685 Asp Lys Ile Val Lys Gln Asp Gly Ala Ser Ser Val Gln His Gln Pro 690 695 700 Cys Cys Thr Val Pro Met Asn Ala Lys Gly Ser Gln Asp Ile Thr Ser 705 710 715 720 Val Ile Lys Ala Leu Gly Gln Leu Ile Ser His Gln Val Pro Leu Ser 725 730 735 Val Gln Pro Phe Ile Asp Gly Leu Lys Arg Glu Leu Thr Leu Cys Gln 740 745 750 Leu Thr Ser Gln Gln Leu Ala Ala His Ala Asn Val Asp Ser Lys Phe 755 760 765 Glu Ser Asn Gln Asp His Leu Leu Gln Gly Glu Val 770 775 780 <210> 10 <211> 6015 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 atg tca tta cca gac aat gct tct aac cac ctt tct gcc aac cag aaa 48 Met Ser Leu Pro Asp Asn Ala Ser Asn His Leu Ser Ala Asn Gln Lys 1 5 10 15 ggc gca tct cag gca agt aaa acc agt aag caa agc aaa atc gcc att 96 Gly Ala Ser Gln Ala Ser Lys Thr Ser Lys Gln Ser Lys Ile Ala Ile 20 25 30 gtc ggt tta gcc act ctg tat cca gac gct aaa acc ccg caa gaa ttt 144 Val Gly Leu Ala Thr Leu Tyr Pro Asp Ala Lys Thr Pro Gln Glu Phe 35 40 45 tgg cag aat ttg ctg gat aaa cgc gac tct cgc agc acc tta act aac 192 Trp Gln Asn Leu Leu Asp Lys Arg Asp Ser Arg Ser Thr Leu Thr Asn 50 55 60 gaa aaa ctc ggc gct aac agc caa gat tat caa ggt gtg caa ggc caa 240 Glu Lys Leu Gly Ala Asn Ser Gln Asp Tyr Gln Gly Val Gln Gly Gln 65 70 75 80 tct gac cgt ttt tat tgt aat aaa ggc ggc tac att gag aac ttc agc 288 Ser Asp Arg Phe Tyr Cys Asn Lys Gly Gly Tyr Ile Glu Asn Phe Ser 85 90 95 ttt aat gct gca ggc tac aaa ttg ccg gag caa agc tta aat ggc ttg 336 Phe Asn Ala Ala Gly Tyr Lys Leu Pro Glu Gln Ser Leu Asn Gly Leu 100 105 110 gac gac agc ttc ctt tgg gcg ctc gat act agc cgt aac gca cta att 384 Asp Asp Ser Phe Leu Trp Ala Leu Asp Thr Ser Arg Asn Ala Leu Ile 115 120 125 gat gct ggt att gat atc aac ggc gct gat tta agc cgc gca ggt gta 432 Asp Ala Gly Ile Asp Ile Asn Gly Ala Asp Leu Ser Arg Ala Gly Val 130 135 140 gtc atg ggc gcg ctg tcg ttc cca act acc cgc tca aac gat ctg ttt 480 Val Met Gly Ala Leu Ser Phe Pro Thr Thr Arg Ser Asn Asp Leu Phe 145 150 155 160 ttg cca att tat cac agc gcc gtt gaa aaa gcc ctg caa gat aaa cta 528 Leu Pro Ile Tyr His Ser Ala Val Glu Lys Ala Leu Gln Asp Lys Leu 165 170 175 ggc gta aag gca ttt aag cta agc cca act aat gct cat acc gct cgc 576 Gly Val Lys Ala Phe Lys Leu Ser Pro Thr Asn Ala His Thr Ala Arg 180 185 190 gcg gca aat gag agc agc cta aat gca gcc aat ggt gcc att gcc cat 624 Ala Ala Asn Glu Ser Ser Leu Asn Ala Ala Asn Gly Ala Ile Ala His 195 200 205 aac agc tca aaa gtg gtg gcc gat gca ctt ggc ctt ggc ggc gca caa 672 Asn Ser Ser Lys Val Val Ala Asp Ala Leu Gly Leu Gly Gly Ala Gln 210 215 220 cta agc cta gat gct gcc tgt gct agt tcg gtt tac tca tta aag ctt 720 Leu Ser Leu Asp Ala Ala Cys Ala Ser Ser Val Tyr Ser Leu Lys Leu 225 230 235 240 gcc tgc gat tac cta agc act ggc aaa gcc gat atc atg cta gca ggc 768 Ala Cys Asp Tyr Leu Ser Thr Gly Lys Ala Asp Ile Met Leu Ala Gly 245 250 255 gca gta tct ggc gcg gat cct ttc ttt att aat atg gga ttc tca atc 816 Ala Val Ser Gly Ala Asp Pro Phe Phe Ile Asn Met Gly Phe Ser Ile 260 265 270 ttc cac gcc tac cca gac cat ggt atc tca gta ccg ttt gat gcc agc 864 Phe His Ala Tyr Pro Asp His Gly Ile Ser Val Pro Phe Asp Ala Ser 275 280 285 agt aaa ggt ttg ttt gct ggc gaa ggc gct ggc gta tta gtg ctt aaa 912 Ser Lys Gly Leu Phe Ala Gly Glu Gly Ala Gly Val Leu Val Leu Lys 290 295 300 cgt ctt gaa gat gcc gag cgc gac aat gac aaa atc tat gcg gtt gtt 960 Arg Leu Glu Asp Ala Glu Arg Asp Asn Asp Lys Ile Tyr Ala Val Val 305 310 315 320 agc ggc gta ggt cta tca aac gac ggt aaa ggc cag ttt gta tta agc 1008 Ser Gly Val Gly Leu Ser Asn Asp Gly Lys Gly Gln Phe Val Leu Ser 325 330 335 cct aat cca aaa ggt cag gtg aag gcc ttt gaa cgt gct tat gct gcc 1056 Pro Asn Pro Lys Gly Gln Val Lys Ala Phe Glu Arg Ala Tyr Ala Ala 340 345 350 agt gac att gag cca aaa gac att gaa gtg att gag tgc cac gca aca 1104 Ser Asp Ile Glu Pro Lys Asp Ile Glu Val Ile Glu Cys His Ala Thr 355 360 365 ggc aca ccg ctt ggc gat aaa att gag ctc act tca atg gaa acc ttc 1152 Gly Thr Pro Leu Gly Asp Lys Ile Glu Leu Thr Ser Met Glu Thr Phe 370 375 380 ttt gaa gac aag ctg caa ggc acc gat gca ccg tta att ggc tca gct 1200 Phe Glu Asp Lys Leu Gln Gly Thr Asp Ala Pro Leu Ile Gly Ser Ala 385 390 395 400 aag tct aac tta ggc cac cta tta act gca gcg ggc atg ccg ggg atc 1248 Lys Ser Asn Leu Gly His Leu Leu Thr Ala Ala Gly Met Pro Gly Ile 405 410 415 atg aag atg atc ttc gcc atg aaa gaa ggt tac ctg ccg cca agt atc 1296 Met Lys Met Ile Phe Ala Met Lys Glu Gly Tyr Leu Pro Pro Ser Ile 420 425 430 aat att agt gat gct atc gct tcg ccg aaa aaa ctc ttc ggt aaa cca 1344 Asn Ile Ser Asp Ala Ile Ala Ser Pro Lys Lys Leu Phe Gly Lys Pro 435 440 445 acc ctg cct agc atg gtt caa ggc tgg cca gat aag cca tcg aat aat 1392 Thr Leu Pro Ser Met Val Gln Gly Trp Pro Asp Lys Pro Ser Asn Asn 450 455 460 cat ttt ggt gta aga acc cgt cac gca ggc gta tcg gta ttt ggc ttt 1440 His Phe Gly Val Arg Thr Arg His Ala Gly Val Ser Val Phe Gly Phe 465 470 475 ggt ggc tgt aac gcc cat ctg ttg ctt gag tca tac aac ggc aaa gga 1488 Gly Gly Cys Asn Ala His Leu Leu Leu Glu Ser Tyr Asn Gly Lys Gly 480 485 490 495 aca gta aag gca gaa gcc act caa gta ccg cgt caa gct gag ccg cta 1536 Thr