CN108587995A - 用于生产鼠李糖脂的细胞和方法 - Google Patents

用于生产鼠李糖脂的细胞和方法 Download PDF

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CN108587995A
CN108587995A CN201810413325.0A CN201810413325A CN108587995A CN 108587995 A CN108587995 A CN 108587995A CN 201810413325 A CN201810413325 A CN 201810413325A CN 108587995 A CN108587995 A CN 108587995A
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S·沙费尔
M·韦塞尔
A·蒂森胡森
N·施泰因
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Abstract

本发明涉及用于生产鼠李糖脂的细胞和核酸及其用途,并且还涉及用于生产鼠李糖脂的方法。

Description

用于生产鼠李糖脂的细胞和方法
本申请是申请日为2011年7月20日,申请号为201180037007.5,题 目为“用于生产鼠李糖脂的细胞和方法”的专利申请的分案申请。
发明领域
本发明涉及用于生产鼠李糖脂的细胞和核酸以及其用途,并且还涉及 用于生产鼠李糖脂的方法。
现有技术
表面活性剂目前基本上以石油化学原料为基础生产。由于可预见的石 油化学原料的短缺以及对基于可再生原料或生物可降解的产品增加的需 求,使用基于可再生原料的表面活性剂是合适的替代选择。
鼠李糖脂由一个(单鼠李糖脂)或两个鼠李糖基(双鼠李糖脂)以及一个或 两个3-羟基脂肪酸残基组成(参见,Handbook of Hydrocarbon and Lipid Microbiology,2010,第3037-51页)。它们具有表面活性的性能,这在各种各 样的作为表面活性剂的应用中是必需的(参见Leitermann et al.,2009)。
目前,这些脂类利用不同的人-和动物-致病菌的野生型分离菌,特别是 假单胞菌属(Pseudomonas)和伯克氏菌属(Burkholderia)的代表菌来产生(参 见,Handbook ofHydrocarbon and Lipid Microbiology,2010,第3037-51页)。 这些生产生物体能够导致疾病的事实大大降低了客户对常规生产的鼠李糖 脂的接受度。而且,由于增加的资本支出和可能的额外工作步骤,更高的 安全要求也对生产成本产生了影响。
尽管,借助这些生产生物体在一定程度上可以实现高产物滴定度以及 高时空和/或碳收率,但这需要使用植物油作为唯一的或共同的底物(参见, Handbook ofHydrocarbon and Lipid Microbiology,2010,第3037-51页)。然而, 与其他碳源相比,植物油是相对昂贵的原料,所述其他碳源诸如例如葡萄 糖、蔗糖或多糖诸如例如淀粉、纤维素和半纤维素,甘油,CO,CO2或CH4。 而且,鼠李糖脂由于它们的表面活性剂特性区分它们自身,因为它们对发 酵过程中的大量泡沫是敏感的。如果应用亲脂性底物,特别会这样。该问题在使用水溶性底物诸如例如葡萄糖、蔗糖、多糖(粉、纤维素和半纤维素) 或甘油时会被显著缓解。最后,由野生型分离菌产生的鼠李糖脂的性能仅 在有限程度上被影响。到目前为止,这仅通过优化工艺管理(pH、氧供给、 介质组成、给料策略、氮气供给、温度、底物的选择等)发生。然而,对某 些产物特性诸如例如不同鼠李糖脂种类的比例(鼠李糖和3-羟基脂肪酸基的 数目)或3-羟基脂肪酸基的链长度和饱和度的非常具体的影响会是可取的, 以便能够调节与应用有关的产物性能。
如果鼠李糖脂在家用、清洁、美容、食品加工、药物、植物保护和其 他应用中被大范围地用作表面活性剂,鼠李糖脂必须看起来能与目前应用 的表面活性剂竞争。这些是高容量化学品,其能够以非常低的成本生产, 对客户没有明显的健康风险,并且具有清楚定义的和可调节的产品规格。 因此鼠李糖脂也必须能够以尽可能低的成本生产,对客户没有健康风险并 且具有尽可能明确的性能。
尽管鼠李糖脂已经基于方便的碳源诸如例如葡萄糖或甘油在GRAS生 物体(一般被视为安全的)中被产生,但在这种情况下这些仅仅是单鼠李糖脂 (Ochsner etal.Appl.Environ.Microbiol.1995.61(9):3503-3506)。
另一方面,Cha et al.in Bioresour Technol.2008.99(7):2192-9描述了通 过导入来自绿脓假单胞菌(Pseudomonas aeruginosa)的rhlA和rhlB基因,在 恶臭假单胞菌(P.putida)中生产来自大豆油的单鼠李糖脂。
因此,对廉价以及安全(从健康的角度)地生产具有明确的和可调节性能 的单-和双鼠李糖脂存在日益增加的需求。例如通过减少单鼠李糖脂富集的 单独酶活性的平衡供给,可以实现这种调节。然而,该调节还可以通过例 如利用具有某些性能的酶来实现,所述性能例如与底物特异性相关的性能, 并因此例如与并入鼠李糖脂的羟基脂肪酸的链长度相关。
因此,本发明的目的是提供利用安全的生产宿主从易获得的碳源生产 鼠李糖脂的可能性。
发明内容
出人意料地,已经发现,下文描述的细胞和其中应用这些细胞的方法 有助于解决本发明阐述的目的。
因此,本发明涉及能够形成鼠李糖脂的细胞,其与它们的野生型相比, 具有基因产物rhlA,rhlB和rhlC同源物的基因产物至少一种增加的活性。
本发明还涉及利用作为生物催化剂的前述细胞和简单的碳源生产鼠李 糖脂的方法。
本发明的优势在于能够应用非致病性并且培养简单的生物体。
另外的优势在于作为唯一的或共同底物的油的使用不是必需的。
另一优势在于借助于本发明,可以生产具有明确的和可调节性能的鼠 李糖脂。
本发明的另一优势在于可以生产双鼠李糖脂。
另外的优势在于与这些活性没有增强的细胞相比,可以生产更高时空 和碳收率的鼠李糖脂。
细胞,优选分离的细胞对实现开始时提及的目的作出了贡献,所述细 胞能够形成至少一种通式(I)的鼠李糖脂或其盐,
其中
m=2、1或0,特别为1或0,
n=1或0,特别为1,
R1和R2=彼此独立地为具有2-24个、优选5-13个碳原子的相同或不同 的有机基团,特别任选地分支的,任选地取代的、特别是羟基取代的,任 选地不饱和的,特别任选地单-、双-或三-不饱和的烃基,优选地选自戊烯 基、庚烯基、壬烯基、十一碳烯基和十三碳烯基以及(CH2)o-CH3的烃基, 其中o=1-23,优选为4-12;
其特征在于它已被进行遗传学修饰,从而使得与其野生型相比,所述 细胞中的酶E1、E2和E3中的至少一种的活性是增加的,其中酶E1能够催 化3-羟基链烷酰-ACP经由3-羟基链烷酰-3-羟基链烷酸-ACP转化为羟基链 烷酰-3-羟基链烷酸;酶E2是鼠李糖基转移酶I,并能够催化dTDP-鼠李糖 和3-羟基链烷酰-3-羟基链烷酸酯转化为α-L-吡喃鼠李糖基-3-羟基链烷酰-3- 羟基链烷酸酯;以及酶E3是鼠李糖基转移酶II,并能够催化dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基链烷酰-3-羟基链烷酸酯转化为α-L-吡喃鼠李糖 基-(1-2)-α-L-吡喃鼠李糖基-3-羟基链烷酰-3-羟基链烷酸酯,其中这些酶E1、 E2和E3优选选自以下一组:
至少一种酶E1,其选自:
酶E1a,其具有多肽序列Seq ID No.2或具有与参照序列Seq ID No.2 相比其中高达25%、优选高达20%、特别优选高达15%、特别是高达10、 9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、取代或其组合 而被修饰,并仍具有至少10%、优选50%、特别优选80%、特别是超过90% 的具有参照序列Seq ID No.2的酶的酶活性,其中酶E1a的酶活性被理解为 意指优选将3-羟基癸酰基-ACP经由3-羟基癸酰基-3-羟基癸酸-ACP转化为 羟基癸酰基-3-羟基癸酸的能力,
酶E1b,其具有多肽序列Seq ID No.18或具有与参照序列Seq ID No.18 相比其中高达25%、优选高达20%、特别优选高达15%、特别是高达10、 9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、取代或其组合 而被修饰,并仍具有至少10%、优选50%、特别优选80%、特别是超过90% 的具有参照序列Seq ID No.18的酶的酶活性,其中酶E1b的酶活性被理解为 意指优选将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四烷 酸-ACP转化为羟基十四烷酰基-3-羟基十四烷酸的能力,
酶E1c,其具有多肽序列Seq ID No.78或具有与参照序列Seq ID No.78 相比其中高达25%、优选高达20%、特别优选高达15%、特别是高达10、 9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、取代或其组合 而被修饰,并且仍具有至少10%、优选50%、特别优选80%、特别是超过 90%的具有参照序列Seq ID No.78的酶的酶活性,其中酶E1c的酶活性被理 解为意指优选将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十 四烷酸-ACP转化为羟基十四烷酰基-3-羟基十四烷酸的能力,
酶E1d,其具有多肽序列Seq ID No.80,或者其具有的多肽序列与参照 序列Seq IDNo.80相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.80的酶的酶活性,其中酶E1d的酶活性被理解为意指优选将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰 基-3-羟基十四烷酸-ACP转化为羟基十四烷酰基-3-羟基十四烷酸的能力,以 及
酶E1e,其具有多肽序列Seq ID No.82,或者其具有的多肽序列与参照 序列Seq IDNo.82相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.82的酶的酶活性,其中E1e的酶 活性被理解为意指优选将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基 -3-羟基十四烷酸-ACP转化为羟基十四烷酰基-3-羟基十四烷酸的能力;
至少一种酶E2,其具有选自以下的多肽序列:
酶E2a,其具有多肽序列Seq ID No.4,或者其具有的多肽序列与参照序 列Seq IDNo.4相比其中高达25%、优选高达20%、特别优选高达15%、特 别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.4的酶的酶活性,其中E2a的酶 活性被理解为意指优选将dTDP-鼠李糖和3-羟基癸酰基-3-羟基癸酸转化为 α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸的能力,
酶E2b,其具有多肽序列Seq ID No.20,或者其具有的多肽序列与参照 序列Seq IDNo.20相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.20的酶的酶活性,其中E2b的酶 活性被理解为意指优选将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷 酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸的能力,
酶E2c,其具有多肽序列Seq ID No.84,或者其具有的多肽序列与参照 序列Seq IDNo.84相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.84的酶的酶活性,其中E2c的酶 活性被理解为意指优选将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷 酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸的能力,
酶E2d,其具有多肽序列Seq ID No.86,或者其具有的多肽序列与参照 序列Seq IDNo.86相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.86的酶的酶活性,其中E2d的酶 活性被理解为意指优选将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷 酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸的能力,以 及
酶E2e,其具有多肽序列Seq ID No.88,或者其具有的多肽序列与参照 序列Seq IDNo.88相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.88的酶的酶活性,其中E2e的酶 活性被理解为意指优选将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷 酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸的能力;以 及,
至少一种酶E3,其选自:
酶E3a,其具有多肽序列Seq ID No.6,或者其具有的多肽序列与参照 序列Seq IDNo.6相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.6的酶的酶活性,其中E3a的酶 活性被理解为意指优选将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基癸酰基 -3-羟基癸酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基癸酰 基-3-羟基癸酸的能力,
酶E3b,其具有多肽序列Seq ID No.22,或者其具有的多肽序列与参照 序列Seq IDNo.22相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.22的酶的酶活性,其中E3b的酶 活性被理解为意指优选将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷 酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3- 羟基十四烷酰基-3-羟基-十四烷酸的能力,
酶E3c,其具有多肽序列Seq ID No.90,或者其具有的多肽序列与参照 序列Seq IDNo.90相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.90的酶的酶活性,其中E3c的酶 活性被理解为意指优选将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷 酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3- 羟基十四烷酰基-3-羟基-十四烷酸的能力,以及
酶E3d,其具有多肽序列Seq ID No.92,或者其具有的多肽序列与参照 序列Seq IDNo.92相比其中高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且仍具有至少10%、优选50%、特别优选80%、 特别是超过92%的具有参照序列Seq ID No.92的酶的酶活性,其中E3d的酶 活性被理解为意指优选将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷 酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3- 羟基十四烷酰基-3-羟基-十四烷酸的能力。
对于一般概况,对照图1。
本文的细胞“野生型”指其基因组以通过进化天然形成的状态存在的 细胞。该术语用于完整细胞以及单独的基因。因此,术语“野生型”特别 不包括其基因序列已通过重组方法被人为地至少部分修饰的那些细胞或那 些基因。
结合本发明,术语“鼠李糖脂”被理解为意指通式(I)的化合物或其盐。
显而易见地,上文实际指出的酶E1a至E3b的活性仅是从上述酶的宽广 活性谱中的具体的示例性选择;对提及的相应的酶活性,在给定酶的情况 下,都有可利用的可靠的测量方法。因此,显而易见的是,底物具有无支 链、饱和C10-烃基的酶同样(即便任选地具有降低的活性)会转化那些含有 C6-或C16-烃基的底物,其任选地还可以是分支的或不饱和的。
术语“增加的酶活性”优选被理解为意指增加的细胞内活性。
下文增加细胞中的酶活性的实施方案适用于酶E1至E3的活性增加,以 及适用于其活性可以任选地被增加的所有随后提及的酶。
原则上,可以通过以下实现酶活性的增加:增加编码酶的一个基因序 列或多个基因序列的拷贝数,利用强启动子或改良的核糖体结合位点,例 如通过转录调节子消弱基因表达的负性调节,或者放大基因表达的正性调 节,修饰基因的密码子使用,以多种方式增加mRNA或酶的半衰期,修饰 基因表达的调节或者利用编码具有增加的活性的合适酶的基因或等位基 因,以及任选地合用这些措施。按照本发明,利用含有所需的基因、该基 因的等位基因或其部分以及任选地含有使基因的表达成为可能的启动子的 载体,例如通过转化、转导、接合或这些方法的组合来产生遗传学修饰的 细胞。特别通过将基因或等位基因整合在细胞的染色体或染色体外复制的 载体中实现异源表达。
DE-A-100 31 999给出了增加细胞中酶活性可能性的一般概况,如通过 丙酮酸羧化酶所示例的,其并入本文作为参考,以及其与增加细胞中酶活 性可能性有关的公开内容组成了本发明公开内容的一部分。
借助1-维和2-维蛋白凝胶分离以及利用合适的分析软件对凝胶中蛋白 浓度的随后光学鉴定,上文和所有随后提及的酶或基因的表达是可检测的。 如果酶活性的增加仅基于对应基因表达的增加,酶活性增加的量可以通过 比较野生型和遗传学修饰的细胞间的1-维或2-维蛋白分离以简单的方法来 确定。用于制备棒状杆菌情况下的蛋白凝胶以及用于鉴定蛋白的常规方法 是Hermann等描述的操作方法(Electrophoresis,22:1712.23(2001))。蛋白浓 度同样可以通过利用待检测蛋白的特异性抗体的蛋白免疫印迹杂交(Sambrook et al.,Molecular Cloning:a laboratory manual,2nd Ed.Cold SpringHarbor Laboratory Press,Cold Spring Harbor,N.Y.USA,1989)以及随后利用 用于浓度测定的合适软件的光学分析(Lohaus and Meyer(1989)Biospektrum, 5:32-39;Lottspeich(1999)Angewandte Chemie 111:2630-2647)来分析。 DNA-结合蛋白的活性可以通过DNA条带位移分析(也称为凝胶阻滞) (Wilson et al.(2001)Journal ofBacteriology,183:2151-2155)来测量。DNA- 结合蛋白对其他基因表达的作用可以通过多种已充分描述的报告基因分析 方法(Sambrook et al.,Molecular Cloning:alaboratory manual,2nd Ed.Cold Spring Harbor Laboratory Press,Cold SpringHarbor,N.Y.USA,1989)来检测。 可以按照多种描述的方法(Donahue et al.(2000)Journal of Bacteriology 182 (19):5624-5627;Ray et al.(2000)Journal ofBacteriology 182(8):2277-2284; Freedberg et al.(1973)Journal of Bacteriology115(3):816-823)来确定细胞内 的酶活性。如果在下文的实施方案中,没有指明测定某种酶活性的实践方 法,优选通过在Hermann et al.,Electophoresis,22:1712-23(2001),Lohaus et al.,Biospektrum 5:32-39(1998),Lottspeich,Angewandte Chemie 111:2630-2647(1999)和Wilson et al.,Journal of Bacteriology 183:2151-2155 (2001)中描述的方法进行酶活性增加的测定以及酶活性降低的测定。
如果酶活性的增加是通过内源性基因的突变实现的,这种突变可以是 通过诸如例如UV照射或诱变化学品的常规方法随机产生的,或者是通过 诸如缺失、插入和/或核苷酸交换的基因工程选择性产生的。修饰的细胞通 过这些突变获得。特别优选的酶突变体还特别是那些不再被反馈-、产物- 或底物抑制的酶,或者至少与野生型酶相比被抑制的程度降低的酶。
如果酶活性的增加是通过酶合成增加实现的,相应基因的拷贝数是增 加的,或者位于结构基因上游的启动子和调节区或核糖体结合位点被突变。 并入结构基因上游的表达盒以相同的方式发挥作用。另外地,通过诱导型 启动子,在任何所需位置适时增加表达是可能的。然而,此外,“增强子” 也可以被指定到酶基因作为调节序列,其通过RNA聚合酶和DNA间的改 善的相互作用同样引起基因表达的增加。作为延长mRNA生命期的措施的 结果,表达同样被改善。而且,通过阻止酶蛋白的降解,酶活性同样被增 加。基因或基因构建体在本文呈现为具有不同拷贝数的质粒或者被整合在 染色体中并扩增。可选地,所关注基因的过表达还可以通过改变培养基组 成和培养管理来实现。对此,本领域的技术人员可以找到指南,特别是在 Martin et al.(Bio/Technology 5,137-146(1987))、在Guerrero etal.(Genes 138, 35-41(1994))、Tsuchiya and Morinaga(Bio/Technology 6,428-430(1988))、在 Eikmanns et al.(Genes 102,93-98(1991))、在EP-A-0 472 869、在US 4,601,893、在Schwarzer and Pühler(Bio/Technology 9,84-87(1991))、在Reinscheid etal.(Applied and Environmental Microbiology 60,126-132 (1994))、在LaBarre etal.(Journal of Bacteriology 175,1001-1007(1993))、在 WO-A-96/15246、在Malumbreset al.(Genes 134,15-24(1993))、在 JP-A-10-229891、在Jensen and Hammer(Biotechnology and Bioengineering 58, 191-195(1998))以及在众所周知的遗传学和分子生物学教科书中找到。上文 描述的措施如同突变那样也会导致遗传学修饰的细胞。
例如,附加体质粒(episomal plasmid)被用来增加相应的基因的表达。对 本领域技术人员而言,合适的质粒或载体原则上是可用于该目的的所有实 施方案。例如,这种质粒和载体可以从Novagen,Promega,New England Biolabs,Clontech或Gibco BRL公司的手册中获取。此外优选的质粒和载体 可以在Glover,D.M.(1985)DNA cloning:apractical approach,Vol.I-III,IRL Press Ltd.,Oxford;Rodriguez,R.L.andDenhardt,D.T(eds)(1988)Vectors:a survey of molecular cloning vectors andtheir uses,179-204,Butterworth, Stoneham;Goeddel,D.V.(1990)Systems forheterologous gene expression, Methods Enzymol.185,3-7;Sambrook,J.;Fritsch,E.F.and Maniatis,T.(1989), Molecular cloning:a laboratory manual,2nd ed.,ColdSpring Harbor Laboratory Press,New York中找到。
然后通过接合或转化,将含待扩增的基因的质粒载体转化为所需的菌 株。接合的方法在例如et al.,Applied and Environmental Microbiology 60:756-759(1994)中有描述。转化的方法在例如Thierbach et al.,Applied Microbiology andBiotechnology 29:356-362(1988),Dunican and Shivnan, Bio/Technology 7:1067-1070(1989)和Tauch et al.,FEMS Microbiology Let-ters 123:343-347(1994)中有描述。通过“交叉(cross-over)”事件的同源 重组后,产生的菌株包含至少两个拷贝的所关注的基因。
在上文使用的表述的情况下以及在下文实施方案中,“相比其野生型增 加的酶Ex活性”优选一直被理解为意指相应的酶Ex活性增加的系数至少为 2,特别优选至少为10,更优选至少为100,仍更优选为至少1,000,以及 最优选为至少10,000。此外,具有“相比其野生型增加的酶Ex活性”的本 发明的细胞还特别包括以下细胞:其野生型不含有该酶Ex活性或者至少没 有可检测的该酶Ex活性,并且其仅在例如通过过表达增加了酶活性后,才 显示可检测的该酶Ex活性。在这种情况下,术语“过表达”或下文实施方 案中使用的表述“增加表达”还包括以下情形:其中起始细胞例如野生型 细胞不表达酶Ex或者至少没有可检测的酶Ex表达,酶Ex可检测的的合成 只有通过重组方法才被诱导。
不导致给定多肽特性和功能明显变化的给定多肽序列的氨基酸基变化 对本领域技术人员是已知的。因此,例如,“保守性氨基酸”可以互换;这 种合适的氨基酸取代的实例有:丙氨酸取代丝氨酸;精氨酸取代赖氨酸; 天冬酰胺取代谷氨酰胺或组氨酸;天冬氨酸取代谷氨酸;半胱氨酸取代丝 氨酸;谷氨酰胺取代天冬酰胺;谷氨酸取代天冬氨酸;甘氨酸取代脯氨酸; 组氨酸取代天冬酰胺或谷氨酰胺;异亮氨酸取代亮氨酸或缬氨酸;亮氨酸 取代甲硫氨酸或缬氨酸;赖氨酸取代精氨酸或谷氨酰胺或谷氨酸;甲硫氨 酸取代亮氨酸或异亮氨酸;苯丙氨酸取代甲硫氨酸或亮氨酸或酪氨酸;丝 氨酸取代苏氨酸;苏氨酸取代丝氨酸;色氨酸取代酪氨酸;酪氨酸取代色 氨酸或苯丙氨酸;缬氨酸取代异亮氨酸或亮氨酸。同样已知的是,例如以 氨基酸插入或缺失的形式的变化,特别在多肽的N-或C-末端的变化往往不 会对多肽的功能有明显影响。
可以通过破坏含酶活性的细胞来确定酶的活性,破坏的方式是本领域 技术人员已知的,例如借助球磨机、弗氏压碎器或借助超声波粉碎器,以 及随后通过在13,000rpm和4℃下离心10分钟分离细胞、细胞碎片和破坏 助剂诸如例如玻璃珠。利用产生的无细胞的粗提取物,然后进行酶测定和 随后的LC-ESI-MS产物检测。可选地,可以以本领域技术人员已知的方式 通过色谱分析法(诸如镍-氮三乙酸亲和色谱分析、链霉亲和素亲和色谱分析、凝胶过滤色谱分析或离子交换色谱分析)对酶进行富集或另外纯化至同 质性。
然后利用如上文所述获得的样品以下述方式确定酶E1的活性:标准测 定含100μM大肠杆菌(E.coli)ACP、1mMβ-巯基乙醇、200μM丙二酰-辅 酶A、40μM辛酰基-辅酶A(对于E1a)或十二酰基-辅酶A(对于E1b)、100μM NADPH、2μg大肠杆菌FabD、2μg结核分支杆菌(Mycobacterium tuberculosis)FabH、1μg大肠杆菌FabG、0.1M磷酸钠缓冲液,pH 7.0和5μg 的酶E1,终体积为120μL。ACP、β-巯基乙醇和磷酸钠缓冲液在37℃下预 孵育30分钟以完全还原ACP。通过添加酶E1启动反应。2ml已被用HCl 酸化至pH为2.0的水用来终止反应,并随后用2ml氯仿/甲醇(2:1(v:v))萃 取两次。通过离心(16,100g,5min,RT)产生分相。取出底层的有机相,在真 空离心机中完全脱水,并将沉淀物溶解到50μl的甲醇中。未溶解的成分通 过离心(16,100g,5min,RT)沉淀,并且通过LC-ESI-MS分析样品。通过相 应的质量示踪(mass traces)和二级质谱(MS2spectra)的分析进行产物的鉴定。
然后利用如上文所述获得的样品如下确定酶E2的活性:标准测定可以 由185μl10mM tris-HCl(pH 7.5)、10μl 125mM dTDP-鼠李糖和50μl蛋白 粗提取物(约1mg总蛋白)或在溶液中的纯化的蛋白(5μg纯化的蛋白)组成。 通过添加10μl 10mM的3-羟基癸酰基-3-羟基癸酸(对于E2a)或3-羟基-十四 烷酰-3-羟基十四烷酸(对于E2b)的乙醇溶液启动反应,并在30℃下摇动(600 rpm)孵育1小时。随后,反应用1ml丙酮进行处理。未溶解的成分通过离 心(16,100g,5min,RT)沉淀,并且通过LC-ESI-MS分析样品。通过相应的 质量示踪和二级质谱的分析进行产物的鉴定。
然后利用如上文所述获得的样品如下确定酶E3的活性:标准测定可以 由185μl10mM tris-HCl(pH 7.5)、10μl 125mM dTDP-鼠李糖和50μl蛋白 粗提取物(约1mg总蛋白)或在溶液中的纯化的蛋白(5μg纯化的蛋白)组成。 通过添加10μl 10mM的α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸(对于 E3a)或α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸(对于E3b)的乙 醇溶液启动反应,并在30℃下摇动(600rpm)孵育1小时。随后,反应用1ml 丙酮进行处理。未溶解的成分通过离心(16,100g,5min,RT)沉淀,并且通过 LC-ESI-MS分析样品。通过相应的质量示踪和二级质谱的分析进行产物的 鉴定。
本发明的细胞优选是具有增加的下述酶组合活性的那些:
E1、E2、E3、E1E2、E1E3、E2E3和E1E2E3
在组合中,E2、E2E3和E1E2E3,特别是E1E2E3是特别优选的。
在具有增加的酶组合E1E2E3活性的本发明的细胞的优选实施方案中,n 优选=1。
本发明的细胞可以是原核细胞或真核细胞。这些可以是哺乳动物细胞 (诸如例如来自人的细胞)、植物细胞或微生物诸如酵母、真菌或细菌,其中 微生物是特别优选的,并且细菌和酵母是最优选的。
合适的细菌、酵母或真菌特别是那些作为细菌、酵母或真菌株保藏在 DeutscheSammlung von Mikroorganismen und Zellkulturen(德国微生物和细 胞培养物保藏中心(German Collection of Microorganisms and Cell Cultures)) GmbH(DSMZ),Brunswick,Germany的细菌、酵母或真菌。按照本发明合适 的细菌属于http:// www.dsmz.de/species/bacteria.htm列出的属,按照本发明 合适的酵母属于http:// www.dsmz.de/species/yeasts.htm列出的那些属,以及 按照本发明合适的真菌是http:// www.dsmz.de/species/fungi.htm列出的那些。
按照本发明优选的细胞是以下属的那些:曲霉属(Aspergillus)、棒状杆 菌属(Corynebacterium)、短杆菌属(Brevibacterium)、芽孢杆菌属(Bacillus)、 不动杆菌属(Acinetobacter)、产碱杆菌属(Alcaligenes)、乳杆菌属 (Lactobacillus)、副球菌属(Paracoccus)、乳球菌属(Lactococcus)、念珠菌属 (Candida)、毕赤酵母属(Pichia)、汉逊酵母属(Hansenula)、克鲁维酵母菌属 (Kluyveromyces)、酵母菌属(Saccharomyces)、埃希氏杆菌属(Escherichia)、 单胞发酵菌属(Zymomonas)、耶罗威亚酵母属(Yarrowia)、甲基杆菌属 (Methylobacterium)、青枯菌属(Ralstonia)、假单胞菌属(Pseudomonas)、红螺 菌属(Rhodospirillum)、红杆菌属(Rhodobacter)、伯克氏菌属(Burkholderia)、 梭菌属(Clostridium)和贪铜菌属(Cupriavidus),其中构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillus niger)、协腹产碱杆菌(Alcaligenes latus)、巨大 芽胞杆菌(Bacillusmegaterium)、枯草芽孢杆菌(Bacillus subtilis)、黄色短杆 菌(Brevibacteriumflavum)、乳糖发酵短杆菌(Brevibacterium lactofermentum)、 须芒草伯克氏菌(Burkholderia andropogonis)、巴西伯克氏菌(B.brasilensis)、 克里多尼亚伯克氏菌(B.caledonica)、卡瑞苯西思伯克氏菌(B.caribensis)、 石竹伯克氏菌(B.caryophylli)、内生霉菌伯克氏菌(B.fungorum)、唐菖蒲伯 克氏菌(B.gladioli)、格氏伯克氏菌(B.glathei)、荚壳伯克氏菌(B.glumae)、 草伯克氏菌(B.graminis)、医院伯克氏菌(B.hospita)、B.kururiensis、吩嗪伯 克氏菌(B.phenazinium)、瘤状伯克氏菌(B.phymatum)、植物令伯克氏菌(B. phytofirmans)、植物伯克氏菌(B.plantarii)、甘蔗伯克氏菌(B.sacchari)、新 加坡伯克氏菌(B.singaporensis)、水下伯克氏菌(B.sordidicolai)、栖土伯克氏 菌(B.terricola)、热带伯克氏菌(B.tropica)、肿块伯克氏菌(B.tuberum)、乌 汶伯克氏菌(B.ubonensis)、乌拉姆伯克氏菌(B.unamae)、B.xenovorans、洋 葱伯克氏菌(B.anthina)、吡咯伯克氏菌(B.pyrrocinia)、泰国伯克氏菌(B. thailandensis)、布兰克念珠菌(Candida blankii)、皱褶念珠菌(Candida rugosa)、谷氨酸棒杆菌(Corynebacterium glutamicum)、高效棒状杆菌(Corynebacteriumefficiens)、大肠杆菌(Escherichia coli)、多形汉逊酵母 (Hansenula polymorpha)、乳酸克鲁维酵母(Kluveromyces lactis)、扭脱甲基杆 菌(Methylobacterium extorquens)、善变副球菌(Paracoccus versutus)、阿根廷 假单胞菌(Pseudomonas argentinensis)、淤泥假单胞菌(P.borbori)、香茅醇假 单胞菌(P.citronellolis)、淡黄假单胞菌(P.flavescens)、门多萨假单胞菌(P. mendocina)、硝基还原假单胞菌(P.nitroreducens)、食油假单胞菌(P. oleovorans)、类产碱假单胞菌(P.pseudoalcaligenes)、食树脂假单孢菌(P. resinovorans)、稻草假单胞菌(P.straminea)、桔黄假单胞菌(P.aurantiaca)、 致金假单胞菌(P.aureofaciens)、绿针假单胞菌(P.chlororaphis)、草莓假单胞 菌(P.fragi)、隆德假单胞菌(P.lundensis)、腐臭假单胞菌(P.taetrolens)、南 极假单胞菌(P.antarctica)、产氮假单胞菌(P.azotoformans)、福德假单胞菌 ('P.blatchfordae')、油菜假单胞菌(P.brassicacearum)、布氏假单胞菌(P. brenneri)、雪松素假单胞菌(P.cedrina)、皱褶假单胞皱褶假单胞(P. corrugata)、荧光假单胞菌(P.fluorescens)、杰萨假单胞菌(P.gessardii)、黎 巴嫩假单胞菌(P.libanensis)、孟氏假单胞菌(P.mandelii)、边缘假单胞菌(P. marginalis)、地中海假单胞菌(P.mediterranea)、南方假单胞菌(P.meridiana)、米氏假单胞菌(P.migulae)、霉味假单胞菌(P.mucidolens)、东方假单胞菌(P.orientalis)、西洋参假单胞菌(P.panacis)、溶解蛋白假单胞菌(P.proteolytica)、 罗得西亚假单胞菌(P.rhodesiae)、类黄假单胞菌(P.synxantha)、赛维瓦尔假 单胞菌(P.thivervalensis)、托拉氏假单胞菌(P.tolaasii)、韦龙氏假单胞菌(P. veronii)、脱氮假单胞菌(P.denitrificans)、百日咳假单胞菌(P.pertucinogena)、 P.cremoricolorata、黄褐假单胞菌(P.fulva)、蒙氏假单胞菌(P.monteilii)、摩 氏假单胞菌(P.mosselii)、副黄假单胞菌(P.parafulva)、恶臭假单胞菌、巴利 阿里假单胞菌(P.balearica)、斯氏假单胞菌(P.stutzeri)、扁桃假单胞菌(P. amygdali)、洋榛假单胞菌(P.avellanae)、番木瓜假单胞菌(P.caricapapayae)、 菊苣假单胞菌(P.cichorii)、晕斑假单胞菌(P.coronafaciens)、天仙果假单胞 菌(P.ficuserectae)、向日葵假单胞菌('P.helianthi')、苦楝假单胞菌(P.meliae)、 萨氏假单胞菌(P.savastanoi)、丁香假单胞菌(P.syringae)、番茄假单胞菌(P. tomato)、绿黄假单胞菌(P.viridiflava)、松香假单胞菌(P.abietaniphila)、嗜 酸红假单胞菌(P.acidophila)、伞菌假单胞菌(P.agarici)、嗜碱性假单胞菌(P. alcaliphila)、解碱假单胞菌(P.alkanolytica)、淀粉假单胞菌(P.amyloderamosa)、铁角蕨假单胞菌(P.asplenii)、固氮假单胞菌(P.azotifigens)、 大麻假单胞菌(P.cannabina)、隐居假单胞菌(P.coenobios)、结冰假单胞菌(P. congelans)、贡斯坦蒂尼假单胞菌(P.costantinii)、克罗斯韦假单胞菌(P. cruciviae)、德里假单胞菌(P.delhiensis)、外囊假单胞菌(P.excibis)、极端假 单胞菌(P.extremorientalis)、弗雷德里克斯堡假单胞菌(P.frederiksbergensis)、 褐鞘假单胞菌(P.fuscovaginae)、石花菜假单胞菌(P.gelidicola)、格氏假单胞 菌(P.grimontii),籼稻假单胞菌(P.indica)、杰氏假单胞菌(P.jessenii)、晋州 假单胞菌(P.jinjuensis)、基尔假单胞菌(P.kilonensis)、P.knackmussii、韩国 丛毛假单胞菌(P.koreensis)、林氏假单胞菌(P.lini)、藤黄假单胞菌(P.lutea)、 摩拉维亚假单胞菌(P.moraviensis)、耳炎假单胞菌(P.otitidis)、海绵假单胞 菌(P.pachastrellae)、P.palleroniana、罂粟茎黑条斑病假单胞菌(P. papaveris)、烂泥假单胞菌(P.peli)、腐卵假单胞菌(P.perolens)、梨孢假单胞 菌(P.poae)、浦项假单胞菌(P.pohangensis)、嗜冷假单胞菌(P.psychrophila)、 耐冷假单胞菌(P.psychrotolerans)、P.rathonis、爬虫假单胞菌(P.reptilivora)、 树脂假单胞菌(P.resiniphila)、根际假单胞菌(P.rhizosphaerae)、浅红假单胞 菌(P.rubescens)、P.salomonii、P.segitis、败血症假单胞菌(P.septica)、猿 猴假单胞菌(P.simiae)、猪假单胞菌(P.suis)、耐热假单胞菌(P. thermotolerans)、铜绿假单胞菌(P.aeruginosa)、山黄麻假单胞菌(P.tremae)、 平凡假单胞菌(P.trivialis)、P.turbinellae、P.tuticorinensis、阴城假单胞菌(P. umsongensis)、温哥华假单胞菌(P.vancouverensis)、P.vranovensis、黄色海 假单胞菌(P.xanthomarina)、真养雷氏菌(Ralstonia eutropha)、深红红螺菌 (Rhodospirillum rubrum)、类球红杆菌(Rhodobacter sphaeroides)、酿酒酵母 (Saccharomyces cerevisiae)、解脂耶罗维亚酵母(Yarrowia lipolytica)和运动发 酵单胞菌(Zymomonas mobilis),特别是恶臭假单胞菌、大肠杆菌和泰国伯克 氏菌是特别优选的。
按照本发明的优选细胞能够如同野生型不形成鼠李糖脂或不形成可检 测量的鼠李糖脂,以及如同野生型还优选没有酶E1、E2和E3的活性或者没 有可检测的E1、E2和E3的活性。
如果本发明的细胞是能够如同野生型形成具有C6-C16的单-链烷酸酯链 长度的聚羟基链烷酸酯的细胞,本发明是有利的。这类细胞例如是伯克氏 菌属,泰国伯克氏菌(Burkholderia thailandensis);假单胞菌属,恶臭假单胞 菌、绿脓假单胞菌、食油假单胞菌、斯氏假单胞菌、荧光假单胞菌 (Pseudomonas fluorescens)、香茅醇假单胞菌、食树脂假单孢菌(Pseudomonas resinovorans)、睾丸酮丛毛单胞菌(Comamonas testosterone)、嗜水气单胞菌 (Aeromonas hydrophila)、钩虫贪铜菌(Cupriavidus necator)、协腹产碱杆菌和 真养雷氏菌。在这种情况下,按照本发明优选的细胞是遗传学修饰的,从 而使得与其野生型相比,它们能够形成较少的聚羟基链烷酸酯。.