Val Lys Ala Glu Ala Thr Gln Val Pro Arg Gln Ala Glu Pro Leu 500 505 510 aaa gtg gtt ggc ctt gcc tcg cac ttt ggg cct ctt agc agc att aat 1584 Lys Val Val Gly Leu Ala Ser His Phe Gly Pro Leu Ser Ser Ile Asn 515 520 525 gca ctc aac aat gct gtg acc caa gat ggg aat ggc ttt atc gaa ctg 1632 Ala Leu Asn Asn Ala Val Thr Gln Asp Gly Asn Gly Phe Ile Glu Leu 530 535 540 ccg aaa aag cgc tgg aaa ggc ctt gaa aag cac agt gaa ctg tta gct 1680 Pro Lys Lys Arg Trp Lys Gly Leu Glu Lys His Ser Glu Leu Leu Ala 545 550 555 gaa ttt ggc tta gca tct gcg cca aaa ggt gct tat gtt gat aac ttc 1728 Glu Phe Gly Leu Ala Ser Ala Pro Lys Gly Ala Tyr Val Asp Asn Phe 560 565 570 575 gag ctg gac ttt tta cgc ttt aaa ctg ccg cca aac gaa gat gac cgt 1776 Glu Leu Asp Phe Leu Arg Phe Lys Leu Pro Pro Asn Glu Asp Asp Arg 580 585 590 ttg atc tca cag cag cta atg cta atg cga gta aca gac gaa gcc att 1824 Leu Ile Ser Gln Gln Leu Met Leu Met Arg Val Thr Asp Glu Ala Ile 595 600 605 cgt gat gcc aag ctt gag ccg ggg caa aaa gta gct gta tta gtg gca 1872 Arg Asp Ala Lys Leu Glu Pro Gly Gln Lys Val Ala Val Leu Val Ala 610 615 620 atg gaa act gag ctt gaa ctg cat cag ttc cgc ggc cgg gtt aac ttg 1920 Met Glu Thr Glu Leu Glu Leu His Gln Phe Arg Gly Arg Val Asn Leu 625 630 635 cat act caa tta gcg caa agt ctt gcc gcc atg ggc gtg agt tta tca 1968 His Thr Gln Leu Ala Gln Ser Leu Ala Ala Met Gly Val Ser Leu Ser 640 645 650 655 acg gat gaa tac caa gcg ctt gaa gcc atc gcc atg gac agc gtg ctt 2016 Thr Asp Glu Tyr Gln Ala Leu Glu Ala Ile Ala Met Asp Ser Val Leu 660 665 670 gat gct gcc aag ctc aat cag tac acc agc ttt att ggt aat att atg 2064 Asp Ala Ala Lys Leu Asn Gln Tyr Thr Ser Phe Ile Gly Asn Ile Met 675 680 685 gcg tca cgc gtg gcg tca cta tgg gac ttt aat ggc cca gcc ttc act 2112 Ala Ser Arg Val Ala Ser Leu Trp Asp Phe Asn Gly Pro Ala Phe Thr 690 695 700 att tca gca gca gag caa tct gtg agc cgc tgt atc gat gtg gcg caa 2160 Ile Ser Ala Ala Glu Gln Ser Val Ser Arg Cys Ile Asp Val Ala Gln 705 710 715 aac ctc atc atg gag gat aac cta gat gcg gtg gtg att gca gcg gtc 2208 Asn Leu Ile Met Glu Asp Asn Leu Asp Ala Val Val Ile Ala Ala Val 720 725 730 735 gat ctc tct ggt agc ttt gag caa gtc att ctt aaa aat gcc att gca 2256 Asp Leu Ser Gly Ser Phe Glu Gln Val Ile Leu Lys Asn Ala Ile Ala 740 745 750 cct gta gcc att gag cca aac ctc gaa gca agc ctt aat cca aca tca 2304 Pro Val Ala Ile Glu Pro Asn Leu Glu Ala Ser Leu Asn Pro Thr Ser 755 760 765 gca agc tgg aat gtc ggt gaa ggt gct ggc gcg gtc gtg ctt gtt aaa 2352 Ala Ser Trp Asn Val Gly Glu Gly Ala Gly Ala Val Val Leu Val Lys 770 775 780 aat gaa gct aca tcg ggc tgc tca tac ggc caa att gat gca ctt ggc 2400 Asn Glu Ala Thr Ser Gly Cys Ser Tyr Gly Gln Ile Asp Ala Leu Gly 785 790 795 ttt gct aaa act gcc gaa aca gcg ttg gct acc gac aag cta ctg agc 2448 Phe Ala Lys Thr Ala Glu Thr Ala Leu Ala Thr Asp Lys Leu Leu Ser 800 805 810 815 caa act gcc aca gac ttt aat aag gtt aaa gtg att gaa act atg gca 2496 Gln Thr Ala Thr Asp Phe Asn Lys Val Lys Val Ile Glu Thr Met Ala 820 825 830 gcg cct gct agc caa att caa tta gcg cca ata gtt agc tct caa gtg 2544 Ala Pro Ala Ser Gln Ile Gln Leu Ala Pro Ile Val Ser Ser Gln Val 835 840 845 act cac act gct gca gag cag cgt gtt ggt cac tgc ttt gct gca gcg 2592 Thr His Thr Ala Ala Glu Gln Arg Val Gly His Cys Phe Ala Ala Ala 850 855 860 ggt atg gca agc cta tta cac ggc tta ctt aac tta aat act gta gcc 2640 Gly Met Ala Ser Leu Leu His Gly Leu Leu Asn Leu Asn Thr Val Ala 865 870 875 caa acc aat aaa gcc aat tgc gcg ctt atc aac aat atc agt gaa aac 2688 Gln Thr Asn Lys Ala Asn Cys Ala Leu Ile Asn Asn Ile Ser Glu Asn 880 885 890 895 caa tta tca cag ctg ttg att agc caa aca gcg agc gaa caa caa gca 2736 Gln Leu Ser Gln Leu Leu Ile Ser Gln Thr Ala Ser Glu Gln Gln Ala 900 905 910 tta acc gcg cgt tta agc aat gag ctt aaa tcc gat gct aaa cac caa 2784 Leu Thr Ala Arg Leu Ser Asn Glu Leu Lys Ser Asp Ala Lys His Gln 915 920 925 ctg gtt aag caa gtc acc tta ggt ggc cgt gat atc tac cag cat att 2832 Leu Val Lys Gln Val Thr Leu Gly Gly Arg Asp Ile Tyr Gln His Ile 930 935 940 gtt gat aca ccg ctt gca agc ctt gaa agc att act cag aaa ttg gcg 2880 Val Asp Thr Pro Leu Ala Ser Leu Glu Ser Ile Thr Gln Lys Leu Ala 945 950 955 caa gcg aca gca tcg aca gtg gtc aac caa gtt aaa cct att aag gcc 2928 Gln Ala Thr Ala Ser Thr Val Val Asn Gln Val Lys Pro Ile Lys Ala 960 965 970 975 gct ggc tca gtc gaa atg gct aac tca ttc gaa acg gaa agc tca gca 2976 Ala Gly Ser Val Glu Met Ala Asn Ser Phe Glu Thr Glu Ser Ser Ala 980 985 990 gag cca caa ata aca att gca gca caa cag act gca aac att ggc gtc 3024 Glu Pro Gln Ile Thr Ile Ala Ala Gln Gln Thr Ala Asn Ile Gly Val 995 1000 1005 acc gct cag gca acc aaa cgt gaa tta ggt acc cca cca atg aca aca 3072 Thr Ala Gln Ala Thr Lys Arg Glu Leu Gly Thr Pro Pro Met Thr Thr 1010 1015 1020 aat acc att gct aat aca gca aat aat tta gac aag act ctt gag act 3120 Asn Thr Ile Ala Asn Thr Ala Asn Asn Leu Asp Lys Thr Leu Glu Thr 1025 1030 1035 gtt gct ggc aat act gtt gct agc aag gtt ggc tct ggc gac ata gtc 3168 Val Ala Gly Asn Thr Val Ala Ser Lys Val Gly Ser Gly Asp Ile Val 1040 1045 1050 1055 aat ttt caa cag aac caa caa ttg