这种细胞在例如De Eugenio et al.,Environ Microbiol.2010.12(1):207-21和Rehm et al.,Appl Environ Microbiol.2001.67(7):3102-9中有描述。
这种与其野生型相比能够形成较少聚羟基链烷酸酯的细胞的特征特别 在于:与其野生型相比,它具有降低的酶E9或E10至少一种的活性。
其中E9代表聚羟基链烷酸酯合成酶,EC:2.3.1.-,特别具有多肽序列Seq IDNo.30或Seq ID No.32,或者其具有的多肽序列与相应的参照序列Seq ID No.30或Seq IDNo.32相比其中有高达25%、优选高达20%、特别优选高 达15%、特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺 失、插入、取代或其组合而被修饰,并且其仍具有至少10%、优选50%、 特别优选80%、特别是超过90%的具有相应的参照序列Seq ID No.30或Seq ID No.32的酶的酶活性,其中酶E9的酶活性被理解为意指优选将3-羟基链 烷酰-辅酶A转化为聚-3-羟基链烷酸,特别是将3-羟基十四烷酰基-辅酶A 转化为聚-3-羟基十四烷酸的能力,以及
E10代表3-羟基链烷酰-ACP:辅酶A转移酶,特别具有多肽序列Seq ID No.34或SeqID No.36,或者其具有的多肽序列与相应的参照序列Seq ID No.34或Seq ID No.36相比其中有高达25%、优选高达20%、特别优选高 达15%、特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺 失、插入、取代或其组合而被修饰,并且其仍具有至少10%、优选50%、 特别优选80%、特别是超过90%的具有相应的参照序列Seq ID No.34或Seq IDNo.36的酶的酶活性,其中酶E10的酶活性被理解为意指将3-羟基链烷酰 -ACP转化为3-羟基链烷酰-辅酶A,特别是将3-羟基链烷酰-ACP转化为3- 羟基十四烷酰基-辅酶A的能力。
对于一般概况,对照图1。
然后通过首先将560μl的100mM tris/HCl,pH 7.5、20μl的35mM在 DMSO中的DTNB以及20μl的41mM 3-羟基癸酰基-辅酶A混合,利用如 上文所述对于酶E1-E3获得的样品来确定酶E9的活性。随后,加入在100μl tris/HCl,pH 7.5中的5μg纯化的酶E9,以及随后在分光光度计中连续记录 412nm处(通过将5,5’-二硫基双(2-硝基苯甲酸)(DTNB)添加到游离SH基团 导致的)消光随着时间的增加(ΔE/min),记录1分钟。
然后利用如上文所述对于酶E1-E3获得的样品来确定酶E10的活性。标 准测定含3mM MgCl2、40μM羟基癸酰基-辅酶A和在50mM tris-HCl,pH 7.5中的20μM大肠杆菌ACP,总体积为200μl。通过添加在50μl tris/HCl, pH 7.5中的5μg纯化的酶启动反应,并在30℃下孵育1小时。通过添加 50%(w/v)的三氯乙酸和10mg/ml的BSA(30μl)终止反应。通过记录在412 nm处的消光随着时间的增加通过分光光度法确定释放的辅酶A,该消光的 增加是通过将5,5’-二硫基双(2-硝基苯甲酸)添加至游离SH基团导致的。
因此,使用的表述“降低的酶Ex活性”优选被理解为意指活性降低的系 数为至少0.5,特别优选至少0.1,更优选至少0.01,甚至更优选至少0.001 以及最优选至少0.0001。表述“降低的活性”还包括没有可检测的活性(“零活 性”)。例如可以通过选择性突变或者通过本领域技术人员已知的降低某一酶 活性的其他措施实现某一酶活性的降低。
用于降低微生物中酶活性的方法是本领域技术人员已知的。
特别地,本文提供了分子生物学技术。本领域技术人员在例如Dubeau etal.2009.BMC Microbiology 9:263;Singh&Microbiology.2008. 154:797-809或Lee et al.FEMS Microbiol Lett.2009.297(1):38-48中找到了 修饰和降低蛋白表达以及伴随的特别针对假单胞菌和伯克氏菌的酶活性降 低,特别是中断特定基因的指南。
按照本发明优选的细胞的特征在于通过修饰包含所述核酸序列之一的 基因实现酶活性的降低,其中所述修饰选自包含以下、优选由以下组成的 组:基因中外源DNA的插入、至少部分基因的缺失、基因序列中的点突变、 RNA干扰(siRNA)、反义RNA或调节序列诸如例如启动子和终止子以及在 基因侧翼的核糖体结合位点的修饰(插入、缺失或点突变)。
在这种情况下,外源DNA被理解为意指对于基因(不是对于生物体)是 “外源”的任何DNA序列,即内源性DNA序列也能够在这种情况下作为 “外源DNA”。
在这种情况下,特异优选的是,基因被选择性标志基因的插入中断, 因此外源性DNA是选择性标志基因,其中优选地,插入通过基因座位的同 源重组来实现。
在本发明的细胞的优选实施方案中,所关注的细胞是恶臭假单胞菌细 胞,其与野生型相比具有降低的聚羟基链烷酸酯合成。这种细胞例如在Ren et al.,JournalApplied Microbiology and Biotechnology 1998Jun,49(6):743-50 中被描述为GPp121、GPp122、GPp123和GPp124,在Huisman et al.,J Biol Chem.1991Feb 5;266(4):2191-8中被描述为GPp104,以及在De Eugenio et al.,Environ Microbiol.2010.12(1):207-21中被描述为KT42C1,以及在 Ouyang et al.Macromol Biosci.2007.7(2):227-33中被描述为KTOY01和 KTOY02,并且是按照本发明优选的细胞。
在本发明的细胞能够形成m=1的鼠李糖脂的情形下,优选的是由R1和R2限定的以下基团
来自3-羟基辛酰基-3-羟基辛酸、3-羟基辛酰基-3-羟基癸酸、3-羟基癸酰基 -3-羟基辛酸、3-羟基辛酰基-3-羟基癸烯酸、3-羟基癸烯酰基-3-羟基辛酸、 3-羟基辛酰基-3-羟基十二烷酸、3-羟基十二酰基-3-羟基辛酸、3-羟基辛酰基 -3-羟基十二碳烯酸、3-羟基十二酰基-3-羟基辛酸、3-羟基癸酰基-3-羟基癸 酸、3-羟基癸酰基-3-羟基癸烯酸、3-羟基癸烯酰基-3-羟基癸酸、3-羟基癸烯 酰基-3-羟基癸烯酸、3-羟基癸酰基-3-羟基十二烷酸、3-羟基十二酰基-3-羟 基癸酸、3-羟基癸酰基-3-羟基十二碳烯酸、3-羟基癸酰基-3-羟基十四碳烯 酸、3-羟基十四烷酰基-3-羟基癸烯酸、3-羟基十二酰基-3-羟基癸酸、3-羟基 癸酰基-3-羟基十四烷酸、3-羟基十四烷酰基-3-羟基癸酸、3-羟基癸酰基-3- 羟基十四碳烯酸、3-羟基十四烯酰基-3-羟基癸酸、3-羟基十二酰基-3-羟基 十二烷酸、3-羟基十二酰基-3-羟基十二烷酸、3-羟基十二酰基-3-羟基十二 碳烯酸、3-羟基十二酰基-3-羟基十四烷酸、3-羟基十四烷酰基-3-羟基十二 烷酸、3-羟基十四烷酰基-3-羟基十四烷酸、3-羟基十六烷酰基-3-羟基十四 烷酸、3-羟基十四烷酰基-3-羟基十六烷酸或3-羟基十六烷酰基-3-羟基十六 烷酸。
对本领域技术人员显而易见的是,本发明的细胞还能够形成具有通式(I) 的不同鼠李糖脂的混合物。
在这种情况下,优选的是,本发明的细胞能够形成具有通式(I)的鼠李 糖脂的混合物,其特征在于在形成的鼠李糖脂超过80%的重量、优选超过 90%重量、特别优选超过90%重量、特别优选超过95%重量中,n=1,以及 通过R1和R2限定的基团在低于形成的鼠李糖脂10%的重量、优选低于5% 的重量、特别优选低于2%的重量中来自3-羟基癸酰基-3-羟基辛酸或3-羟 基辛酰基-3-羟基癸酸,
其中指出的重量百分比涉及形成的所有具有通式(I)的鼠李糖脂之和。
如果本发明的细胞关于E1至E3另外地被遗传学修饰是有利的,从而使 得它与其野生型相比,具有增加的至少一种酶活性,如在下文指定的每一 种情形中,所述酶选自:
至少一种酶E4,即dTTP:α-D-葡萄糖-1-磷酸胸苷转移酶,EC 2.7.7.24, 特别是具有多肽序列Seq ID No.10的酶,或者该酶具有的多肽序列与参照 序列Seq ID No.10相比其中有高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且其仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.10的酶的酶活性,其中酶E4的 酶活性被理解为意指将α-D-葡萄糖-1-磷酸和dTTP转化为dTDP-葡萄糖的 能力,
至少一种酶E5,即dTTP-葡萄糖-4,6-水解酶,EC 4.2.1.46,特别是具有 多肽序列Seq ID No.12的酶,或者该酶具有的多肽序列与参照序列Seq ID No.12相比其中有高达25%、优选高达20%、特别优选高达15%、特别是 高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、取代 或其组合而被修饰,并且其仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.12的酶的酶活性,其中酶E5的 酶活性被理解为意指将dTDP-葡萄糖转化为dTDP-4-脱氢-6-脱氧-D-葡萄糖 的能力,
至少一种酶E6,即dTDP-4-脱氢鼠李糖-3,5-差向异构酶,EC 5.1.3.13, 特别是具有多肽序列Seq ID No.14的酶,或者该酶具有的多肽序列与参照 序列Seq ID No.14相比其中有高达25%、优选高达20%、特别优选高达15%、 特别是高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、 取代或其组合而被修饰,并且其仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.14的酶的酶活性,其中酶E6的 酶活性被理解为意指将dTDP-4-脱氢-6-脱氧-D-葡萄糖转化为dTDP-4-脱氢 -6-脱氧-L-甘露糖的能力,以及
至少一种酶E7,即dTDP-4-脱氢鼠李糖还原酶,EC 1.1.1.133,特别是具 有多肽序列Seq ID No.16的酶,或者该酶具有的多肽序列与参照序列Seq ID No.16相比其中有高达25%、优选高达20%、特别优选高达15%、特别是 高达10、9、8、7、6、5、4、3、2、1%的氨基酸基通过缺失、插入、取代 或其组合而被修饰,并且其仍具有至少10%、优选50%、特别优选80%、 特别是超过90%的具有参照序列Seq ID No.16的酶的酶活性,其中酶E7的 酶活性被理解为意指将dTDP-4-脱氢-6-脱氧-L-甘露糖转化为dTDP-6-脱氧 -L-甘露糖的能力。
通过孵育α-D-葡萄糖-1-磷酸(1.3mM)与dTTP(5mM)和在50μl磷酸 钠缓冲液、pH8.5中的5μg纯化的酶E4,以及在30℃下孵育5、10和20 分钟后通过添加20μl氯仿终止反应,利用如上文所获得的对于酶E1至E3的样品确定酶E4的活性。然后将混合物涡旋并以16,000g在室温下离心5 分钟。将水相转移至新的反应容器,并用80μl的水再次萃取有机相。合并两水相,并通过HPLC进行分析。此处使用Phenosphere ODS2柱(250x 4.6 mm;Phenomenex,Torrance,USA)或Spheresorb ODS2柱(250x 4.6mm; Waters,Milford,USA)。利用0.5MKH2PO4(洗脱液A),以1ml min–1的流 速进行分析物的洗脱15分钟,随后以0.7ml min–1的流速,用高达80%的 洗脱液A和20%甲醇线性梯度洗脱14分钟的时段。然后将从ODS2柱洗脱下的分析物注射入Phenosphere SAX离子交换柱(250x 4.6mm;Phenomenex, Torrance,USA),并且利用1ml min–1的流速和线性甲酸铵梯度(在25分钟2 至600mM)洗脱分析物。然后利用光电二极管阵列检测器(DAD)通过其UV 吸收进行dTDP-葡萄糖的定量。胸苷的吸收最大值在267nm处。通过确证 的核苷酸糖(Sigma-Aldrich,Munich,USA)进行校准。
通过孵育dTDP-α-D-葡萄糖(1.3mM)与在50μl磷酸钠缓冲液、pH 8.5 中的5μg纯化的酶E5,以及在30℃下孵育5、10和20分钟后通过添加20 μl氯仿终止反应,利用如上文所获得的对于酶E1至E3的样品确定酶E5的 活性。然后将混合物涡旋并在室温下以16,000g离心5分钟。将水相转移 至新的反应容器,并用80μl的水再次萃取有机相。合并两水相,并通过HPLC进行分析。此处使用Phenosphere ODS2柱(250x 4.6mm;Phenomenex, Torrance,USA)或Spheresorb ODS2柱(250x 4.6mm;Waters,Milford,USA)。 利用0.5M KH2PO4(洗脱液A),以1ml min–1的流速进行分析物的洗脱15 分钟,随后以0.7ml min–1的流速,用高达80%的洗脱液A和20%甲醇的线 性梯度洗脱14分钟的时段。然后将从ODS2柱洗脱下的分析物注射入 Phenosphere SAX离子交换柱(250x 4.6mm;Phenomenex,Torrance,USA), 并且利用1mlmin–1的流速和线性甲酸铵梯度(在25分钟,2至600mM)洗 脱分析物。然后利用光电二极管阵列检测器(DAD)通过其UV吸收进行 dTDP-葡萄糖和dTDP-4-脱氢-6-脱氧-D-葡萄糖的定量。胸苷的吸收最大值 在267nm处。通过确证的核苷酸糖(Sigma-Aldrich,Munich,USA)进行校准。
通过首先在30℃下孵育dTDP-α-D-葡萄糖(1.3mM)与在50μl磷酸钠 缓冲液、pH8.5中的5μg纯化的酶E5 10分钟,利用如上文获得的对于酶 E1至E3的样品确定酶E6的活性。随后,添加0.5μg纯化的酶E6,在30℃ 下孵育5、10和20分钟后,通过添加20μl氯仿终止反应。然后涡旋混合 物并在室温下以16,000g离心5分钟。将水相转移至新的反应容器,并用80μl的水再次萃取有机相。合并两水相,并通过HPLC进行分析。此处使 用PhenosphereODS2柱(250x 4.6mm;Phenomenex,Torrance,USA)或 Spheresorb ODS2柱(250x 4.6mm;Waters,Milford,USA)。利用0.5M KH2PO4(洗脱液A),以1ml min–1的流速进行分析物的洗脱15分钟,随后 以0.7ml min–1的流速,用高达80%的洗脱液A和20%甲醇的线性梯度洗脱14分钟的时段。然后将从ODS2柱洗脱下的分析物注射入Phenosphere SAX 离子交换柱(250x 4.6mm;Phenomenex,Torrance,USA),并且利用1ml min–1的流速和线性甲酸铵梯度(在25分钟,2至600mM)洗脱分析物。然后利用 光电二极管阵列检测器(DAD)通过其UV吸收进行dTDP-葡萄糖、dTDP-4- 脱氢-6-脱氧-D-葡萄糖和dTDP-6-脱氧-L-甘露糖的定量。胸苷的吸收最大值 在267nm处。通过确证的核苷酸糖(Sigma-Aldrich,Munich,USA)进行校准。
通过首先在30℃下孵育dTDP-α-D-葡萄糖(1.3mM)与在50μl磷酸钠 缓冲液、pH8.5中的5μg纯化的酶E5 10分钟,利用如上文所述获得的对于 酶E1至E3的样品确定酶E7的活性。随后,添加5μg纯化的酶E6和0.5μg 纯化的酶E7以及NADPH(10mM),在30℃下孵育5、10和20分钟后, 通过添加20μl氯仿终止反应。然后涡旋混合物并在室温下以16,000g离心 5分钟。将水相转移至新的反应容器,并用80μl的水再次萃取有机相。合 并两水相,并通过HPLC进行分析。此处使用Phenosphere ODS2柱(250x 4.6 mm;Phenomenex,Torrance,USA)或Spheresorb ODS2柱(250x 4.6mm; Waters,Milford,USA)。利用0.5M KH2PO4(洗脱液A),以1ml min–1的流 速进行分析物的洗脱15分钟,随后以0.7ml min–1的流速,用高达80%的洗脱液A和20%甲醇的线性梯度洗脱14分钟的时段。然后将从ODS2柱洗 脱下的分析物注射入Phenosphere SAX离子交换柱(250x 4.6mm; Phenomenex,Torrance,USA),并且利用1mlmin–1的流速和线性甲酸铵梯度 (在25分钟,2至600mM)洗脱分析物。然后利用光电二极管阵列检测器 (DAD)通过其UV吸收进行dTDP-葡萄糖、dTDP-4-脱氢-6-脱氧-D-葡萄糖、 dTDP-6-脱氧-L-甘露糖和dTDP-4-脱氢-6-脱氧-L-甘露糖的定量。胸苷的吸 收最大值在267nm处。通过确证的核苷酸糖(Sigma-Aldrich,Munich,USA) 进行校准。
按照本发明的细胞是优选的,其具有增加的下述酶组合的活性:
E4E5,E4E6,E4E7,E5E6,E5E7,E6E7,E4E5E6,E4E5E7,E5E6E7,E4E6E7, E4E5E6E7,其中组合E4E5E6E7是特别优选的。
如果本发明的细胞在脂肪酸生物合成中被遗传学修饰从而使得导致脂 酰-ACP和丙二酰-辅酶A转化为3-酮脂酰-ACP和/或3-酮脂酰-ACP转化为 (R)-3-羟基链烷酰-ACP的酶反应增加,按照本发明可以是有利的。另外地 或可选地,按照本发明可以是有利的,如果本发明的细胞在脂肪酸生物合 成中被遗传学修饰从而使得导致(R)-3-羟基链烷酰-ACP转化为反式-2-烯酰 -ACP和/或反式-2-烯酰-ACP转化为脂酰-ACP的酶反应减弱。
如果本发明的细胞在脂肪酸β-氧化中被遗传学修饰,从而使得导致脂 酰-辅酶A转化为反式-2-烯酰-辅酶A和/或反式-2-烯酰-辅酶A转化为(S)-3- 羟基链烷酰-辅酶A的酶反应增加可以是正好一样有利的。另外地或可选地, 按照本发明可以是有利的,如果本发明的细胞在脂肪酸β-氧化中被遗传学 修饰,从而使得导致(S)-3-羟基链烷酰-辅酶A转化为3-酮脂酰-辅酶A和/ 或3-酮脂酰-辅酶A转化为脂酰-辅酶A和乙酰-辅酶A的酶反应被消弱。
对于一般概况,对照图1。
因为本发明的细胞可以有利地用于生产鼠李糖脂,以及因为这些脂类 随后任选地被纯化,如果本发明的细胞相比其野生型具有增加的至少酶E8的活性是有利的,所述酶E8催化通式(I)的鼠李糖脂从细胞输送至周围介质 中。
优选地,在这种情况下,蛋白E8选自:
酶E8,其具有肽序列Seq ID No.8、Seq ID No.24、Seq ID No.26或Seq IDNo.28,或者其具有的多肽序列与相应的参照序列Seq ID No.8、Seq ID No.24、Seq IDNo.26或Seq ID No.28相比其中有高达25%、优选高达20%、 特别优选高达15%、特别是高达10、9、8、7、6、5、4、3、2、1%的氨基 酸基通过缺失、插入、取代或其组合而被修饰,并且其仍具有至少50%、 优选65%、特别优选80%、特别是超过90%的具有相应的参照序列SeqID No.8、Seq ID No.24、Seq ID No.26或Seq ID No.28的酶的酶活性,其中酶 E8的酶活性被理解为意指将通式(I)的鼠李糖脂从细胞输送至周围介质中的 能力。
本发明的细胞的进一步优选的实施方案的特征在于它含有至少一种下 述本发明的核酸或载体。
本发明的细胞可以有利地用于生产鼠李糖脂。因此本发明的另一主题 是本发明的细胞用于生产具有通式(I)的化合物的用途。
本发明的另一主题是用于生产具有通式(I)的鼠李糖脂的方法,其中
m=2、1或0,特别为1或0,
n=1或0,特别为1,
R1和R2=彼此独立地为具有2-24、优选5-13个碳原子的相同或不同 的有机基团,特别任选地分支的,任选地取代的、特别是羟基取代的,任 选地不饱和的、特别任选地单-、双-或三-不饱和的烃基,优选地选自戊烯 基、庚烯基、壬烯基、十一碳烯基和十三碳烯基以及(CH2)o-CH3的烃基, 其中o=1-23,优选为4-12;
所述方法包括以下处理步骤:
I)使本发明的细胞与含碳源的介质接触
II)在使细胞从碳源形成鼠李糖脂成为可能的条件下培养细胞以及
III)任选地分离形成的鼠李糖脂。
为了生产上述产物的目的,可以将本发明的遗传学修饰的细胞在分批 工艺(分批培养)或在分批补料工艺(补料工艺)或重复的分批补料工艺(重复 的补料工艺)中与营养介质连续或间断接触,并由此培养。半连续工艺也是 可能的,如在GB-A-1009370中描述的。已知培养方法的概述描述于Chmiel (“Bioprozesstechnik 1.Einführung in dieBioverfahrenstechnik”[Bioprocess Technology 1.Introduction to the BioprocessTechnique](Gustav Fischer Verlag, Stuttgart,1991))教科书中或Storhas(“Bioreaktoren und periphere Einrichtungen”[Bioreactors and PeripheralDevices],Vieweg Verlag, Brunswick/Wiesbaden,1994)教科书中。
使用的培养基必需以合适的方式满足相应的菌株的需求。例如,对不 同酵母株的培养基的描述包含在“Nonconventional yeast in biotechnology” (Ed.Klaus Wolf,Springer-Verlag Berlin,1996)中。
使用的碳源可以是碳水化合物,诸如例如葡萄糖、蔗糖、阿拉伯糖、 木糖、乳糖、果糖、麦芽糖、糖蜜、淀粉、纤维素和半纤维素,植物和动 物油和脂,诸如例如大豆油、红花油、花生油、大麻籽油、麻风树油、椰 子脂、葫芦油、亚麻籽油、玉米油、婴粟油、月见草油、橄榄油、棕榈仁 油、棕榈油、油菜籽油、芝麻油、向日葵油、葡萄籽油、胡桃油、麦胚油 和椰子油,脂肪酸,诸如例如辛酸、癸酸、月桂酸、肉豆蔻酸、棕榈酸、 棕榈油酸、硬脂酸、花生四烯酸、二十二酸、油酸、亚油酸、亚麻酸、γ- 亚麻酸和其甲酯或乙酯以及脂肪酸混合物,含有上述脂肪酸的单-、双-和三 甘油酯,醇类,诸如例如甘油、乙醇和甲醇,烃类,诸如甲烷,含碳的气 体和气体混合物,诸如CO、CO2、合成气或烟道气,氨基酸类诸如L-谷氨 酸或L-缬氨酸,或者有机酸诸如例如乙酸。这些物质可以单独地被利用或 以混合物被利用。利用碳水化合物特别是单糖、寡糖或多糖作为碳源在US 6,01,494和US 6,136,576中有描述,以及利用烃类特别是烷烃、烯烃和炔烃 以及衍生于其的一元羧酸和衍生于这些一元羧酸的单-、双-和三甘油酯,以 及利用甘油和乙酸是特别优选的。含甘油与辛酸、癸酸、月桂酸、肉豆蔻酸、棕榈酸、棕榈油酸、硬脂酸、花生四烯酸、二十二酸、油酸、亚油酸、 亚麻酸和/或γ-亚麻酸的酯化产物的单-、双-和三甘油酯是尤其特别优选的。
本发明很大的一个优势在于本发明的细胞能够从最简单的碳源诸如葡 糖糖、蔗糖或甘油形成鼠李糖脂,从而使得在本发明的方法中在介质中供 应较长链的碳源不是必需的。因此在本发明的方法的步骤I)中介质不含有 或者不含有可检测量的链长度大于6个碳原子的羧酸或者从其衍生的酯或 甘油酯,即缺乏可用性的情况下,它是有利的。
使用的氮源可以是含有机氮的化合物诸如蛋白胨、酵母膏、肉膏、麦 芽膏、玉米浸渍水、大豆粉和尿素或者无机化合物诸如硫酸铵、氯化铵、 磷酸铵、碳酸铵和硝酸铵、氨、氢氧化铵或氨水。氮源可以单独被利用或 作为混合物被利用。
使用的磷源可以是磷酸、磷酸二氢钾或磷酸氢二钾或者对应的含钠盐。 培养基还必须含有金属盐,诸如例如硫酸镁或硫酸铁,其对于生长是必须 的。最后,必需的生长促进剂诸如氨基酸和维生素可以另外地应用到上述 物质中。而且,合适的前体可以添加至培养基中。所述原料可以在培养期 间以合适的方式以单一批次或进料的形式添加到培养物中。.
诸如氢氧化钠、氢氧化钾、氨或氨水的碱性化合物或者诸如磷酸或硫 酸的酸性化合物适合用于控制培养物的pH。消泡剂诸如例如脂肪酸聚乙二 醇酯可以用于控制泡沫产生。合适的选择性作用物质诸如例如抗生素可以 添加至介质用于维持质粒的稳定性。为了维持有氧条件,氧气或者含氧气 的气体混合物诸如例如空气被掺入培养中。
培养温度通常超过20℃、优选超过25℃,它还可以超过40℃,其中 有利地,培养温度不超过95℃、特别优选不超过90℃以及最优选不超过 80℃。
在本发明的方法的步骤III)中,由细胞形成的鼠李糖脂可以任选地从细 胞和/或营养介质中分离,其中对于分离,本领域技术人员已知的所有用于 从复杂组分中分离低分子量物质的方法都是可以的,诸如例如过滤、萃取、 吸附(色谱分析)或者结晶。
而且,产物相含有生物质残留物和多种杂质,诸如油、脂肪酸和其他 营养介质成分。优选在无溶剂工艺中进行杂质的分离。因此,例如,产物 相可以用水稀释以便于pH的调整。然后可以通过利用酸或碱降低或升高 pH将鼠李糖脂转化为水溶形式,使产物和水相均质化。潜在地,通过在较 高温度例如在60-90℃下孵育以及不停的混合可以辅助鼠李糖脂在水相中 的溶解。通过随后利用碱或酸升高或降低pH,可以将鼠李糖脂再次转化为 非水溶形式,从而使得它们能够容易地从水相中分离。然后用水洗涤产物 相一次或几次以去除水溶性杂质。
例如,可以通过借助合适溶剂、有利地借助有机溶剂的萃取将油残留 物分离出去。烷烃诸如例如正己烷优选用作溶剂。
作为上述无溶剂工艺的替代,利用合适的溶剂例如酯诸如例如乙酸乙 酯或乙酸丁酯可以实现产物从水相的分离。所述的萃取步骤可以以任何所 需的顺序进行。
在这种情况下,优选应用溶剂,特别是有机溶剂。优选将正戊醇作为 溶剂。例如进行蒸馏以去除溶剂。随后,冻干的产物可以进一步被纯化, 例如借助色谱分析法。作为举例,在这一点可以提到借助合适溶剂的沉淀、 借助合适溶剂的萃取、例如借助环糊精或环糊精衍生物的络合、结晶、借 助于色谱分析法的纯化或分离、或者将鼠李糖脂转化为易于分离的衍生物。
可以利用本发明的方法生产的鼠李糖脂同样是本发明的主题,特别还 是上述的鼠李糖脂混合物,其可以利用本发明的方法生产。
可以利用本发明的方法生产的鼠李糖脂和混合物可以有利地应用在清 洁剂、化妆品或药物制剂以及植物保护制剂中。
因此,本发明的另一主题是利用本发明的方法获得的鼠李糖脂用于生 产化妆品、皮肤或药物制剂、用于生产植物保护制剂以及用于生产护理和 清洁剂以及表面活性剂浓缩物的用途。
术语“护理剂(care agents)”在此处被理解为意指满足以下目的的制剂: 维持物品在其原始形式、减少或避免外部影响力(例如,时间、光照、温度、 压力、污染、与其他接触该物品的反应化合物的化学反应)的作用,诸如例 如老化、污染、材料疲劳,甚或改善所需的物品积极特性。对于最后一点, 例如,可以提到改善的头发光泽或者所考虑物品更大的弹性。
“植物保护制剂”被理解为意指它们的制备性质明显用于植物保护的 那些制剂,如果来自由除草剂、杀菌剂、杀虫剂、杀螨剂、杀线虫剂、抗 鸟类破坏的包含物质、植物营养剂和改善土壤结构剂组成的种类的至少一 种化合物包含在制剂中,特别如此。
按照本发明,利用本发明的方法生产的鼠李糖脂优选在用于家政、工 业、特别用于硬面、皮革或纺织品的护理和清洁剂中使用。
分离的核酸提供了对实现目的的贡献,所述核酸在每种情况下含有选 自[A1-G1]、[A2-G2]和[A3-G3]三组的至少一个序列,其中
组[A1-G1]由以下序列组成:
A1a)Seq ID No.1的序列,其中该序列编码蛋白,其能够将3-羟基癸酰 基-ACP经由3-羟基癸酰基-3-羟基癸酰基-ACP转化为3-羟基癸酰基-3-羟基 癸酸,
B1a)无内含子的序列,其来源于A1a)的序列,并与Seq ID No.1的序 列编码相同的蛋白或肽,
C1a)编码包含Seq ID No.2的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将3-羟基癸酰基-ACP经由3-羟基癸酰基-3-羟基癸酰基 -ACP转化为3-羟基癸酰基-3-羟基癸酸,
D1a)与组A1a)-C1a)之一、特别优选组A1a)的序列至少有70%、特别 优选至少有90%、更优选至少有95%以及最优选至少有99%相同性的序列, 其中该序列优选编码蛋白或肽,其能够将3-羟基癸酰基-ACP经由3-羟基癸 酰基-3-羟基癸酰基-ACP转化为3-羟基癸酰基-3-羟基癸酸,
E1a)与组A1a)-D1a)之一、特别优选组A1a)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将3-羟基癸酰基-ACP经由3-羟基癸酰基-3-羟基癸 酰基-ACP转化为3-羟基癸酰基-3-羟基癸酸,
F1a)组A1a)-E1a)之一、特别优选组A1a)的序列的衍生物,其通过至 少一个碱基、优选至少2个碱基、更优选至少5个碱基和最优选至少10个 碱基,但优选不超过100个碱基、特别优选不超过50个碱基和最优选不超 过25个碱基的取代、添加、倒位和/或缺失获得的,其中该衍生物优选编码 蛋白或肽,其能够将3-羟基癸酰基-ACP经由3-羟基癸酰基-3-羟基癸酰基 -ACP转化为3-羟基癸酰基-3-羟基癸酸,
G1a)组A1a)-F1a)之一的序列的互补序列,特别优选组A1a)的序列的 互补序列,
A1b)Seq ID No.17的序列,其中该序列编码蛋白,其能够将3-羟基十 四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟 基十四烷酰基-3-羟基十四烷酸,
B1b)无内含子的序列,其来源于A1b)的序列,并与Seq ID No.17的序 列编码相同的蛋白或肽,
C1b)编码包含Seq ID No.18的氨基酸序列的蛋白或肽的序列,以及所 述蛋白或肽优选能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3- 羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
D1b)与组A1b)-C1b)之一、特别优选组A1b)的序列至少有70%、特别 优选至少有90%、更优选至少有95%以及最优选至少有99%相同性的序列, 其中该序列优选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3- 羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基 十四烷酸,
E1b)与组A1b)-D1b)之一、特别优选组A1b)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基 -3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
F1b)通过组A1b)-E1b)之一、特别优选组A1b)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四 烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,以及
G1b)组A1b)-F1b)之一、特别优选组A1b)的序列的互补序列,以及
A1c)Seq ID No.77的序列,其中该序列编码蛋白,其能够将3-羟基十 四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟 基十四烷酰基-3-羟基十四烷酸,
B1c)无内含子的序列,其来源于A1c)的序列,并与Seq ID No.77的序 列编码相同的蛋白或肽,
C1c)编码包含Seq ID No.78的氨基酸序列的蛋白或肽的序列,以及所 述蛋白或肽优选能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3- 羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
D1c)与组A1c)-C1c)之一、特别优选组A1c)的序列至少有70%、特别 优选至少有90%、更优选至少有95%以及最优选至少有99%相同性的序列, 其中该序列优选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3- 羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基 十四烷酸,
E1c)与组A1c)-D1c)之一、特别优选组A1c)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基 -3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
F1c)通过组A1c)-E1c)之一、特别优选组A1c)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四 烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,以及
G1c)组A1c)-F1c)之一、特别优选组A1c)的序列的互补序列,以及
A1d)Seq ID No.79序列,其中该序列编码蛋白,其能够将3-羟基十四 烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟基 十四烷酰基-3-羟基十四烷酸,
B1d)无内含子的序列,其来源于A1d)的序列,并与Seq ID No.79的序 列编码相同的蛋白或肽,
C1d)编码包含Seq ID No.80的氨基酸序列的蛋白或肽的序列,以及所 述蛋白或肽优选能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3- 羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
D1d)与组A1d)-C1d)之一、特别优选组A1d)的序列至少有70%、特别 优选至少有90%、更优选至少有95%以及最优选至少有99%相同性的序列, 其中该序列优选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3- 羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基 十四烷酸,
E1d)与组A1d)-C1d)之一、特别优选组A1d)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基 -3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
F1d)通过组A1d)-E1d)之一、特别优选组A1d)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四 烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,以及
G1d)组A1d)-F1d)之一、特别优选组A1d)的序列的互补序列,以及
A1e)Seq ID No.81的序列,其中该序列编码蛋白,其能够将3-羟基十 四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟 基十四烷酰基-3-羟基十四烷酸,
B1e)无内含子的序列,其来源于A1e)的序列,并与Seq ID No.81的序 列编码相同的蛋白或肽,
C1e)编码包含Seq ID No.82的氨基酸序列的蛋白或肽的序列,以及所 述蛋白或肽优选能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3- 羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
D1e)与组A1e)-C1e)之一、特别优选组A1e)的序列至少有70%、特别 优选至少有90%、更优选至少有95%以及最优选至少有99%相同性的序列, 其中该序列优选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3- 羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基 十四烷酸,
E1e)与组A1e)-D1e)之一、特别优选组A1e)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基 -3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
F1e)通过组A1e)-E1e)之一、特别优选组A1e)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四 烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,和
G1e)组A1e)-F1e)之一、特别优选组A1e)的序列的互补序列,以及
组[A2-G2],其由以下序列组成:
A2a)Seq ID No.3的序列,其中该序列编码蛋白,其能够将dTDP-鼠李 糖和3-羟基癸酰基-3-羟基癸酸转化为α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟 基癸酸,
B2a)无内含子序列,其来源于A2a)的序列,并与Seq ID No.3序列编 码相同的蛋白或肽,
C2a)编码包含Seq ID No.4的氨基酸序列的蛋白或肽的序列,以及所 述蛋白或肽优选能够将dTDP-鼠李糖和3-羟基癸酰基-3-羟基癸酸转化为 α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸,
D2a)与组A2a)-C2a)之一、特别优选组A2a)的序列至少有80%、特别 优选至少有90%、更优选至少有95%以及最优选至少有99%相同性的序列, 其中该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基癸酰基-3- 羟基癸酸转化为α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸,
E2a)与组A2a)-D2a)之一、特别优选组A2a)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基癸酰基-3-羟基癸酸转化 为α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸,
F2a)通过组A2a)-E2a)之一、特别优选组A2a)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和3-羟基癸酰基-3-羟基癸酸转化为α-L-吡喃 鼠李糖基-3-羟基癸酰基-3-羟基癸酸,
G2a)组A2a)-F2a)之一、特别优选组A2a)的序列的互补序列,
A2b)Seq ID No.19的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基 十四烷酰基-3-羟基十四烷酸,
B2b)无内含子的序列,其来源于A2b)的序列,并与Seq ID No.19的序 列编码相同的蛋白或肽,
C2b)编码包含Seq ID No.20的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化 为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
D2b)与组A2b)-C2b)之一、特别优选组A2b)的序列有至少70%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3- 羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷 酸,
E2b)与组A2b)-D2b)之一、特别优选组A2b)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四 烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
F2b)通过组A2b)-E2b)之一、特别优选组A2b)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为 α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,和
G2b)组A2b)-F2b)之一、特别优选组A2b)的序列的互补序列,
A2c)Seq ID No.83的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基 十四烷酰基-3-羟基十四烷酸,
B2c)无内含子的序列,其来源于A2c)的序列,并与Seq ID No.83的序 列编码相同的蛋白或肽,
C2c)编码包含Seq ID No.84的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化 为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
D2c)与组A2c)-C2c)之一、特别优选组A2c)的序列有至少70%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3- 羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷 酸,
E2c)与组A2c)-D2c)之一、特别优选组A2c)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四 烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
F2c)通过组A2c)-E2c)之一、特别优选组A2c)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为 α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,和
G2c)组A2c)-F2c)之一、特别优选组A2c)的序列的互补序列,
A2d)Seq ID No.85的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基 十四烷酰基-3-羟基十四烷酸,
B2d)无内含子的序列,其来源于A2d)的序列,并与Seq ID No.85的序 列编码相同的蛋白或肽,
C2d)编码包含Seq ID No.86的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化 为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
D2d)与组A2d)-C2d))之一、特别优选组A2d)的序列有至少70%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3- 羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷 酸,
E2d)与组A2d)-D2d)之一、特别优选组A2d)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四 烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
F2d)通过组A2d)-E2d)之一、特别优选组A2d)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为 α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,和
G2d)组A2d)-F2d)之一、特别优选组A2d)的序列的互补序列,以及
A2e)Seq ID No.87的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基 十四烷酰基-3-羟基十四烷酸,
B2e)无内含子的序列,其来源于A2e)的序列,并与Seq ID No.87的序 列编码相同的蛋白或肽,
C2e)编码包含Seq ID No.88的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化 为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
D2e)与组A2e)-C2e)之一、特别优选组A2e)的序列有至少70%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3- 羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷 酸,
E2e)与组A2e)-D2e)之一、特别优选组A2e)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四 烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
F2e)通过组A2e)-E2e)之一、特别优选组A2e)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为 α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,和
G2e)组A2e)-F2e)之一、特别优选组A2e)的序列的互补序列,以及
组[A3-G3],其由下述序列组成:
A3a)Seq ID No.5的序列,其中该序列编码蛋白,其能够将dTDP-鼠李 糖和α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸转化为α-L-吡喃鼠李糖基 -(1–2)-α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基-癸酸,
B3a)无内含子的序列,其来源于A3a)的序列,并与Seq ID No.5的序 列编码相同的蛋白或肽,
C3a)编码包含Seq ID No.6的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基 癸酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟 基-癸酸,
D3a)与组A3a)-C3a)之一、特别优选组A3a)的序列有至少80%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3- 羟基癸酰基-3-羟基癸酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基 -3-羟基癸酰基-3-羟基癸酸,
E3a)与组A3a)-D3a)之一、特别优选组A3a)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基癸酰 基-3-羟基癸酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基癸 酰基-3-羟基-癸酸,
F3a)通过组A3a)-E3a)之一、特别优选组A3a)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸 酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基- 癸酸,
G3a)组A3a)-F3a)之一、特别优选组A3a)的序列的互补序列,
A3b)Seq ID No.21的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基-十四烷酸转化为α-L- 吡喃鼠李糖基-(1-2)-α-L-吡喃鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
B3b)无内含子的序列,其来源于A3b)的序列,并与Seq ID No.21的序 列编码相同的蛋白或肽,
C3b)编码包含Seq ID No.22的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3- 羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十 四烷酰-3-羟基十四烷酸,
D3b)与组A3b)-C3b)之一、特别优选组A3b)的序列有至少60%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3- 羟基十四烷酰基-3-羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃 鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
E3b)与组A3b)-D3b)之一、特别优选组A3b)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四 烷酰基-3-羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1-2)-α-L-吡喃鼠李糖基 -3-羟基十四烷酰-3-羟基十四烷酸,
F3b)通过组A3b)-E3b)之一、特别优选组A3b)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟 基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十四 烷酰-3-羟基十四烷酸,以及
G3b)组A3b)-F3b)之一、特别优选组A3b)的序列的互补序列,
A3c)Seq ID No.89的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基-十四烷酸转化为α-L- 吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
B3c)无内含子的序列,其来源于A3c)的序列,并与Seq ID No.89的序 列编码相同的蛋白或肽,
C3c)编码包含Seq ID No.90的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3- 羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十 四烷酰-3-羟基十四烷酸,
D3c)与组A3c)-C3c)之一、特别优选组A3c)的序列有至少60%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3- 羟基十四烷酰基-3-羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃 鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
E3c)与组A3c)-D3c)之一、特别优选组A3c)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四 烷酰基-3-羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基 -3-羟基十四烷酰-3-羟基十四烷酸,
F3c)通过组A3c)-E3c)之一、特别优选组A3c)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟 基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十四 烷酰-3-羟基十四烷酸,和
G3c)组A3c)-F3c)之一、特别优选组A3c)的序列的互补序列,以及
A3d)Seq ID No.91的序列,其中该序列编码蛋白,其能够将dTDP-鼠 李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基-十四烷酸转化为α-L- 吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
B3d)无内含子的序列,其来源于A3d)的序列,并与Seq ID No.91的序 列编码相同的蛋白或肽,
C3d)编码包含Seq ID No.92的氨基酸序列的蛋白或肽的序列,所述蛋 白或肽优选能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3- 羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十 四烷酰-3-羟基十四烷酸,
D3d)与组A3d)-C3d)之一、特别优选组A3d)的序列有至少60%、特别 优选至少90%、更优选至少95%以及最优选至少99%相同性的序列,其中 该序列优选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3- 羟基十四烷酰基-3-羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃 鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
E3d)与组A3d)-D3d)之一、特别优选组A3d)的序列的互补链杂交的序 列,或者考虑到遗传密码子的简并性,会与其杂交的序列,其中该序列优 选编码蛋白或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四 烷酰基-3-羟基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基 -3-羟基十四烷酰-3-羟基十四烷酸,
F3d)通过组A3d)-E3d)之一、特别优选组A3d)的序列的至少一个碱基、 优选至少2个碱基、更优选至少5个碱基和最优选至少10个碱基,但优选 不超过100个碱基、特别优选不超过50个碱基和最优选不超过25个碱基 的取代、添加、倒位和/或缺失获得的衍生物,其中该衍生物优选编码蛋白 或肽,其能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟 基-十四烷酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基十四 烷酰-3-羟基十四烷酸,和
G3d)组A3d)-F3d)之一、特别优选组A3d)的序列的互补序列。
本文借助已知的方法确定“核苷酸相同性”或“氨基酸相同性”。一般 地,考虑到特殊需求,使用特定的带算法的计算机程序。
目前用于确定相同性的优选方法产生待比较序列间的最大相同性。用 于确定相同性的计算机程序包括但不限于GCG程序包,包括:GAP(Deveroy, J.et al.,Nucleic AcidResearch 12(1984),第387页,Genetics Computer Group University of Wisconsin,Medicine(Wi))以及BLASTP、BLASTN和FASTA (Altschul,S.et al.,Journal of MolecularBiology 215(1990),第403-410页)。 BLAST程序可以从美国国家生物技术信息中心(NCBI)获得,以及从另外的 来源(BLAST handbook,Altschul S.et al.,NCBI NLM NIH BethesdaND 22894;Altschul S.et al.,上述)获得。
已知的Smith-Waterman算法也可以用于确定核苷酸相同性。
当使用BLASTN程序(Altschul,S.et al.,Journal of Molecular Biology 215(1990),第403-410页)时,用于确定“核苷酸相同性”的优选参数为:
期望阈值: 10
字长: 28
匹配分值: 1
错配分值: -2
空位罚分: 线性罚分
上述参数在核苷酸序列比较中是缺省参数。
GAP程序也适合使用上述参数。
当使用BLASTP程序(Altschul,S.et al.,Journal of Molecular Biology 215(1990),第403-410页)时,用于确定“氨基酸相同性”的优选参数为:
期望阈值: 10
字长: 3
矩阵: BLOSUM62
空位罚分: 存在罚分:11;延伸罚分:1
组成性调整(Compositional adjustments):条件组成性分数矩阵调整(Conditional compositional score matrix adjustment)
上述参数在氨基酸序列比较中是缺省参数。
GAP程序也适合使用上述参数。
按照上述算法的60%相同性意味着与本发明有关的60%相同性。这适 用于更高的相同性。
特征“与序列的互补链杂交的序列,或者考虑到遗传密码子的简并性, 会与其杂交的序列”指在优选严格的条件下,与参照序列的互补链杂交的 序列,或者考虑到遗传密码子的简并性,会与其杂交的序列。例如,按照 Boehringer公司(Mannheim)的地高辛标记试剂盒的操作方案,可以在2x SSC中于68℃下进行杂交。例如,优选的杂交条件是在7%SDS、1%BSA、 1mM EDTA、250mM磷酸钠缓冲液(pH 7.2)中于65℃下孵育过夜,然后在 65℃下用2x SSC、0.1%SDS洗涤。
按照本发明分离的DNA的衍生物,其按照替代选择F1)、F2)或F3)可 以通过按照组A1)-E1)、A2)-E2)和A3)-E3)之一的序列的一个或多个碱基的 取代、添加、倒位和/或缺失来获得,特别包括在它们编码的蛋白中导致保 守性氨基酸交换的那些序列,诸如例如导致甘氨酸交换为丙氨酸或者天冬 氨酸交换为谷氨酸。这种功能中立的突变被描述为有义突变,并不导致多 肽活性的根本改变。此外,已知多肽N-和/或C-末端的变化不显著消弱其功能或者甚至可以稳定该功能,因此其中碱基连接在含本发明的核酸的序列 3’-端或5’-端的DNA序列也相应地包括在本发明中。本领域的技术人员特 别在Ben-Bassat et al.(Journal of Bacteriology 169:751-757(1987))、O'Regan et al.(Gene 77:237-251(1989))、Sahin-Toth et al.(Protein Sciences 3:240-247 (1994))、Hochuli et al.(Bio/Technology 6:1321-1325(1988))和已知的遗传和 分子生物学教科书中找到这方面的信息。
本发明的核酸优选是载体,特别是表达载体或基因过表达盒。合适的 载体是本领域技术人员已知的所有载体,其通常应用于将DNA包含入宿主 细胞中。这些载体既可以自主复制,因为它们具有复制起始点,诸如例如2μ 质粒或ARS(自主复制序列)的那些,或者整合入染色体(非复制质粒)中。载 体还可以被理解为意指根本没有复制起始点的线性DNA片段,诸如例如基 因插入物或基因过表达盒。基因过表达盒通常由标志物、欲过表达的基因以及与基因表达相关的调节区诸如例如启动子和终止子组成。优选的载体 选自质粒和表达盒,诸如例如大肠杆菌酵母穿梭质粒;表达载体、基因插 入物或基因过表达盒是特别优选的,特别是下述的载体Seq ID No.38,Seq ID No.40,Seq ID No.42,Seq ID No.45和Seq ID No.47。
按照本发明的载体的优选实施方案,组[A1-G1]、[A2-G2]和[A3-G3]的 序列在至少一个组成型或可调节的启动子控制下,所述启动子适用于这些 DNA序列编码的多肽在微生物的细胞中表达,优选细菌、酵母或真菌细胞, 其中构巢曲霉、黑曲霉、协腹产碱杆菌、巨大芽胞杆菌、枯草芽孢杆菌、 黄色短杆菌、乳糖发酵短杆菌、须芒草伯克氏菌、巴西伯克氏菌、克里多 尼亚伯克氏菌、卡瑞苯西思伯克氏菌、石竹伯克氏菌、内生霉菌伯克氏菌、 唐菖蒲伯克氏菌、格氏伯克氏菌、荚壳伯克氏菌、草伯克氏菌、医院伯克 氏菌、B.kururiensis、吩嗪伯克氏菌、瘤状伯克氏菌、植物令伯克氏菌、植 物伯克氏菌、甘蔗伯克氏菌、新加坡伯克氏菌、水下伯克氏菌、栖土伯克 氏菌、热带伯克氏菌、肿块伯克氏菌、乌汶伯克氏菌、乌拉姆伯克氏菌、 B.xenovorans、洋葱伯克氏菌、吡咯伯克氏菌、泰国伯克氏菌、布兰克念珠菌、皱褶念珠菌、谷氨酸棒杆菌、高效棒状杆菌、大肠杆菌、多形汉逊酵 母、乳酸克鲁维酵母、扭脱甲基杆菌、善变副球菌、阿根廷假单胞菌、淤 泥假单胞菌、香茅醇假单胞菌、淡黄假单胞菌、门多萨假单胞菌、硝基还 原假单胞菌、食油假单胞菌、类产碱假单胞菌、食树脂假单孢菌、稻草假 单胞菌、桔黄假单胞菌、致金假单胞菌、绿针假单胞菌、草莓假单胞菌、 隆德假单胞菌、腐臭假单胞菌、南极假单胞菌、产氮假单胞菌、福德假单 胞菌、油菜假单胞菌、布氏假单胞菌、雪松素假单胞菌、皱褶假单胞皱褶 假单胞、荧光假单胞菌、杰萨假单胞菌、黎巴嫩假单胞菌、孟氏假单胞菌、 边缘假单胞菌、地中海假单胞菌、南方假单胞菌、米氏假单胞菌、霉味假 单胞菌、东方假单胞菌、西洋参假单胞菌、溶解蛋白假单胞菌、罗得西亚 假单胞菌、类黄假单胞菌、赛维瓦尔假单胞菌、托拉氏假单胞菌、韦龙氏 假单胞菌、脱氮假单胞菌、百日咳假单胞菌、P.cremoricolorata、黄褐假单 胞菌、蒙氏假单胞菌、摩氏假单胞菌、副黄假单胞菌、恶臭假单胞菌、巴 利阿里假单胞菌、斯氏假单胞菌、扁桃假单胞菌、洋榛假单胞菌、番木瓜 假单胞菌、菊苣假单胞菌、晕斑假单胞菌、天仙果假单胞菌、向日葵假单胞菌、苦楝假单胞菌、萨氏假单胞菌、丁香假单胞菌、番茄假单胞菌、绿 黄假单胞菌、松香假单胞菌、嗜酸红假单胞菌、伞菌假单胞菌、嗜碱性假 单胞菌、解碱假单胞菌、淀粉假单胞菌、铁角蕨假单胞菌、固氮假单胞菌、 大麻假单胞菌、隐居假单胞菌、结冰假单胞菌、贡斯坦蒂尼假单胞菌、克 罗斯韦假单胞菌、德里假单胞菌、外囊假单胞菌、极端假单胞菌、弗雷德 里克斯堡假单胞菌、褐鞘假单胞菌、石花菜假单胞菌、格氏假单胞菌,籼稻 假单胞菌、杰氏假单胞菌、晋州假单胞菌、基尔假单胞菌、P.knackmussii、 韩国丛毛假单胞菌、林氏假单胞菌、藤黄假单胞菌、摩拉维亚假单胞菌、 耳炎假单胞菌、海绵假单胞菌、P.palleroniana、罂粟茎黑条斑病假单胞菌、 烂泥假单胞菌、腐卵假单胞菌、梨孢假单胞菌、浦项假单胞菌、嗜冷假单 胞菌、耐冷假单胞菌、P.rathonis、爬虫假单胞菌、树脂假单胞菌、根际假 单胞菌、浅红假单胞菌、P.salomonii、P.segitis、败血症假单胞菌、猿猴假 单胞菌、猪假单胞菌、耐热假单胞菌、铜绿假单胞菌、山黄麻假单胞菌、 平凡假单胞菌、P.turbinellae、P.tuticorinensis、阴城假单胞菌、温哥华假 单胞菌、P.vranovensis、黄色海假单胞菌、真养雷氏菌、深红红螺菌、类球 红杆菌、酿酒酵母、解脂耶罗维亚酵母、运动发酵单胞菌,特别是恶臭假 单胞菌、大肠杆菌和泰国伯克氏菌是特别优选的。组成型启动子的实例有 lac,lacUV5,tac,trc(在每个例子中,在本发明的细胞中缺乏LacI阻遏子)、 Ltet-O1(在本发明的细胞中缺乏TetR阻遏子)、T5和gap。诱导型启动子的 实例有lac,lacUV5,tac,trc(在每个例子中,在本发明的细胞中存在LacI阻 遏子)、Ltet-O1(在本发明的细胞中存在TetR阻遏子)、T5(联合lac操纵子, 和在本发明的细胞中存在LacI阻遏子)、SP6和T7(存在编码同源RNA聚 合酶的基因,其表达本身受调节)。除了启动子外,本发明的载体还应该优 选包含核糖体结合位点以及终止子。此处特别优选的是,本发明的核酸被 并入包含启动子、核糖体结合位点和终止子的载体的表达盒中。除了上述 的结构元件外,载体另外可以包含本领域技术人员已知的选择基因。
所有指出的百分比(%)是质量百分比,如果没有另外指明的话。
在下文呈现的实施例中,通过举例的方式描述了本发明,而不是意欲 将本发明限制在实施例中提及的实施方案中,本发明的应用范围源于整个 说明书和权利要求书。
附图简述
图1:脂肪酸生物合成、脂肪酸β-氧化以及这些代谢途径与鼠李糖脂(酶 E1、E2和E3)和聚羟基链烷酸酯(酶E9和E10)生物合成的联系。显示了脂肪 酸生物合成、脂肪酸β-氧化、鼠李糖脂生物合成和聚羟基链烷酸酯生物合成 中的碳流向。没有显示辅酶的消耗和形成、氧化还原当量以及核苷酸。
图2:在CMP培养基中培养48h、72h和96h后,重组菌株恶臭假单 胞菌KT2440pBBR1MCS-2和pBBR1MCS-2::ABC以及GPp104 pBBR1MCS-2和pBBR1MCS-2::ABC的双鼠李糖脂形成(mg/l/OD 600nm)。 通过HPLC对鼠李糖脂浓度进行分析。
图3:在CMP培养基中培养48h、72h和96h后,重组菌株恶臭假单 胞菌KT2440pBBR1MCS-2,pBBR1MCS-2::AB和pBBR1MCS-2::ABM以及 GPp104pBBR1MCS-2,pBBR1MCS-2::AB和pBBR1MCS-2::ABM的单鼠李 糖脂形成(峰面积/OD 600nm)。通过HPLC对鼠李糖脂浓度进行分析。
实施例
1.用于在恶臭假单胞菌中异源表达绿脓假单胞菌1707基因rhlA和rhlB的 载体pBBR1MCS-2::AB的构建
为了异源表达绿脓假单胞菌DSM1707基因rhlA和rhlB,构建了质粒 pBBR1MCS-2::AB(Seq ID No.38)。为此,通过GeneArt AG公司(Regensburg) 合成了合成性操纵子rhlAB(Seq ID No.37),并间克隆(intercloned)在商品化 载体pMA(GeneArt AG)中。合成的基础是已知的绿脓假单胞菌DSM1707 的基因组序列。从载体pMA::AB开始,通过BglII和XbaI从载体切割下合 成性操纵子,并随后连接在用BamHI和XbaI切割的表达载体pBBR1MCS-2(Seq ID No.49)中(描述在Kovach et al.,1995:Four new derivatives of the broadhost range cloning vector pBBR1MCS carrying different antibiotic-resistancecassettes.Gene,166:175-176中)。产生的质粒 pBBR1MCS-2::AB(Seq ID No.38)大小为7422个碱基对。以本领域技术人员 已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL,Karlsruhe)的 连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2(Seq ID No.49)和pBBR1MCS-2::AB 进行恶臭假单胞菌KT2440和GPp104的转化。分离并分析了10个克隆的 质粒DNA。获得的携带质粒的菌株被命名为恶臭假单胞菌KT2440 pBBR1MCS-2、恶臭假单胞菌GPp104pBBR1MCS-2、恶臭假单胞菌KT2440pBBR1MCS-2::AB和恶臭假单胞菌GPp104pBBR1MCS-2::AB。
2.用于在恶臭假单胞菌中异源表达绿脓假单胞菌DSM1707基因rhlA、rhlB 和rhlC的载体pBBR1MCS-2::ABC的构建
为了异源表达绿脓假单胞菌DSM1707基因rhlA、rhlB和rhlC,构建 了质粒pBBR1MCS-2::ABC(Seq ID No.40)。为此,通过GeneArt AG公司 (Regensburg)合成了合成性操纵子rhlABC(Seq ID No.39),并间克隆在商品 化载体pMA(GeneArt AG)中。合成的基础是已知的绿脓假单胞菌DSM1707 的基因组序列。从载体pMA::ABC开始,通过BglII和XbaI从载体切割下 合成性操纵子,并随后连接在用BamHI和XbaI切割的表达载体 pBBR1MCS-2(Seq ID No.49)中(Kovach et al.,1995:Four new derivatives of the broad hostrange cloning vector pBBR1MCS carrying different antibiotic-resistancecassettes.Gene,166:175-176。产生的质粒 pBBR1MCS-2::ABC(Seq ID No.40)的大小为8409个碱基对。以本领域技术 人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL,Karlsruhe) 的连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::ABC进行恶臭假单胞菌KT2440 和GPp104的转化。分离并分析每10个克隆的质粒DNA。获得的携带质粒 的菌株被命名为恶臭假单胞菌KT2440pBBR1MCS-2::ABC和恶臭假单胞菌 GPp104pBBR1MCS-2::ABC。
3.用于在恶臭假单胞菌中异源表达绿脓假单胞菌DSM1707基因rhlA、rhlB 和pa1131的载体pBBR1MCS-2::ABM的构建
为了异源表达绿脓假单胞菌DSM1707基因rhlA,rhlB和pa1131,构建 了质粒pBBR1MCS-2::ABM(Seq ID No.42)。为此,通过GeneArt AG公司 (Regensburg)合成了合成性操纵子rhlAB-pa1131(Seq ID No.41),并间克隆在 商品化载体pMA(GeneArt AG)中。合成的基础是已知的绿脓假单胞菌 DSM1707的基因组序列。从载体pMA::ABM开始,通过BglII和XbaI从载 体切割下合成性操纵子,并随后连接在用BamHI和XbaI切割的表达载体pBBR1MCS-2(Seq ID No.49)中(Kovach et al.,1995:Four new derivatives of thebroad host range cloning vector pBBR1MCS carrying different antibiotic-resistance cassettes.Gene,166:175-176)。产生的质粒 pBBR1MCS-2::ABM(Seq IDNo.42)的大小为8702个碱基对。以本领域技术 人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL,Karlsruhe) 的连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::ABM进行恶臭假单胞菌KT2440 和GPp104的转化。分离并分析每10个克隆的质粒DNA。获得的携带质粒 的菌株被命名为恶臭假单胞菌KT2440pBBR1MCS-2::ABM和恶臭假单胞 菌GPp104pBBR1MCS-2::ABM。
4.通过重组恶臭假单胞菌菌株产生的鼠李糖脂的定量
将重组菌株恶臭假单胞菌KT2440pBBR1MCS-2、恶臭假单胞菌 KT2440pBBR1MCS-2::AB、恶臭假单胞菌KT2440pBBR1MCS-2::ABC、恶 臭假单胞菌KT2440pBBR1MCS-2::ABM、恶臭假单胞菌GPp104 pBBR1MCS-2、恶臭假单胞菌GPp104pBBR1MCS-2::AB、恶臭假单胞菌GPp104pBBR1MCS-2::ABC和恶臭假单胞菌GPp104pBBR1MCS-2::ABM 培养在LB琼脂卡那霉素(50μg/ml)平板上。
对于鼠李糖脂的产生,使用下文指定为CMP培养基的培养基。该培养 基由2%(w/v)葡萄糖、0.007%(w/v)KH2PO4、0.11%Na2HPO4x 2H2O、0.2% (w/v)NaNO3、0.04%(w/v)MgSO4x H2O、0.01%(w/v)CaCl2x 2H2O和0.2% (v/v)微量元素溶液组成。微量元素溶液由0.2%(w/v)FeSO4x 7H2O、0.15% (w/v)MnSO4x H2O和0.06%(w/v)(NH4)MO7O24x 4H2O组成。用NaOH将 培养基的pH调整到6.7,并随后通过高压灭菌器(121℃,20min)对培养基进 行灭菌。在培养期间不需要调整pH。
为了研究鼠李糖脂在摇动瓶中的产生,首先准备预培养物。为此,使 用在LB琼脂平板上新划线的菌株的接种环,并将10ml LB培养基接种在 100ml锥形烧瓶中。所有重组的恶臭假单胞菌菌株在LB培养基中,将50 μg/ml卡那霉素添加在LB培养基中。在30℃和200rpm下,培养菌株过夜。
预培养物用于接种在250ml的锥形烧瓶中的50ml CMP培养基(起始 OD600 0.1)。在200rpm和30℃下,培养培养物至多120h。在24h的间隔, 从培养瓶中移出1ml肉汤样品。如下进行用于下述色谱分析的样品制备:
利用移液管(Combitip),将1ml丙酮加入2ml反应容器中,然后立即 封闭反应容器以使蒸发最小化。随后加入1ml肉汤。将肉汤/丙酮混合物涡 旋后,在13,000rpm下离心3min,然后将800μl的上清转移至HPLC容器 中。
为了检测和定量鼠李糖脂,使用蒸发光散射检测器(Sedex LT-ELSD Model85LT)。通过Agilent Technologies 1200系列(Santa Clara,California)和 Zorbax SB-C8快速分辨柱(4.6x 150mm,3.5μm,Agilent)进行实际的测量。 注射体积为5μl,以及该方法的运行时间为20min。作为流动相,使用水 性0.1%TFA(三氟乙酸,溶液A)和甲醇(溶液B)。柱温度为40℃。ELSD(检 测器温度为60℃)和DAD(二极管阵列,210nm)作为检测器。该方法中使用 梯度为:
t[min] 溶液B vol.% 流速[ml/min]
0.00 70% 1.00
15.00 100% 1.00
15.01 70% 1.00
20.00 70% 1.00
尽管恶臭假单胞菌KT2440pBBR1MCS-2和GPp104pBBR1MCS-2没 有产生鼠李糖脂,在重组菌株恶臭假单胞菌KT2440pBBR1MCS-2::AB、恶 臭假单胞菌KT2440pBBR1MCS-2::ABC、恶臭假单胞菌KT2440 pBBR1MCS-2::ABM、恶臭假单胞菌GPp104pBBR1MCS-2::AB、恶臭假单 胞菌GPp104pBBR1MCS-2::ABC和恶臭假单胞菌GPp104 pBBR1MCS-2::ABM中,可检测到不同鼠李糖脂种类的形成(图2和3)。
通过将pBBR1MCS-2::AB和pBBR1MCS-2::ABM合并入恶臭假单胞 菌,可能生成单鼠李糖脂(图3)。因为不存在单鼠李糖脂的参考资料,所以 通过相应的质量示踪和LC-MS中的二级质谱的分析进行产物的鉴定。
如果rhlC(pBBR1MCS-2::ABC)被另外地合并入菌株中,产生单-和双鼠 李糖脂(图2)。
对通过恶臭假单胞菌pBBR1MCS-2::AB和恶臭假单胞菌 pBBR1MCS-2::ABM形成的鼠李糖脂的直接比较显示,铜绿假单胞菌p3111 与铜绿假单胞菌rhlAB的共表达导致鼠李糖脂生物合成的改善(图3)。尽管 菌株恶臭假单胞菌KT2440pBBR1MCS-2::AB和恶臭假单胞菌GPp104 pBBR1MCS-2::AB在120小时后产生了约39个(恶臭假单胞菌KT2440 pBBR1MCS-2::AB)和23个(恶臭假单胞菌GPp104pBBR1MCS-2::AB)峰 面积鼠李糖脂/OD 600nm,菌株恶臭假单胞菌KT2440pBBR1MCS-2::ABM 和恶臭假单胞菌GPp104pBBR1MCS-2::ABM在120小时后形成了约50个 (恶臭假单胞菌KT2440pBBR1MCS-2::ABM)以及62个(恶臭假单胞菌GPp104pBBR1MCS-2::ABM)峰面积鼠李糖脂/OD 600nm。
如果比较菌株恶臭假单胞菌KT2440pBBR1MCS-2::ABM和恶臭假单 胞菌GPp104pBBR1MCS-2::ABM的单鼠李糖脂合成,可能在PHA-阴性的 突变恶臭假单胞菌GPp104pBBR1MCS-2::ABM中检测到62个峰面积/OD 600nm(120h培养),以及用恶臭假单胞菌KT2440pBBR1MCS-2::ABM, 检测到50个面积/OD 600nm的单鼠李糖脂(图3)。
对在菌株恶臭假单胞菌KT2440和GPp104中的双鼠李糖脂形成的比较 分析也表明,在恶臭假单胞菌GPp104的PHA-阴性菌株背景中有更多的双 鼠李糖脂形成。恶臭假单胞菌GPp104pBBR1MCS-2::ABC形成了平均113 mg/l/OD 600nm的双鼠李糖脂(96h),而用恶臭假单胞菌KT2440 pBBR1MCS-2::ABC,在96h后,仅能够检测到55mg/l/OD 600nm的双鼠 李糖脂(图2)。
因此可能表明,利用PHA合成减弱的菌株背景导致鼠李糖脂生物合成 的改善。
5.用于在恶臭假单胞菌中异源表达绿脓假单胞菌DSM1707基因rhlA、 rhlB、pa1131和rhlC的载体pBBR1MCS-2::ABMC的构建
为了异源表达绿脓假单胞菌DSM1707基因rhlA、rhlB、pa1131和rhlC, 构建了质粒pBBR1MCS-2::ABMC(Seq ID No.51)。为此,通过GeneArt AG 公司(Regensburg)合成了合成性操纵子rhlAB-pa1131-rhlC(Seq ID No.50), 并间克隆在商品化载体pMA(GeneArt AG)中。合成的基础是已知的绿脓假 单胞菌DSM1707的基因组序列。从载体pMA::ABMC开始,通过BglII和 XbaI从载体切割下合成性操纵子,并随后连接在用BamHI和XbaI切割的 表达载体pBBR1MCS-2(Seq ID No.49)中(Kovach et al.,1995:Four new derivatives of thebroad-host-range cloning vector pBBR1MCS carrying different antibiotic-resistance cassettes.Gene,166:175-176)。产生的质粒 pBBR1MCS-2::ABMC(Seq IDNo.51)的大小为9663个碱基对。以本领域技 术人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL, Karlsruhe)的连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::ABMC进行恶臭假单胞菌KT2440 和GPp104的转化。分离并分析每10个克隆的质粒DNA。获得的携带质粒 的菌株被命名为恶臭假单胞菌KT2440pBBR1MCS-2::ABMC和恶臭假单胞 菌GPp104pBBR1MCS-2::ABMC。
6.通过重组恶臭假单胞菌株和铜绿假单胞菌株产生的鼠李糖脂的定量比较
将重组菌株恶臭假单胞菌GPp104pBBR1MCS-2和恶臭假单胞菌 GPp104pBBR1MCS-2::ABMC以及铜绿假单胞菌DSM 19880培养在LB 琼脂卡那霉素(50μg/ml;恶臭假单胞菌)和LB琼脂(铜绿假单胞菌)平板上。
为了产生鼠李糖脂,使用下文被指定为CMP培养基的培养基。该培养 基由2%(w/v)葡萄糖、0.007%(w/v)KH2PO4、0.11%Na2HPO4x 2H2O、0.2% (w/v)NaNO3、0.04%(w/v)MgSO4x H2O、0.01%(w/v)CaCl2x 2H2O和0.2% (v/v)的微量元素溶液组成。微量元素溶液由0.2%(w/v)FeSO4x 7H2O、 0.15%(w/v)MnSO4x H2O和0.06%(w/v)(NH4)MO7O24x 4H2O组成。利用 NaOH将培养基的pH调整至6.7,并随后通过高压灭菌器(121℃,20min) 对培养基进行灭菌。在培养期间不需要调整pH。
为了研究鼠李糖脂在摇动瓶中的产生,首先准备预培养物。为此,使 用在LB琼脂平板上新划线的菌株的接种环,并将10ml LB培养基接种在 100ml锥形烧瓶中。将重组的恶臭假单胞菌菌株培养在LB培养基中,将 50μg/ml卡那霉素添加在LB培养基中。将铜绿假单胞菌培养在LB培养基 中。在30℃和200rpm下,培养菌株过夜。
预培养物用于接种在250ml锥形烧瓶中的50ml CMP培养基(起始 OD600 0.1)。在200rpm和30℃下,培养培养物至多120h。在24h的间隔, 从培养瓶中移出1ml肉汤样品。如下进行用于下述色谱分析的样品制备:
利用移液管(Combitip),将1ml丙酮加入2ml反应容器中,然后立即 封闭反应容器以使蒸发最小化。随后加入1ml肉汤。将肉汤/丙酮混合物涡 旋后,在13,000rpm下离心3min,然后将800μl的上清转移至HPLC容器。
为了检测形成的产物,将5μl注射入Accela UPLC单元(Thermo Scientific,Dreieich)。利用半UPLC柱“Pursuit XRs ULTRA(C8,2.8μm,2.1x 100mm)(Varian,Darmstadt)分析待研究的物质。在40℃下,通过由流动相 A1(H2O,0.1%(v/v)TFA)和流动相B1(methanol,0.1%(v/v)TFA)组成的梯 度,利用0.3ml/min的流速,在25分钟内进行分离。梯度的时间进程如下:
时间[min] 流动相A1[%] 流动相B1[%]
0 30 70
15 0 100
25 0 100
25.01 30 70
32 30 70
通过波长范围为200-600nm的DAD检测器和质量进行检测,选择性 利用高分辨率FT-ICR LTQ-FT FT-ICR LTQ-FT质谱仪(Thermo Scientific, Dreieich),扫描范围为m/e100-1000。通过ESI(电喷雾电离)进行电离。借 助FT-ICR质量分析仪,利用分辨率R=100000以及质量精确度≤2ppm, 确定准确的质量和经验化学式。通过分析相应的质量示踪和二级质谱进行 产物的鉴定。为了能够比较菌株,对比了相应物质的峰面积。
菌株恶臭假单胞菌GPp104 pBBR1MCS-2根本没有形成鼠李糖脂。恶 臭假单胞菌BR1MCS-2::ABMC和铜绿假单胞菌DSM 19880形成了鼠李糖 脂,其中恶臭假单胞菌GPp104pBBR1MCS-2::ABMC形成的双-和单鼠李 糖脂的比例例如为4:1,用铜绿假单胞菌DSM 19880形成的双-和单鼠李糖 脂的比例例如为2:1。而且,相比铜绿假单胞菌DSM 19880,菌株恶臭假单 胞菌GPp104 pBBR1MCS-2::ABMC没有形成或者仅形成极少的具有以下基 团的鼠李糖脂:通过R1和R2确定的来自3-羟基辛酰基-3-羟基癸酸或3-羟 基癸酰基-3-羟基-辛酸的基团。
7.用于恶臭假单胞菌中异源表达的载体pBBR1MCS-2::rfbBDAC和 pBBR1MCS-2::ABC_rfbBDAC的构建
在Trenzyme GmbH公司(Konstanz),从臭假单胞菌KT2440的染色体 DNA开始扩增鼠李糖生物合成操纵子rfbBDAC。为此,使用以下引物:
RL1:5’-TATATATAGAATTCGCGTCATCTGTCTACGACAACAC-3’
(Seq ID No.48)
RL2:5’-TATATATAGAATTCGGCTGCGCTACCGCAGCCCTTC-3’(Seq ID No.43)
将获得的PCR产物间克隆在Trenzyme′s短吻鳄(alligator)克隆系统中并 在大肠杆菌DH5α(New England Biolabs;Frankfurt)中转化。分析不同候选物 的载体并测序。进行成功和无误的DNA测序后,通过EcoRI切割载体, 并分离靶片段rfbBDAC。为了进一步的间克隆(inter-cloning),以相同的方式 切割载体pBBR1MCS-2(Kovach et al.,1995:Fournew derivatives of the broad-host-range cloning vecoter pBBR1MCS carryingdifferent antibiotic-resistance cassettes.Gene,166:175-176)。切割的靶片段(rfbBDAC) 和切割的载体(pBBR1MCS-2)通过常规的连接法合并。产生的载体 pBBR1MCS-2::rfbBDAC(Seq ID No.45)也转化在大肠杆菌DH5α(New England Biolabs;Frankfurt)中。关于质粒的成功摄取,研究一些转化株的候 选物。
载体pBBR1MCS-2::rfbBDAC作为PCR的基体。使用了以下寡核苷酸:
RL_XbaI-fw:5’-TATATATATCTAGAATTAATGCAGCTGGCACGAC -3’(Seq ID No.44)
RL_Xba_rev:5’-GGCCGCTCTAGAACTAGTGGA-3’(Seq ID No.46)
利用New England Biolabs(Frankfurt)聚合酶的PhusionTM高保真预混合 物进行PCR。它以本领域技术人员已知的方式进行。将靶序列(lac启动子 和rfbBDAC)间克隆在Trenzyme短吻鳄(alligator)克隆系统中。选择大肠杆菌 DH5α(New England Biolabs;Frankfurt)转化株,以及分离不同候选物的质粒 DNA并测序。检验并研究序列的正确性后,利用XbaI切割载体。通过常规 连接方法,将靶片段连接入同样用XbaI切割的pBBR1MCS-2::ABC(见上文) 中。获得的靶载体pBBR1MCS-2::ABC_rfbBDAC(Seq ID No.47)的大小为12249个碱基对。对载体的插入物进行测序。以本领域技术人员已知的方式 实施PCR、通过琼脂糖凝胶电泳校验PCR的成功扩增、进行DNA的溴化 乙锭染色、确定PCR片段大小、纯化PCR产物以及确定DNA浓度。
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::ABC_rfbBDAC进行恶臭假单胞菌 KT2440和GPp104的转化。分离并分析每10个克隆的质粒DNA。获得的 携带质粒的菌株被命名为恶臭假单胞菌KT2440pBBR1MCS-2::ABC_rfbBDAC和恶臭假单胞菌GPp104 pBBR1MCS-2::ABC_rfbBDAC。
8.通过过表达或不过表达rfbBDAC操纵子的重组恶臭假单胞菌菌株产生 的鼠李糖脂的定量
将重组菌株恶臭假单胞菌KT2440pBBR1MCS-2、恶臭假单胞菌 KT2440pBBR1MCS-2::ABC、恶臭假单胞菌KT2440 pBBR1MCS-2::ABC_rfbBDAC、恶臭假单胞菌GPp104pBBR1MCS-2、恶 臭假单胞菌GPp104pBBR1MCS-2::ABC和恶臭假单胞菌GPp104 pBBR1MCS-2::ABC_rfbBDAC培养在LB琼脂卡那霉素(50μg/ml)平板上。
为了生产鼠李糖脂,使用下文被指定为CMP培养基的培养基。该培养 基由2%(w/v)葡萄糖、0.007%(w/v)KH2PO4、0.11%Na2HPO4x 2H2O、0.2% (w/v)NaNO3、0.04%(w/v)MgSO4x H2O、0.01%(w/v)CaCl2x 2H2O和0.2% (v/v)微量元素溶液组成。微量元素溶液由0.2%(w/v)FeSO4x 7H2O、0.15% (w/v)MnSO4x H2O和0.06%(w/v)(NH4)MO7O24x 4H2O组成。利用NaOH 将培养基的pH调整为6.7,并随后通过高压灭菌器(121℃,20min)对培养基 进行灭菌。在培养期间不需要调整pH。
为了研究鼠李糖脂在摇动瓶中的产生,首先准备预培养物。为此,使 用在LB琼脂平板上新划线的菌株的接种环,并将10ml LB培养基接种在 100ml锥形烧瓶中。所有重组的恶臭假单胞菌菌株都培养在LB培养基中, 将50μg/ml卡那霉素添加在LB培养基中。在30℃和200rpm下,培养恶 臭假单胞菌菌株过夜。
预培养物用于接种在250ml的锥形烧瓶中的50ml CMP培养基(起始 OD600 0.1)。在200rpm和30℃下,将培养物培养至多120h。在24h的间 隔,从培养瓶中移出1ml肉汤样品。如下进行用于以下色谱分析的样品制 备:
利用移液管(Combitip),将1ml丙酮加入2ml反应容器中,然后立即 封闭反应容器以使蒸发最小化。随后加入1ml肉汤。将肉汤/丙酮混合物涡 旋后,在13,000rpm下离心3min,然后将800μl的上清转移至HPLC容器 中。
为了检测和定量鼠李糖脂,使用蒸发光散射检测器(Sedex LT-ELSD Model85LT)。通过Agilent Technologies 1200系列(Santa Clara,California)和 Zorbax SB-C8快速分辨柱(4.6x 150mm,3.5μm,Agilent)进行实际的测量。 注射体积为5μl,以及该方法的运行时间为20min。作为流动相,使用水 性0.1%TFA(三氟乙酸,溶液A)和甲醇(溶液B)。柱温度为40℃。ELSD(检 测器温度为60℃)和DAD(二极管阵列,210nm)作为检测器。该方法中使用 梯度为:
t[min] 溶液B vol.% 流速[ml/min]
0.00 70% 1.00
15.00 100% 1.00
15.01 70% 1.00
20.00 70% 1.00
尽管恶臭假单胞菌KT2440pBBR1MCS-2和GPp104pBBR1MCS-2不 产生鼠李糖脂,在重组菌株恶臭假单胞菌KT2440pBBR1MCS-2::ABC、恶 臭假单胞菌KT2440pBBR1MCS-2::ABC_rfbBDAC、恶臭假单胞菌GPp104 pBBR1MCS-2::ABC和恶臭假单胞菌GPp104pBBR1MCS-2::ABC_rfbBDAC 中,可检测到鼠李糖脂的形成。
相比恶臭假单胞菌KT2440pBBR1MCS-2::ABC,恶臭假单胞菌KT2440 pBBR1MCS-2::ABC_rfbBDAC显示了双-和单鼠李糖脂的形成增加;相比恶 臭假单胞菌GPp104pBBR1MCS-2::ABC,恶臭假单胞菌GPp104 pBBR1MCS-2::ABC_rfbBDAC显示了双-和单鼠李糖脂的形成增加。这清楚 地显示了rfbBDAC表达的扩增对单-和双鼠李糖脂形成的积极影响。
如果比较菌株恶臭假单胞菌KT2440pBBR1MCS-2::ABC_rfbBDAC和 恶臭假单胞菌GPp104pBBR1MCS-2::ABC_rfbBDAC的单-和双鼠李糖脂 的生物合成,在PHA-阴性的突变恶臭假单胞菌GPp104 pBBR1MCS-2::ABC_rfbBDAC中检测到增加的单-和双鼠李糖脂合成。
如上文已经描述的,使用PHA合成被灭活的菌株背景,鼠李糖脂生物 合成增加。
9.重组大肠杆菌W3110pBBR1MCS-2::ABC和大肠杆菌W3110 pBBR1MCS-2::ABC_rfbBDAC的产生
如以前所述的(Miller JH.A Short Course in Bacterial Genetics:ALaboratory Manual and Handbook for Escherichia coli and Related Bacteria.Plainview,NY:Cold Spring Harbor Lab.Press;1992),通过电穿孔法进行大肠 杆菌W3110的转化。分离并分析每10个克隆的质粒DNA。获得的携带质 粒的菌株被命名为大肠杆菌W3110pBBR1MCS-2::ABC_rfbBDAC和大肠 杆菌W3110pBBR1MCS-2::ABC_rfbBDAC。
10.通过过表达和不过表达rfbBDAC操纵子的重组大肠杆菌菌株产生的鼠 李糖脂的定量
将重组菌株大肠杆菌W3110pBBR1MCS-2、大肠杆菌W3110 pBBR1MCS-2::ABC和大肠杆菌W3110pBBR1MCS-2::ABC_rfbBDAC培养 在LB琼脂卡那霉素(50μg/ml)平板上。
为了产生鼠李糖脂,使用下文被指定为CMP培养基的培养基。该培养 基由2%(w/v)葡萄糖、0.007%(w/v)KH2PO4、0.11%Na2HPO4x 2H2O、0.2% (w/v)NaNO3、0.04%(w/v)MgSO4x H2O、0.01%(w/v)CaCl2x 2H2O和0.2% (v/v)的微量元素溶液组成。微量元素溶液由0.2%(w/v)FeSO4x 7H2O、 0.15%(w/v)MnSO4x H2O和0.06%(w/v)(NH4)MO7O24x 4H2O组成。利用 NaOH将培养基的pH调整至6.7,并随后通过高压灭菌器(121℃,20min) 对培养基进行灭菌。在培养期间不需要调整pH。
为了研究鼠李糖脂在摇动瓶中的产生,首先准备预培养物。为此,使 用在LB琼脂平板上新划线的菌株的接种环,并将10ml LB培养基接种在 100ml锥形烧瓶中。所有重组的大肠杆菌菌株培养在LB培养基中,将50 μg/ml卡那霉素添加在LB培养基中。在37℃和200rpm下,培养大肠杆菌 菌株过夜。
预培养物用于接种在250ml的锥形烧瓶中的50ml CMP培养基(起始 OD600 0.1)。在200rpm和30℃下,将培养物培养至多120h。在24h的间 隔,从培养瓶中移出1ml肉汤样品。如下进行用于以下色谱分析的样品制 备:
利用移液管(Combitip),将1ml丙酮加入2ml反应容器中,然后立即 封闭反应容器以使蒸发最小化。随后加入1ml肉汤。将肉汤/丙酮混合物涡 旋后,在13,000rpm下离心3min,然后将800μl的上清转移至HPLC容器 中。
为了检测和定量鼠李糖脂,使用蒸发光散射检测器(Sedex LT-ELSD Model85LT)。通过Agilent Technologies 1200系列(Santa Clara,California)和 Zorbax SB-C8快速分辨柱(4.6x 150mm,3.5μm,Agilent)进行实际的测量。 注射体积为5μl,以及该方法的运行时间为20min。水性0.1%TFA(三氟乙 酸,溶液A)和甲醇(溶液B)用作流动相。柱温度为40℃。ELSD(检测器温 度为60℃)和DAD(二极管阵列,210nm)作为检测器。该方法中使用梯度为:
t[min] 溶液B vol.% 流速[ml/min]
0.00 70% 1.00
15.00 100% 1.00
15.01 70% 1.00
20.00 70% 1.00
尽管大肠杆菌W3110pBBR1MCS-2不产生鼠李糖脂,在重组菌株大肠 杆菌W3110pBBR1MCS-2::ABC和大肠杆菌W3110 pBBR1MCS-2::ABC_rfbBDAC中,可检测到单-和双鼠李糖脂的形成,其中 大肠杆菌W3110pBBR1MCS-2::ABC_rfbBDAC形成的单-和双鼠李糖脂明 显多于大肠杆菌W3110pBBR1MCS-2::ABC。这表明,绿脓假单胞菌 DSM1707的rhlABC的异源表达导致了单-和双鼠李糖脂在大肠杆菌中的形 成。这还表明,rfbBDAC表达的增强对单-和双鼠李糖脂形成的积极影响。 11.用于在恶臭假单胞菌中异源表达绿脓假单胞菌DSM1707基因rhlA、 rhlB和rhlC以及泰国伯克氏菌E264基因BTH_II1077、BT_II1080和 BT_II1081的载体pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081的构建
为了在恶臭假单胞菌中异源表达绿脓假单胞菌DSM1707基因rhlA, rhlB和rhlC以及泰国伯克氏菌E264基因BTH_II1077,BT_II1080和 BT_II1081,构建了质粒pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081(Seq ID No.69)。为此,通过DNA 2.0公司(Menlo Park,CA,USA)合成了合成性 操纵子BTH_II1077,BT_II1080和BT_II1081(Seq ID No.70),并将其间克隆 在商品化载体pJ294(DNA 2.0;Menlo Park,CA,USA)中。合成的基础是菌株 泰国伯克氏菌E264的基因组序列。从载体pJ294-BTH_II1077- II1080-II1081开始,通过XbaI从该载体切割下合成性操纵子,并随后将其 连接在同样用XbaI切割的载体pBBR1MCS-2::ABC(Seq ID No.40)中。获得 的靶载体pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081(Seq IDNo.69)的 大小为13768个碱基对。对载体的插入物进行测序。以本领域技术人员已 知的方式实施PCR、通过琼脂糖凝胶电泳校验PCR的成功扩增、进行DNA 的溴化乙锭染色、确定PCR片段大小、纯化PCR产物以及确定DNA浓度。
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081 (Seq ID No.69)进行恶臭假单胞菌KT2440和GPp104的转化。分离并分析每 10个克隆的质粒DNA。获得的携带质粒的菌株被命名为恶臭假单胞菌 KT2440pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081和恶臭假单胞菌 GPp104pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081。
12.通过过表达和不过表达泰国伯克氏菌E264基因BTH_II1077、BT_II1080 和BT_II1081的重组恶臭假单胞菌菌株产生的鼠李糖脂的定量
将在实施例1、2和11中产生的重组菌株恶臭假单胞菌菌株恶臭假单 胞菌KT2440pBBR1MCS-2::AB、恶臭假单胞菌KT2440 pBBR1MCS-2::AB-BTH_II1077-II1080-II1081、恶臭假单胞菌GPp104 pBBR1MCS-2::AB、恶臭假单胞菌GPp104pBBR1MCS-2::AB-BTH_II1077- II1080-II1081、恶臭假单胞菌KT2440pBBR1MCS-2::ABC、恶臭假单胞菌KT2440pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081、恶臭假单胞菌 GPp104pBBR1MCS-2::ABC和恶臭假单胞菌GPp104 pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081培养在LB琼脂卡那霉素(50 μg/ml)平板上。
为了产生鼠李糖脂,使用下文被指定为M9培养基的培养基。该培养基 由2%(w/v)葡萄糖、0.3%(w/v)KH2PO4、0.679%Na2HPO4、0.05%(w/v)NaCl、 0.2%(w/v)NH4Cl、0.049%(w/v)MgSO4x 7H2O和0.1%(v/v)微量元素溶液 组成。微量元素溶液由1.78%(w/v)FeSO4x7H2O、0.191%(w/v)MnCl2x 7 H2O、3.65%(w/v)HCl、0.187%(w/v)ZnSO4x 7H2O、0.084%(v/v)Na EDTA x 2H2O、0.03%(v/v)H3BO3、0.025%(w/v)Na2MoO4x 2H2O和0.47%(w/v)CaCl2x 2H2O组成。利用NH4OH将培养基的pH调整至7.4,随后通过高 压灭菌器(121℃,20min)对培养基进行灭菌。在培养期间不需要调整pH。
为了研究鼠李糖脂在摇动瓶中的产生,首先准备预培养物。为此,使 用在LB琼脂平板上新划线的菌株的接种环,并将10ml LB培养基接种在 100ml锥形烧瓶中。所有重组的恶臭假单胞菌菌株培养在LB培养基中,将 50μg/ml卡那霉素添加在LB培养基中。在37℃和200rpm下,培养恶臭 假单胞菌菌株过夜。
预培养物用于接种在250ml的锥形烧瓶中的50ml M9培养基(+50 μg/ml卡那霉素)(起始OD600 0.1)。在200rpm和30℃下,培养培养物。在 24h的间隔,从培养瓶中移出1ml肉汤样品。如在实施例4中所述,进行 用于以下色谱分析的样品制备以及色谱分析本身。
重组菌株恶臭假单胞菌KT2440pBBR1MCS-2::AB-BTH_II1077- II1080-II1081和恶臭假单胞菌GPp104pBBR1MCS-2::AB-BTH_II1077- II1080-II1081显示比菌株恶臭假单胞菌KT2440pBBR1MCS-2::AB和恶臭 假单胞菌GPp104pBBR1MCS-2::AB形成了明显增多的单鼠李糖脂。这证 实了来自泰国伯克氏菌E264的BTH_II1077-II1080-II1081的扩增增加了单 鼠李糖脂在含绿脓假单胞菌DSM1707基因rhlAB的恶臭假单胞菌菌株中的 形成。
还显示,重组菌株恶臭假单胞菌KT2440 pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081和恶臭假单胞菌GPp104 pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081比菌株恶臭假单胞菌 KT2440pBBR1MCS-2::ABC和恶臭假单胞菌GPp104pBBR1MCS-2::ABC 形成了明显更多的单-和双鼠李糖脂。这证明了来自泰国伯克氏菌E264的 BTH_II1077-II1080-II1081的扩增增加了单-和双鼠李糖脂在含绿脓假单胞 菌DSM1707基因rhlABC的恶臭假单胞菌菌株中的形成。
最后显示,相比菌株恶臭假单胞菌KT2440,在恶臭假单胞菌GPp104 菌株背景中聚羟基丁酸酯形成的减少导致了鼠李糖脂形成的增加,因为菌 株恶臭假单胞菌KT2440pBBR1MCS-2::AB、恶臭假单胞菌KT2440 pBBR1MCS-2::ABC、恶臭假单胞菌KT2440pBBR1MCS-2::AB-BTH_II1077-II1080-II1081和恶臭假单胞菌KT2440 pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081能够形成比相应的对照菌株 恶臭假单胞菌GPp104pBBR1MCS-2::AB、恶臭假单胞菌GPp104 pBBR1MCS-2::ABC,恶臭假单胞菌GPp104 pBBR1MCS-2::AB-BTH_II1077-II1080-II1081和恶臭假单胞菌GPp104 pBBR1MCS-2::ABC-BTH_II1077-II1080-II1081明显减少的单-()和单-和双 鼠李糖脂()。
13.用于在恶臭假单胞菌中异源表达绿脓假单胞菌DSM1707基因rhlA、 rhlB、pa1131和rhlC的载体pBBR1MCS-2::ABCM的构建
为了异源表达绿脓假单胞菌DSM1707基因rhlA,rhlB,pa1131和rhlC, 构建了质粒pBBR1MCS-2::ABCM(Seq ID No.58)。为此,从含有以下寡核 苷酸的菌株绿脓假单胞菌PAO1(DSM 1707)的基因组DNA开始,扩增基因 pa1131(Seq ID No.59):
MFS2.0_xbaI_fw:5’-AGGAAATCTAGATGAGAGGCCGGCAAGGATAC-3’ (Seq ID No.60)
MFS2.0_XbaI_rev:5’-CCAGGTTCTAGACGCCAGGATTGAACAGTACC-3’ (Seq IDNo.61)。
利用来自New England Biolabs(Frankfurt)聚合酶的PhusionTM高保真预 混合物进行PCR产物(1483个碱基对)的扩增。利用XbaI切割PCR产物并 通过Fast Link连接试剂盒(Epicentre Technologies;Madison,WI,USA)将其连 接在同样用XbaI切割的载体pBBR1MCS-2::ABC(Seq ID No.40)中。获得的 靶载体pBBR1MCS-2::ABCM(Seq ID No.58)的大小为9892个碱基对。对载 体的插入物进行测序。按照生产商的说明书,通过DNeasy血液和组织试剂 盒(Qiagen;Hilden)分离染色体DNA。以本领域技术人员已知的方式实施 PCR、通过琼脂糖凝胶电泳校验PCR的成功扩增、进行DNA的溴化乙锭 染色、确定PCR片段大小、纯化PCR产物以及确定DNA浓度。.
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::ABCM进行恶臭假单胞菌KT2440 和GPp104的转化。分离并分析每10个克隆的质粒DNA。获得的携带质粒 的菌株被命名为恶臭假单胞菌KT2440pBBR1MCS-2::ABCM和恶臭假单 胞菌GPp104pBBR1MCS-2::ABCM。
14.通过过表达或不过表达绿脓假单胞菌DSM1707pa1131基因的重组恶 臭假单胞菌菌株产生的鼠李糖脂的定量
将在实施例2和13中产生的重组菌株恶臭假单胞菌菌株恶臭假单胞菌KT2440pBBR1MCS-2::ABC、恶臭假单胞菌KT2440pBBR1MCS-2::ABCM、 恶臭假单胞菌KT2440pBBR1MCS-2::ABC和恶臭假单胞菌GPp104 pBBR1MCS-2::ABCM培养在LB琼脂卡那霉素(50μg/ml)平板上。随后如实 施例12所述的,进行用于生产鼠李糖脂的培养。
如实施例4中所述的,进行用于以下色谱分析的样品制备以及色谱分 析本身。
结果显示在下表中。
48小时孵育后,通过过表达或不过表达绿脓假单胞菌基因pa1131的恶臭假 单胞菌菌株形成的双-和单鼠李糖脂
结果显示,在两种菌株背景(KT2440:野生型和具有灭活的聚羟基丁酸 酯形成的GPp104)中过表达铜绿假单胞菌基因pa1131导致双-和单鼠李糖脂 形成增加。结果还显示,聚羟基丁酸酯形成在GPp104中的减少一般导致鼠 李糖脂形成增加。
15.用于在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA和rhlB的载体pEC-XT99A::AB的构建
为了在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA和rhlB,构建了质粒pEC-XT99A::AB(Seq ID No.52)。为此,通过 GeneArt AG公司(Regensburg)合成了合成性操纵子rhlAB(Seq ID No.37),并 将其间克隆在商品化的载体pMA(GeneArtAG)中。合成的基础是已知的绿 脓假单胞菌DSM1707的基因组序列。从载体pMA::AB开始,通过BglII 和XbaI从载体切割下合成性操纵子,并随后将其连接入利用BamHI和XbaI 切割的表达载体pEC-XT99A(美国专利7118904)中。产生的质粒 pEC-XT99A::AB(Seq ID No.52)的大小为9793个碱基对。以本领域技术人 员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL,Karlsruhe) 的连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Liebl et al.,FEMS Microbiol.Lett.53:299–303(1989)),利 用载体pEC-XT99A::AB进行谷氨酸棒杆菌ATCC13032的转化。在LBHIS 琼脂平板(18.5g/l的脑心浸出液肉汤、0.5M山梨醇、5g/l的Bacto胰蛋白 胨、2.5g/l的Bacto酵母膏、5g/l的NaCl和18g/l的Bacto琼脂,补加有 5mg/l的四环素)上进行转化株的选择。将平板在33℃孵育两天。获得的携 带质粒的菌株被命名为谷氨酸棒杆菌pEC-XT99A::AB。
16.用于在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA、rhlB和rhlC的载体pEC-XT99A::ABC的构建
为了在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA,rhlB和rhlC,构建了质粒pEC-XT99A::ABC(Seq ID No.53)。为此,通 过GeneArt AG公司(Regensburg)合成了合成性操纵子rhlABC(Seq ID No.39),并将其间克隆在商品化的载体pMA(GeneArt AG)中。合成的基础 是已知的绿脓假单胞菌DSM1707的基因组序列。从载体pMA::ABC开始, 通过BglII和XbaI从载体切割下合成性操纵子,并随后将其连接入利用BamHI和XbaI切割的表达载体pEC-XT99A(美国专利7118904)。产生的质 粒pEC-XT99A::ABC(Seq ID No.53)的大小为10780个碱基对。以本领域技 术人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL, Karlsruhe)的连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Liebl et al.,FEMS Microbiol.Lett.53:299–303(1989)),利 用载体pEC-XT99A::ABC进行谷氨酸棒杆菌ATCC13032的转化。在LBHIS 琼脂平板(18.5g/l的脑心浸出液肉汤、0.5M山梨醇、5g/l的Bacto胰蛋白 胨、2.5g/l的Bacto酵母膏、5g/l的NaCl和18g/l的Bacto琼脂,补加有5 mg/l的四环素)上进行转化株的选择。将平板在33℃孵育两天。获得的携 带质粒的菌株被命名为谷氨酸棒杆菌pEC-XT99A::ABC。
17.用于在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA、rhlB和pa1131的载体pEC-XT99A::ABM的构建
为了在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA,rhlB和pa1131,构建了质粒载体pEC-XT99A::ABM(Seq ID No.54)。 为此,通过GeneArt AG公司(Regensburg)合成了合成性操纵子rhlABM(Seq ID No.41),并将其间克隆在商品化的载体pMA(GeneArt AG)中。合成的基 础是已知的绿脓假单胞菌DSM1707的基因组序列。从载体pMA::ABM开 始,通过BglII和XbaI从载体切割下合成性操纵子,并随后将其连接入利 用BamHI和XbaI切割的表达载体pEC-XT99A(美国专利7118904)。产生的 质粒pEC-XT99A::ABM(Seq ID No.54)的大小为11073个碱基对。以本领域 技术人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL, Karlsruhe)的连接和转化。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Liebl et al.,FEMS Microbiol.Lett.53:299–303(1989)),利 用载体pEC-XT99A::ABM,进行谷氨酸棒杆菌ATCC13032的转化。在 LBHIS琼脂平板(18.5g/l的脑心浸出液肉汤、0.5M山梨醇、5g/l的Bacto 胰蛋白胨、2.5g/l的Bacto酵母膏、5g/l的NaCl和18g/l的Bacto琼脂, 补加有5mg/l的四环素)上进行转化株的选择。将平板在33℃孵育两天。 获得的携带质粒的菌株被命名为谷氨酸棒杆菌pEC-XT99A::ABM。
18.用于在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA、rhlB、pa1131和rhlC的载体pEC-XT99A::ABCM的构建
为了在谷氨酸棒杆菌中异源表达来自绿脓假单胞菌DSM1707的基因 rhlA,rhlB,pa1131和rhlC,构建了质粒pEC-XT99A::ABCM(Seq ID No.55)。 为此,从菌株绿脓假单胞菌PAO1(DSM 1707)的基因组DNA开始,利用以 下的寡核苷酸扩增基因pa1131(Seq IDNo.59):
MFS2.0_xbaI_fw:5’-AGGAAATCTAGATGAGAGGCCGGCAAGGATAC-3’(Seq ID No.60)
MFS2.0_XbaI_rev:5’-CCAGGTTCTAGACGCCAGGATTGAACAGTACC-3’ (Seq IDNo.61)。
利用来自New England Biolabs(Frankfurt)聚合酶的PhusionTM高保真预 混合物进行PCR产物(1483个碱基对)的扩增。利用XbaI切割PCR产物并 通过Fast Link连接试剂盒(Epicentre Technologies;Madison,WI,USA)将其连 接在同样用XbaI切割的载体pBBR1MCS-2::ABC(Seq ID No.40)中。获得的 靶载体pEC-XT99A::ABCM(Seq ID No.55)的大小为12263个碱基对。对载 体的插入物进行测序。按照生产商的说明书,通过DNeasy血液和组织试剂 盒(Qiagen;Hilden)分离染色体DNA。以本领域技术人员已知的方式实施 PCR、通过琼脂糖凝胶电泳校验PCR的成功扩增、进行DNA的溴化乙锭 染色、确定PCR片段大小、纯化PCR产物以及确定DNA浓度。
如以前所述的(Liebl et al.,FEMS Microbiol.Lett.53:299–303(1989)),利 用载体pEC-XT99A::ABCM进行谷氨酸棒杆菌ATCC13032的转化。在 LBHIS琼脂平板(18.5g/l的脑心浸出液肉汤、0.5M山梨醇、5g/l的Bacto 胰蛋白胨、2.5g/l的Bacto酵母膏、5g/l的NaCl和18g/l的Bacto琼脂, 补加有5mg/l的四环素)上进行转化株的选择。将平板在33℃孵育两天。 获得的携带质粒的菌株被命名为谷氨酸棒杆菌pEC-XT99A::ABCM。
19.用于在谷氨酸棒杆菌中异源表达的载体pVWEX1::rfbBDAC的构建
为了在谷氨酸棒杆菌中lac启动子的控制下,异源表达来自恶臭假单胞 菌的基因rfbBDAC,构建了载体pVWEX1::rfbBDAC(Seq ID No.57)。为此, 利用XbaI消化载体pBBR1MCS-2::rfbBDAC(Seq ID No.45),并将含有来自 恶臭假单胞菌KT2440的基因rfbBDAC和lac启动子的片段(3840bp)连接 入用XbaI消化的载体pVWEX1(Seq ID No.56)。产生的质粒 pVWEX1::rfbBDAC(Seq ID No.57)的大小为12311个碱基对。以本领域技 术人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL, Karlsruhe)的连接和转化。通过DNA序列分析来检查插入物的确实性。
如以前所述的(Liebl et al.,FEMS Microbiol.Lett.53:299–303(1989)),利 用载体pVWEX1::rfbBDAC进行谷氨酸棒杆菌ATCC13032pEC-XT99A、 ATCC13032pEC-XT99A::AB、ATCC13032pEC-XT99A::ABM、ATCC13032 pEC-XT99A::ABC和ATCC13032pEC-XT99A::ABCM的转化。在LBHIS 琼脂平板(18.5g/l的脑心浸出液肉汤、0.5M山梨醇、5g/l的Bacto胰蛋白胨、2.5g/l的Bacto酵母膏、5g/l的NaCl和18g/l的Bacto琼脂,补加有5 mg/l的四环素和25mg/l的卡那霉素)上进行转化株的选择。将平板在33℃ 孵育两天。获得的携带质粒的菌株被命名为谷氨酸棒杆菌pEC-XT99A pVWEX1::rfbBDAC、谷氨酸棒杆菌pEC-XT99A::ABpVWEX1::rfbBDAC、 谷氨酸棒杆菌pEC-XT99A::ABM pVWEX1::rfbBDAC、谷氨酸棒杆菌 pEC-XT99A::ABC pVWEX1::rfbBDAC和谷氨酸棒杆菌 pEC-XT99A::ABCM pVWEX1::rfbBDAC。
20.通过重组谷氨酸棒杆菌菌株产生的鼠李糖脂的定量
将在实施例15-19中产生的重组菌株谷氨酸棒杆菌菌株:谷氨酸棒杆菌 pEC-XT99A、谷氨酸棒杆菌pEC-XT99A::AB、谷氨酸棒杆菌 pEC-XT99A::ABC、谷氨酸棒杆菌pEC-XT99A::ABM、谷氨酸棒杆菌 pEC-XT99A::ABCM、谷氨酸棒杆菌pEC-XT99A pVWEX1::rfbBDAC、谷氨 酸棒杆菌pEC-XT99A::AB pVWEX1::rfbBDAC、谷氨酸棒杆菌 pEC-XT99A::ABM pVWEX1::rfbBDAC、谷氨酸棒杆菌pEC-XT99A::ABC pVWEX1::rfbBDAC和谷氨酸棒杆菌pEC-XT99A::ABCM pVWEX1::rfbBDAC培养在使用了5mg/l四环素以及5mg/l四环素和25mg/l卡那霉素的LBHIS琼脂平板上。为了研究鼠李糖脂在摇动瓶中的产生, 首先准备预培养物。为此,使用在LBHIS琼脂平板上新划线的菌株的接种 环,并将10ml LBHIS培养基(18.5g/l的脑心浸出液肉汤、0.5M山梨醇、5 g/l的Bacto胰蛋白胨、2.5g/l的Bacto酵母膏和5g/l的NaCl,补加有5mg/l 四环素或5mg/l四环素和25mg/l卡那霉素)接种在100ml锥形烧瓶中。在 33℃和200rpm下,培养菌株过夜。第二天早上,将50ml CGXII培养基(含 5mg/l四环素或者5mg/l四环素和25mg/l卡那霉素)接种在含有带1ml预 培养物的挡板的500ml锥形烧瓶中(起始OD600 0.1)。
CGXII培养基:
●20g/l的(NH4)2SO4(Merck)
●5g/l的尿素(Merck)
●1g/l的KH2PO4(Merck)
●1g/l的K2HPO4(Merck)
●0.25g/l的MgSO4·7H2O(Merck)
●10mg/l的CaCl2(Merck)
●42g/l的MOPS(Roth)
●0.2mg/l的生物素(Merck)
●1ml/l的微量盐溶液
●利用NaOH调整至pH 7
●高压灭菌后,加入1ml/l的原儿茶酸(30g/l,溶解在稀释的NaOH中, 灭菌过滤的)和40g/l的葡萄糖(Merck)
微量盐溶液:
●10g/l的FeSO4·7H2O(Merck)
●10g/l的MnSO4·H2O(Merck)
●1g/l的ZnSO4·7H2O(Merck)
●0.2g/l的CuSO4·5H2O(Merck)
●20mg/l的NiCl2·6H2O(Merck)
●利用HCl溶解酸化至pH为1
将培养物在200rpm和33℃下培养直至光密度(600nm)为0.4–0.6。在 该光密度,通过添加IPTG(异丙基-β-D-硫代半乳糖吡喃糖苷;1mM终浓度) 诱导培养物。随后的表达同样在33℃和200rpm下进行72h。在24h的间 隔,从培养瓶中移出1ml肉汤样品。如实施例4中所述,进行用于以下色 谱分析的样品制备和色谱分析本身。
尽管谷氨酸棒杆菌pEC-XT99A不产生鼠李糖脂,在重组菌株谷氨酸棒 杆菌pEC-XT99A::AB、谷氨酸棒杆菌pEC-XT99A::ABC、谷氨酸棒杆菌 pEC-XT99A::ABM和谷氨酸棒杆菌pEC-XT99A::ABCM中,可检测到鼠李 糖脂的形成。借助于参考资料,据显示,谷氨酸棒杆菌pEC-XT99A::AB和 谷氨酸棒杆菌pEC-XT99A::ABM仅形成单鼠李糖脂,而谷氨酸棒杆菌pEC-XT99A::ABC、谷氨酸棒杆菌pEC-XT99A::ABM和谷氨酸棒杆菌 pEC-XT99A::ABCM能够形成双鼠李糖脂和单鼠李糖脂。此外,据显示, 谷氨酸棒杆菌pEC-XT99A::ABM和谷氨酸棒杆菌pEC-XT99A::ABCM相比 相应的没有扩增来自绿脓假单胞菌的pa1131基因的参照菌株谷氨酸棒杆菌 pEC-XT99A::AB和谷氨酸棒杆菌pEC-XT99A::ABC,能够形成更多的单鼠 李糖脂或双鼠李糖脂以及单鼠李糖脂。
而且,据显示,菌株谷氨酸棒杆菌pEC-XT99A::AB pVWEX1::rfbBDAC、谷氨酸棒杆菌pEC-XT99A::ABM pVWEX1::rfbBDAC、 谷氨酸棒杆菌pEC-XT99A::ABC pVWEX1::rfbBDAC和谷氨酸棒杆菌 pEC-XT99A::ABCM pVWEX1::rfbBDAC相比没有扩增来自恶臭假单胞菌 的rfbBDA基因的菌株谷氨酸棒杆菌pEC-XT99A::ABM、谷氨酸棒杆菌 pEC-XT99A::ABC和谷氨酸棒杆菌pEC-XT99A::ABCM,形成明显更多的 单-鼠李糖脂(谷氨酸棒杆菌pEC-XT99A::ABpVWEX1::rfbBDAC和谷氨酸 棒杆菌pEC-XT99A::ABM pVWEX1::rfbBDAC)或者单-和双鼠李糖脂(谷氨 酸棒杆菌pEC-XT99A::ABC pVWEX1::rfbBDAC和谷氨酸棒杆菌 pEC-XT99A::ABCM pVWEX1::rfbBDAC)。
21.携带质粒pBBR1MCS-2、pBBR1MCS-2::AB、pBBR1MCS-2::ABC、 pBBR1MCS-2::ABM和pBBR1MCS-2::ABCM的假单胞菌菌株的构建