gct caa caa gct cac ctc gcc ttt 3216 Asn Phe Gln Gln Asn Gln Gln Leu Ala Gln Gln Ala His Leu Ala Phe 1060 1065 1070 ctt gaa agc cgc agt gcg ggt atg aag gtg gct gat gct tta ttg aag 3264 Leu Glu Ser Arg Ser Ala Gly Met Lys Val Ala Asp Ala Leu Leu Lys 1075 1080 1085 caa cag cta gct caa gta aca ggc caa act atc gat aat cag gcc ctc 3312 Gln Gln Leu Ala Gln Val Thr Gly Gln Thr Ile Asp Asn Gln Ala Leu 1090 1095 1100 gat act caa gcc gtc gat act caa aca agc gag aat gta gcg att gcc 3360 Asp Thr Gln Ala Val Asp Thr Gln Thr Ser Glu Asn Val Ala Ile Ala 1105 1110 1115 gca gaa tca cca gtt caa gtt aca aca cct gtt caa gtt aca aca cct 3408 Ala Glu Ser Pro Val Gln Val Thr Thr Pro Val Gln Val Thr Thr Pro 1120 1125 1130 1135 gtt caa atc agt gtt gtg gag tta aaa cca gat cac gct aat gtg cca 3456 Val Gln Ile Ser Val Val Glu Leu Lys Pro Asp His Ala Asn Val Pro 1140 1145 1150 cca tac acg ccg cca gtg cct gca tta aag ccg tgt atc tgg aac tat 3504 Pro Tyr Thr Pro Pro Val Pro Ala Leu Lys Pro Cys Ile Trp Asn Tyr 1155 1160 1165 gcc gat tta gtt gag tac gca gaa ggc gat atc gcc aag gta ttt ggc 3552 Ala Asp Leu Val Glu Tyr Ala Glu Gly Asp Ile Ala Lys Val Phe Gly 1170 1175 1180 agt gat tat gcc att atc gac agc tac tcg cgc cgc gta cgt cta ccg 3600 Ser Asp Tyr Ala Ile Ile Asp Ser Tyr Ser Arg Arg Val Arg Leu Pro 1185 1190 1195 acc act gac tac ctg ttg gta tcg cgc gtg acc aaa ctt gat gcg acc 3648 Thr Thr Asp Tyr Leu Leu Val Ser Arg Val Thr Lys Leu Asp Ala Thr 1200 1205 1210 1215 atc aat caa ttt aag cca tgc tca atg acc act gag tac gac atc cct 3696 Ile Asn Gln Phe Lys Pro Cys Ser Met Thr Thr Glu Tyr Asp Ile Pro 1220 1225 1230 gtt gat gcg ccg tac tta gta gac gga caa atc cct tgg gcg gta gca 3744 Val Asp Ala Pro Tyr Leu Val Asp Gly Gln Ile Pro Trp Ala Val Ala 1235 1240 1245 gta gaa tca ggc caa tgt gac ttg atg ctt att agc tat ctc ggt atc 3792 Val Glu Ser Gly Gln Cys Asp Leu Met Leu Ile Ser Tyr Leu Gly Ile 1250 1255 1260 gac ttt gag aac aaa ggc gag cgg gtt tat cga cta ctc gat tgt acc 3840 Asp Phe Glu Asn Lys Gly Glu Arg Val Tyr Arg Leu Leu Asp Cys Thr 1265 1270 1275 ctc acc ttc cta ggc gac ttg cca cgt ggc gga gat acc cta cgt tac 3888 Leu Thr Phe Leu Gly Asp Leu Pro Arg Gly Gly Asp Thr Leu Arg Tyr 1280 1285 1290 1295 gac att aag atc aat aac tat gct cgc aac ggc gac acc ctg ctg ttc 3936 Asp Ile Lys Ile Asn Asn Tyr Ala Arg Asn Gly Asp Thr Leu Leu Phe 1300 1305 1310 ttc ttc tcg tat gag tgt ttt gtt ggc gac aag atg atc ctc aag atg 3984 Phe Phe Ser Tyr Glu Cys Phe Val Gly Asp Lys Met Ile Leu Lys Met 1315 1320 1325 gat ggc ggc tgc gct ggc ttc ttc act gat gaa gag ctt gcc gac ggt 4032 Asp Gly Gly Cys Ala Gly Phe Phe Thr Asp Glu Glu Leu Ala Asp Gly 1330 1335 1340 aaa ggc gtg att cgc aca gaa gaa gag att aaa gct cgc agc cta gtg 4080 Lys Gly Val Ile Arg Thr Glu Glu Glu Ile Lys Ala Arg Ser Leu Val 1345 1350 1355 caa aag caa cgc ttt aat ccg tta cta gat tgt cct aaa acc caa ttt 4128 Gln Lys Gln Arg Phe Asn Pro Leu Leu Asp Cys Pro Lys Thr Gln Phe 1360 1365 1370 1375 agt tat ggt gat att cat aag cta tta act gct gat att gag ggt tgt 4176 Ser Tyr Gly Asp Ile His Lys Leu Leu Thr Ala Asp Ile Glu Gly Cys 1380 1385 1390 ttt ggc cca agc cac agt ggc gtc cac cag ccg tca ctt tgt ttc gca 4224 Phe Gly Pro Ser His Ser Gly Val His Gln Pro Ser Leu Cys Phe Ala 1395 1400 1405 tct gaa aaa ttc ttg atg att gaa caa gtc agc aag gtt gat cgc act 4272 Ser Glu Lys Phe Leu Met Ile Glu Gln Val Ser Lys Val Asp Arg Thr 1410 1415 1420 ggc ggt act tgg gga ctt ggc tta att gag ggt cat aag cag ctt gaa 4320 Gly Gly Thr Trp Gly Leu Gly Leu Ile Glu Gly His Lys Gln Leu Glu 1425 1430 1435 gca gac cac tgg tac ttc cca tgt cat ttc aag ggc gac caa gtg atg 4368 Ala Asp His Trp Tyr Phe Pro Cys His Phe Lys Gly Asp Gln Val Met 1440 1445 1450 1455 gct ggc tcg cta atg gct gaa ggt tgt ggc cag tta ttg cag ttc tat 4416 Ala Gly Ser Leu Met Ala Glu Gly Cys Gly Gln Leu Leu Gln Phe Tyr 1460 1465 1470 atg ctg cac ctt ggt atg cat acc caa act aaa aat ggt cgt ttc caa 4464 Met Leu His Leu Gly Met His Thr Gln Thr Lys Asn Gly Arg Phe Gln 1475 1480 1485 cct ctt gaa aac gcc tca cag caa gta cgc tgt cgc ggt caa gtg ctg 4512 Pro Leu Glu Asn Ala Ser Gln Gln Val Arg Cys Arg Gly Gln Val Leu 1490 1495 1500 cca caa tca ggc gtg cta act tac cgt atg gaa gtg act gaa atc ggt 4560 Pro Gln Ser Gly Val Leu Thr Tyr Arg Met Glu Val Thr Glu Ile Gly 1505 1510 1515 ttc agt cca cgc cca tat gct aaa gct aac atc gat atc ttg ctt aat 4608 Phe Ser Pro Arg Pro Tyr Ala Lys Ala Asn Ile Asp Ile Leu Leu Asn 1520 1525 1530 1535 ggc aaa gcg gta gtg gat ttc caa aac cta ggg gtg atg ata aaa gag 4656 Gly Lys Ala Val Val Asp Phe Gln Asn Leu Gly Val Met Ile Lys Glu 1540 1545 1550 gaa gat gag tgt act cgt tat cca ctt ttg act gaa tca aca acg gct 4704 Glu Asp Glu Cys Thr Arg Tyr Pro Leu Leu Thr Glu Ser Thr Thr Ala 1555 1560 1565 agc act gca caa gta aac gct caa aca agt gcg aaa aag gta tac aag 4752 Ser Thr Ala Gln Val Asn Ala Gln Thr Ser Ala Lys Lys Val Tyr Lys 1570 1575 1580 cca gca tca gtc aat gcg cca tta atg gca caa att cct gat ctg act 4800 Pro Ala Ser Val Asn Ala Pro Leu Met Ala Gln Ile Pro Asp Leu Thr 1585 1590 1595 aaa gag cca aac aag ggc gtt att ccg att tcc