通过电穿孔法,将质粒pBBR1MCS-2、pBBR1MCS-2::AB、 pBBR1MCS-2::ABC、pBBR1MCS-2::ABM和pBBR1MCS-2::ABCM合并入 荧光假单胞菌DSM 50090、荧光假单胞菌DSM9958、恶臭假单胞菌DSM 6899、恶臭假单胞菌DSM 50204、恶臭假单胞菌50194、油菜假单胞菌DSM 13227、斯氏假单胞菌DSM 10701、斯氏假单胞菌DSM 4166和黄褐假单胞 菌DSM17717中。如以前所述(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),进行假单胞菌菌株的转化。在营养琼脂平板(5g/l蛋白胨、 3g/l肉膏、15g/l琼脂、pH 7,补加有50mg/l卡那霉素)上进行转化株的选 择。将平板在30℃,更确切地说在28℃下孵育2天。获得的携带质粒的菌 株被命名为荧光假单胞菌DSM 50090pBBR1MCS-2、荧光假单胞菌DSM 9958pBBR1MCS-2、恶臭假单胞菌DSM 6899pBBR1MCS-2、恶臭假单胞 菌DSM50204pBBR1MCS-2、恶臭假单胞菌50194pBBR1MCS-2、油菜假 单胞菌DSM 13227pBBR1MCS-2、斯氏假单胞菌DSM 10701 pBBR1MCS-2、斯氏假单胞菌DSM 4166pBBR1MCS-2、黄褐假单胞菌DSM 17717pBBR1MCS-2、荧光假单胞菌DSM 50090pBBR1MCS-2::AB、荧光 假单胞菌DSM9958pBBR1MCS-2::AB、恶臭假单胞菌DSM 6899 pBBR1MCS-2::AB、恶臭假单胞菌DSM50204pBBR1MCS-2::AB、恶臭假 单胞菌50194pBBR1MCS-2::AB、油菜假单胞菌DSM 13227pBBR1MCS-2::AB、斯氏假单胞菌DSM 10701pBBR1MCS-2::AB、斯氏假 单胞菌DSM4166pBBR1MCS-2::AB、黄褐假单胞菌DSM 17717 pBBR1MCS-2::AB、荧光假单胞菌DSM50090pBBR1MCS-2::ABC、荧光假 单胞菌DSM 9958pBBR1MCS-2::ABC、恶臭假单胞菌DSM6899 pBBR1MCS-2::ABC、恶臭假单胞菌DSM 50204pBBR1MCS-2::ABC、恶臭 假单胞菌50194pBBR1MCS-2::ABC、油菜假单胞菌DSM 13227 pBBR1MCS-2::ABC、斯氏假单胞菌DSM10701pBBR1MCS-2::ABC、斯氏 假单胞菌DSM 4166pBBR1MCS-2::ABC、黄褐假单胞菌DSM17717 pBBR1MCS-2::ABC、荧光假单胞菌DSM 50090pBBR1MCS-2::ABCM、荧 光假单胞菌DSM9958pBBR1MCS-2::ABCM、恶臭假单胞菌DSM 6899 pBBR1MCS-2::ABCM、恶臭假单胞菌DSM50204pBBR1MCS-2::ABCM、 恶臭假单胞菌50194pBBR1MCS-2::ABCM、油菜假单胞菌DSM13227 pBBR1MCS-2::ABCM、斯氏假单胞菌DSM 10701pBBR1MCS-2::ABCM, 斯氏假单胞菌DSM4166pBBR1MCS-2::ABCM、黄褐假单胞菌DSM 17717 pBBR1MCS-2::ABCM、荧光假单胞菌DSM50090pBBR1MCS-2::ABM、荧 光假单胞菌DSM 9958pBBR1MCS-2::ABM、恶臭假单胞菌DSM6899 pBBR1MCS-2::ABM、恶臭假单胞菌DSM 50204pBBR1MCS-2::ABM、恶 臭假单胞菌50194pBBR1MCS-2::ABM、油菜假单胞菌DSM 13227 pBBR1MCS-2::ABM、斯氏假单胞菌DSM10701pBBR1MCS-2::ABM、斯 氏假单胞菌DSM 4166pBBR1MCS-2::ABM和黄褐假单胞菌DSM17717 pBBR1MCS-2::ABM。
22.通过重组假单胞菌菌株产生的鼠李糖脂的定量
将在实施例21中产生的重组菌株假单胞菌菌株荧光假单胞菌DSM 50090、荧光假单胞菌DSM 9958pBBR1MCS-2、恶臭假单胞菌DSM 6899 pBBR1MCS-2、恶臭假单胞菌DSM50204pBBR1MCS-2、恶臭假单胞菌 50194pBBR1MCS-2、油菜假单胞菌DSM 13227pBBR1MCS-2、斯氏假单 胞菌DSM 10701pBBR1MCS-2、斯氏假单胞菌DSM 4166pBBR1MCS-2、 黄褐假单胞菌DSM 17717pBBR1MCS-2、荧光假单胞菌DSM 50090 pBBR1MCS-2::AB、荧光假单胞菌DSM9958pBBR1MCS-2::AB、恶臭假单 胞菌DSM 6899pBBR1MCS-2::AB、恶臭假单胞菌DSM 50204pBBR1MCS-2::AB、恶臭假单胞菌50194pBBR1MCS-2::AB、油菜假单胞菌 DSM13227pBBR1MCS-2::AB、斯氏假单胞菌DSM 10701 pBBR1MCS-2::AB、斯氏假单胞菌DSM4166pBBR1MCS-2::AB、黄褐假 单胞菌DSM 17717pBBR1MCS-2::AB,荧光假单胞菌DSM 50090pBBR1MCS-2::ABC、荧光假单胞菌DSM 9958pBBR1MCS-2::ABC、恶臭 假单胞菌DSM6899pBBR1MCS-2::ABC、恶臭假单胞菌DSM 50204 pBBR1MCS-2::ABC、恶臭假单胞菌50194pBBR1MCS-2::ABC、油菜假单 胞菌DSM 13227pBBR1MCS-2::ABC、斯氏假单胞菌DSM10701 pBBR1MCS-2::ABC、斯氏假单胞菌DSM 4166pBBR1MCS-2::ABC、黄褐 假单胞菌DSM17717pBBR1MCS-2::ABC、荧光假单胞菌DSM 50090 pBBR1MCS-2::ABCM、荧光假单胞菌DSM9958pBBR1MCS-2::ABCM、 恶臭假单胞菌DSM 6899pBBR1MCS-2::ABCM、恶臭假单胞菌DSM50204pBBR1MCS-2::ABCM、恶臭假单胞菌50194pBBR1MCS-2::ABCM、 油菜假单胞菌DSM13227pBBR1MCS-2::ABCM、斯氏假单胞菌DSM 10701pBBR1MCS-2::ABCM、斯氏假单胞菌DSM4166 pBBR1MCS-2::ABCM、黄褐假单胞菌DSM 17717pBBR1MCS-2::ABCM、 荧光假单胞菌DSM50090pBBR1MCS-2::ABM、荧光假单胞菌DSM 9958 pBBR1MCS-2::ABM、恶臭假单胞菌DSM6899pBBR1MCS-2::ABM、恶臭 假单胞菌DSM 50204pBBR1MCS-2::ABM、恶臭假单胞菌50194pBBR1MCS-2::ABM、油菜假单胞菌DSM 13227pBBR1MCS-2::ABM、斯 氏假单胞菌DSM10701pBBR1MCS-2::ABM、斯氏假单胞菌DSM 4166 pBBR1MCS-2::ABM和黄褐假单胞菌DSM17717pBBR1MCS-2::ABM培 养在LB琼脂卡那霉素(50μg/ml)平板上。随后如实施例12所述进行用于产 生鼠李糖脂的培养。如实施例4所述进行用于下文的色谱分析的样品制备 和色谱分析自身。
尽管假单胞菌菌株荧光假单胞菌DSM 50090、荧光假单胞菌DSM 9958 pBBR1MCS-2、恶臭假单胞菌DSM 6899pBBR1MCS-2、恶臭假单胞菌 DSM 50204pBBR1MCS-2、恶臭假单胞菌50194pBBR1MCS-2、油菜假单 胞菌DSM 13227pBBR1MCS-2、斯氏假单胞菌DSM10701pBBR1MCS-2、 斯氏假单胞菌DSM 4166pBBR1MCS-2、黄褐假单胞菌DSM 17717pBBR1MCS-2没有产生鼠李糖脂,在重组菌株荧光假单胞菌DSM 50090 pBBR1MCS-2::AB、荧光假单胞菌DSM 9958pBBR1MCS-2::AB、恶臭假 单胞菌DSM 6899pBBR1MCS-2::AB、恶臭假单胞菌DSM 50204 pBBR1MCS-2::AB、恶臭假单胞菌50194pBBR1MCS-2::AB、油菜假单胞 菌DSM13227pBBR1MCS-2::AB、斯氏假单胞菌DSM 10701 pBBR1MCS-2::AB、斯氏假单胞菌DSM4166pBBR1MCS-2::AB、黄褐假 单胞菌DSM 17717pBBR1MCS-2::AB、荧光假单胞菌DSM 50090pBBR1MCS-2::ABM、荧光假单胞菌DSM 9958pBBR1MCS-2::ABM、恶臭 假单胞菌DSM6899pBBR1MCS-2::ABM、恶臭假单胞菌DSM 50204 pBBR1MCS-2::ABM、恶臭假单胞菌50194pBBR1MCS-2::ABM、油菜假单 胞菌DSM 13227pBBR1MCS-2::ABM、斯氏假单胞菌DSM10701 pBBR1MCS-2::ABM、斯氏假单胞菌DSM 4166pBBR1MCS-2::ABM和黄褐 假单胞菌DSM17717pBBR1MCS-2::ABM中,可以检测到单鼠李糖脂的形 成,以及在菌株荧光假单胞菌DSM50090pBBR1MCS-2::ABC、荧光假单 胞菌DSM 9958pBBR1MCS-2::ABC、恶臭假单胞菌DSM6899 pBBR1MCS-2::ABC、恶臭假单胞菌DSM 50204pBBR1MCS-2::ABC、恶臭 假单胞菌50194pBBR1MCS-2::ABC、油菜假单胞菌DSM 13227 pBBR1MCS-2::ABC、斯氏假单胞菌DSM10701pBBR1MCS-2::ABC、斯氏 假单胞菌DSM 4166pBBR1MCS-2::ABC、黄褐假单胞菌DSM17717 pBBR1MCS-2::ABC、荧光假单胞菌DSM 50090pBBR1MCS-2::ABCM、荧 光假单胞菌DSM9958pBBR1MCS-2::ABCM、恶臭假单胞菌DSM 6899 pBBR1MCS-2::ABCM、恶臭假单胞菌DSM50204pBBR1MCS-2::ABCM、 恶臭假单胞菌50194pBBR1MCS-2::ABCM、油菜假单胞菌DSM13227 pBBR1MCS-2::ABCM、斯氏假单胞菌DSM 10701pBBR1MCS-2::ABCM、 斯氏假单胞菌DSM4166pBBR1MCS-2::ABCM和黄褐假单胞菌DSM 17717pBBR1MCS-2::ABCM中,可以检测到单-和双鼠李糖脂的形成。
而且,相比相应的没有铜绿假单胞菌基因pa1131的参照菌株荧光假单 胞菌DSM50090pBBR1MCS-2::AB、荧光假单胞菌DSM 9958 pBBR1MCS-2::AB、恶臭假单胞菌DSM6899pBBR1MCS-2::AB、恶臭假 单胞菌DSM 50204pBBR1MCS-2::AB、恶臭假单胞菌50194pBBR1MCS-2::AB、油菜假单胞菌DSM 13227pBBR1MCS-2::AB、斯氏假 单胞菌DSM10701pBBR1MCS-2::AB、斯氏假单胞菌DSM 4166 pBBR1MCS-2::AB和黄褐假单胞菌DSM17717pBBR1MCS-2::AB以及没有 来自绿脓假单胞菌的pa1131基因扩增的荧光假单胞菌DSM50090 pBBR1MCS-2::ABC、荧光假单胞菌DSM 9958pBBR1MCS-2::ABC、恶臭 假单胞菌DSM6899pBBR1MCS-2::ABC、恶臭假单胞菌DSM 50204 pBBR1MCS-2::ABC、恶臭假单胞菌50194pBBR1MCS-2::ABC、油菜假单 胞菌DSM 13227pBBR1MCS-2::ABC、斯氏假单胞菌DSM10701 pBBR1MCS-2::ABC、斯氏假单胞菌DSM 4166pBBR1MCS-2::ABC和黄褐 假单胞菌DSM17717pBBR1MCS-2::ABC,重组假单胞菌菌株荧光假单胞 菌DSM 50090pBBR1MCS-2::ABM、荧光假单胞菌DSM 9958 pBBR1MCS-2::ABM、恶臭假单胞菌DSM 6899pBBR1MCS-2::ABM、恶臭假单胞菌DSM 50204pBBR1MCS-2::ABM、恶臭假单胞菌50194 pBBR1MCS-2::ABM、油菜假单胞菌DSM 13227pBBR1MCS-2::ABM、斯 氏假单胞菌DSM 10701pBBR1MCS-2::ABM、斯氏假单胞菌DSM 4166 pBBR1MCS-2::ABM、黄褐假单胞菌DSM 17717pBBR1MCS-2::ABM形成 了较少的单鼠李糖脂,以及重组假单胞菌菌株荧光假单胞菌DSM 50090 pBBR1MCS-2::ABCM、荧光假单胞菌DSM 9958pBBR1MCS-2::ABCM、 恶臭假单胞菌DSM 6899pBBR1MCS-2::ABCM、恶臭假单胞菌DSM 50204pBBR1MCS-2::ABCM、恶臭假单胞菌50194pBBR1MCS-2::ABCM、 油菜假单胞菌DSM 13227pBBR1MCS-2::ABCM、斯氏假单胞菌DSM 10701pBBR1MCS-2::ABCM、斯氏假单胞菌DSM 4166 pBBR1MCS-2::ABCM和黄褐假单胞菌DSM 17717pBBR1MCS-2::ABCM 形成了较少的单-和双鼠李糖脂。
23.用于在恶臭假单胞菌中异源表达来自绿脓假单胞菌PAO1、绿脓假单胞 菌PA7、绿脓假单胞菌1和泰国伯克氏菌E264的可选rhlA、rhlB和rhlC 基因的载体pBBR1MCS-2::ABPAO1-C1和pBBR1MCS-2::ABPA7-CE264的 构建
为了异源表达来自绿脓假单胞菌PAO1和绿脓假单胞菌PA7的基因 rhlA、rhlB和rhlC,首先构建质粒pBBR1MCS-2::ABPAO1(Seq ID No.62)和 pBBR1MCS-2::ABPA7(Seq IDNo.63)。为此,通过DNA 2.0公司(Menlo Park, CA,U.S.A)(Regensburg)合成了合成性操纵子rhlABPAO1(Seq ID No.64)和 rhlABPA7(Seq ID No.65),并间克隆在商品化载体pJ294(DNA 2.0)中。合成 的基础是已知的菌株绿脓假单胞菌PAO1和绿脓假单胞菌PA7的基因组序 列。从载体pJ294::ABPAO1和pJ294::ABPA7开始,通过KpnI和XbaI从载 体切割下合成性操纵子,并随后连接在用KpnI和XbaI切割的表达载体 pBBR1MCS-2(Seq ID No.49)(Kovachet al.,1995:Four new derivatives of the broad-host-range cloning vectorpBBR1MCS carrying different antibiotic-resistance cassettes.Gene,166:175-176)中。产生的质粒 pBBR1MCS-2::ABPAO1(Seq ID No.62)和pBBR1MCS-2::ABPA7(Seq IDNo.63)的大小为7332和7354个碱基对。以本领域技术人员已知的方式进行 化学感受态大肠杆菌DH5α细胞(Gibco-BRL,Karlsruhe)的连接和转化。通过 DNA序列分析来检验插入物的确实性。
在第二步中,产生了质粒pBBR1MCS-2::ABPAO1-C1(Seq ID No.66) 和pBBR1MCS-2::ABPA7-CE264(Seq ID No.67)。为此,通过DNA 2.0公司 (Menlo Park,CA,U.S.A)合成了来自绿脓假单胞菌1(Seq ID No.68)和泰国伯 克氏菌E264(Seq ID No.76)的rhlC基因,并间克隆在商品化载体pJ294 (DNA 2.0)中。合成的基础是已知的菌株绿脓假单胞菌1和泰国伯克氏菌 E264的基因组序列。从载体pJ294::C1和pJ294::CE264开始,通过Xba和 SacI从载体切割下rhlC基因,并随后连接在同样用Xba和SacI切割的载 体pBBR1MCS-2::ABPAO1(Seq ID No.62)和pBBR1MCS-2::ABPA7(Seq ID No.63)中。产生的质粒pBBR1MCS-2::ABPAO1-C1(Seq ID No.66)和 pBBR1MCS-2::ABPA7-CE264(Seq ID No.67)的大小为8325和8335个碱基 对。以本领域技术人员已知的方式进行化学感受态大肠杆菌DH5α细胞(Gibco-BRL,Karlsruhe)的连接和转化。通过DNA序列分析来检验插入物的 确实性。通过DNA序列分析来检验插入物的确实性。
如以前所述的(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2、pBBR1MCS-2::ABPAO1-C1和 pBBR1MCS-2::ABPA7-CE264进行恶臭假单胞菌KT2440和GPp104的转 化。分离并分析每10个克隆的质粒DNA。获得的携带质粒的菌株被命名 为恶臭假单胞菌KT2440pBBR1MCS-2、恶臭假单胞菌KT2440 pBBR1MCS-2::ABPAO1-C1、恶臭假单胞菌KT2440 pBBR1MCS-2::ABPA7-CE264、恶臭假单胞菌GPp104pBBR1MCS-2、恶臭 假单胞菌GPp104pBBR1MCS-2::ABPAO1-C1和恶臭假单胞菌GPp104pBBR1MCS-2::ABPA7-CE264。
24.通过具有来自绿脓假单胞菌PAO1、绿脓假单胞菌PA7、绿脓假单胞 菌1和泰国伯克氏菌E264的可选rhlA、rhlB和rhlC基因的重组恶臭假单 胞菌菌株产生的鼠李糖脂的定量
将实施例23产生的重组菌株恶臭假单胞菌菌株培养在LB琼脂卡那霉 素(50μg/ml)平板上。随后如实施例12所述进行用于产生鼠李糖脂的培养。 如实施例4所述进行用于下文的色谱分析的样品制备和色谱分析自身。
尽管菌株恶臭假单胞菌KT2440pBBR1MCS-2和恶臭假单胞菌GPp104 pBBR1MCS-2不能产生单-和双鼠李糖脂,菌株恶臭假单胞菌KT2440 pBBR1MCS-2::ABPAO1-C1、恶臭假单胞菌KT2440 pBBR1MCS-2::ABPA7-CE264、恶臭假单胞菌GPp104 pBBR1MCS-2::ABPAO1-C1和恶臭假单胞菌GPp104 pBBR1MCS-2::ABPA7-CE264形成了单鼠李糖脂以及双鼠李糖脂。据显示, 聚羟基丁酸酯形成消减的菌株(恶臭假单胞菌GPp104 pBBR1MCS-2::ABPAO1-C1和恶臭假单胞菌GPp104 pBBR1MCS-2::ABPA7-CE264)比聚羟基丁酸酯形成没有消减的菌株(恶臭假 单胞菌KT2440pBBR1MCS-2::ABPAO1-C1和恶臭假单胞菌KT2440 pBBR1MCS-2::ABPA7-CE264)能够产生更多的单-和双鼠李糖脂。
25.用于在恶臭假单胞菌和大肠杆菌中过表达恶臭假单胞菌rfbBDAC操纵 子的载体pBBR1MCS-2::AB_rfbBDAC、pBBR1MCS-2::ABM_rfbBDAC和 pBBR1MCS-2::ABMC_rfbBDAC的构建
为了构建用于在恶臭假单胞菌和大肠杆菌中过表达恶臭假单胞菌 rfbBDAC操纵子的载体pBBR1MCS-2::AB_rfbBDAC、 pBBR1MCS-2::ABM_rfbBDAC和pBBR1MCS-2::ABMC_rfbBDAC,首先通 过PCR扩增恶臭假单胞菌rfbBDAC操纵子。载体pBBR1MCS-2::rfbBDAC(Seq ID No.45)作为PCR的基体。使用以下寡核苷酸:
RL_AgeI-fw:5’-TATATATAACCGGTATTAATGCAGCTGGCACGAC-3’ (Seq ID No.71)
RL_AgeI_rev:5’-GGCCGACCGGTACTAGTGGA-3’(Seq ID No.72)
利用New England Biolabs(Frankfurt)聚合酶的PhusionTM高保真预混合 物进行PCR。它以本领域技术人员已知的方式进行。将靶序列(lac启动子 和rfbBDAC)间克隆在Trenzyme短吻鳄(alligator)克隆系统中。选择大肠杆菌 DH5α(New England Biolabs;Frankfurt)转化株,以及分离不同候选物的质粒 DNA并对其进行测序。核对并检验序列的正确性后,利用AgeI切割载体。 通过常规的连接方法,将靶片段连接入同样用AgeI切割的载体 pBBR1MCS-2::AB(Seq ID No.38)、pBBR1MCS-2::ABM(Seq ID No.42)和 pBBR1MCS-2::ABMC(Seq ID No.51)中。产生的载体 pBBR1MCS-2::AB_rfbBDAC(Seq ID No.73)、pBBR1MCS-2::ABM_rfbBDAC(Seq ID No.74)和 pBBR1MCS-2::ABMC_rfbBDAC(Seq IDNo.75)的大小为11960、13289和 14250个碱基对。对载体的插入物进行测序。以本领域技术人员已知的方式 实施PCR、通过琼脂糖凝胶电泳校验PCR的成功扩增、进行DNA的溴化 乙锭染色、确定PCR片段大小、纯化PCR产物以及确定DNA浓度。
如以前所述(Iwasaki et al.Biosci.Biotech.Biochem.1994. 58(5):851-854),利用载体pBBR1MCS-2::AB_rfbBDAC、 pBBR1MCS-2::ABM_rfbBDAC和pBBR1MCS-2::ABMC_rfbBDAC进行恶 臭假单胞菌KT2440的转化。分离并分析每10个克隆的质粒DNA。获得的 携带质粒的菌株被命名为恶臭假单胞菌KT2440 pBBR1MCS-2::AB_rfbBDAC、恶臭假单胞菌KT2440 pBBR1MCS-2::ABM_rfbBDAC和恶臭假单胞菌KT2440 pBBR1MCS-2::ABMC_rfbBDAC。
26.通过重组恶臭假单胞菌KT2440pBBR1MCS-2::AB_rfbBDAC、恶臭假 单胞菌KT2440pBBR1MCS-2::ABM_rfbBDAC、恶臭假单胞菌KT2440 pBBR1MCS-2::ABC_rfbBDAC、恶臭假单胞菌KT2440 pBBR1MCS-2::ABMC_rfbBDAC、恶臭假单胞菌KT2440 pBBR1MCS-2::AB、恶臭假单胞菌KT2440pBBR1MCS-2::ABM、恶臭假单 胞菌KT2440pBBR1MCS-2::ABC和恶臭假单胞菌KT2440 pBBR1MCS-2::ABMC产生的鼠李糖脂的定量
将在实施例2、7和25中产生的重组菌株恶臭假单胞菌菌株培养在LB 琼脂-卡那霉素(50μg/ml)平板上。随后如实施例12所述进行用于生产鼠李 糖脂的培养。如实施例4所述进行用于下文色谱分析的样品制备和色谱分 析本身。
据显示,恶臭假单胞菌KT2440pBBR1MCS-2::AB_rfbBDAC、恶臭假 单胞菌KT2440pBBR1MCS-2::ABM_rfbBDAC、恶臭假单胞菌KT2440 pBBR1MCS-2::AB和恶臭假单胞菌KT2440pBBR1MCS-2::ABM能够形成 单鼠李糖脂,而恶臭假单胞菌KT2440pBBR1MCS-2::ABMC_rfbBDAC、 恶臭假单胞菌KT2440pBBR1MCS-2::ABC_rfbBDAC、恶臭假单胞菌KT2440pBBR1MCS-2::ABC和恶臭假单胞菌KT2440 pBBR1MCS-2::ABMC能够形成单-和双鼠李糖脂。
此外,据显示,恶臭假单胞菌KT2440pBBR1MCS-2::ABM_rfbBDAC、 恶臭假单胞菌KT2440pBBR1MCS-2::ABM、KT2440 pBBR1MCS-2::ABMC_rfbBDAC和KT2440pBBR1MCS-2::ABMC相比对应 的没有绿脓假单胞菌基因pa1131扩增的对照菌株恶臭假单胞菌KT2440pBBR1MCS-2::AB_rfbBDAC、恶臭假单胞菌KT2440pBBR1MCS-2::AB、 KT2440pBBR1MCS-2::ABC_rfbBDAC和KT2440pBBR1MCS-2::ABC能够 形成更多的单-和双鼠李糖脂。
最后,据显示,恶臭假单胞菌KT2440pBBR1MCS-2::AB_rfbBDAC、 恶臭假单胞菌KT2440pBBR1MCS-2::ABM_rfbBDAC、恶臭假单胞菌 KT2440pBBR1MCS-2::ABC_rfbBDAC、恶臭假单胞菌KT2440 pBBR1MCS-2::ABMC_rfbBDAC相比相应的没有恶臭假单胞菌基因 rfbBDAC扩增的对照菌株恶臭假单胞菌KT2440pBBR1MCS-2::AB、恶臭 假单胞菌KT2440pBBR1MCS-2::ABM、恶臭假单胞菌KT2440 pBBR1MCS-2::ABC、恶臭假单胞菌KT2440pBBR1MCS-2::ABMC能够形 成更多的单鼠李糖脂(恶臭假单胞菌KT2440pBBR1MCS-2::AB_rfbBDAC 和恶臭假单胞菌KT2440pBBR1MCS-2::ABM_rfbBDAC)以及单-和双鼠李 糖脂(恶臭假单胞菌KT2440pBBR1MCS-2::ABC_rfbBDAC和恶臭假单胞 菌KT2440pBBR1MCS-2::ABMC_rfbBDAC)。
27.重组大肠杆菌W3110pBBR1MCS-2::AB、大肠杆菌W3110 pBBR1MCS-2::ABM、大肠杆菌W3110pBBR1MCS-2::ABC、大肠杆菌 W3110pBBR1MCS-2::ABCM、大肠杆菌W3110pBBR1MCS-2::AB_rfbBDAC、大肠杆菌W3110 pBBR1MCS-2::ABM_rfbBDAC、大肠杆菌W3110pBBR1MCS-2::ABC_rfbBDAC和大肠杆菌W3110 pBBR1MCS-2::ABCM_rfbBDAC的产生
如以前所述(Miller JH.AShort Course in Bacterial Genetics:A LaboratoryManual and Handbook for Escherichia coil and Related Bacteria.Plainview,NY:Cold Spring Harbor Lab.Press;1992),通过电穿孔法进行大肠杆菌W3110 的转化。分离并分析每10个克隆的质粒DNA。获得的携带质粒的菌株命 名为大肠杆菌W3110pBBR1MCS-2::AB、大肠杆菌W3110 pBBR1MCS-2::ABM、大肠杆菌W3110pBBR1MCS-2::ABC,大肠杆菌W3110pBBR1MCS-2::ABCM、大肠杆菌W3110 pBBR1MCS-2::AB_rfbBDAC、大肠杆菌W3110pBBR1MCS-2::ABM_rfbBDAC、大肠杆菌W3110 pBBR1MCS-2::ABC_rfbBDAC和大肠杆菌W3110pBBR1MCS-2::ABCM_rfbBDAC。
28.通过重组大肠杆菌W3110pBBR1MCS-2::AB、大肠杆菌W3110 pBBR1MCS-2::ABM、大肠杆菌W3110pBBR1MCS-2::ABC、大肠杆菌 W3110pBBR1MCS-2::ABCM、大肠杆菌W3110pBBR1MCS-2::AB_rfbBDAC、大肠杆菌W3110 pBBR1MCS-2::ABM_rfbBDAC、大肠杆菌W3110pBBR1MCS-2::ABC_rfbBDAC和大肠杆菌W3110 pBBR1MCS-2::ABCM_rfbBDAC产生的鼠李糖脂的定量
将实施例27中产生的重组大肠杆菌菌株培养在LB琼脂卡那霉素(50 μg/ml)平板上。随后如实施例10所述进行用于生产鼠李糖脂的培养。如实 施例4所述进行用于下文色谱分析的样品制备和色谱分析本身。
据显示,大肠杆菌W3110pBBR1MCS-2::AB、大肠杆菌W3110 pBBR1MCS-2::ABM、大肠杆菌W3110pBBR1MCS-2::AB_rfbBDAC和大肠 杆菌W3110pBBR1MCS-2::ABM_rfbBDAC能够形成单鼠李糖脂,而大肠杆 菌W3110pBBR1MCS-2::ABC、大肠杆菌W3110pBBR1MCS-2::ABCM、大肠杆菌W3110pBBR1MCS-2::ABC_rfbBDAC和大肠杆菌W3110 pBBR1MCS-2::ABCM_rfbBDAC能够形成单-和双鼠李糖脂。此外,据显示, 大肠杆菌W3110pBBR1MCS-2::ABM和大肠杆菌W3110 pBBR1MCS-2::ABM_rfbBDAC相比没有绿脓假单胞菌基因pa1131扩增的 大肠杆菌W3110pBBR1MCS-2::AB和大肠杆菌W3110 pBBR1MCS-2::AB_rfbBDAC形成更多的单鼠李糖脂。
此外,据显示,大肠杆菌W3110pBBR1MCS-2::ABCM和大肠杆菌 W3110pBBR1MCS-2::ABCM_rfbBDAC相比没有绿脓假单胞菌基因pa1131 扩增的大肠杆菌W3110pBBR1MCS-2::ABC和大肠杆菌W3110 pBBR1MCS-2::ABC_rfbBDAC形成更多的单-和双鼠李糖脂。此外,据显示,大肠杆菌W3110pBBR1MCS-2::ABM和大肠杆菌W3110 pBBR1MCS-2::ABM_rfbBDAC相比没有绿脓假单胞菌基因pa1131扩增的 大肠杆菌W3110pBBR1MCS-2::AB和大肠杆菌W3110pBBR1MCS-2::AB_rfbBDAC形成更多的单鼠李糖脂。
最后,据显示,大肠杆菌W3110pBBR1MCS-2::AB_rfbBDAC、大肠杆 菌W3110pBBR1MCS-2::ABM_rfbBDAC、大肠杆菌W3110 pBBR1MCS-2::ABC_rfbBDAC和大肠杆菌W3110 pBBR1MCS-2::ABCM_rfbBDAC相比相应的没有恶臭假单胞菌基因 rfbBDAC扩增的对照菌株大肠杆菌W3110pBBR1MCS-2::AB、大肠杆菌 W3110pBBR1MCS-2::ABM、大肠杆菌W3110pBBR1MCS-2::ABC和大肠 杆菌W3110pBBR1MCS-2::ABCM能够形成更多的单鼠李糖脂(大肠杆菌 W3110pBBR1MCS-2::AB_rfbBDAC、大肠杆菌W3110 pBBR1MCS-2::ABM_rfbBDAC)以及单-和双鼠李糖脂(大肠杆菌W3110 pBBR1MCS-2::ABC_rfbBDAC和大肠杆菌W3110pBBR1MCS-2::ABCM_rfbBDAC)。
序列表
<110> 赢创德固赛有限公司
<120> 用于生产鼠李糖脂的细胞和方法
<130> 201000205
<160> 92
<170> PatentIn version 3.5
<210> 1
<211> 888
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(888)
<400> 1
atg cgg cgc gaa agt ctg ttg gta tcg gtt tgc aag ggc ctg cgg gta 48
Met Arg Arg Glu Ser Leu Leu Val Ser Val Cys Lys Gly Leu Arg Val
1 5 10 15
cat gtc gag cgc gtt ggg cag gat ccc ggg cgc agc acg gtg atg ctg 96
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Ser Thr Val Met Leu
20 25 30
gtc aac ggc gcg atg gcg acc acc gcc tcg ttc gcc cgg acc tgc aag 144
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
tgc ctg gcc gaa cat ttc aac gtg gtg ctg ttc gac ctg ccc ttc gcc 192
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
ggg cag tcg cgt cag cac aac ccg cag cgg ggg ttg atc acc aag gac 240
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
gac gag gtg gaa atc ctc ctg gcg ctg atc gag cgc ttc gag gtc aat 288
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Glu Val Asn
85 90 95
cac ctg gtc tcc gcg tcc tgg ggc ggt atc tcc acg ctg ctg gcg ctg 336
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
tcg cgc aat ccg cgc ggc atc cgc agc tcg gtg gtg atg gca ttc gcc 384
Ser Arg Asn Pro Arg Gly Ile Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
cct gga ctg aac cag gcg atg ctc gac tac gtc ggg cgg gcg cag gcg 432
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Ala
130 135 140
ctg atc gag ctg gac gac aag tcg gcg atc ggc cat ctg ctc aac gag 480
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
acc gtc ggc aaa tac ctg ccg ccg cgc ctg aaa gcc agc aac cat cag 528
Thr Val Gly Lys Tyr Leu Pro Pro Arg Leu Lys Ala Ser Asn His Gln
165 170 175
cac atg gct tcg ctg gcc acc ggc gaa tac gag cag gcg cgc ttt cac 576
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
atc gac cag gtg ctg gcg ctc aac gat cgg ggc tac ctg gct tgc ctg 624
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ala Cys Leu
195 200 205
gag cgg atc cag agc cac gtg cat ttc atc aac ggc agc tgg gac gaa 672
Glu Arg Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
tac acc acc gcc gag gac gcc cgc cag ttc cgc gac tac ctg ccg cac 720
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
tgc agt ttc tcg cgg gtg gag ggc acc ggg cat ttc ctc gac ctg gag 768
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
tcc aag ctg gcc gcg gta cgc gtg cac cgc gcc ctg ctc gag cac ctg 816
Ser Lys Leu Ala Ala Val Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
ctg aag caa ccg gag ccg cag cgg gcg gaa cgc gcg gcg gga ttc cac 864
Leu Lys Gln Pro Glu Pro Gln Arg Ala Glu Arg Ala Ala Gly Phe His
275 280 285
gag atg gcc atc ggc tac gcc tga 888
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 2
<211> 295
<212> PRT
<213> Pseudomonas aeruginosa
<400> 2
Met Arg Arg Glu Ser Leu Leu Val Ser Val Cys Lys Gly Leu Arg Val
1 5 10 15
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Ser Thr Val Met Leu
20 25 30
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Glu Val Asn
85 90 95
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
Ser Arg Asn Pro Arg Gly Ile Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Ala
130 135 140
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
Thr Val Gly Lys Tyr Leu Pro Pro Arg Leu Lys Ala Ser Asn His Gln
165 170 175
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ala Cys Leu
195 200 205
Glu Arg Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
Ser Lys Leu Ala Ala Val Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
Leu Lys Gln Pro Glu Pro Gln Arg Ala Glu Arg Ala Ala Gly Phe His
275 280 285
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 3
<211> 1281
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(1281)
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atg cac gcc atc ctc atc gcc atc ggc tcg gcc ggc gac gta ttt ccc 48
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
ttc atc ggc ctg gcc cgg acc ctg aaa ttg cgc ggg cac cgc gtg agc 96
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
ctc tgc acc atc ccg gtg ttt cgc gac gcg gtg gag cag cac ggc atc 144
Leu Cys Thr Ile Pro Val Phe Arg Asp Ala Val Glu Gln His Gly Ile
35 40 45
gcg ttc gtc ccg ctg agc gac gaa ctg acc tac cgc cgg acc atg ggc 192
Ala Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
gat ccg cgc ctg tgg gac ccc aag acg tcc ttc ggc gtg ctc tgg caa 240
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
acc atc gcc ggg atg atc gag ccg gtc tac gag tac gtc tcg gcg cag 288
Thr Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Ser Ala Gln
85 90 95
cgc cat gac gac atc gtg gtg gtc ggc tcg ctc tgg gcg ctg ggc gca 336
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
cgc atc gct cac gag aag tac ggg att ccc tac ctg tcc gcg cag gtc 384
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Ala Gln Val
115 120 125
tcg cca tcg acc ttg ttg tcg gcg cac ctg ccg ccg gta cac ccc aag 432
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Lys
130 135 140
ttc aac gtg ccc gag cag atg ccg ctg gcg atg cgc aag ctg ctc tgg 480
Phe Asn Val Pro Glu Gln Met Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
cgc tgc atc gag cgc ttc aag ctg gat cgc acc tgc gcg ccg gat atc 528
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Asp Ile
165 170 175
aac gcg gtg cgg cgc aag gtc ggc ctg gag acg ccg gtg aag cgc atc 576
Asn Ala Val Arg Arg Lys Val Gly Leu Glu Thr Pro Val Lys Arg Ile
180 185 190
ttc acc caa tgg atg cat tcg ccg cag ggc gtg gtc tgc ctg ttc ccg 624
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Val Cys Leu Phe Pro
195 200 205
gcc tgg ttc gcg ccg ccc cag cag gat tgg ccg caa ccc ctg cac atg 672
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
acc ggc ttc ccg ctg ttc gac ggc agt atc ccg ggg acc ccg ctc gac 720
Thr Gly Phe Pro Leu Phe Asp Gly Ser Ile Pro Gly Thr Pro Leu Asp
225 230 235 240
gac gaa ctg caa cgc ttt ctc gat cag ggc agc cgg ccg ctg gtg ttc 768
Asp Glu Leu Gln Arg Phe Leu Asp Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
acc cag ggc tcg acc gaa cac ctg cag ggc gac ttc tac gcc atg gcc 816
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
ctg cgc gcg ctg gaa cgc ctc ggc gcg cgt ggg atc ttc ctc acc ggc 864
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
gcc ggc cag gaa ccg ctg cgc ggc ttg ccg aac cac gtg ctg cag cgc 912
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Asn His Val Leu Gln Arg
290 295 300
gcc tac gcg cca ctg gga gcc ttg ctg cca tcg tgc gcc ggg ctg gtc 960
Ala Tyr Ala Pro Leu Gly Ala Leu Leu Pro Ser Cys Ala Gly Leu Val
305 310 315 320
cat ccg ggc ggt atc ggc gcc atg agc ctg gcc ttg gcg gcg ggg gtg 1008
His Pro Gly Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
ccg cag gtg ctg ctg ccc tgc gcc cac gac cag ttc gac aat gcc gaa 1056
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
cgg ctg gtc cgg ctc ggc tgc ggg atg cgc ctg ggc gtg cca ttg cgc 1104
Arg Leu Val Arg Leu Gly Cys Gly Met Arg Leu Gly Val Pro Leu Arg
355 360 365
gag cag gag ttg cgc ggg gcg ctg tgg cgc ttg ctc gag gac ccg gcc 1152
Glu Gln Glu Leu Arg Gly Ala Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
atg gcg gcg gcc tgt cgg cgt ttc atg gaa ttg tca caa ccg cac agt 1200
Met Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
atc gct tgc ggt aaa gcg gcc cag gtg gtc gaa cgt tgt cat agg gag 1248
Ile Ala Cys Gly Lys Ala Ala Gln Val Val Glu Arg Cys His Arg Glu
405 410 415
ggg gat gcg cga tgg ctg aag gct gcg tcc tga 1281
Gly Asp Ala Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 4
<211> 426
<212> PRT
<213> Pseudomonas aeruginosa
<400> 4
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
Leu Cys Thr Ile Pro Val Phe Arg Asp Ala Val Glu Gln His Gly Ile
35 40 45
Ala Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
Thr Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Ser Ala Gln
85 90 95
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Ala Gln Val
115 120 125
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Lys
130 135 140
Phe Asn Val Pro Glu Gln Met Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Asp Ile
165 170 175
Asn Ala Val Arg Arg Lys Val Gly Leu Glu Thr Pro Val Lys Arg Ile
180 185 190
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Val Cys Leu Phe Pro
195 200 205
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
Thr Gly Phe Pro Leu Phe Asp Gly Ser Ile Pro Gly Thr Pro Leu Asp
225 230 235 240
Asp Glu Leu Gln Arg Phe Leu Asp Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Asn His Val Leu Gln Arg
290 295 300
Ala Tyr Ala Pro Leu Gly Ala Leu Leu Pro Ser Cys Ala Gly Leu Val
305 310 315 320
His Pro Gly Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
Arg Leu Val Arg Leu Gly Cys Gly Met Arg Leu Gly Val Pro Leu Arg
355 360 365
Glu Gln Glu Leu Arg Gly Ala Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
Met Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
Ile Ala Cys Gly Lys Ala Ala Gln Val Val Glu Arg Cys His Arg Glu
405 410 415
Gly Asp Ala Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 5
<211> 978
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(978)
<400> 5
atg gac cgg ata gac atg ggc gtg ctg gtg gta ctg ttc aat cct ggc 48
Met Asp Arg Ile Asp Met Gly Val Leu Val Val Leu Phe Asn Pro Gly
1 5 10 15
gac gac gac ctg gaa cac ctt ggc gaa ctg gcg gcg gcg ttt ccg caa 96
Asp Asp Asp Leu Glu His Leu Gly Glu Leu Ala Ala Ala Phe Pro Gln
20 25 30
ctg cgc ttc ctt gcc gtc gac aac tca ccg cac agc gat ccg cag cgc 144
Leu Arg Phe Leu Ala Val Asp Asn Ser Pro His Ser Asp Pro Gln Arg
35 40 45
aat gcc cgg ctg cgc ggg caa ggc atc gcc gtg ctg cac cac ggc aac 192
Asn Ala Arg Leu Arg Gly Gln Gly Ile Ala Val Leu His His Gly Asn
50 55 60
cgg cag ggc atc gcc ggc gcc ttc aac cag gga ctc gac gcg cta ttc 240
Arg Gln Gly Ile Ala Gly Ala Phe Asn Gln Gly Leu Asp Ala Leu Phe
65 70 75 80
cgg cgt ggc gtg cag ggt gtg ctg ctg ctc gac cag gac tcc cgt ccc 288
Arg Arg Gly Val Gln Gly Val Leu Leu Leu Asp Gln Asp Ser Arg Pro
85 90 95
ggc ggc gcc ttc ctc gcc gcc cag tgg cgc aac ctg cag gcg cgc aac 336
Gly Gly Ala Phe Leu Ala Ala Gln Trp Arg Asn Leu Gln Ala Arg Asn
100 105 110
ggt cag gcc tgc ctg ctc ggc cca cgg atc ttc gac cgg ggt gac cgg 384
Gly Gln Ala Cys Leu Leu Gly Pro Arg Ile Phe Asp Arg Gly Asp Arg
115 120 125
cgc ttc ctg ccg gcc atc cat ctc gac gga ctg acg ctc agg caa ttg 432
Arg Phe Leu Pro Ala Ile His Leu Asp Gly Leu Thr Leu Arg Gln Leu
130 135 140
tct ctg gac ggc ctg acg acc ccg cag cgc acc tcg ttc ctg atc tcc 480
Ser Leu Asp Gly Leu Thr Thr Pro Gln Arg Thr Ser Phe Leu Ile Ser
145 150 155 160
tcc ggc tgc ctg ctg acc cgc gag gcc tac cag cgc ctc ggc cac ttc 528
Ser Gly Cys Leu Leu Thr Arg Glu Ala Tyr Gln Arg Leu Gly His Phe
165 170 175
gac gag gaa ctg ttc atc gac cac gtg gac acc gaa tac agc ctg cgc 576
Asp Glu Glu Leu Phe Ile Asp His Val Asp Thr Glu Tyr Ser Leu Arg
180 185 190
gcc cag gcg ctg gac gtg ccc ctg tac gtc gac ccg cgg ctg gtc ctc 624
Ala Gln Ala Leu Asp Val Pro Leu Tyr Val Asp Pro Arg Leu Val Leu
195 200 205
gag cac cgc atc ggc acg cgc aag acc cgc cgc ctc ggc ggt ctc agc 672
Glu His Arg Ile Gly Thr Arg Lys Thr Arg Arg Leu Gly Gly Leu Ser
210 215 220
ctc agc gcg atg aac cac gcc ccg ctg cgc cgc tac tac ctg gcg cgc 720
Leu Ser Ala Met Asn His Ala Pro Leu Arg Arg Tyr Tyr Leu Ala Arg
225 230 235 240
aac ggc ctg ctg gtc ctg cgc cgc tac gcc cgg tcc tcg ccg ctg gcc 768
Asn Gly Leu Leu Val Leu Arg Arg Tyr Ala Arg Ser Ser Pro Leu Ala
245 250 255
ctg ctg gcg aac ctg ccg acc ctg acc cag ggc ctc gcg gtg ctc ctg 816
Leu Leu Ala Asn Leu Pro Thr Leu Thr Gln Gly Leu Ala Val Leu Leu
260 265 270
ctc gaa cgc gac aag ctg ctc aag ctg cgc tgc ctg ggc tgg ggc ctg 864
Leu Glu Arg Asp Lys Leu Leu Lys Leu Arg Cys Leu Gly Trp Gly Leu
275 280 285
tgg gac ggc ctg cgg gga cgc ggc ggc gcg ctg gag acc aac cgc ccg 912
Trp Asp Gly Leu Arg Gly Arg Gly Gly Ala Leu Glu Thr Asn Arg Pro
290 295 300
cgc ctg ctg aag cgc ctc gcc ggc ccg gcc gtg gcg tcc gta gct tcc 960
Arg Leu Leu Lys Arg Leu Ala Gly Pro Ala Val Ala Ser Val Ala Ser
305 310 315 320
ggc aag gcc aag gcc tag 978
Gly Lys Ala Lys Ala
325
<210> 6
<211> 325
<212> PRT
<213> Pseudomonas aeruginosa
<400> 6
Met Asp Arg Ile Asp Met Gly Val Leu Val Val Leu Phe Asn Pro Gly
1 5 10 15
Asp Asp Asp Leu Glu His Leu Gly Glu Leu Ala Ala Ala Phe Pro Gln
20 25 30
Leu Arg Phe Leu Ala Val Asp Asn Ser Pro His Ser Asp Pro Gln Arg
35 40 45
Asn Ala Arg Leu Arg Gly Gln Gly Ile Ala Val Leu His His Gly Asn
50 55 60
Arg Gln Gly Ile Ala Gly Ala Phe Asn Gln Gly Leu Asp Ala Leu Phe
65 70 75 80
Arg Arg Gly Val Gln Gly Val Leu Leu Leu Asp Gln Asp Ser Arg Pro
85 90 95
Gly Gly Ala Phe Leu Ala Ala Gln Trp Arg Asn Leu Gln Ala Arg Asn
100 105 110
Gly Gln Ala Cys Leu Leu Gly Pro Arg Ile Phe Asp Arg Gly Asp Arg
115 120 125
Arg Phe Leu Pro Ala Ile His Leu Asp Gly Leu Thr Leu Arg Gln Leu
130 135 140
Ser Leu Asp Gly Leu Thr Thr Pro Gln Arg Thr Ser Phe Leu Ile Ser
145 150 155 160
Ser Gly Cys Leu Leu Thr Arg Glu Ala Tyr Gln Arg Leu Gly His Phe
165 170 175
Asp Glu Glu Leu Phe Ile Asp His Val Asp Thr Glu Tyr Ser Leu Arg
180 185 190
Ala Gln Ala Leu Asp Val Pro Leu Tyr Val Asp Pro Arg Leu Val Leu
195 200 205
Glu His Arg Ile Gly Thr Arg Lys Thr Arg Arg Leu Gly Gly Leu Ser
210 215 220
Leu Ser Ala Met Asn His Ala Pro Leu Arg Arg Tyr Tyr Leu Ala Arg
225 230 235 240
Asn Gly Leu Leu Val Leu Arg Arg Tyr Ala Arg Ser Ser Pro Leu Ala
245 250 255
Leu Leu Ala Asn Leu Pro Thr Leu Thr Gln Gly Leu Ala Val Leu Leu
260 265 270
Leu Glu Arg Asp Lys Leu Leu Lys Leu Arg Cys Leu Gly Trp Gly Leu
275 280 285
Trp Asp Gly Leu Arg Gly Arg Gly Gly Ala Leu Glu Thr Asn Arg Pro
290 295 300
Arg Leu Leu Lys Arg Leu Ala Gly Pro Ala Val Ala Ser Val Ala Ser
305 310 315 320
Gly Lys Ala Lys Ala
325
<210> 7
<211> 1269
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(1269)
<400> 7
gtg tcc acg acc agc ctc tgc ccc tcc gcc acg cgg gaa cac ggt ccc 48
Val Ser Thr Thr Ser Leu Cys Pro Ser Ala Thr Arg Glu His Gly Pro
1 5 10 15
ggc gcg aaa cgc gtc ctg cct ctg ctg ttc ctc acc tgc ctg ctg gat 96
Gly Ala Lys Arg Val Leu Pro Leu Leu Phe Leu Thr Cys Leu Leu Asp
20 25 30
gcc gct ggc gtc ggc ctg atc gtg ccc ctg ctg ccg acg ctg atc ggc 144
Ala Ala Gly Val Gly Leu Ile Val Pro Leu Leu Pro Thr Leu Ile Gly
35 40 45
agc gtg gcg ccg ctg gcg gtc cgc gac gcg gcc acc tgg ggc gcc gcc 192
Ser Val Ala Pro Leu Ala Val Arg Asp Ala Ala Thr Trp Gly Ala Ala
50 55 60
ctg gtg atg acc ttc gcg ctg ctg caa ttg ttc ttt tcg ccg gtc ctc 240
Leu Val Met Thr Phe Ala Leu Leu Gln Leu Phe Phe Ser Pro Val Leu
65 70 75 80
ggc agc ctc agc gac cgc ttc gga cgc cgc ccc gtc ctg gtc ctg gcg 288
Gly Ser Leu Ser Asp Arg Phe Gly Arg Arg Pro Val Leu Val Leu Ala
85 90 95
atg ctc ggc ttc gcc ctc agc tat ctg ctg ctg gcg ctg gcc gac agc 336
Met Leu Gly Phe Ala Leu Ser Tyr Leu Leu Leu Ala Leu Ala Asp Ser
100 105 110
ctc tgg atg ctg ttc ctc ggt cgc gcg ctg gcc ggg ctc acc ggc gcc 384
Leu Trp Met Leu Phe Leu Gly Arg Ala Leu Ala Gly Leu Thr Gly Ala
115 120 125
agc gtg gcc acc gcg atg gcc tgc gcg gct gac ctc ggc acg cac ggg 432
Ser Val Ala Thr Ala Met Ala Cys Ala Ala Asp Leu Gly Thr His Gly
130 135 140
cag cgc acc cgg cac ttc ggc tgg ctg tac gcc ggc ctc gcc ctg ggc 480
Gln Arg Thr Arg His Phe Gly Trp Leu Tyr Ala Gly Leu Ala Leu Gly
145 150 155 160
atg atc ctc ggc ccc gcc ctc ggt ggg ctg ctg gcg gtg cac ggc acg 528
Met Ile Leu Gly Pro Ala Leu Gly Gly Leu Leu Ala Val His Gly Thr
165 170 175
acg ctg ccg ctg ttg ctg gcc gcc ggc ctg tgc ctg ctc aac gcc ctg 576
Thr Leu Pro Leu Leu Leu Ala Ala Gly Leu Cys Leu Leu Asn Ala Leu
180 185 190
ctc gcc ggc ctg ttc ctc gag gaa acc ctg ccc ccg acg cga cgc cgc 624
Leu Ala Gly Leu Phe Leu Glu Glu Thr Leu Pro Pro Thr Arg Arg Arg
195 200 205
cgc ctg gac ccg agg cgg atg aat gcc ttg cgc tcg atc agc ggc ctg 672
Arg Leu Asp Pro Arg Arg Met Asn Ala Leu Arg Ser Ile Ser Gly Leu
210 215 220
gct cgg caa ccg ggg gtc gga cgc ctg ctg gcg gtg ctt gcc ctg gta 720
Ala Arg Gln Pro Gly Val Gly Arg Leu Leu Ala Val Leu Ala Leu Val
225 230 235 240
ttc ctc ggc ttg cag gcg gtg atg gtg gtc tgg ccg ttc ttc gtg atc 768
Phe Leu Gly Leu Gln Ala Val Met Val Val Trp Pro Phe Phe Val Ile
245 250 255
gag aag ttt cac tgg agc agc gcc tgg atc ggc tac tcg ctg gcc ctc 816
Glu Lys Phe His Trp Ser Ser Ala Trp Ile Gly Tyr Ser Leu Ala Leu
260 265 270
tac ggc gtg ctc gcg gtg ctc gcc cag acc ctc ggc gtg aac ctc tgc 864
Tyr Gly Val Leu Ala Val Leu Ala Gln Thr Leu Gly Val Asn Leu Cys
275 280 285
aag cgg cgc ctg gac gac gcc cgc ctg ctg cgc ctg ggc ctc gcc ctg 912
Lys Arg Arg Leu Asp Asp Ala Arg Leu Leu Arg Leu Gly Leu Ala Leu
290 295 300
caa ggc tgc ggc ctg ctg ctg ttc gcc ctg gtc gac tcg tca ttc tgg 960
Gln Gly Cys Gly Leu Leu Leu Phe Ala Leu Val Asp Ser Ser Phe Trp
305 310 315 320
ctg gtc tgc gcg ctg ctg ccc ttc gcg ctc ggc agc ctc gcc acc ccg 1008
Leu Val Cys Ala Leu Leu Pro Phe Ala Leu Gly Ser Leu Ala Thr Pro
325 330 335
gcc atg cag ggg ctg ctc tcg gcc cgc gtg ccg gtc gac cgc cag ggc 1056
Ala Met Gln Gly Leu Leu Ser Ala Arg Val Pro Val Asp Arg Gln Gly
340 345 350
gag ttg cag ggc gtg ctg agc agc ctg atg agc ctc gcc gcg atc gtc 1104
Glu Leu Gln Gly Val Leu Ser Ser Leu Met Ser Leu Ala Ala Ile Val
355 360 365
ggt ccg ccg ctg atg agc ggc ctg ttc cac tgg ggc agc ggt ccg ctc 1152
Gly Pro Pro Leu Met Ser Gly Leu Phe His Trp Gly Ser Gly Pro Leu
370 375 380
gcg ccg ctg ccc ctg gcc ggc gcg cca ttc ctc gcc ggc gcc ctt ctc 1200
Ala Pro Leu Pro Leu Ala Gly Ala Pro Phe Leu Ala Gly Ala Leu Leu
385 390 395 400
gtt ctg gcc ggg ctg gtc ctg gcc tgg caa ctt cga cct acg gga gaa 1248
Val Leu Ala Gly Leu Val Leu Ala Trp Gln Leu Arg Pro Thr Gly Glu
405 410 415
gaa cga tca tgg acc gga tag 1269
Glu Arg Ser Trp Thr Gly
420
<210> 8
<211> 422
<212> PRT
<213> Pseudomonas aeruginosa
<400> 8
Val Ser Thr Thr Ser Leu Cys Pro Ser Ala Thr Arg Glu His Gly Pro
1 5 10 15
Gly Ala Lys Arg Val Leu Pro Leu Leu Phe Leu Thr Cys Leu Leu Asp
20 25 30
Ala Ala Gly Val Gly Leu Ile Val Pro Leu Leu Pro Thr Leu Ile Gly
35 40 45
Ser Val Ala Pro Leu Ala Val Arg Asp Ala Ala Thr Trp Gly Ala Ala
50 55 60
Leu Val Met Thr Phe Ala Leu Leu Gln Leu Phe Phe Ser Pro Val Leu
65 70 75 80
Gly Ser Leu Ser Asp Arg Phe Gly Arg Arg Pro Val Leu Val Leu Ala
85 90 95
Met Leu Gly Phe Ala Leu Ser Tyr Leu Leu Leu Ala Leu Ala Asp Ser
100 105 110
Leu Trp Met Leu Phe Leu Gly Arg Ala Leu Ala Gly Leu Thr Gly Ala
115 120 125
Ser Val Ala Thr Ala Met Ala Cys Ala Ala Asp Leu Gly Thr His Gly
130 135 140
Gln Arg Thr Arg His Phe Gly Trp Leu Tyr Ala Gly Leu Ala Leu Gly
145 150 155 160
Met Ile Leu Gly Pro Ala Leu Gly Gly Leu Leu Ala Val His Gly Thr
165 170 175
Thr Leu Pro Leu Leu Leu Ala Ala Gly Leu Cys Leu Leu Asn Ala Leu
180 185 190
Leu Ala Gly Leu Phe Leu Glu Glu Thr Leu Pro Pro Thr Arg Arg Arg
195 200 205
Arg Leu Asp Pro Arg Arg Met Asn Ala Leu Arg Ser Ile Ser Gly Leu
210 215 220
Ala Arg Gln Pro Gly Val Gly Arg Leu Leu Ala Val Leu Ala Leu Val
225 230 235 240
Phe Leu Gly Leu Gln Ala Val Met Val Val Trp Pro Phe Phe Val Ile
245 250 255
Glu Lys Phe His Trp Ser Ser Ala Trp Ile Gly Tyr Ser Leu Ala Leu
260 265 270
Tyr Gly Val Leu Ala Val Leu Ala Gln Thr Leu Gly Val Asn Leu Cys
275 280 285
Lys Arg Arg Leu Asp Asp Ala Arg Leu Leu Arg Leu Gly Leu Ala Leu
290 295 300
Gln Gly Cys Gly Leu Leu Leu Phe Ala Leu Val Asp Ser Ser Phe Trp
305 310 315 320
Leu Val Cys Ala Leu Leu Pro Phe Ala Leu Gly Ser Leu Ala Thr Pro
325 330 335
Ala Met Gln Gly Leu Leu Ser Ala Arg Val Pro Val Asp Arg Gln Gly
340 345 350
Glu Leu Gln Gly Val Leu Ser Ser Leu Met Ser Leu Ala Ala Ile Val
355 360 365
Gly Pro Pro Leu Met Ser Gly Leu Phe His Trp Gly Ser Gly Pro Leu
370 375 380
Ala Pro Leu Pro Leu Ala Gly Ala Pro Phe Leu Ala Gly Ala Leu Leu
385 390 395 400
Val Leu Ala Gly Leu Val Leu Ala Trp Gln Leu Arg Pro Thr Gly Glu
405 410 415
Glu Arg Ser Trp Thr Gly
420
<210> 9
<211> 882
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(882)
<400> 9
atg gct cgt aaa gga att att ctg gcc ggc ggt tcg ggt aca cgc ctg 48
Met Ala Arg Lys Gly Ile Ile Leu Ala Gly Gly Ser Gly Thr Arg Leu
1 5 10 15
cat ccg gcc aca ctt tcg gtt tcg aag cag ctg ctg ccg gtg tat gac 96
His Pro Ala Thr Leu Ser Val Ser Lys Gln Leu Leu Pro Val Tyr Asp
20 25 30
aaa ccg atg atc tac tac ccg ctg agc acc ctg ctg ctc gct ggt atc 144
Lys Pro Met Ile Tyr Tyr Pro Leu Ser Thr Leu Leu Leu Ala Gly Ile
35 40 45
cgg gac atc ctg atc att tcc acc ccg cag gac acc ccg cgc ttc gaa 192
Arg Asp Ile Leu Ile Ile Ser Thr Pro Gln Asp Thr Pro Arg Phe Glu
50 55 60
cag ctg ctg ggc gat ggc agc cag tgg ggc ctg aac ctg tca tac gca 240
Gln Leu Leu Gly Asp Gly Ser Gln Trp Gly Leu Asn Leu Ser Tyr Ala
65 70 75 80
ata caa cca agc ccg gat ggc ttg gcg caa gcg ttc acc atc ggc gct 288
Ile Gln Pro Ser Pro Asp Gly Leu Ala Gln Ala Phe Thr Ile Gly Ala
85 90 95
gac ttc atc ggt aac gac cct tct gcg ttg gtt ctc ggt gac aat att 336
Asp Phe Ile Gly Asn Asp Pro Ser Ala Leu Val Leu Gly Asp Asn Ile
100 105 110
ttc tac ggc cat gac ttc cag gca ctg cta ttg aac gca gat aaa cgt 384
Phe Tyr Gly His Asp Phe Gln Ala Leu Leu Leu Asn Ala Asp Lys Arg
115 120 125
gaa tcc ggt gct tca gta ttc gct tat cat gtt cat gac cca gaa cgc 432
Glu Ser Gly Ala Ser Val Phe Ala Tyr His Val His Asp Pro Glu Arg
130 135 140
tat ggc gta gcg gag ttt gac gat agc ggt cgc gta ttg tcg ctg gaa 480
Tyr Gly Val Ala Glu Phe Asp Asp Ser Gly Arg Val Leu Ser Leu Glu
145 150 155 160
gaa aaa ccg gca gtt cca aag tct agc tat gcg gtc acc ggc ctg tat 528
Glu Lys Pro Ala Val Pro Lys Ser Ser Tyr Ala Val Thr Gly Leu Tyr
165 170 175
ttc tat gac aat cag gta gtc aat ctg gct cgc gag ctg aag cct tcc 576
Phe Tyr Asp Asn Gln Val Val Asn Leu Ala Arg Glu Leu Lys Pro Ser
180 185 190
cca cgt ggc gag ctg gaa atc acc gac ctc aac aac ctt tac ttg cag 624
Pro Arg Gly Glu Leu Glu Ile Thr Asp Leu Asn Asn Leu Tyr Leu Gln
195 200 205
cag cag cag ttg cag gtc gaa atc atg ggc cgt ggc tat gcg tgg ctc 672
Gln Gln Gln Leu Gln Val Glu Ile Met Gly Arg Gly Tyr Ala Trp Leu
210 215 220
gac acc ggc acg cac gac agt ctg ctg gag gct agc cag tac atc gca 720
Asp Thr Gly Thr His Asp Ser Leu Leu Glu Ala Ser Gln Tyr Ile Ala
225 230 235 240
acc atg gag cgc cgt cag ggc ttg aaa gtc gcc tgc cct gag gaa att 768
Thr Met Glu Arg Arg Gln Gly Leu Lys Val Ala Cys Pro Glu Glu Ile
245 250 255
tgc tac cgc gct ggc tgg atc aac gct gag caa ctc gag tgc ctg gct 816
Cys Tyr Arg Ala Gly Trp Ile Asn Ala Glu Gln Leu Glu Cys Leu Ala
260 265 270
caa cca ctg ctg aaa aac ggt tat ggc aag tat ctg cag aac ttg ctg 864
Gln Pro Leu Leu Lys Asn Gly Tyr Gly Lys Tyr Leu Gln Asn Leu Leu
275 280 285
aaa gag aag gtg ttc tga 882
Lys Glu Lys Val Phe
290
<210> 10
<211> 293
<212> PRT
<213> Pseudomonas putida
<400> 10
Met Ala Arg Lys Gly Ile Ile Leu Ala Gly Gly Ser Gly Thr Arg Leu
1 5 10 15
His Pro Ala Thr Leu Ser Val Ser Lys Gln Leu Leu Pro Val Tyr Asp
20 25 30
Lys Pro Met Ile Tyr Tyr Pro Leu Ser Thr Leu Leu Leu Ala Gly Ile
35 40 45
Arg Asp Ile Leu Ile Ile Ser Thr Pro Gln Asp Thr Pro Arg Phe Glu
50 55 60
Gln Leu Leu Gly Asp Gly Ser Gln Trp Gly Leu Asn Leu Ser Tyr Ala
65 70 75 80
Ile Gln Pro Ser Pro Asp Gly Leu Ala Gln Ala Phe Thr Ile Gly Ala
85 90 95
Asp Phe Ile Gly Asn Asp Pro Ser Ala Leu Val Leu Gly Asp Asn Ile
100 105 110
Phe Tyr Gly His Asp Phe Gln Ala Leu Leu Leu Asn Ala Asp Lys Arg
115 120 125
Glu Ser Gly Ala Ser Val Phe Ala Tyr His Val His Asp Pro Glu Arg
130 135 140
Tyr Gly Val Ala Glu Phe Asp Asp Ser Gly Arg Val Leu Ser Leu Glu
145 150 155 160
Glu Lys Pro Ala Val Pro Lys Ser Ser Tyr Ala Val Thr Gly Leu Tyr
165 170 175
Phe Tyr Asp Asn Gln Val Val Asn Leu Ala Arg Glu Leu Lys Pro Ser
180 185 190
Pro Arg Gly Glu Leu Glu Ile Thr Asp Leu Asn Asn Leu Tyr Leu Gln
195 200 205
Gln Gln Gln Leu Gln Val Glu Ile Met Gly Arg Gly Tyr Ala Trp Leu
210 215 220
Asp Thr Gly Thr His Asp Ser Leu Leu Glu Ala Ser Gln Tyr Ile Ala
225 230 235 240
Thr Met Glu Arg Arg Gln Gly Leu Lys Val Ala Cys Pro Glu Glu Ile
245 250 255
Cys Tyr Arg Ala Gly Trp Ile Asn Ala Glu Gln Leu Glu Cys Leu Ala
260 265 270
Gln Pro Leu Leu Lys Asn Gly Tyr Gly Lys Tyr Leu Gln Asn Leu Leu
275 280 285
Lys Glu Lys Val Phe
290
<210> 11
<211> 1101
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(1101)
<400> 11
atg att cta gta aca ggc gga gcc ggc ttc atc ggc tca aat ttc gta 48
Met Ile Leu Val Thr Gly Gly Ala Gly Phe Ile Gly Ser Asn Phe Val
1 5 10 15
ctg caa tgg tgt gcg cac aat gag gaa ccc gtc ctc aac ctc gac gcc 96
Leu Gln Trp Cys Ala His Asn Glu Glu Pro Val Leu Asn Leu Asp Ala
20 25 30
ctg acc tac gca ggc aac ctg gcc aac ctg cag ccg ctg gaa ggc aac 144
Leu Thr Tyr Ala Gly Asn Leu Ala Asn Leu Gln Pro Leu Glu Gly Asn
35 40 45
cct cag cat cgc ttt gtg caa ggc aat att tgc gat gct gcg ctt ctg 192
Pro Gln His Arg Phe Val Gln Gly Asn Ile Cys Asp Ala Ala Leu Leu
50 55 60
acc aag ctg ttc gca gag cac cgc ccg cgc gcc gtg gtt cac ttc gcg 240
Thr Lys Leu Phe Ala Glu His Arg Pro Arg Ala Val Val His Phe Ala
65 70 75 80
gcg gaa tcc cat gta gac cgc tca atc acc ggc ccc gaa gcg ttt gtc 288
Ala Glu Ser His Val Asp Arg Ser Ile Thr Gly Pro Glu Ala Phe Val
85 90 95
gaa acc aac gtg atg ggc acg ttt cgc ttg ctt gaa gcc gcc cgg gcg 336
Glu Thr Asn Val Met Gly Thr Phe Arg Leu Leu Glu Ala Ala Arg Ala
100 105 110
cat tgg aat agt ttg gaa ggt gca gag aag gag gcc ttc cgt ttc ctc 384
His Trp Asn Ser Leu Glu Gly Ala Glu Lys Glu Ala Phe Arg Phe Leu
115 120 125
cat gtc tct acc gac gaa gtc tac ggc aca cta ggg cca aac gac ccg 432
His Val Ser Thr Asp Glu Val Tyr Gly Thr Leu Gly Pro Asn Asp Pro
130 135 140
gcg ttc acc gaa acc acg ccg tac gcg ccg aac agc cca tac tcc gcc 480
Ala Phe Thr Glu Thr Thr Pro Tyr Ala Pro Asn Ser Pro Tyr Ser Ala
145 150 155 160
agc aag gca gcc agc gac cat ctg gta cgc tcg tat ttc cat acc tac 528
Ser Lys Ala Ala Ser Asp His Leu Val Arg Ser Tyr Phe His Thr Tyr
165 170 175
ggc atg ccg gta ctc act acc aac tgc tcc aac aat tac ggg ccg ctc 576
Gly Met Pro Val Leu Thr Thr Asn Cys Ser Asn Asn Tyr Gly Pro Leu
180 185 190
cac ttc ccg gaa aaa ctg atc ccg ctg atg atc gtc aac gca ctc gcc 624
His Phe Pro Glu Lys Leu Ile Pro Leu Met Ile Val Asn Ala Leu Ala
195 200 205
ggt aag gcg ctg cct gtc tat ggc gac ggc cag caa atc cgc gac tgg 672
Gly Lys Ala Leu Pro Val Tyr Gly Asp Gly Gln Gln Ile Arg Asp Trp
210 215 220
ctg tat gtc gaa gat cac tgc tcg ggc atc cgt cgc gta ctg gaa gcc 720
Leu Tyr Val Glu Asp His Cys Ser Gly Ile Arg Arg Val Leu Glu Ala
225 230 235 240
ggt gcg ttc ggc gag acg tac aat att ggc ggc tgg aat gaa aaa gcc 768
Gly Ala Phe Gly Glu Thr Tyr Asn Ile Gly Gly Trp Asn Glu Lys Ala
245 250 255
aac att gac att gtg cgt aca ctc tgc agc ctt ctc gac gag atg gca 816
Asn Ile Asp Ile Val Arg Thr Leu Cys Ser Leu Leu Asp Glu Met Ala
260 265 270
cct gcg gca tcg cgc cag gta atc aat cag aag acc ggc gag cct gtc 864
Pro Ala Ala Ser Arg Gln Val Ile Asn Gln Lys Thr Gly Glu Pro Val
275 280 285
gaa cag tat gca gaa ctc atc gcc tac gta acc gac cgc cca ggc cat 912
Glu Gln Tyr Ala Glu Leu Ile Ala Tyr Val Thr Asp Arg Pro Gly His
290 295 300
gac cgc cgt tat gcc atc gat gca cgc aag atc gag cgg gag ctc ggc 960
Asp Arg Arg Tyr Ala Ile Asp Ala Arg Lys Ile Glu Arg Glu Leu Gly
305 310 315 320
tgg aaa cct gcc gaa acc ttc gag acg ggc att cga aag aca gtc gct 1008
Trp Lys Pro Ala Glu Thr Phe Glu Thr Gly Ile Arg Lys Thr Val Ala
325 330 335
tgg tac ttg gcc aac cag aaa tgg gta aaa ggt gtc atg gac ggc agc 1056
Trp Tyr Leu Ala Asn Gln Lys Trp Val Lys Gly Val Met Asp Gly Ser
340 345 350
tac cgt gac tgg gtg gca caa caa tac ggg gca aat aaa gcg tga 1101
Tyr Arg Asp Trp Val Ala Gln Gln Tyr Gly Ala Asn Lys Ala
355 360 365
<210> 12
<211> 366
<212> PRT
<213> Pseudomonas putida
<400> 12
Met Ile Leu Val Thr Gly Gly Ala Gly Phe Ile Gly Ser Asn Phe Val
1 5 10 15
Leu Gln Trp Cys Ala His Asn Glu Glu Pro Val Leu Asn Leu Asp Ala
20 25 30
Leu Thr Tyr Ala Gly Asn Leu Ala Asn Leu Gln Pro Leu Glu Gly Asn
35 40 45
Pro Gln His Arg Phe Val Gln Gly Asn Ile Cys Asp Ala Ala Leu Leu
50 55 60
Thr Lys Leu Phe Ala Glu His Arg Pro Arg Ala Val Val His Phe Ala
65 70 75 80
Ala Glu Ser His Val Asp Arg Ser Ile Thr Gly Pro Glu Ala Phe Val
85 90 95
Glu Thr Asn Val Met Gly Thr Phe Arg Leu Leu Glu Ala Ala Arg Ala
100 105 110
His Trp Asn Ser Leu Glu Gly Ala Glu Lys Glu Ala Phe Arg Phe Leu
115 120 125
His Val Ser Thr Asp Glu Val Tyr Gly Thr Leu Gly Pro Asn Asp Pro
130 135 140
Ala Phe Thr Glu Thr Thr Pro Tyr Ala Pro Asn Ser Pro Tyr Ser Ala
145 150 155 160
Ser Lys Ala Ala Ser Asp His Leu Val Arg Ser Tyr Phe His Thr Tyr
165 170 175
Gly Met Pro Val Leu Thr Thr Asn Cys Ser Asn Asn Tyr Gly Pro Leu
180 185 190
His Phe Pro Glu Lys Leu Ile Pro Leu Met Ile Val Asn Ala Leu Ala
195 200 205
Gly Lys Ala Leu Pro Val Tyr Gly Asp Gly Gln Gln Ile Arg Asp Trp
210 215 220
Leu Tyr Val Glu Asp His Cys Ser Gly Ile Arg Arg Val Leu Glu Ala
225 230 235 240
Gly Ala Phe Gly Glu Thr Tyr Asn Ile Gly Gly Trp Asn Glu Lys Ala
245 250 255
Asn Ile Asp Ile Val Arg Thr Leu Cys Ser Leu Leu Asp Glu Met Ala
260 265 270
Pro Ala Ala Ser Arg Gln Val Ile Asn Gln Lys Thr Gly Glu Pro Val
275 280 285
Glu Gln Tyr Ala Glu Leu Ile Ala Tyr Val Thr Asp Arg Pro Gly His
290 295 300
Asp Arg Arg Tyr Ala Ile Asp Ala Arg Lys Ile Glu Arg Glu Leu Gly
305 310 315 320
Trp Lys Pro Ala Glu Thr Phe Glu Thr Gly Ile Arg Lys Thr Val Ala
325 330 335
Trp Tyr Leu Ala Asn Gln Lys Trp Val Lys Gly Val Met Asp Gly Ser
340 345 350
Tyr Arg Asp Trp Val Ala Gln Gln Tyr Gly Ala Asn Lys Ala
355 360 365
<210> 13
<211> 549
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(549)
<400> 13
atg caa gcc att ccg ctg gat atc ccc gaa gtc gtg ctg ttt acc ccc 48
Met Gln Ala Ile Pro Leu Asp Ile Pro Glu Val Val Leu Phe Thr Pro
1 5 10 15
aag gtt ttt ggc gac gaa cgt ggt ttc ttc tac gag agc ttc aac gcc 96
Lys Val Phe Gly Asp Glu Arg Gly Phe Phe Tyr Glu Ser Phe Asn Ala
20 25 30
cgt gtt ttc agc gaa gtg acc ggc ctg cag ccc gac ttc gta caa gac 144
Arg Val Phe Ser Glu Val Thr Gly Leu Gln Pro Asp Phe Val Gln Asp
35 40 45
aac cac tcg cgc tcg gta aaa ggc gtg ctc cgt ggc ctg cac tat cag 192
Asn His Ser Arg Ser Val Lys Gly Val Leu Arg Gly Leu His Tyr Gln
50 55 60
ctg gca cct cac gcc cag ggc aag ctg gtg cgt gtg gtg caa ggc gaa 240
Leu Ala Pro His Ala Gln Gly Lys Leu Val Arg Val Val Gln Gly Glu
65 70 75 80
gtc ttc gat gtt gcg gtg gat atc cgt cgc tcg tcc aca acc ttc ggt 288