cat gtt gaa gca cca 4848 Lys Glu Pro Asn Lys Gly Val Ile Pro Ile Ser His Val Glu Ala Pro 1600 1605 1610 1615 att acg cca gac tac ccg aac cgt gta cct gat aca gtg cca ttc acg 4896 Ile Thr Pro Asp Tyr Pro Asn Arg Val Pro Asp Thr Val Pro Phe Thr 1620 1625 1630 ccg tat cac atg ttt gag ttt gct aca ggc aat atc gaa aac tgt ttc 4944 Pro Tyr His Met Phe Glu Phe Ala Thr Gly Asn Ile Glu Asn Cys Phe 1635 1640 1645 ggg cca gag ttc tca atc tat cgc ggc atg atc cca cca cgt aca cca 4992 Gly Pro Glu Phe Ser Ile Tyr Arg Gly Met Ile Pro Pro Arg Thr Pro 1650 1655 1660 tgc ggt gac tta caa gtg acc aca cgt gtg att gaa gtt aac ggt aag 5040 Cys Gly Asp Leu Gln Val Thr Thr Arg Val Ile Glu Val Asn Gly Lys 1665 1670 1675 cgt ggc gac ttt aaa aag cca tca tcg tgt atc gct gaa tat gaa gtg 5088 Arg Gly Asp Phe Lys Lys Pro Ser Ser Cys Ile Ala Glu Tyr Glu Val 1680 1685 1690 1695 cct gca gat gcg tgg tat ttc gat aaa aac agc cac ggc gca gtg atg 5136 Pro Ala Asp Ala Trp Tyr Phe Asp Lys Asn Ser His Gly Ala Val Met 1700 1705 1710 cca tat tca att tta atg gag atc tca ctg caa cct aac ggc ttt atc 5184 Pro Tyr Ser Ile Leu Met Glu Ile Ser Leu Gln Pro Asn Gly Phe Ile 1715 1720 1725 tca ggt tac atg ggc aca acc cta ggc ttc cct ggc ctt gag ctg ttc 5232 Ser Gly Tyr Met Gly Thr Thr Leu Gly Phe Pro Gly Leu Glu Leu Phe 1730 1735 1740 ttc cgt aac tta gac ggt agc ggt gag tta cta cgt gaa gta gat tta 5280 Phe Arg Asn Leu Asp Gly Ser Gly Glu Leu Leu Arg Glu Val Asp Leu 1745 1750 1755 cgt ggt aaa acc atc cgt aac gac tca cgt tta tta tca aca gtg atg 5328 Arg Gly Lys Thr Ile Arg Asn Asp Ser Arg Leu Leu Ser Thr Val Met 1760 1765 1770 1775 gcc ggc act aac atc atc caa agc ttt agc ttc gag cta agc act gac 5376 Ala Gly Thr Asn Ile Ile Gln Ser Phe Ser Phe Glu Leu Ser Thr Asp 1780 1785 1790 ggt gag cct ttc tat cgc ggc act gcg gta ttt ggc tat ttt aaa ggt 5424 Gly Glu Pro Phe Tyr Arg Gly Thr Ala Val Phe Gly Tyr Phe Lys Gly 1795 1800 1805 gac gca ctt aaa gat cag cta ggc cta gat aac ggt aaa gtc act cag 5472 Asp Ala Leu Lys Asp Gln Leu Gly Leu Asp Asn Gly Lys Val Thr Gln 1810 1815 1820 cca tgg cat gta gct aac ggc gtt gct gca agc act aag gtg aac ctg 5520 Pro Trp His Val Ala Asn Gly Val Ala Ala Ser Thr Lys Val Asn Leu 1825 1830 1835 ctt gat aag agc tgc cgt cac ttt aat gcg cca gct aac cag cca cac 5568 Leu Asp Lys Ser Cys Arg His Phe Asn Ala Pro Ala Asn Gln Pro His 1840 1845 1850 1855 tat cgt cta gcc ggt ggt cag ctg aac ttt atc gac agt gtt gaa att 5616 Tyr Arg Leu Ala Gly Gly Gln Leu Asn Phe Ile Asp Ser Val Glu Ile 1860 1865 1870 gtt gat aat ggc ggc acc gaa ggt tta ggt tac ttg tat gcc gag cgc 5664 Val Asp Asn Gly Gly Thr Glu Gly Leu Gly Tyr Leu Tyr Ala Glu Arg 1875 1880 1885 acc att gac cca agt gat tgg ttc ttc cag ttc cac ttc cac caa gat 5712 Thr Ile Asp Pro Ser Asp Trp Phe Phe Gln Phe His Phe His Gln Asp 1890 1895 1900 ccg gtt atg cca ggc tcc tta ggt gtt gaa gca att att gaa acc atg 5760 Pro Val Met Pro Gly Ser Leu Gly Val Glu Ala Ile Ile Glu Thr Met 1905 1910 1915 caa gct tac gct att agt aaa gac ttg ggc gca gat ttc aaa aat cct 5808 Gln Ala Tyr Ala Ile Ser Lys Asp Leu Gly Ala Asp Phe Lys Asn Pro 1920 1925 1930 1935 aag ttt ggt cag att tta tcg aac atc aag tgg aag tat cgc ggt caa 5856 Lys Phe Gly Gln Ile Leu Ser Asn Ile Lys Trp Lys Tyr Arg Gly Gln 1940 1945 1950 atc aat ccg ctg aac aag cag atg tct atg gat gtc agc att act tca 5904 Ile Asn Pro Leu Asn Lys Gln Met Ser Met Asp Val Ser Ile Thr Ser 1955 1960 1965 atc aaa gat gaa gac ggt aag aaa gtc atc aca ggt aat gcc agc ttg 5952 Ile Lys Asp Glu Asp Gly Lys Lys Val Ile Thr Gly Asn Ala Ser Leu 1970 1975 1980 agt aaa gat ggt ctg cgc ata tac gag gtc ttc gat ata gct atc agc 6000 Ser Lys Asp Gly Leu Arg Ile Tyr Glu Val Phe Asp Ile Ala Ile Ser 1985 1990 1995 atc gaa gaa tct gta 6015 Ile Glu Glu Ser Val 2000 <210> 11 <211> 2005 <212> PRT <400> 1 Met Ser Leu Pro Asp Asn Ala Ser Asn His Leu Ser Ala Asn Gln Lys 1 5 10 15 Gly Ala Ser Gln Ala Ser Lys Thr Ser Lys Gln Ser Lys Ile Ala Ile 20 25 30 Val Gly Leu Ala Thr Leu Tyr Pro Asp Ala Lys Thr Pro Gln Glu Phe 35 40 45 Trp Gln Asn Leu Leu Asp Lys Arg Asp Ser Arg Ser Thr Leu Thr Asn 50 55 60 Glu Lys Leu Gly Ala Asn Ser Gln Asp Tyr Gln Gly Val Gln Gly Gln 65 70 75 80 Ser Asp Arg Phe Tyr Cys Asn Lys Gly Gly Tyr Ile Glu Asn Phe Ser 85 90 95 Phe Asn Ala Ala Gly Tyr Lys Leu Pro Glu Gln Ser Leu Asn Gly Leu 100 105 110 Asp Asp Ser Phe Leu Trp Ala Leu Asp Thr Ser Arg Asn Ala Leu Ile 115 120 125 Asp Ala Gly Ile Asp Ile Asn Gly Ala Asp Leu Ser Arg Ala Gly Val 130 135 140 Val Met Gly Ala Leu Ser Phe Pro Thr Thr Arg Ser Asn Asp Leu Phe 145 150 155 160 Leu Pro Ile Tyr His Ser Ala Val Glu Lys Ala Leu Gln Asp Lys Leu 165 170 175 Gly Val Lys Ala Phe Lys Leu Ser Pro Thr Asn Ala His Thr Ala Arg 180 185 190 Ala Ala Asn Glu Ser Ser Leu Asn Ala Ala Asn Gly Ala Ile Ala His 195 200 205 Asn Ser Ser Lys Val Val Ala Asp Ala Leu Gly Leu Gly Gly Ala Gln 210 215 220 Leu Ser Leu Asp Ala Ala Cys Ala Ser Ser Val Tyr Ser Leu Lys Leu 225 230 235 240 Ala Cys Asp Tyr Leu Ser Thr Gly Lys Ala Asp Ile Met Leu Ala Gly 245 250 255 Ala Val Ser Gly Ala Asp Pro Phe Phe Ile Asn Met Gly Phe Ser Ile 260 265 270 Phe His Ala Tyr Pro