Val Phe Asp Val Ala Val Asp Ile Arg Arg Ser Ser Thr Thr Phe Gly
85 90 95
aaa tgg gta ggt gcg gtg ttg tcg gcc gag aac aag aac cag ctg tgg 336
Lys Trp Val Gly Ala Val Leu Ser Ala Glu Asn Lys Asn Gln Leu Trp
100 105 110
atc ccg cca ggg ttc gca cac ggg ttc gtc acg ttg agt gaa acc gca 384
Ile Pro Pro Gly Phe Ala His Gly Phe Val Thr Leu Ser Glu Thr Ala
115 120 125
gag ttc ctc tac aag acc acc gac ttc tac tcg ccg cag tgc gag cgc 432
Glu Phe Leu Tyr Lys Thr Thr Asp Phe Tyr Ser Pro Gln Cys Glu Arg
130 135 140
tgc att gcc tgg aat gat ccg gca gtg ggt atc gaa tgg ccc atc gac 480
Cys Ile Ala Trp Asn Asp Pro Ala Val Gly Ile Glu Trp Pro Ile Asp
145 150 155 160
tcc gta cca agc ttg tct ggc aag gac cag ctt ggg gtc gca ttg gct 528
Ser Val Pro Ser Leu Ser Gly Lys Asp Gln Leu Gly Val Ala Leu Ala
165 170 175
gac gcc gaa ctg ttc gac taa 549
Asp Ala Glu Leu Phe Asp
180
<210> 14
<211> 182
<212> PRT
<213> Pseudomonas putida
<400> 14
Met Gln Ala Ile Pro Leu Asp Ile Pro Glu Val Val Leu Phe Thr Pro
1 5 10 15
Lys Val Phe Gly Asp Glu Arg Gly Phe Phe Tyr Glu Ser Phe Asn Ala
20 25 30
Arg Val Phe Ser Glu Val Thr Gly Leu Gln Pro Asp Phe Val Gln Asp
35 40 45
Asn His Ser Arg Ser Val Lys Gly Val Leu Arg Gly Leu His Tyr Gln
50 55 60
Leu Ala Pro His Ala Gln Gly Lys Leu Val Arg Val Val Gln Gly Glu
65 70 75 80
Val Phe Asp Val Ala Val Asp Ile Arg Arg Ser Ser Thr Thr Phe Gly
85 90 95
Lys Trp Val Gly Ala Val Leu Ser Ala Glu Asn Lys Asn Gln Leu Trp
100 105 110
Ile Pro Pro Gly Phe Ala His Gly Phe Val Thr Leu Ser Glu Thr Ala
115 120 125
Glu Phe Leu Tyr Lys Thr Thr Asp Phe Tyr Ser Pro Gln Cys Glu Arg
130 135 140
Cys Ile Ala Trp Asn Asp Pro Ala Val Gly Ile Glu Trp Pro Ile Asp
145 150 155 160
Ser Val Pro Ser Leu Ser Gly Lys Asp Gln Leu Gly Val Ala Leu Ala
165 170 175
Asp Ala Glu Leu Phe Asp
180
<210> 15
<211> 903
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(903)
<400> 15
gtg aaa atc ctg ctg ttg ggg aaa aac ggg caa gta ggc tgg gag cta 48
Val Lys Ile Leu Leu Leu Gly Lys Asn Gly Gln Val Gly Trp Glu Leu
1 5 10 15
cag cgc gcc ttg gcg ccg ctg ggt gag gtc att gcg ctg gat cgt cag 96
Gln Arg Ala Leu Ala Pro Leu Gly Glu Val Ile Ala Leu Asp Arg Gln
20 25 30
ggg gcc gag ggc tta tgt ggc gac ttg tcc aac ctg gac ggc ttg gcc 144
Gly Ala Glu Gly Leu Cys Gly Asp Leu Ser Asn Leu Asp Gly Leu Ala
35 40 45
gct acg att cgt cag ctg gcg ccg gac gtg atc gtc aac gct gct gcc 192
Ala Thr Ile Arg Gln Leu Ala Pro Asp Val Ile Val Asn Ala Ala Ala
50 55 60
tac act gca gtg gat aaa gct gag agc gat cag gca ctg gct gca atg 240
Tyr Thr Ala Val Asp Lys Ala Glu Ser Asp Gln Ala Leu Ala Ala Met
65 70 75 80
atc aat gcc gcg gct cct gct gta tta gca cgt gaa aca gca gct ttg 288
Ile Asn Ala Ala Ala Pro Ala Val Leu Ala Arg Glu Thr Ala Ala Leu
85 90 95
ggc gcc tgg ttg att cac tat tcc acc gat tat gta ttt gac ggc agc 336
Gly Ala Trp Leu Ile His Tyr Ser Thr Asp Tyr Val Phe Asp Gly Ser
100 105 110
ggc agt cag cgc tgg gag gaa act gcg cct acc ggc ccc ctt tcg gtc 384
Gly Ser Gln Arg Trp Glu Glu Thr Ala Pro Thr Gly Pro Leu Ser Val
115 120 125
tac ggc cgg acc aag ctg gaa ggc gag cat gcc att ctc gcc agc ggc 432
Tyr Gly Arg Thr Lys Leu Glu Gly Glu His Ala Ile Leu Ala Ser Gly
130 135 140
gcc aag gcc gtg gta ctg cgc acc agc tgg gtg tat gct gcg cgc ggg 480
Ala Lys Ala Val Val Leu Arg Thr Ser Trp Val Tyr Ala Ala Arg Gly
145 150 155 160
cac aat ttt gcc aag acc atg ctg cgc ctg gcg gcg gag cgt gag acg 528
His Asn Phe Ala Lys Thr Met Leu Arg Leu Ala Ala Glu Arg Glu Thr
165 170 175
ttg agc gtg gta gca gac caa ttt ggc gca ccc acg ggc gct gac ctg 576
Leu Ser Val Val Ala Asp Gln Phe Gly Ala Pro Thr Gly Ala Asp Leu
180 185 190
atc gcc gac gtt act gca cac atc ctg cgg caa atc ttc aat ggg caa 624
Ile Ala Asp Val Thr Ala His Ile Leu Arg Gln Ile Phe Asn Gly Gln
195 200 205
gac aac cgt cac ctg gca ggg att tac cac ttg gct gcg tcc ggt gaa 672
Asp Asn Arg His Leu Ala Gly Ile Tyr His Leu Ala Ala Ser Gly Glu
210 215 220
acc tct tgg cat ggt ttt gct cag ttc gtg ctg gcg cat gct caa cgc 720
Thr Ser Trp His Gly Phe Ala Gln Phe Val Leu Ala His Ala Gln Arg
225 230 235 240
act ggc gta gcg ctg aaa gtg aca gct gat aag gtt gcc gca atc agc 768
Thr Gly Val Ala Leu Lys Val Thr Ala Asp Lys Val Ala Ala Ile Ser
245 250 255
acc gaa gct tat cca gta cct gca cca cgt ccg cgc aac tcg cgc ctg 816
Thr Glu Ala Tyr Pro Val Pro Ala Pro Arg Pro Arg Asn Ser Arg Leu
260 265 270
gca ctg ggc aaa ctg gaa aac acg ttc aat ttc aaa atg ccg ctt tgg 864
Ala Leu Gly Lys Leu Glu Asn Thr Phe Asn Phe Lys Met Pro Leu Trp
275 280 285
gag caa ggc gtg caa cgt atg ctg gac gaa atc cag taa 903
Glu Gln Gly Val Gln Arg Met Leu Asp Glu Ile Gln
290 295 300
<210> 16
<211> 300
<212> PRT
<213> Pseudomonas putida
<400> 16
Val Lys Ile Leu Leu Leu Gly Lys Asn Gly Gln Val Gly Trp Glu Leu
1 5 10 15
Gln Arg Ala Leu Ala Pro Leu Gly Glu Val Ile Ala Leu Asp Arg Gln
20 25 30
Gly Ala Glu Gly Leu Cys Gly Asp Leu Ser Asn Leu Asp Gly Leu Ala
35 40 45
Ala Thr Ile Arg Gln Leu Ala Pro Asp Val Ile Val Asn Ala Ala Ala
50 55 60
Tyr Thr Ala Val Asp Lys Ala Glu Ser Asp Gln Ala Leu Ala Ala Met
65 70 75 80
Ile Asn Ala Ala Ala Pro Ala Val Leu Ala Arg Glu Thr Ala Ala Leu
85 90 95
Gly Ala Trp Leu Ile His Tyr Ser Thr Asp Tyr Val Phe Asp Gly Ser
100 105 110
Gly Ser Gln Arg Trp Glu Glu Thr Ala Pro Thr Gly Pro Leu Ser Val
115 120 125
Tyr Gly Arg Thr Lys Leu Glu Gly Glu His Ala Ile Leu Ala Ser Gly
130 135 140
Ala Lys Ala Val Val Leu Arg Thr Ser Trp Val Tyr Ala Ala Arg Gly
145 150 155 160
His Asn Phe Ala Lys Thr Met Leu Arg Leu Ala Ala Glu Arg Glu Thr
165 170 175
Leu Ser Val Val Ala Asp Gln Phe Gly Ala Pro Thr Gly Ala Asp Leu
180 185 190
Ile Ala Asp Val Thr Ala His Ile Leu Arg Gln Ile Phe Asn Gly Gln
195 200 205
Asp Asn Arg His Leu Ala Gly Ile Tyr His Leu Ala Ala Ser Gly Glu
210 215 220
Thr Ser Trp His Gly Phe Ala Gln Phe Val Leu Ala His Ala Gln Arg
225 230 235 240
Thr Gly Val Ala Leu Lys Val Thr Ala Asp Lys Val Ala Ala Ile Ser
245 250 255
Thr Glu Ala Tyr Pro Val Pro Ala Pro Arg Pro Arg Asn Ser Arg Leu
260 265 270
Ala Leu Gly Lys Leu Glu Asn Thr Phe Asn Phe Lys Met Pro Leu Trp
275 280 285
Glu Gln Gly Val Gln Arg Met Leu Asp Glu Ile Gln
290 295 300
<210> 17
<211> 1041
<212> DNA
<213> Burkholderia thailandensis
<220>
<221> CDS
<222> (1)..(1041)
<400> 17
atg cgc ggt tcc ggc gag tgg gta gcc gct gcg gcg cgc gtg agg cag 48
Met Arg Gly Ser Gly Glu Trp Val Ala Ala Ala Ala Arg Val Arg Gln
1 5 10 15
ggc ggg cag atc gcg cgg gag ggc ggc tac gtg gag gcg tcc atc aag 96
Gly Gly Gln Ile Ala Arg Glu Gly Gly Tyr Val Glu Ala Ser Ile Lys
20 25 30
ggc gcg gga tca gct cat ttg ccg agc aga tgc ggg aga tac gcc atg 144
Gly Ala Gly Ser Ala His Leu Pro Ser Arg Cys Gly Arg Tyr Ala Met
35 40 45
cct atc gag aaa cag gtg gta gcg ctg ccg agc gga ctg aag gtc cac 192
Pro Ile Glu Lys Gln Val Val Ala Leu Pro Ser Gly Leu Lys Val His
50 55 60
gtc gag cgt cat gtg ttc gat ccg gcc ttc gag acg gtc atc ctc gtg 240
Val Glu Arg His Val Phe Asp Pro Ala Phe Glu Thr Val Ile Leu Val
65 70 75 80
aac ggc gcg ctg gcg acg acc gcg tcg ttc ggc cag acg att cgc tac 288
Asn Gly Ala Leu Ala Thr Thr Ala Ser Phe Gly Gln Thr Ile Arg Tyr
85 90 95
ctg ggc gaa cgc gtg aac gcg gtg tgc ttc gac ttg ccg tac gcg ggc 336
Leu Gly Glu Arg Val Asn Ala Val Cys Phe Asp Leu Pro Tyr Ala Gly
100 105 110
cag tcg cgc cag cac aat ccg ggc gag tac att ctg acg aag gac gac 384
Gln Ser Arg Gln His Asn Pro Gly Glu Tyr Ile Leu Thr Lys Asp Asp
115 120 125
gag gtg gag att ctg ctg cac ctg gcc gag cgg ttc gag ccg agt ttc 432
Glu Val Glu Ile Leu Leu His Leu Ala Glu Arg Phe Glu Pro Ser Phe
130 135 140
ctg ctg tcg gtg tcg tgg ggc ggg gtg gcg tcg ctg ttc gcg ctg gcg 480
Leu Leu Ser Val Ser Trp Gly Gly Val Ala Ser Leu Phe Ala Leu Ala
145 150 155 160
cgg ggg tgc gcg agc gtg cgg cgg gcg gtg atc gcg tcg ttc tcg ccg 528
Arg Gly Cys Ala Ser Val Arg Arg Ala Val Ile Ala Ser Phe Ser Pro
165 170 175
ttc ctg aac gac gcg atg acg gat tac gtg acg cgc gcg cgc gat cac 576
Phe Leu Asn Asp Ala Met Thr Asp Tyr Val Thr Arg Ala Arg Asp His
180 185 190
atc gcg gcg ggg gag aac ctg aag gcg gcg cag ttg ctc aac gac acg 624
Ile Ala Ala Gly Glu Asn Leu Lys Ala Ala Gln Leu Leu Asn Asp Thr
195 200 205
gtg ggg cgc tac ctg ccg cgg atc atg aag ctg tac aac tac cgg tat 672
Val Gly Arg Tyr Leu Pro Arg Ile Met Lys Leu Tyr Asn Tyr Arg Tyr
210 215 220
ctg acg aag ctg ccg cgc acc gag cag gac cag gtg gcg ttc cac gtc 720
Leu Thr Lys Leu Pro Arg Thr Glu Gln Asp Gln Val Ala Phe His Val
225 230 235 240
gac cag atc ctg tcg atg cgg ccg gag cag tac ctg ccg gaa ttc cgc 768
Asp Gln Ile Leu Ser Met Arg Pro Glu Gln Tyr Leu Pro Glu Phe Arg
245 250 255
cag atc ggc tgc gcg gtg aag ttc atc aac ggc gag ctg gac gag tac 816
Gln Ile Gly Cys Ala Val Lys Phe Ile Asn Gly Glu Leu Asp Glu Tyr
260 265 270
acg acg gcg tcg gac gtg cgg cgg ctg gcg gcc tac gtg cgg cgc gcg 864
Thr Thr Ala Ser Asp Val Arg Arg Leu Ala Ala Tyr Val Arg Arg Ala
275 280 285
gag ttc gcg acg atc cgg cag gcg ggg cac ttc ctg gac ctc gag ggg 912
Glu Phe Ala Thr Ile Arg Gln Ala Gly His Phe Leu Asp Leu Glu Gly
290 295 300
cgt cag cag cag gag cag ctt cgc gcg gcg atc ctg ggc ttc ttc ggc 960
Arg Gln Gln Gln Glu Gln Leu Arg Ala Ala Ile Leu Gly Phe Phe Gly
305 310 315 320
gac gag cgg gcg agc gcg gcg cgc gac gac gcg cag gac gag acg ctc 1008
Asp Glu Arg Ala Ser Ala Ala Arg Asp Asp Ala Gln Asp Glu Thr Leu
325 330 335
gcg ccg ctc ggt cag ttg ccg gcg ctg tcg tag 1041
Ala Pro Leu Gly Gln Leu Pro Ala Leu Ser
340 345
<210> 18
<211> 346
<212> PRT
<213> Burkholderia thailandensis
<400> 18
Met Arg Gly Ser Gly Glu Trp Val Ala Ala Ala Ala Arg Val Arg Gln
1 5 10 15
Gly Gly Gln Ile Ala Arg Glu Gly Gly Tyr Val Glu Ala Ser Ile Lys
20 25 30
Gly Ala Gly Ser Ala His Leu Pro Ser Arg Cys Gly Arg Tyr Ala Met
35 40 45
Pro Ile Glu Lys Gln Val Val Ala Leu Pro Ser Gly Leu Lys Val His
50 55 60
Val Glu Arg His Val Phe Asp Pro Ala Phe Glu Thr Val Ile Leu Val
65 70 75 80
Asn Gly Ala Leu Ala Thr Thr Ala Ser Phe Gly Gln Thr Ile Arg Tyr
85 90 95
Leu Gly Glu Arg Val Asn Ala Val Cys Phe Asp Leu Pro Tyr Ala Gly
100 105 110
Gln Ser Arg Gln His Asn Pro Gly Glu Tyr Ile Leu Thr Lys Asp Asp
115 120 125
Glu Val Glu Ile Leu Leu His Leu Ala Glu Arg Phe Glu Pro Ser Phe
130 135 140
Leu Leu Ser Val Ser Trp Gly Gly Val Ala Ser Leu Phe Ala Leu Ala
145 150 155 160
Arg Gly Cys Ala Ser Val Arg Arg Ala Val Ile Ala Ser Phe Ser Pro
165 170 175
Phe Leu Asn Asp Ala Met Thr Asp Tyr Val Thr Arg Ala Arg Asp His
180 185 190
Ile Ala Ala Gly Glu Asn Leu Lys Ala Ala Gln Leu Leu Asn Asp Thr
195 200 205
Val Gly Arg Tyr Leu Pro Arg Ile Met Lys Leu Tyr Asn Tyr Arg Tyr
210 215 220
Leu Thr Lys Leu Pro Arg Thr Glu Gln Asp Gln Val Ala Phe His Val
225 230 235 240
Asp Gln Ile Leu Ser Met Arg Pro Glu Gln Tyr Leu Pro Glu Phe Arg
245 250 255
Gln Ile Gly Cys Ala Val Lys Phe Ile Asn Gly Glu Leu Asp Glu Tyr
260 265 270
Thr Thr Ala Ser Asp Val Arg Arg Leu Ala Ala Tyr Val Arg Arg Ala
275 280 285
Glu Phe Ala Thr Ile Arg Gln Ala Gly His Phe Leu Asp Leu Glu Gly
290 295 300
Arg Gln Gln Gln Glu Gln Leu Arg Ala Ala Ile Leu Gly Phe Phe Gly
305 310 315 320
Asp Glu Arg Ala Ser Ala Ala Arg Asp Asp Ala Gln Asp Glu Thr Leu
325 330 335
Ala Pro Leu Gly Gln Leu Pro Ala Leu Ser
340 345
<210> 19
<211> 1482
<212> DNA
<213> Burkholderia thailandensis
<220>
<221> CDS
<222> (1)..(1482)
<400> 19
atg gat gcg ggc cgg atc ggg ctg cat gac gcg gcg gcg gcg ggg cgc 48
Met Asp Ala Gly Arg Ile Gly Leu His Asp Ala Ala Ala Ala Gly Arg
1 5 10 15
atc ggc atg acg gaa gcg ttc gcg tca cgg gcc cgt tgt tcg gcg gcg 96
Ile Gly Met Thr Glu Ala Phe Ala Ser Arg Ala Arg Cys Ser Ala Ala
20 25 30
gcc ctg gcg gca ggc gga cga gcc ccg gcc ggc gat gga cga tcc ggt 144
Ala Leu Ala Ala Gly Gly Arg Ala Pro Ala Gly Asp Gly Arg Ser Gly
35 40 45
tcg aac agg gct gcg gcg aac ggt tcc gtc gac tgc cgc gcg gga tgg 192
Ser Asn Arg Ala Ala Ala Asn Gly Ser Val Asp Cys Arg Ala Gly Trp
50 55 60
aat gac gag gcg atg gct aaa gta atc gtg acg gcg atc ggg tcg gcg 240
Asn Asp Glu Ala Met Ala Lys Val Ile Val Thr Ala Ile Gly Ser Ala
65 70 75 80
ggc gac gtg cac ccg ttg ctg ggg gtg agc cgg gcg ctg tcg gcg cgg 288
Gly Asp Val His Pro Leu Leu Gly Val Ser Arg Ala Leu Ser Ala Arg
85 90 95
ggc cac gag gtg gtg ttc tgc acg cat gcg ccg ttc gag gcg gcg gtg 336
Gly His Glu Val Val Phe Cys Thr His Ala Pro Phe Glu Ala Ala Val
100 105 110
cgc gcg agc ggc ttc gcg ttc gtg ccg gtg ggc acg gcc gag gac tac 384
Arg Ala Ser Gly Phe Ala Phe Val Pro Val Gly Thr Ala Glu Asp Tyr
115 120 125
gtg cgg gcg atg gcg gac ccg gcg ctg tgg gat ccg cgc acg tcg ttc 432
Val Arg Ala Met Ala Asp Pro Ala Leu Trp Asp Pro Arg Thr Ser Phe
130 135 140
aag acg ctg tgg cgg gtg atc gcg ccg gtg gtg agg ccg cac ttc gag 480
Lys Thr Leu Trp Arg Val Ile Ala Pro Val Val Arg Pro His Phe Glu
145 150 155 160
gtg ctg cgc gcg ctg agc gac gcg gac acg gtg ctg gtg ggc acg ctg 528
Val Leu Arg Ala Leu Ser Asp Ala Asp Thr Val Leu Val Gly Thr Leu
165 170 175
tgg gcg ttc tcg gcg cgg ctg atg cag gag cgc ttc ggc acg cgg tac 576
Trp Ala Phe Ser Ala Arg Leu Met Gln Glu Arg Phe Gly Thr Arg Tyr
180 185 190
gtg tcg gtg cag gtg tcg ccg tcg acg ctg ctg tcg gcg cat gcg ccg 624
Val Ser Val Gln Val Ser Pro Ser Thr Leu Leu Ser Ala His Ala Pro
195 200 205
ccg acg cac aag cgg ctg acg atc ccg aag ggc ctg ccg ctg gcg gtg 672
Pro Thr His Lys Arg Leu Thr Ile Pro Lys Gly Leu Pro Leu Ala Val
210 215 220
aag gcg ggg ctg atg acg ctg atc gag cgg cag gtg ctg gac cgg gtg 720
Lys Ala Gly Leu Met Thr Leu Ile Glu Arg Gln Val Leu Asp Arg Val
225 230 235 240
tgc ggc ccg gag ctg aac gcg gcg cgg cag gcg ctg ggc ctg gcg ccg 768
Cys Gly Pro Glu Leu Asn Ala Ala Arg Gln Ala Leu Gly Leu Ala Pro
245 250 255
gcg aag cgg atc ctg ggc cgg tgg ctg cat tcg acg gac ggg gtg ctg 816
Ala Lys Arg Ile Leu Gly Arg Trp Leu His Ser Thr Asp Gly Val Leu
260 265 270
tgc ctg ttt ccg tcg tgg ttc gcg ccg gcg cag ccg gac tgg ccg gcg 864
Cys Leu Phe Pro Ser Trp Phe Ala Pro Ala Gln Pro Asp Trp Pro Ala
275 280 285
aac cac ctg caa agc ggg ttt ccg ctg ttc aac gac gcg ggt ccg gcg 912
Asn His Leu Gln Ser Gly Phe Pro Leu Phe Asn Asp Ala Gly Pro Ala
290 295 300
cag gcg gat gcg gag ctg gag gcg ttc gtc gcg tcg ggc gag gcg ccg 960
Gln Ala Asp Ala Glu Leu Glu Ala Phe Val Ala Ser Gly Glu Ala Pro
305 310 315 320
gtg gtg ttc acg gcg ggc tcg acg ctg gtg gac ggc cgc acg tat gag 1008
Val Val Phe Thr Ala Gly Ser Thr Leu Val Asp Gly Arg Thr Tyr Glu
325 330 335
cac gcg gtg acg cag gtg ctg cag gcc acg ggg gtg cgg ggg att ctg 1056
His Ala Val Thr Gln Val Leu Gln Ala Thr Gly Val Arg Gly Ile Leu
340 345 350
ctc gcg ccg gat gcg ccg gat gcg ccg gcg gca tcg gac ggg gcg gcg 1104
Leu Ala Pro Asp Ala Pro Asp Ala Pro Ala Ala Ser Asp Gly Ala Ala
355 360 365
ctg ctc aag cgc cgc tac gtg ccg ctc gcg gcg ttg ctg ccg cgc tgc 1152
Leu Leu Lys Arg Arg Tyr Val Pro Leu Ala Ala Leu Leu Pro Arg Cys
370 375 380
cgg gcg ctg gtg cac cac ggg ggg atc ggg acg gcg tcg ctc gcg tac 1200
Arg Ala Leu Val His His Gly Gly Ile Gly Thr Ala Ser Leu Ala Tyr
385 390 395 400
gcg gcg ggg gtg ccg cag gtg gtg acg ccg ttc gcg cac gac cag ttc 1248
Ala Ala Gly Val Pro Gln Val Val Thr Pro Phe Ala His Asp Gln Phe
405 410 415
gac aac gcg cag cgg gtg gcg gcg agc ggc tgc ggg gtg cgg ctg gac 1296
Asp Asn Ala Gln Arg Val Ala Ala Ser Gly Cys Gly Val Arg Leu Asp
420 425 430
gcg ccg gtg cgc ggc gag ccg ctc gcg cgg gcg ctg gcg cag gtg ctg 1344
Ala Pro Val Arg Gly Glu Pro Leu Ala Arg Ala Leu Ala Gln Val Leu
435 440 445
ggc gac gcg gcg atg gcg gcg cgc tgc gcg cag gtg cgc gcg cgg atg 1392
Gly Asp Ala Ala Met Ala Ala Arg Cys Ala Gln Val Arg Ala Arg Met
450 455 460
gcg gcg gag ccg aac ggc tgc gac gcg gcg gcg cgc ttc atc gag cgc 1440
Ala Ala Glu Pro Asn Gly Cys Asp Ala Ala Ala Arg Phe Ile Glu Arg
465 470 475 480
ttc gcg ccg ggc gtc gcg gcg cgg cgg gcg cag ccg gca tga 1482
Phe Ala Pro Gly Val Ala Ala Arg Arg Ala Gln Pro Ala
485 490
<210> 20
<211> 493
<212> PRT
<213> Burkholderia thailandensis
<400> 20
Met Asp Ala Gly Arg Ile Gly Leu His Asp Ala Ala Ala Ala Gly Arg
1 5 10 15
Ile Gly Met Thr Glu Ala Phe Ala Ser Arg Ala Arg Cys Ser Ala Ala
20 25 30
Ala Leu Ala Ala Gly Gly Arg Ala Pro Ala Gly Asp Gly Arg Ser Gly
35 40 45
Ser Asn Arg Ala Ala Ala Asn Gly Ser Val Asp Cys Arg Ala Gly Trp
50 55 60
Asn Asp Glu Ala Met Ala Lys Val Ile Val Thr Ala Ile Gly Ser Ala
65 70 75 80
Gly Asp Val His Pro Leu Leu Gly Val Ser Arg Ala Leu Ser Ala Arg
85 90 95
Gly His Glu Val Val Phe Cys Thr His Ala Pro Phe Glu Ala Ala Val
100 105 110
Arg Ala Ser Gly Phe Ala Phe Val Pro Val Gly Thr Ala Glu Asp Tyr
115 120 125
Val Arg Ala Met Ala Asp Pro Ala Leu Trp Asp Pro Arg Thr Ser Phe
130 135 140
Lys Thr Leu Trp Arg Val Ile Ala Pro Val Val Arg Pro His Phe Glu
145 150 155 160
Val Leu Arg Ala Leu Ser Asp Ala Asp Thr Val Leu Val Gly Thr Leu
165 170 175
Trp Ala Phe Ser Ala Arg Leu Met Gln Glu Arg Phe Gly Thr Arg Tyr
180 185 190
Val Ser Val Gln Val Ser Pro Ser Thr Leu Leu Ser Ala His Ala Pro
195 200 205
Pro Thr His Lys Arg Leu Thr Ile Pro Lys Gly Leu Pro Leu Ala Val
210 215 220
Lys Ala Gly Leu Met Thr Leu Ile Glu Arg Gln Val Leu Asp Arg Val
225 230 235 240
Cys Gly Pro Glu Leu Asn Ala Ala Arg Gln Ala Leu Gly Leu Ala Pro
245 250 255
Ala Lys Arg Ile Leu Gly Arg Trp Leu His Ser Thr Asp Gly Val Leu
260 265 270
Cys Leu Phe Pro Ser Trp Phe Ala Pro Ala Gln Pro Asp Trp Pro Ala
275 280 285
Asn His Leu Gln Ser Gly Phe Pro Leu Phe Asn Asp Ala Gly Pro Ala
290 295 300
Gln Ala Asp Ala Glu Leu Glu Ala Phe Val Ala Ser Gly Glu Ala Pro
305 310 315 320
Val Val Phe Thr Ala Gly Ser Thr Leu Val Asp Gly Arg Thr Tyr Glu
325 330 335
His Ala Val Thr Gln Val Leu Gln Ala Thr Gly Val Arg Gly Ile Leu
340 345 350
Leu Ala Pro Asp Ala Pro Asp Ala Pro Ala Ala Ser Asp Gly Ala Ala
355 360 365
Leu Leu Lys Arg Arg Tyr Val Pro Leu Ala Ala Leu Leu Pro Arg Cys
370 375 380
Arg Ala Leu Val His His Gly Gly Ile Gly Thr Ala Ser Leu Ala Tyr
385 390 395 400
Ala Ala Gly Val Pro Gln Val Val Thr Pro Phe Ala His Asp Gln Phe
405 410 415
Asp Asn Ala Gln Arg Val Ala Ala Ser Gly Cys Gly Val Arg Leu Asp
420 425 430
Ala Pro Val Arg Gly Glu Pro Leu Ala Arg Ala Leu Ala Gln Val Leu
435 440 445
Gly Asp Ala Ala Met Ala Ala Arg Cys Ala Gln Val Arg Ala Arg Met
450 455 460
Ala Ala Glu Pro Asn Gly Cys Asp Ala Ala Ala Arg Phe Ile Glu Arg
465 470 475 480
Phe Ala Pro Gly Val Ala Ala Arg Arg Ala Gln Pro Ala
485 490
<210> 21
<211> 966
<212> DNA
<213> Burkholderia thailandensis
<220>
<221> CDS
<222> (1)..(966)
<400> 21
atg acg atc ctg ggg gcg ctg gtg att ctg tac gac ccg acg gac gag 48
Met Thr Ile Leu Gly Ala Leu Val Ile Leu Tyr Asp Pro Thr Asp Glu
1 5 10 15
cag ttg tcg ggg ctg gag gcg ctc gcg cgc gac agc gac gcg ctc gtg 96
Gln Leu Ser Gly Leu Glu Ala Leu Ala Arg Asp Ser Asp Ala Leu Val
20 25 30
gtc gtg gac aac acg ccg cac gag cac gcg gcg gcg cgc gag cgg gtg 144
Val Val Asp Asn Thr Pro His Glu His Ala Ala Ala Arg Glu Arg Val
35 40 45
cgt gcg ctg tcg gcg cgg acg aac acg gtg tgg cga cac cac ggc aac 192
Arg Ala Leu Ser Ala Arg Thr Asn Thr Val Trp Arg His His Gly Asn
50 55 60
cgg ggc ggg gtc gcg ggc ggg tac aac gcg ggg ctg tcg gtg ctg ttc 240
Arg Gly Gly Val Ala Gly Gly Tyr Asn Ala Gly Leu Ser Val Leu Phe
65 70 75 80
gcg cag ggc gtc gag gcg gtc gcg ctg ttc gac cag gac tcg acg gtg 288
Ala Gln Gly Val Glu Ala Val Ala Leu Phe Asp Gln Asp Ser Thr Val
85 90 95
ccg gcc ggg tac ttc gag cgg atg cgc gag gcg tgc gcg caa ctg ggt 336
Pro Ala Gly Tyr Phe Glu Arg Met Arg Glu Ala Cys Ala Gln Leu Gly
100 105 110
gag caa ccg ggc gcg cac gcg ggc gcg ttc atc gcg ggc ccg cgg atc 384
Glu Gln Pro Gly Ala His Ala Gly Ala Phe Ile Ala Gly Pro Arg Ile
115 120 125
tac gac gcg aac gag cag cgc ttc ctg ccg gag ctg atg acg agc ggg 432
Tyr Asp Ala Asn Glu Gln Arg Phe Leu Pro Glu Leu Met Thr Ser Gly
130 135 140
gtg acg gtg cgc cgc gtg cgg gtg gag ggc gag acg gcg ccg cag cgc 480
Val Thr Val Arg Arg Val Arg Val Glu Gly Glu Thr Ala Pro Gln Arg
145 150 155 160
tgc gcg ttc ctg atc tcg tcg ggc agc gtg att tcg cgg gcc gcg tac 528
Cys Ala Phe Leu Ile Ser Ser Gly Ser Val Ile Ser Arg Ala Ala Tyr
165 170 175
gcg cgg ctc ggt cga ttc gac gag gcg ctg ttc atc gat cac gtc gac 576
Ala Arg Leu Gly Arg Phe Asp Glu Ala Leu Phe Ile Asp His Val Asp
180 185 190
acc gag tat tgc ctg cgc gcg ctc gcg cac aac gtg ccg ctg tac gtg 624
Thr Glu Tyr Cys Leu Arg Ala Leu Ala His Asn Val Pro Leu Tyr Val
195 200 205
gtg ccg ccg ctc gtg ctg acg cac cgg atc ggc gcg cgg cgc cgg cac 672
Val Pro Pro Leu Val Leu Thr His Arg Ile Gly Ala Arg Arg Arg His
210 215 220
aag gtg ggg ccg ttc gag ctg acg gcg atg cat cac ggg tgg ttg cgc 720
Lys Val Gly Pro Phe Glu Leu Thr Ala Met His His Gly Trp Leu Arg
225 230 235 240
cga tac tac ggc gcg cgc aac gcg atg caa ctg ggg ctg cag tac ggc 768
Arg Tyr Tyr Gly Ala Arg Asn Ala Met Gln Leu Gly Leu Gln Tyr Gly
245 250 255
ttg cgg ttt ccg gtg gcg ctg gtg ccg aat ctg ctg acg ata tgg cag 816
Leu Arg Phe Pro Val Ala Leu Val Pro Asn Leu Leu Thr Ile Trp Gln
260 265 270
gtg atc cag gtg gtg ctg tgc gag cgg gag aag ggc gcg aag ctg cgc 864
Val Ile Gln Val Val Leu Cys Glu Arg Glu Lys Gly Ala Lys Leu Arg
275 280 285
ggg atc gcg ctg ggc gtg ctc gac ggc ctg ttc ggg cgg ctg gga tcg 912
Gly Ile Ala Leu Gly Val Leu Asp Gly Leu Phe Gly Arg Leu Gly Ser
290 295 300
ttc gac gat gcg cgc gcg ggc gcg gcg gcg cgc gag ccg gtg cgg cag 960
Phe Asp Asp Ala Arg Ala Gly Ala Ala Ala Arg Glu Pro Val Arg Gln
305 310 315 320
gaa tga 966
Glu
<210> 22
<211> 321
<212> PRT
<213> Burkholderia thailandensis
<400> 22
Met Thr Ile Leu Gly Ala Leu Val Ile Leu Tyr Asp Pro Thr Asp Glu
1 5 10 15
Gln Leu Ser Gly Leu Glu Ala Leu Ala Arg Asp Ser Asp Ala Leu Val
20 25 30
Val Val Asp Asn Thr Pro His Glu His Ala Ala Ala Arg Glu Arg Val
35 40 45
Arg Ala Leu Ser Ala Arg Thr Asn Thr Val Trp Arg His His Gly Asn