Asp His Gly Ile Ser Val Pro Phe Asp Ala Ser 275 280 285 Ser Lys Gly Leu Phe Ala Gly Glu Gly Ala Gly Val Leu Val Leu Lys 290 295 300 Arg Leu Glu Asp Ala Glu Arg Asp Asn Asp Lys Ile Tyr Ala Val Val 305 310 315 320 Ser Gly Val Gly Leu Ser Asn Asp Gly Lys Gly Gln Phe Val Leu Ser 325 330 335 Pro Asn Pro Lys Gly Gln Val Lys Ala Phe Glu Arg Ala Tyr Ala Ala 340 345 350 Ser Asp Ile Glu Pro Lys Asp Ile Glu Val Ile Glu Cys His Ala Thr 355 360 365 Gly Thr Pro Leu Gly Asp Lys Ile Glu Leu Thr Ser Met Glu Thr Phe 370 375 380 Phe Glu Asp Lys Leu Gln Gly Thr Asp Ala Pro Leu Ile Gly Ser Ala 385 390 395 400 Lys Ser Asn Leu Gly His Leu Leu Thr Ala Ala Gly Met Pro Gly Ile 405 410 415 Met Lys Met Ile Phe Ala Met Lys Glu Gly Tyr Leu Pro Pro Ser Ile 420 425 430 Asn Ile Ser Asp Ala Ile Ala Ser Pro Lys Lys Leu Phe Gly Lys Pro 435 440 445 Thr Leu Pro Ser Met Val Gln Gly Trp Pro Asp Lys Pro Ser Asn Asn 450 455 460 His Phe Gly Val Arg Thr Arg His Ala Gly Val Ser Val Phe Gly Phe 465 470 475 480 Gly Gly Cys Asn Ala His Leu Leu Leu Glu Ser Tyr Asn Gly Lys Gly 485 490 495 Thr Val Lys Ala Glu Ala Thr Gln Val Pro Arg Gln Ala Glu Pro Leu 500 505 510 Lys Val Val Gly Leu Ala Ser His Phe Gly Pro Leu Ser Ser Ile Asn 515 520 525 Ala Leu Asn Asn Ala Val Thr Gln Asp Gly Asn Gly Phe Ile Glu Leu 530 535 540 Pro Lys Lys Arg Trp Lys Gly Leu Glu Lys His Ser Glu Leu Leu Ala 545 550 555 560 Glu Phe Gly Leu Ala Ser Ala Pro Lys Gly Ala Tyr Val Asp Asn Phe 565 570 575 Glu Leu Asp Phe Leu Arg Phe Lys Leu Pro Pro Asn Glu Asp Asp Arg 580 585 590 Leu Ile Ser Gln Gln Leu Met Leu Met Arg Val Thr Asp Glu Ala Ile 595 600 605 Arg Asp Ala Lys Leu Glu Pro Gly Gln Lys Val Ala Val Leu Val Ala 610 615 620 Met Glu Thr Glu Leu Glu Leu His Gln Phe Arg Gly Arg Val Asn Leu 625 630 635 640 His Thr Gln Leu Ala Gln Ser Leu Ala Ala Met Gly Val Ser Leu Ser 645 650 655 Thr Asp Glu Tyr Gln Ala Leu Glu Ala Ile Ala Met Asp Ser Val Leu 660 665 670 Asp Ala Ala Lys Leu Asn Gln Tyr Thr Ser Phe Ile Gly Asn Ile Met 675 680 685 Ala Ser Arg Val Ala Ser Leu Trp Asp Phe Asn Gly Pro Ala Phe Thr 690 695 700 Ile Ser Ala Ala Glu Gln Ser Val Ser Arg Cys Ile Asp Val Ala Gln 705 710 715 720 Asn Leu Ile Met Glu Asp Asn Leu Asp Ala Val Val Ile Ala Ala Val 725 730 735 Asp Leu Ser Gly Ser Phe Glu Gln Val Ile Leu Lys Asn Ala Ile Ala 740 745 750 Pro Val Ala Ile Glu Pro Asn Leu Glu Ala Ser Leu Asn Pro Thr Ser 755 760 765 Ala Ser Trp Asn Val Gly Glu Gly Ala Gly Ala Val Val Leu Val Lys 770 775 780 Asn Glu Ala Thr Ser Gly Cys Ser Tyr Gly Gln Ile Asp Ala Leu Gly 785 790 795 800 Phe Ala Lys Thr Ala Glu Thr Ala Leu Ala Thr Asp Lys Leu Leu Ser 805 810 815 Gln Thr Ala Thr Asp Phe Asn Lys Val Lys Val Ile Glu Thr Met Ala 820 825 830 Ala Pro Ala Ser Gln Ile Gln Leu Ala Pro Ile Val Ser Ser Gln Val 835 840 845 Thr His Thr Ala Ala Glu Gln Arg Val Gly His Cys Phe Ala Ala Ala 850 855 860 Gly Met Ala Ser Leu Leu His Gly Leu Leu Asn Leu Asn Thr Val Ala 865 870 875 880 Gln Thr Asn Lys Ala Asn Cys Ala Leu Ile Asn Asn Ile Ser Glu Asn 885 890 895 Gln Leu Ser Gln Leu Leu Ile Ser Gln Thr Ala Ser Glu Gln Gln Ala 900 905 910 Leu Thr Ala Arg Leu Ser Asn Glu Leu Lys Ser Asp Ala Lys His Gln 915 920 925 Leu Val Lys Gln Val Thr Leu Gly Gly Arg Asp Ile Tyr Gln His Ile 930 935 940 Val Asp Thr Pro Leu Ala Ser Leu Glu Ser Ile Thr Gln Lys Leu Ala 945 950 955 960 Gln Ala Thr Ala Ser Thr Val Val Asn Gln Val Lys Pro Ile Lys Ala 965 970 975 Ala Gly Ser Val Glu Met Ala Asn Ser Phe Glu Thr Glu Ser Ser Ala 980 985 990 Glu Pro Gln Ile Thr Ile Ala Ala Gln Gln Thr Ala Asn Ile Gly Val 995 1000 1005 Thr Ala Gln Ala Thr Lys Arg Glu Leu Gly Thr Pro Pro Met Thr Thr 1010 1015 1020 Asn Thr Ile Ala Asn Thr Ala Asn Asn Leu Asp Lys Thr Leu Glu Thr 1025 1030 1035 1040 Val Ala Gly Asn Thr Val Ala Ser Lys Val Gly Ser Gly Asp Ile Val 1045 1050 1055 Asn Phe Gln Gln Asn Gln Gln Leu Ala Gln Gln Ala His Leu Ala Phe 1060 1065 1070 Leu Glu Ser Arg Ser Ala Gly Met Lys Val Ala Asp Ala Leu Leu Lys 1075 1080 1085 Gln Gln Leu Ala Gln Val Thr Gly Gln Thr Ile Asp Asn Gln Ala Leu 1090 1095 1100 Asp Thr Gln Ala Val Asp Thr Gln Thr Ser Glu Asn Val Ala Ile Ala 1105 1110 1115 1120 Ala Glu Ser Pro Val Gln Val Thr Thr Pro Val Gln Val Thr Thr Pro 1125 1130 1135 Val Gln Ile Ser Val Val Glu Leu Lys Pro Asp His Ala Asn Val Pro 1140 1145 1150 Pro Tyr Thr Pro Pro Val Pro Ala Leu Lys Pro Cys Ile Trp Asn Tyr 1155 1160 1165 Ala Asp Leu Val Glu Tyr Ala Glu Gly Asp Ile Ala Lys Val Phe Gly 1170 1175 1180 Ser Asp Tyr Ala Ile Ile Asp Ser Tyr Ser Arg Arg Val Arg Leu Pro 1185 1190 1195 1200 Thr Thr Asp Tyr Leu Leu Val Ser Arg Val Thr Lys Leu Asp Ala Thr 1205 1210 1215 Ile Asn Gln Phe Lys Pro Cys Ser Met Thr Thr Glu Tyr Asp Ile Pro 1220 1225 1230 Val Asp Ala Pro Tyr Leu Val Asp Gly Gln Ile Pro Trp Ala Val Ala 1235 1240 1245 Val Glu Ser Gly Gln Cys Asp Leu Met Leu Ile Ser Tyr Leu Gly Ile 1250 1255 1260 Asp Phe Glu Asn Lys Gly Glu Arg Val Tyr Arg Leu Leu Asp Cys Thr 1265 1270 1275 1280 Leu Thr Phe Leu Gly Asp Leu Pro Arg Gly Gly Asp Thr Leu Arg Tyr 1285 1290 1295 Asp Ile Lys Ile Asn Asn Tyr Ala Arg Asn Gly Asp Thr Leu Leu Phe 1300 1305 1310 Phe Phe Ser Tyr Glu Cys Phe Val Gly Asp Lys Met Ile Leu Lys Met 1315 1320 1325 Asp Gly Gly Cys Ala Gly Phe Phe Thr Asp Glu Glu Leu Ala Asp Gly 1330 1335 1340 Lys Gly Val Ile Arg Thr Glu Glu Glu Ile Lys Ala Arg Ser Leu Val 1345 1350 1355 1360 Gln Lys Gln Arg Phe Asn Pro Leu Leu Asp Cys Pro Lys Thr Gln Phe 1365 1370 1375 Ser Tyr Gly Asp Ile His Lys Leu Leu Thr Ala Asp Ile Glu Gly Cys 1380 1385 1390 Phe Gly Pro Ser His Ser Gly Val His Gln Pro Ser Leu Cys Phe Ala 1395 1400 1405 Ser Glu Lys Phe Leu Met Ile Glu Gln Val Ser Lys Val Asp Arg Thr 1410 1415 1420 Gly Gly Thr Trp Gly Leu Gly Leu Ile Glu Gly His Lys Gln Leu Glu 1425 1430 1435 1440 Ala Asp His Trp Tyr Phe Pro Cys His Phe Lys Gly Asp Gln Val Met 1445 1450 1455 Ala Gly Ser Leu Met Ala Glu Gly Cys Gly Gln Leu Leu Gln Phe Tyr 1460 1465 1470 Met Leu His Leu Gly Met His Thr Gln Thr Lys Asn Gly Arg Phe Gln 1475 1480 1485 Pro Leu Glu Asn Ala Ser Gln Gln Val Arg Cys Arg Gly Gln Val Leu 1490 1495 1500 Pro Gln Ser Gly Val Leu Thr Tyr Arg Met Glu Val Thr Glu Ile Gly 1505 1510 1515 1520 Phe Ser Pro Arg Pro Tyr Ala Lys Ala Asn Ile Asp Ile Leu Leu Asn 1525 1530 1535 Gly Lys Ala Val Val Asp Phe Gln Asn Leu Gly Val Met Ile Lys Glu 1540 1545 1550 Glu Asp Glu Cys Thr Arg Tyr Pro Leu Leu Thr Glu Ser Thr Thr Ala 1555 1560 1565 Ser Thr Ala Gln Val Asn Ala Gln Thr Ser Ala Lys Lys Val Tyr Lys 1570 1575 1580 Pro Ala Ser Val Asn Ala Pro Leu Met Ala Gln Ile Pro Asp Leu Thr 1585 1590 1595 1600 Lys Glu Pro Asn Lys Gly Val Ile Pro Ile Ser His Val Glu Ala Pro 1605 1610 1615 Ile Thr Pro Asp Tyr Pro Asn Arg Val Pro Asp Thr Val Pro Phe Thr 1620 1625 1630 Pro Tyr His Met Phe Glu Phe Ala Thr Gly Asn Ile Glu Asn Cys Phe 1635 1640 1645 Gly Pro Glu Phe Ser Ile Tyr Arg Gly Met Ile Pro Pro Arg Thr Pro 1650 1655 1660 Cys Gly Asp Leu Gln Val Thr Thr Arg Val Ile Glu Val Asn Gly Lys 1665 1670 1675 1680 Arg Gly Asp Phe Lys Lys Pro Ser Ser Cys Ile Ala Glu Tyr Glu Val 1685 1690 1695 Pro Ala Asp Ala Trp Tyr Phe Asp Lys Asn Ser His Gly Ala Val Met 1700 1705 1710 Pro Tyr Ser Ile Leu Met Glu Ile Ser Leu Gln Pro Asn Gly Phe Ile 1715 1720 1725 Ser Gly Tyr Met Gly Thr Thr Leu Gly Phe Pro Gly Leu Glu Leu Phe 1730 1735 1740 Phe Arg Asn Leu Asp Gly Ser Gly Glu Leu Leu Arg Glu Val Asp Leu 1745 1750 1755 1760 Arg Gly Lys Thr Ile Arg Asn Asp Ser Arg Leu Leu Ser Thr Val Met 1765 1770 1775 Ala Gly Thr Asn Ile Ile Gln Ser Phe Ser Phe Glu Leu Ser Thr Asp 1780 1785 1790 Gly Glu Pro Phe Tyr Arg Gly Thr Ala Val Phe Gly Tyr Phe Lys Gly 1795 1800 1805 Asp Ala Leu Lys Asp Gln Leu Gly Leu Asp Asn Gly Lys Val Thr Gln 1810 1815 1820 Pro Trp His Val Ala Asn Gly Val Ala Ala Ser Thr Lys Val Asn Leu 1825 1830 1835 1840 Leu Asp Lys Ser Cys Arg His Phe Asn Ala Pro Ala Asn Gln Pro His 1845 1850 1855 Tyr Arg Leu Ala Gly Gly Gln Leu Asn Phe Ile Asp Ser Val Glu Ile 1860 1865 1870 Val Asp Asn Gly Gly Thr Glu Gly Leu Gly Tyr Leu Tyr Ala Glu Arg 1875 1880 1885 Thr Ile Asp Pro Ser Asp Trp Phe Phe Gln Phe His Phe His Gln Asp 1890 1895 1900 Pro Val Met Pro Gly Ser Leu Gly Val Glu Ala Ile Ile Glu Thr Met 1905 1910 1915 1920 Gln Ala Tyr Ala Ile Ser Lys Asp Leu Gly Ala Asp Phe Lys Asn Pro 1925 1930 1935 Lys Phe Gly Gln Ile Leu Ser Asn Ile Lys Trp Lys Tyr Arg Gly Gln 1940 1945 1950 Ile Asn Pro Leu Asn Lys Gln Met Ser Met Asp Val Ser Ile Thr Ser 1955 1960 1965 Ile Lys Asp Glu Asp Gly Lys Lys Val Ile Thr Gly Asn Ala Ser Leu 1970 1975 1980 Ser Lys Asp Gly Leu Arg Ile Tyr Glu Val Phe Asp Ile Ala Ile Ser 1985 1990 1995 2000 Ile Glu Glu Ser Val 2005 <210> 12 <211> 1626 <212> DNA <213> Shewanella putrefaciens SCRC-2874 (FERM BP-1625) <400> 1 atg aat cct aca gca act aac gaa atg ctt tct ccg tgg cca tgg gct 48 Met Asn Pro Thr Ala Thr Asn Glu Met Leu Ser Pro Trp Pro Trp Ala 1 5 10 15 gtg aca gag tca aat atc agt ttt gac gtg caa gtg atg gaa caa caa 96 Val Thr Glu Ser Asn Ile Ser Phe Asp Val Gln Val Met Glu Gln Gln 20 25 30 ctt aaa gat ttt agc cgg gca tgt tac gtg gtc aat cat gcc gac cac 144 Leu Lys Asp Phe Ser Arg Ala Cys Tyr Val Val Asn His Ala Asp His 35 40 45 ggc ttt ggt att gcg caa act gcc gat atc gtg act gaa caa gcg gca 192 Gly Phe Gly Ile Ala Gln Thr Ala Asp Ile Val Thr Glu Gln Ala Ala 50 55 60 aac agc aca gat tta cct gtt agt gct ttt act cct gca tta ggt acc 240 Asn Ser Thr Asp Leu Pro Val Ser Ala Phe Thr Pro Ala Leu Gly Thr 65 70 75 80 gaa agc cta ggc gac aat aat ttc cgc cgc gtt cac ggc gtt aaa tac 288 Glu Ser Leu Gly Asp Asn Asn Phe Arg Arg Val His Gly Val Lys Tyr 85 90 95 gct tat tac gca ggc gct atg gca aac ggt att tca tct gaa gag cta 336 Ala Tyr Tyr Ala Gly Ala Met Ala Asn Gly Ile Ser Ser Glu Glu Leu 100 105 110 gtg att gcc cta ggt caa gct ggc att ttg tgt tcg ttt gga gca gcc 384 Val Ile Ala Leu Gly Gln Ala Gly Ile Leu Cys Ser Phe Gly Ala Ala 115 120 125 ggt ctt att cca agt cgc gtt gaa gcg gca att aac cgt att caa gca 432 Gly Leu Ile Pro Ser Arg Val Glu Ala Ala Ile Asn Arg Ile Gln Ala 130 135 140 gcg ctg cca aat ggc cct tat atg ttt aac ctt atc cat agt cct agc 480 Ala Leu Pro Asn Gly Pro Tyr Met Phe Asn Leu Ile His Ser Pro Ser 145 150 155 160 gag cca gca tta gag cgt ggc agc gta gag cta ttt tta aag cat aag 528 Glu Pro Ala Leu Glu Arg Gly Ser Val Glu Leu Phe Leu Lys His Lys 165 170 175 gta cgc acc gtt gaa gca tca gct ttc tta ggt cta aca cca caa atc 576 Val Arg Thr Val Glu Ala Ser Ala Phe Leu Gly Leu Thr Pro Gln Ile 180 185 190 gtc tat tac cgt gca gca gga ttg agc cga gac gca caa ggt aaa gtt 624 Val Tyr Tyr Arg Ala Ala Gly Leu Ser Arg Asp Ala Gln Gly Lys Val 195 200 205 gtg gtt ggt aac aag gtt atc gct aaa gta agt cgc acc gaa gtg gct 672 Val Val Gly Asn Lys Val Ile Ala Lys Val Ser Arg Thr Glu Val Ala 210 215 220 gaa aag ttt atg atg cca gcg ccc gca aaa atg cta caa aaa cta gtt 720 Glu Lys Phe Met Met Pro Ala Pro Ala Lys Met Leu Gln Lys Leu Val 225 230 235 240 gat gac ggt tca att acc gct gag caa atg gag ctg gcg caa ctt gta 768 Asp Asp Gly Ser Ile Thr Ala Glu Gln Met Glu Leu Ala Gln Leu Val 245 250 255 cct atg gct gac gac atc act gca gag gcc gat tca ggt ggc cat act 816 Pro Met Ala Asp Asp Ile Thr Ala Glu Ala Asp Ser Gly Gly His Thr 260 265 270 gat aac cgt cca tta gta aca ttg ctg cca acc att tta gcg ctg aaa 864 Asp Asn Arg Pro Leu Val Thr Leu Leu Pro Thr Ile Leu Ala Leu Lys 275 280 285 gaa gaa att caa gct aaa tac caa tac gac act cct att cgt gtc ggt 912 Glu Glu Ile Gln Ala Lys Tyr Gln Tyr Asp Thr Pro Ile Arg Val Gly 290 295 300 tgt ggt ggc ggt gtg ggt acg cct gat gca gcg ctg gca acg ttt aac 960 Cys Gly Gly Gly Val Gly Thr Pro Asp Ala Ala Leu Ala Thr Phe Asn 305 310 315 320 atg ggc gcg gcg tat att gtt acc ggc tct atc aac caa gct tgt gtt 1008 Met Gly Ala Ala Tyr Ile Val Thr Gly Ser Ile Asn Gln Ala Cys Val 325 330 335 gaa gcg ggc gca agt gat cac act cgt aaa tta ctt gcc acc act gaa 1056 Glu Ala Gly Ala Ser Asp His Thr Arg Lys Leu Leu Ala Thr Thr Glu 340 345 350 atg gcc gat gtg act atg gca cca gct gca gat atg ttc gag atg ggc 1104 Met Ala Asp Val Thr Met Ala Pro Ala Ala Asp Met Phe Glu Met Gly 355 360 365 gta aaa ctg cag gtg gtt aag cgc ggc acg cta ttc cca atg cgc gct 1152 Val Lys Leu Gln Val Val Lys Arg Gly Thr Leu Phe Pro Met Arg Ala 370 375 380 aac aag cta tat gag atc tac acc cgt tac gat tca atc gaa gcg atc 1200 Asn Lys Leu Tyr Glu Ile Tyr Thr Arg Tyr Asp Ser Ile Glu Ala Ile 385 390 395 400 cca tta gac gag cgt gaa aag ctt gag aaa caa gta ttc cgc tca agc 1248 Pro Leu Asp Glu Arg Glu Lys Leu Glu Lys Gln Val Phe Arg Ser Ser 405 410 415 cta gat gaa ata tgg gca ggt aca gtg gcg cac ttt aac gag cgc gac 1296 Leu Asp Glu Ile Trp Ala Gly Thr Val Ala His Phe Asn Glu Arg Asp 420 425 430 cct aag caa atc gaa cgc gca gag ggt aac cct aag cgt aaa atg gca 1344 Pro Lys Gln Ile Glu Arg Ala Glu Gly Asn Pro Lys Arg Lys Met Ala 435 440 445 ttg att ttc cgt tgg tac tta ggt ctt tct agt cgc tgg tca aac tca 1392 Leu Ile Phe Arg Trp Tyr Leu Gly Leu Ser Ser Arg Trp Ser Asn Ser 450 455 460 ggc gaa gtg ggt cgt gaa atg gat tat caa att tgg gct ggc cct gct 1440 Gly Glu Val Gly Arg Glu Met Asp Tyr Gln Ile Trp Ala Gly Pro Ala 465 470 475 480 ctc ggt gca ttt aac caa tgg gca aaa ggc agt tac tta gat aac tat 1488 Leu Gly Ala Phe Asn Gln Trp Ala Lys Gly Ser Tyr Leu Asp Asn Tyr 485 490 495 caa gac cga aat gcc gtc gat ttg gca aag cac tta atg tac ggc gcg 1536 Gln Asp Arg Asn Ala Val Asp Leu Ala Lys His Leu Met Tyr Gly Ala 500 505 510 gct tac tta aat cgt att aac tcg cta acg gct caa ggc gtt aaa gtg 1584 Ala Tyr Leu Asn Arg Ile Asn Ser Leu Thr Ala Gln Gly Val Lys Val 515 520 525 cca gca cag tta ctt cgc tgg aag cca aac caa aga atg gcc 1626 Pro Ala Gln Leu Leu Arg Trp Lys Pro Asn Gln Arg Met Ala 530 535 540 <210> 13 <211> 542 <212> PRT <400> 1 Met Asn Pro Thr Ala Thr Asn Glu Met Leu Ser Pro Trp Pro Trp Ala 1 5 10 15 Val Thr Glu Ser Asn Ile Ser Phe Asp Val Gln Val Met Glu Gln Gln 20 25 30 Leu Lys Asp Phe Ser Arg Ala Cys Tyr Val Val Asn His Ala Asp His 35 40 45 Gly Phe Gly Ile Ala Gln Thr Ala Asp Ile Val Thr Glu Gln Ala Ala 50 55 60 Asn Ser Thr Asp Leu Pro Val Ser Ala Phe Thr Pro Ala Leu Gly Thr 65 70 75 80 Glu Ser Leu Gly Asp Asn Asn Phe Arg Arg Val His Gly Val Lys Tyr 85 90 95 Ala Tyr Tyr Ala Gly Ala Met Ala Asn Gly Ile Ser Ser Glu Glu Leu 100 105 110 Val Ile Ala Leu Gly Gln Ala Gly Ile Leu Cys Ser Phe Gly Ala Ala 115 120 125 Gly Leu Ile Pro Ser Arg Val Glu Ala Ala Ile Asn Arg Ile Gln Ala 130 135 140 Ala Leu Pro Asn Gly Pro Tyr Met Phe Asn Leu Ile His Ser Pro Ser 145 150 155 160 Glu Pro Ala Leu Glu Arg Gly Ser Val Glu Leu Phe Leu Lys His Lys 165 170 175 Val Arg Thr Val Glu Ala Ser Ala Phe Leu Gly Leu Thr Pro Gln Ile 180 185 190 Val Tyr Tyr Arg Ala Ala Gly Leu Ser Arg Asp Ala Gln Gly Lys Val 195 200 205 Val Val Gly Asn Lys Val Ile Ala Lys Val Ser Arg Thr Glu Val Ala 210 215 220 Glu Lys Phe Met Met Pro Ala Pro Ala Lys Met Leu Gln Lys Leu Val 225 230 235 240 Asp Asp Gly Ser Ile Thr Ala Glu Gln Met Glu Leu Ala Gln Leu Val 245 250 255 Pro Met Ala Asp Asp Ile Thr Ala Glu Ala Asp Ser Gly Gly His Thr 260 265 270 Asp Asn Arg Pro Leu Val Thr Leu Leu Pro Thr Ile Leu Ala Leu Lys 275 280 285 Glu Glu Ile Gln Ala Lys Tyr Gln Tyr Asp Thr Pro Ile Arg Val Gly 290 295 300 Cys Gly Gly Gly Val Gly Thr Pro Asp Ala Ala Leu Ala Thr Phe Asn 305 310 315 320 Met Gly Ala Ala Tyr Ile Val Thr Gly Ser Ile Asn Gln Ala Cys Val 325 330 335 Glu Ala Gly Ala Ser Asp His Thr Arg Lys Leu Leu Ala Thr Thr Glu 340 345 350 Met Ala Asp Val Thr Met Ala Pro Ala Ala Asp Met Phe Glu Met Gly 355 360 365 Val Lys Leu Gln Val Val Lys Arg Gly Thr Leu Phe Pro Met Arg Ala 370 375 380 Asn Lys Leu Tyr Glu Ile Tyr Thr Arg Tyr Asp Ser Ile Glu Ala Ile 385 390 395 400 Pro Leu Asp Glu Arg Glu Lys Leu Glu Lys Gln Val Phe Arg Ser Ser 405 410 415 Leu Asp Glu Ile Trp Ala Gly Thr Val Ala His Phe Asn Glu Arg Asp 420 425 430 Pro Lys Gln Ile Glu Arg Ala Glu Gly Asn Pro Lys Arg Lys Met Ala 435 440 445 Leu Ile Phe Arg Trp Tyr Leu Gly Leu Ser Ser Arg Trp Ser Asn Ser 450 455 460 Gly Glu Val Gly Arg Glu Met Asp Tyr Gln Ile Trp Ala Gly Pro Ala 465 470 475 480 Leu Gly Ala Phe Asn Gln Trp Ala Lys Gly Ser Tyr Leu Asp Asn Tyr 485 490 495 Gln Asp Arg Asn Ala Val Asp Leu Ala Lys His Leu Met Tyr Gly Ala 500 505 510 Ala Tyr Leu Asn Arg Ile Asn Ser Leu Thr Ala Gln Gly Val Lys Val 515 520 525 Pro Ala Gln Leu Leu Arg Trp Lys Pro Asn Gln Arg Met Ala 530 535 540
【図１】 本発明のプラスミドの一態様であるｐＪＲＤ
ＥＰＡ−Ｓの構造を示す。
ＥＰＡ−Ｓの構造を示す。
【手続補正書】
【提出日】平成１２年８月２９日（２０００．８．２
９）
９）
【手続補正１】
【補正対象書類名】明細書
【補正対象項目名】００１３
【補正方法】変更
【補正内容】
【００１３】実施例１ 小型化プラスミドの作製 特開平8-242867に記載のプラスミドpEPAに挿入されたEP
A生合成遺伝子群のうち、EPA生合成に必須であるORF3、
6、7、8および9のサブクローニングを行った。ORF5、
6、7、8および9については、クローニングベクターpBSI
IKS(+)（Stratagene社製）のXbaI-SpeI部位にXbaI-SpeI
断片（23,045-31,443）、XbaI部位にXbaI-XbaI断片（1
2,314-23,045）SpeI部位にSpeI-NheI断片（31,443-32,5
14）を順次サブクローニングを行いΔX4XbNh/pBS を作
製した。ΔX4XbNh/pBS をNotIで処理したものをT4DNAポ
リメラーゼにより平滑末端を作り、それをXhoIで処理し
て DNA断片Aを得た。 また、ORF3については、R/pSTV28
（HpaI断片7,951-9,129を宝酒造製ベクターpSTV28のSma
I部位に挿入したもの）をEcoRI及びPstIで処理して切り
出した断片をpBSIIKS(+)のEcoRI-PstI部位に挿入してR/
pBSを作製した。R/pBSをPstIで処理後T4DNAポリメラー
ゼにより平滑末端を作りXhoIリンカーを導入してからXh
oIでORF3を含む断片を切り出しDNA断片Bを得た。広域宿
主ベクターであるpJRD215（カナマイシン及びストレプ
トマイシン耐性)のXhoI-StuI部位に断片Aをパッカジン
ラムダDNAパッケージングシステム（Promega社製）によ
り導入した後に、XhoI部位に断片BをDNA ライゲーショ
ンキット （宝酒造製）を用いて導入しプラスミドを完
成させた。これをpJRDEPA-Sと命名した（図1）。
A生合成遺伝子群のうち、EPA生合成に必須であるORF3、
6、7、8および9のサブクローニングを行った。ORF5、
6、7、8および9については、クローニングベクターpBSI
IKS(+)（Stratagene社製）のXbaI-SpeI部位にXbaI-SpeI
断片（23,045-31,443）、XbaI部位にXbaI-XbaI断片（1
2,314-23,045）SpeI部位にSpeI-NheI断片（31,443-32,5
14）を順次サブクローニングを行いΔX4XbNh/pBS を作
製した。ΔX4XbNh/pBS をNotIで処理したものをT4DNAポ
リメラーゼにより平滑末端を作り、それをXhoIで処理し
て DNA断片Aを得た。 また、ORF3については、R/pSTV28
（HpaI断片7,951-9,129を宝酒造製ベクターpSTV28のSma
I部位に挿入したもの）をEcoRI及びPstIで処理して切り
出した断片をpBSIIKS(+)のEcoRI-PstI部位に挿入してR/
pBSを作製した。R/pBSをPstIで処理後T4DNAポリメラー
ゼにより平滑末端を作りXhoIリンカーを導入してからXh
oIでORF3を含む断片を切り出しDNA断片Bを得た。広域宿
主ベクターであるpJRD215（カナマイシン及びストレプ
トマイシン耐性)のXhoI-StuI部位に断片Aをパッカジン
ラムダDNAパッケージングシステム（Promega社製）によ
り導入した後に、XhoI部位に断片BをDNA ライゲーショ
ンキット （宝酒造製）を用いて導入しプラスミドを完
成させた。これをpJRDEPA-Sと命名した（図1）。
【手続補正２】
【補正対象書類名】明細書
【補正対象項目名】００２５
【補正方法】変更
【補正内容】
【００２５】
【配列表】 <110> Sagami Chemical Reserach Center, Japan Bioindustry Association, Director-General of Agency of Industrial Science and Technology. <120> A plasmid carrying the eicosapentaenoic acid synthesis gene claste r and a transgenic cyanobacterium producing eicosapentaenoic acid. <130> SO18226 <160> 13
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ テーマコート゛(参考） (Ｃ１２Ｎ 1/12 (Ｃ１２Ｐ 7/64 Ｃ１２Ｒ 1:89） Ｃ１２Ｒ 1:89） (Ｃ１２Ｐ 7/64 Ｃ１２Ｎ 15/00 ＺＮＡＡ Ｃ１２Ｒ 1:89） Ｃ１２Ｒ 1:89） (72)発明者 湯 玲子 神奈川県横浜市緑区霧が丘３−24−４− 108 (72)発明者 山田 章子 神奈川県相模原市若松６−１−27−Ｂ− 305 (72)発明者 松永 是 東京都小金井市本町４−20−15 (72)発明者 竹山 春子 東京都府中市幸町２−40−Ｃ−106 (72)発明者 倉根 隆一郎 茨城県つくば市東１丁目１番３ 工業技術 院生命工学工業技術研究所内 Ｆターム(参考） 4B024 AA03 BA07 CA04 DA06 GA14 HA01 4B064 AD90 CA02 CA08 CA19 CC24 DA16 4B065 AA01X AA01Y AA26X AB01 BA02 BA22 CA13
Claims (3)
- 【請求項１】 配列番号２，４，６，８，１０，及び１
２で示される塩基配列によってコードされたイコサペン
タエン酸生合成酵素群をコードする遺伝子群を、広域宿
主ベクターにクローニングして得られるプラスミド。 - 【請求項２】 配列番号３，５，７，９，１１，及び１
３で示される塩基配列で表されるイコサペンタエン酸生
合成酵素群をコードする遺伝子群を、広域宿主ベクター
にクローニングして得られるプラスミド。 - 【請求項３】 請求項１又は２に記載のプラスミドを導
入して得られるイコサペンタエン酸を産生するラン藻。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32916999A JP4221476B2 (ja) | 1999-11-19 | 1999-11-19 | イコサペンタエン酸生合成遺伝子群をクローニングしたプラスミド及びイコサペンタエン酸を産生するラン藻 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32916999A JP4221476B2 (ja) | 1999-11-19 | 1999-11-19 | イコサペンタエン酸生合成遺伝子群をクローニングしたプラスミド及びイコサペンタエン酸を産生するラン藻 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2001145490A true JP2001145490A (ja) | 2001-05-29 |
| JP4221476B2 JP4221476B2 (ja) | 2009-02-12 |
Family
ID=18218426
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP32916999A Expired - Lifetime JP4221476B2 (ja) | 1999-11-19 | 1999-11-19 | イコサペンタエン酸生合成遺伝子群をクローニングしたプラスミド及びイコサペンタエン酸を産生するラン藻 |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4221476B2 (ja) |
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| US11623911B2 (en) | 2013-12-18 | 2023-04-11 | Commonwealth Scientific And Industrial Research Organisation | Lipid comprising docosapentaenoic acid |
| US10793507B2 (en) | 2014-06-27 | 2020-10-06 | Commonwealth Scientific And Industrial Research Organisation | Lipid compositions comprising triacylglycerol with long-chain polyunsaturated fatty acids at the SN-2 position |
| US10005713B2 (en) | 2014-06-27 | 2018-06-26 | Commonwealth Scientific And Industrial Research Organisation | Lipid compositions comprising triacylglycerol with long-chain polyunsaturated fatty acids at the sn-2 position |
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