50 55 60
Arg Gly Gly Val Ala Gly Gly Tyr Asn Ala Gly Leu Ser Val Leu Phe
65 70 75 80
Ala Gln Gly Val Glu Ala Val Ala Leu Phe Asp Gln Asp Ser Thr Val
85 90 95
Pro Ala Gly Tyr Phe Glu Arg Met Arg Glu Ala Cys Ala Gln Leu Gly
100 105 110
Glu Gln Pro Gly Ala His Ala Gly Ala Phe Ile Ala Gly Pro Arg Ile
115 120 125
Tyr Asp Ala Asn Glu Gln Arg Phe Leu Pro Glu Leu Met Thr Ser Gly
130 135 140
Val Thr Val Arg Arg Val Arg Val Glu Gly Glu Thr Ala Pro Gln Arg
145 150 155 160
Cys Ala Phe Leu Ile Ser Ser Gly Ser Val Ile Ser Arg Ala Ala Tyr
165 170 175
Ala Arg Leu Gly Arg Phe Asp Glu Ala Leu Phe Ile Asp His Val Asp
180 185 190
Thr Glu Tyr Cys Leu Arg Ala Leu Ala His Asn Val Pro Leu Tyr Val
195 200 205
Val Pro Pro Leu Val Leu Thr His Arg Ile Gly Ala Arg Arg Arg His
210 215 220
Lys Val Gly Pro Phe Glu Leu Thr Ala Met His His Gly Trp Leu Arg
225 230 235 240
Arg Tyr Tyr Gly Ala Arg Asn Ala Met Gln Leu Gly Leu Gln Tyr Gly
245 250 255
Leu Arg Phe Pro Val Ala Leu Val Pro Asn Leu Leu Thr Ile Trp Gln
260 265 270
Val Ile Gln Val Val Leu Cys Glu Arg Glu Lys Gly Ala Lys Leu Arg
275 280 285
Gly Ile Ala Leu Gly Val Leu Asp Gly Leu Phe Gly Arg Leu Gly Ser
290 295 300
Phe Asp Asp Ala Arg Ala Gly Ala Ala Ala Arg Glu Pro Val Arg Gln
305 310 315 320
Glu
<210> 23
<211> 1557
<212> DNA
<213> Burkholderia thailandensis
<220>
<221> CDS
<222> (1)..(1557)
<400> 23
atg tcg gcg gat cag gcg ggc gtt gcg ccg ccg gcg gcc gcc ccg ctg 48
Met Ser Ala Asp Gln Ala Gly Val Ala Pro Pro Ala Ala Ala Pro Leu
1 5 10 15
cgc ggc gcg aag ctc gcg ctg ctg acg ttc gcg ctg tcg ctc gcg acg 96
Arg Gly Ala Lys Leu Ala Leu Leu Thr Phe Ala Leu Ser Leu Ala Thr
20 25 30
ttc atc gaa gtg ctg gat tcg acg gtg gcg aac gtg gcg gtg ccg gcg 144
Phe Ile Glu Val Leu Asp Ser Thr Val Ala Asn Val Ala Val Pro Ala
35 40 45
atc tcg ggc agc ctc ggg gtg tcg aac agc cag ggc acg tgg gtg atc 192
Ile Ser Gly Ser Leu Gly Val Ser Asn Ser Gln Gly Thr Trp Val Ile
50 55 60
agc tcg tac tcg gtg gcc gcg gcg atc gcg gtg ccg ctg acg ggg tgg 240
Ser Ser Tyr Ser Val Ala Ala Ala Ile Ala Val Pro Leu Thr Gly Trp
65 70 75 80
ctt gcg cgg cgc gtg ggc gag ctg agg ctg ttc gtg gcg tcg gtg atc 288
Leu Ala Arg Arg Val Gly Glu Leu Arg Leu Phe Val Ala Ser Val Ile
85 90 95
ctg ttc acg ctg acg tcg ctg ctg tgc ggg ctc gcg cgg gac ctg gag 336
Leu Phe Thr Leu Thr Ser Leu Leu Cys Gly Leu Ala Arg Asp Leu Glu
100 105 110
gtg ctg gtt gcg tgc cgg gcg ctg cag ggg ctg ttc tcg ggg ccg atg 384
Val Leu Val Ala Cys Arg Ala Leu Gln Gly Leu Phe Ser Gly Pro Met
115 120 125
gtg ccg ctg tcg cag acg atc ctg atg cgc gcg ttc ccg ccg gcg cgg 432
Val Pro Leu Ser Gln Thr Ile Leu Met Arg Ala Phe Pro Pro Ala Arg
130 135 140
cgc acg ctg gcg ctg gcg ctg tgg ggg atg acg gtg ctg ctc gcg ccg 480
Arg Thr Leu Ala Leu Ala Leu Trp Gly Met Thr Val Leu Leu Ala Pro
145 150 155 160
atc ttc ggg ccg gtg gtg ggc ggc tgg ctg atc gac aac ttc tcg tgg 528
Ile Phe Gly Pro Val Val Gly Gly Trp Leu Ile Asp Asn Phe Ser Trp
165 170 175
ccg tgg atc ttc ctg atc aac ctg ccg atc ggg ctg ttc tcg ttc gcg 576
Pro Trp Ile Phe Leu Ile Asn Leu Pro Ile Gly Leu Phe Ser Phe Ala
180 185 190
gtg tgc acg ctg atg ctg cgc ccg cag gcg cag cgc ggc gag gcg agc 624
Val Cys Thr Leu Met Leu Arg Pro Gln Ala Gln Arg Gly Glu Ala Ser
195 200 205
ccg atc gac gcg ccg ggg atc gtg ctg ctg gtg atc ggg gtg ggc tcg 672
Pro Ile Asp Ala Pro Gly Ile Val Leu Leu Val Ile Gly Val Gly Ser
210 215 220
ctg cag gcg atg ctg gac ctg ggg cac gac cgg ggc tgg ttc gat tcg 720
Leu Gln Ala Met Leu Asp Leu Gly His Asp Arg Gly Trp Phe Asp Ser
225 230 235 240
ccg ctg atc acg gcg ctg gcg atc gcg gcg ggg gtg tcg ctc gtg tcg 768
Pro Leu Ile Thr Ala Leu Ala Ile Ala Ala Gly Val Ser Leu Val Ser
245 250 255
ctg ctg atc tgg gag ctg ggc gag gcg cat ccg gtg gtg gat ctg agc 816
Leu Leu Ile Trp Glu Leu Gly Glu Ala His Pro Val Val Asp Leu Ser
260 265 270
ctg ttc cgg gag cgg acc ttc acg ttc tgc gtg gtg atc atc tcg ctg 864
Leu Phe Arg Glu Arg Thr Phe Thr Phe Cys Val Val Ile Ile Ser Leu
275 280 285
ggg atg atg agc ttc tcg gtg gtg ggg gtg gtg ttt ccg ctg tgg ctg 912
Gly Met Met Ser Phe Ser Val Val Gly Val Val Phe Pro Leu Trp Leu
290 295 300
cag gcg gtg atg gga tac acg gcg tac cag gcg ggg ctg gcg acg gcg 960
Gln Ala Val Met Gly Tyr Thr Ala Tyr Gln Ala Gly Leu Ala Thr Ala
305 310 315 320
tcg atg ggg gtg ctg gcg ctg gtg ttc tcg atc ctg gtg ggg ctg tac 1008
Ser Met Gly Val Leu Ala Leu Val Phe Ser Ile Leu Val Gly Leu Tyr
325 330 335
gcg agc cgg gtg gac gcg cgg gtg ctg gtg acg ttc ggg ttc ggg gtg 1056
Ala Ser Arg Val Asp Ala Arg Val Leu Val Thr Phe Gly Phe Gly Val
340 345 350
ttt gcg gcg gtg atg tgg tgg agc acg cac ttc acg ctg tcg atg acg 1104
Phe Ala Ala Val Met Trp Trp Ser Thr His Phe Thr Leu Ser Met Thr
355 360 365
ttc gcg cag gtg gtg acg ccg cgg ctg att cag ggg atg ggg ctg ccg 1152
Phe Ala Gln Val Val Thr Pro Arg Leu Ile Gln Gly Met Gly Leu Pro
370 375 380
tgc ttc ttc ata ccg ctg acg gcg gcg acg ctg tcg cgg gtg ccg gac 1200
Cys Phe Phe Ile Pro Leu Thr Ala Ala Thr Leu Ser Arg Val Pro Asp
385 390 395 400
gag aag ctg gcg gcg gcg tcg agc ctg tcg aac ttc ctg cgg acg ctg 1248
Glu Lys Leu Ala Ala Ala Ser Ser Leu Ser Asn Phe Leu Arg Thr Leu
405 410 415
tcg gcg gcg ttc ggc acg gcg ctg agc gtg acg tgg tgg gac aac cgg 1296
Ser Ala Ala Phe Gly Thr Ala Leu Ser Val Thr Trp Trp Asp Asn Arg
420 425 430
gcg acg tac cac tac gcg gtg gtg tcg caa tcg gtg acg cgc gcc tcg 1344
Ala Thr Tyr His Tyr Ala Val Val Ser Gln Ser Val Thr Arg Ala Ser
435 440 445
gag aac acg cag cgg tac gtg gac gcg ctg cac gcg atg ggg ctg cac 1392
Glu Asn Thr Gln Arg Tyr Val Asp Ala Leu His Ala Met Gly Leu His
450 455 460
ggc gcg cgg gag ctg agc tcg ctg cac cag gtg gtg cgg cag cag gcg 1440
Gly Ala Arg Glu Leu Ser Ser Leu His Gln Val Val Arg Gln Gln Ala
465 470 475 480
tac atg atg gcg acg aac gac atg ttc tac atg gcg agc gcg acg tgc 1488
Tyr Met Met Ala Thr Asn Asp Met Phe Tyr Met Ala Ser Ala Thr Cys
485 490 495
ctg ctg ctg gcg ggg ctg atg tgg ctg acg cgg ccg aag cgg ggc gcg 1536
Leu Leu Leu Ala Gly Leu Met Trp Leu Thr Arg Pro Lys Arg Gly Ala
500 505 510
gcg gcg gcg ctc ggg cac tga 1557
Ala Ala Ala Leu Gly His
515
<210> 24
<211> 518
<212> PRT
<213> Burkholderia thailandensis
<400> 24
Met Ser Ala Asp Gln Ala Gly Val Ala Pro Pro Ala Ala Ala Pro Leu
1 5 10 15
Arg Gly Ala Lys Leu Ala Leu Leu Thr Phe Ala Leu Ser Leu Ala Thr
20 25 30
Phe Ile Glu Val Leu Asp Ser Thr Val Ala Asn Val Ala Val Pro Ala
35 40 45
Ile Ser Gly Ser Leu Gly Val Ser Asn Ser Gln Gly Thr Trp Val Ile
50 55 60
Ser Ser Tyr Ser Val Ala Ala Ala Ile Ala Val Pro Leu Thr Gly Trp
65 70 75 80
Leu Ala Arg Arg Val Gly Glu Leu Arg Leu Phe Val Ala Ser Val Ile
85 90 95
Leu Phe Thr Leu Thr Ser Leu Leu Cys Gly Leu Ala Arg Asp Leu Glu
100 105 110
Val Leu Val Ala Cys Arg Ala Leu Gln Gly Leu Phe Ser Gly Pro Met
115 120 125
Val Pro Leu Ser Gln Thr Ile Leu Met Arg Ala Phe Pro Pro Ala Arg
130 135 140
Arg Thr Leu Ala Leu Ala Leu Trp Gly Met Thr Val Leu Leu Ala Pro
145 150 155 160
Ile Phe Gly Pro Val Val Gly Gly Trp Leu Ile Asp Asn Phe Ser Trp
165 170 175
Pro Trp Ile Phe Leu Ile Asn Leu Pro Ile Gly Leu Phe Ser Phe Ala
180 185 190
Val Cys Thr Leu Met Leu Arg Pro Gln Ala Gln Arg Gly Glu Ala Ser
195 200 205
Pro Ile Asp Ala Pro Gly Ile Val Leu Leu Val Ile Gly Val Gly Ser
210 215 220
Leu Gln Ala Met Leu Asp Leu Gly His Asp Arg Gly Trp Phe Asp Ser
225 230 235 240
Pro Leu Ile Thr Ala Leu Ala Ile Ala Ala Gly Val Ser Leu Val Ser
245 250 255
Leu Leu Ile Trp Glu Leu Gly Glu Ala His Pro Val Val Asp Leu Ser
260 265 270
Leu Phe Arg Glu Arg Thr Phe Thr Phe Cys Val Val Ile Ile Ser Leu
275 280 285
Gly Met Met Ser Phe Ser Val Val Gly Val Val Phe Pro Leu Trp Leu
290 295 300
Gln Ala Val Met Gly Tyr Thr Ala Tyr Gln Ala Gly Leu Ala Thr Ala
305 310 315 320
Ser Met Gly Val Leu Ala Leu Val Phe Ser Ile Leu Val Gly Leu Tyr
325 330 335
Ala Ser Arg Val Asp Ala Arg Val Leu Val Thr Phe Gly Phe Gly Val
340 345 350
Phe Ala Ala Val Met Trp Trp Ser Thr His Phe Thr Leu Ser Met Thr
355 360 365
Phe Ala Gln Val Val Thr Pro Arg Leu Ile Gln Gly Met Gly Leu Pro
370 375 380
Cys Phe Phe Ile Pro Leu Thr Ala Ala Thr Leu Ser Arg Val Pro Asp
385 390 395 400
Glu Lys Leu Ala Ala Ala Ser Ser Leu Ser Asn Phe Leu Arg Thr Leu
405 410 415
Ser Ala Ala Phe Gly Thr Ala Leu Ser Val Thr Trp Trp Asp Asn Arg
420 425 430
Ala Thr Tyr His Tyr Ala Val Val Ser Gln Ser Val Thr Arg Ala Ser
435 440 445
Glu Asn Thr Gln Arg Tyr Val Asp Ala Leu His Ala Met Gly Leu His
450 455 460
Gly Ala Arg Glu Leu Ser Ser Leu His Gln Val Val Arg Gln Gln Ala
465 470 475 480
Tyr Met Met Ala Thr Asn Asp Met Phe Tyr Met Ala Ser Ala Thr Cys
485 490 495
Leu Leu Leu Ala Gly Leu Met Trp Leu Thr Arg Pro Lys Arg Gly Ala
500 505 510
Ala Ala Ala Leu Gly His
515
<210> 25
<211> 1662
<212> DNA
<213> Burkholderia thailandensis
<220>
<221> CDS
<222> (1)..(1662)
<400> 25
atg cgc gcg cgg gcg cgg cgg cgc gcg agc cgg tgc ggc agg aat gaa 48
Met Arg Ala Arg Ala Arg Arg Arg Ala Ser Arg Cys Gly Arg Asn Glu
1 5 10 15
cgg aac ggg ccg cag cgg gat acc gga aag caa gaa gga cgc atc ata 96
Arg Asn Gly Pro Gln Arg Asp Thr Gly Lys Gln Glu Gly Arg Ile Ile
20 25 30
cga atg acg cag aca gca acg caa gca gcc act cgc gcg atg atc gcg 144
Arg Met Thr Gln Thr Ala Thr Gln Ala Ala Thr Arg Ala Met Ile Ala
35 40 45
aca gga agc cgc gcg gcg cgc cgg ctc gcg gca gcc gcg ctc gcg tgg 192
Thr Gly Ser Arg Ala Ala Arg Arg Leu Ala Ala Ala Ala Leu Ala Trp
50 55 60
gcg ctc gcc ggc tgc gtg ccg tcg ggc ttc gag ccg gcg ctc gcg ccg 240
Ala Leu Ala Gly Cys Val Pro Ser Gly Phe Glu Pro Ala Leu Ala Pro
65 70 75 80
cgc acg ccg ggc gac gac gcg ctc gcg cac acg gcg ggg ggc gcc gcg 288
Arg Thr Pro Gly Asp Asp Ala Leu Ala His Thr Ala Gly Gly Ala Ala
85 90 95
cac ggc gca tgg ccg agc ccc gac tgg gtc cgg cag ctc ggc gat ccg 336
His Gly Ala Trp Pro Ser Pro Asp Trp Val Arg Gln Leu Gly Asp Pro
100 105 110
caa ctc gac gcg ctc gtc gac gag gcg ctg cgg cag aac ccg acg ctg 384
Gln Leu Asp Ala Leu Val Asp Glu Ala Leu Arg Gln Asn Pro Thr Leu
115 120 125
cag gcc gcg cag gcg cgc atc ggc gtc gcg cag tcg cag ctg cag cag 432
Gln Ala Ala Gln Ala Arg Ile Gly Val Ala Gln Ser Gln Leu Gln Gln
130 135 140
ttc gaa tcg ctg acg ggg ctc acc gcg acg gcg ggc gcg tcg ctc tcc 480
Phe Glu Ser Leu Thr Gly Leu Thr Ala Thr Ala Gly Ala Ser Leu Ser
145 150 155 160
aag gcg cac gtg ccg cgc tcg ggc ggc acc atc aat acg acg ttc aac 528
Lys Ala His Val Pro Arg Ser Gly Gly Thr Ile Asn Thr Thr Phe Asn
165 170 175
ggc ttg ccg gtg tcg gtg ccg ctc gtc ggc gaa tcg gtg gtg tcg tcg 576
Gly Leu Pro Val Ser Val Pro Leu Val Gly Glu Ser Val Val Ser Ser
180 185 190
tcg tcg ctg ttc gtc ggg ctg aac tat cag ctg gac ctg tgg ggc aag 624
Ser Ser Leu Phe Val Gly Leu Asn Tyr Gln Leu Asp Leu Trp Gly Lys
195 200 205
aac gcg gcg gcc acg cgc ggg ctg ctg tcg atg cgc gat gcg gcg cgc 672
Asn Ala Ala Ala Thr Arg Gly Leu Leu Ser Met Arg Asp Ala Ala Arg
210 215 220
gtg gag gcc gag cag gcg cgg ctc gcg ctg tcg gtg gcg atc gtg acg 720
Val Glu Ala Glu Gln Ala Arg Leu Ala Leu Ser Val Ala Ile Val Thr
225 230 235 240
ctg tac ggc gag ctg gac cgc gcg tat gcg ctg cgc gag ctg ctg cag 768
Leu Tyr Gly Glu Leu Asp Arg Ala Tyr Ala Leu Arg Glu Leu Leu Gln
245 250 255
cag aag cgc cgc gcg agc gag cag gtg gag acg gtg ctg cgc gag cgc 816
Gln Lys Arg Arg Ala Ser Glu Gln Val Glu Thr Val Leu Arg Glu Arg
260 265 270
gcg gcg cgc ggg atc gac aac ggc tac gat gcg gac gac gcg gcg ctc 864
Ala Ala Arg Gly Ile Asp Asn Gly Tyr Asp Ala Asp Asp Ala Ala Leu
275 280 285
aag cgg ggc aag ctg ctc gag cag ctc gcg ctg acc gac gag cag atc 912
Lys Arg Gly Lys Leu Leu Glu Gln Leu Ala Leu Thr Asp Glu Gln Ile
290 295 300
cag ttg cag aag ctg caa ctg ggg gtg ctg agc ggg cgg ggg ccg gag 960
Gln Leu Gln Lys Leu Gln Leu Gly Val Leu Ser Gly Arg Gly Pro Glu
305 310 315 320
cgc ggg ctg tcg ctc gcg cgg ccg aag ctc gcg ccg ctc gcg gac gcg 1008
Arg Gly Leu Ser Leu Ala Arg Pro Lys Leu Ala Pro Leu Ala Asp Ala
325 330 335
ccg ctg ccg gcg cgg ctg ccg gcc ggg ctg ctg ggg cgg cgg ccg gac 1056
Pro Leu Pro Ala Arg Leu Pro Ala Gly Leu Leu Gly Arg Arg Pro Asp
340 345 350
atc gtc gcg gcg cgg ctg cgg gtg gag gcg gcg tac gcg gcg atc gac 1104
Ile Val Ala Ala Arg Leu Arg Val Glu Ala Ala Tyr Ala Ala Ile Asp
355 360 365
ggc acg cgc gcg tcg ttc tac ccg gac gtg aac ctg gcg gcg ctg ggc 1152
Gly Thr Arg Ala Ser Phe Tyr Pro Asp Val Asn Leu Ala Ala Leu Gly
370 375 380
ggg ctg ttc gcg ctc acg ccg gcg tcg ctg ttc aag cac gat gcg ctg 1200
Gly Leu Phe Ala Leu Thr Pro Ala Ser Leu Phe Lys His Asp Ala Leu
385 390 395 400
ggg ggc tcg atc ggt ccg gcg ctg tcg ctg ccg atc ttc gat cgc ggc 1248
Gly Gly Ser Ile Gly Pro Ala Leu Ser Leu Pro Ile Phe Asp Arg Gly
405 410 415
cgg ctg aag gcg aag ctg ggg ggc gac gtg gcg aac gcg gac gtg gcg 1296
Arg Leu Lys Ala Lys Leu Gly Gly Asp Val Ala Asn Ala Asp Val Ala
420 425 430
ctg gcg ctg tac aac cag acg gtg gat gcg gcg ctg ggc gag gtg gcg 1344
Leu Ala Leu Tyr Asn Gln Thr Val Asp Ala Ala Leu Gly Glu Val Ala
435 440 445
cgg cag ttg acg tcg ctg tcg acg gtg gat gcg ctg ctc gag gcg cag 1392
Arg Gln Leu Thr Ser Leu Ser Thr Val Asp Ala Leu Leu Glu Ala Gln
450 455 460
cag cag gcg gtg cgc tcg gcg cag cgg atg gtg gcg ctg gcg cag gac 1440
Gln Gln Ala Val Arg Ser Ala Gln Arg Met Val Ala Leu Ala Gln Asp
465 470 475 480
cgg cac cgg cgg ggg atg ggg atg cgc aag gac gtg aac gtg gcg aag 1488
Arg His Arg Arg Gly Met Gly Met Arg Lys Asp Val Asn Val Ala Lys
485 490 495
ctg acg ctg ctg gac gag cgt gcg cac gtg atc gag ctg cag gcg cgg 1536
Leu Thr Leu Leu Asp Glu Arg Ala His Val Ile Glu Leu Gln Ala Arg
500 505 510
cgg cgg acg ctg cgg gtg ggg ctg atc ggg gcg ctg ggc ggc ggc ttc 1584
Arg Arg Thr Leu Arg Val Gly Leu Ile Gly Ala Leu Gly Gly Gly Phe
515 520 525
gac gcg cgg ccg gcg ggc ggc gcg ccg ctc gcg cag ggc aag ccg ttc 1632
Asp Ala Arg Pro Ala Gly Gly Ala Pro Leu Ala Gln Gly Lys Pro Phe
530 535 540
gcg gcg gcg agc gac agg ccg ccc gat tga 1662
Ala Ala Ala Ser Asp Arg Pro Pro Asp
545 550
<210> 26
<211> 553
<212> PRT
<213> Burkholderia thailandensis
<400> 26
Met Arg Ala Arg Ala Arg Arg Arg Ala Ser Arg Cys Gly Arg Asn Glu
1 5 10 15
Arg Asn Gly Pro Gln Arg Asp Thr Gly Lys Gln Glu Gly Arg Ile Ile
20 25 30
Arg Met Thr Gln Thr Ala Thr Gln Ala Ala Thr Arg Ala Met Ile Ala
35 40 45
Thr Gly Ser Arg Ala Ala Arg Arg Leu Ala Ala Ala Ala Leu Ala Trp
50 55 60
Ala Leu Ala Gly Cys Val Pro Ser Gly Phe Glu Pro Ala Leu Ala Pro
65 70 75 80
Arg Thr Pro Gly Asp Asp Ala Leu Ala His Thr Ala Gly Gly Ala Ala
85 90 95
His Gly Ala Trp Pro Ser Pro Asp Trp Val Arg Gln Leu Gly Asp Pro
100 105 110
Gln Leu Asp Ala Leu Val Asp Glu Ala Leu Arg Gln Asn Pro Thr Leu
115 120 125
Gln Ala Ala Gln Ala Arg Ile Gly Val Ala Gln Ser Gln Leu Gln Gln
130 135 140
Phe Glu Ser Leu Thr Gly Leu Thr Ala Thr Ala Gly Ala Ser Leu Ser
145 150 155 160
Lys Ala His Val Pro Arg Ser Gly Gly Thr Ile Asn Thr Thr Phe Asn
165 170 175
Gly Leu Pro Val Ser Val Pro Leu Val Gly Glu Ser Val Val Ser Ser
180 185 190
Ser Ser Leu Phe Val Gly Leu Asn Tyr Gln Leu Asp Leu Trp Gly Lys
195 200 205
Asn Ala Ala Ala Thr Arg Gly Leu Leu Ser Met Arg Asp Ala Ala Arg
210 215 220
Val Glu Ala Glu Gln Ala Arg Leu Ala Leu Ser Val Ala Ile Val Thr
225 230 235 240
Leu Tyr Gly Glu Leu Asp Arg Ala Tyr Ala Leu Arg Glu Leu Leu Gln
245 250 255
Gln Lys Arg Arg Ala Ser Glu Gln Val Glu Thr Val Leu Arg Glu Arg
260 265 270
Ala Ala Arg Gly Ile Asp Asn Gly Tyr Asp Ala Asp Asp Ala Ala Leu
275 280 285
Lys Arg Gly Lys Leu Leu Glu Gln Leu Ala Leu Thr Asp Glu Gln Ile
290 295 300
Gln Leu Gln Lys Leu Gln Leu Gly Val Leu Ser Gly Arg Gly Pro Glu
305 310 315 320
Arg Gly Leu Ser Leu Ala Arg Pro Lys Leu Ala Pro Leu Ala Asp Ala
325 330 335
Pro Leu Pro Ala Arg Leu Pro Ala Gly Leu Leu Gly Arg Arg Pro Asp
340 345 350
Ile Val Ala Ala Arg Leu Arg Val Glu Ala Ala Tyr Ala Ala Ile Asp
355 360 365
Gly Thr Arg Ala Ser Phe Tyr Pro Asp Val Asn Leu Ala Ala Leu Gly
370 375 380
Gly Leu Phe Ala Leu Thr Pro Ala Ser Leu Phe Lys His Asp Ala Leu
385 390 395 400
Gly Gly Ser Ile Gly Pro Ala Leu Ser Leu Pro Ile Phe Asp Arg Gly
405 410 415
Arg Leu Lys Ala Lys Leu Gly Gly Asp Val Ala Asn Ala Asp Val Ala
420 425 430
Leu Ala Leu Tyr Asn Gln Thr Val Asp Ala Ala Leu Gly Glu Val Ala
435 440 445
Arg Gln Leu Thr Ser Leu Ser Thr Val Asp Ala Leu Leu Glu Ala Gln
450 455 460
Gln Gln Ala Val Arg Ser Ala Gln Arg Met Val Ala Leu Ala Gln Asp
465 470 475 480
Arg His Arg Arg Gly Met Gly Met Arg Lys Asp Val Asn Val Ala Lys
485 490 495
Leu Thr Leu Leu Asp Glu Arg Ala His Val Ile Glu Leu Gln Ala Arg
500 505 510
Arg Arg Thr Leu Arg Val Gly Leu Ile Gly Ala Leu Gly Gly Gly Phe
515 520 525
Asp Ala Arg Pro Ala Gly Gly Ala Pro Leu Ala Gln Gly Lys Pro Phe
530 535 540
Ala Ala Ala Ser Asp Arg Pro Pro Asp
545 550
<210> 27
<211> 1401
<212> DNA
<213> Burkholderia thailandensis
<220>
<221> CDS
<222> (1)..(1401)
<400> 27
atg cgg ccc gaa gcc acc gac acc cga aga cac cga cac caa cgc cac 48
Met Arg Pro Glu Ala Thr Asp Thr Arg Arg His Arg His Gln Arg His
1 5 10 15
ctt cac cgt gta cac gag cga ttc aac cga cac cgc ccc cga gca tcg 96
Leu His Arg Val His Glu Arg Phe Asn Arg His Arg Pro Arg Ala Ser
20 25 30
aag ccc gtc ggg ccg atc cgc gac ggc ttg cgg gcc ggc ccg gcc gtt 144
Lys Pro Val Gly Pro Ile Arg Asp Gly Leu Arg Ala Gly Pro Ala Val
35 40 45
gcc ggc cgg cgc cac cga cat cac gca cgc gaa gac ctt gaa cga tac 192
Ala Gly Arg Arg His Arg His His Ala Arg Glu Asp Leu Glu Arg Tyr
50 55 60
cgc cac cga tac ccc gcg cgc gaa ggc gcc cac cga tcc ggc cgc cct 240
Arg His Arg Tyr Pro Ala Arg Glu Gly Ala His Arg Ser Gly Arg Pro
65 70 75 80
cga cgg cgc gca cgc gca gcc cgt gcc ggc gca cga gcg cgg atc gcc 288
Arg Arg Arg Ala Arg Ala Ala Arg Ala Gly Ala Arg Ala Arg Ile Ala
85 90 95
tcc gcc gcc gga agc cgc ggc gac gct cgc cgc gcg ccg cgc gac gcg 336
Ser Ala Ala Gly Ser Arg Gly Asp Ala Arg Arg Ala Pro Arg Asp Ala
100 105 110
ccg ccg gcg ctt cgc gct gtt ctt cgg gct gct ggc gct ggc cgc gct 384
Pro Pro Ala Leu Arg Ala Val Leu Arg Ala Ala Gly Ala Gly Arg Ala
115 120 125
gac cgc ggg gct cta ctg gtt cgt cgc cgg gcg ctt cag cga gga gac 432
Asp Arg Gly Ala Leu Leu Val Arg Arg Arg Ala Leu Gln Arg Gly Asp
130 135 140
gga cga cgc gta cgt ggc cgg caa cgt ggt gca gat cgc cgc gca gat 480
Gly Arg Arg Val Arg Gly Arg Gln Arg Gly Ala Asp Arg Arg Ala Asp
145 150 155 160
cca ggg gac ggt gac cga cgt gct ggt ggc gga cac gca gca ggt gaa 528
Pro Gly Asp Gly Asp Arg Arg Ala Gly Gly Gly His Ala Ala Gly Glu
165 170 175
ggc ggg gca ggc gct ggt gaa gct cga cga cgc gga cgc gtc ggc ggc 576
Gly Gly Ala Gly Ala Gly Glu Ala Arg Arg Arg Gly Arg Val Gly Gly
180 185 190
gtt cgc gca ggc gcg ggc gca gct cgc gca ggc ggt gcg gca ggt ggc 624
Val Arg Ala Gly Ala Gly Ala Ala Arg Ala Gly Gly Ala Ala Gly Gly
195 200 205
gaa cac gcg gct ctc gat ggg gat gta cga gga gac ggt gaa ggc gcg 672
Glu His Ala Ala Leu Asp Gly Asp Val Arg Gly Asp Gly Glu Gly Ala
210 215 220
cga ggc gga cct gaa gct tgc gca gca ggc gta tcc gga gga act ggc 720
Arg Gly Gly Pro Glu Ala Cys Ala Ala Gly Val Ser Gly Gly Thr Gly
225 230 235 240
gcg gcg aaa gtc gtc gct ggc gaa cgc gca ggc ggc gct ggc ggg ggc 768
Ala Ala Lys Val Val Ala Gly Glu Arg Ala Gly Gly Ala Gly Gly Gly
245 250 255
gca ggc gca gct gga ggc ggc gcg cgc gct ggg cag cga gcg gcc ggt 816
Ala Gly Ala Ala Gly Gly Gly Ala Arg Ala Gly Gln Arg Ala Ala Gly
260 265 270
cga gca gaa ccc ggc ggt gca gca ggc ggc cgc gca gtt caa gct ggc 864
Arg Ala Glu Pro Gly Gly Ala Ala Gly Gly Arg Ala Val Gln Ala Gly
275 280 285
gta ccg gaa cct gag gcg cac gac gat cgt gtc gcc ggt gga cgg cac 912
Val Pro Glu Pro Glu Ala His Asp Asp Arg Val Ala Gly Gly Arg His
290 295 300
ggt cgg tca gcg gtc ggt gca gat cgg tca gca ggt ggg gcc ggg ggt 960
Gly Arg Ser Ala Val Gly Ala Asp Arg Ser Ala Gly Gly Ala Gly Gly
305 310 315 320
gcc gct gat gtc ggt ggt gca gtt gcg gca ggt gtg ggt gga ggc gaa 1008
Ala Ala Asp Val Gly Gly Ala Val Ala Ala Gly Val Gly Gly Gly Glu
325 330 335
ctt caa gga agg gca gat ccg gca cat gcg ggt ggg cca gcc ggt gcg 1056
Leu Gln Gly Arg Ala Asp Pro Ala His Ala Gly Gly Pro Ala Gly Ala
340 345 350
gct cga atc gga cct gta cgg cgc gcg ggt gac gta cca cgg ccg ggt 1104
Ala Arg Ile Gly Pro Val Arg Arg Ala Gly Asp Val Pro Arg Pro Gly
355 360 365
gga ggg ggt ctc ggc ggg cac ggg cag cgc gtt ctc gat gct gcc gtc 1152
Gly Gly Gly Leu Gly Gly His Gly Gln Arg Val Leu Asp Ala Ala Val
370 375 380
gca gaa cgc ggc ggg gaa ctg gat caa ggt ggt gca gcg cct gcc ggt 1200
Ala Glu Arg Gly Gly Glu Leu Asp Gln Gly Gly Ala Ala Pro Ala Gly
385 390 395 400
ggt gat ctc gct gga gcc gtc gga gct ggc ggc gca ccc gct gcg ggt 1248
Gly Asp Leu Ala Gly Ala Val Gly Ala Gly Gly Ala Pro Ala Ala Gly
405 410 415
ggg gct gtc gat gcg cgc gac ggt gga gac gaa ggt gcg tgg cgg ccg 1296
Gly Ala Val Asp Ala Arg Asp Gly Gly Asp Glu Gly Ala Trp Arg Pro
420 425 430
cct gct cga cgg cga cgc gcc gct gcc ggg gct gcg cac gcg ggt gca 1344
Pro Ala Arg Arg Arg Arg Ala Ala Ala Gly Ala Ala His Ala Gly Ala
435 440 445
cga agc gca ggc ggg cga ggc cga ggc cgc ggc ttc ggc agt gat tcg 1392
Arg Ser Ala Gly Gly Arg Gly Arg Gly Arg Gly Phe Gly Ser Asp Ser
450 455 460
gga gaa tga 1401
Gly Glu
465
<210> 28
<211> 466
<212> PRT
<213> Burkholderia thailandensis
<400> 28
Met Arg Pro Glu Ala Thr Asp Thr Arg Arg His Arg His Gln Arg His
1 5 10 15
Leu His Arg Val His Glu Arg Phe Asn Arg His Arg Pro Arg Ala Ser
20 25 30
Lys Pro Val Gly Pro Ile Arg Asp Gly Leu Arg Ala Gly Pro Ala Val
35 40 45
Ala Gly Arg Arg His Arg His His Ala Arg Glu Asp Leu Glu Arg Tyr
50 55 60
Arg His Arg Tyr Pro Ala Arg Glu Gly Ala His Arg Ser Gly Arg Pro
65 70 75 80
Arg Arg Arg Ala Arg Ala Ala Arg Ala Gly Ala Arg Ala Arg Ile Ala
85 90 95
Ser Ala Ala Gly Ser Arg Gly Asp Ala Arg Arg Ala Pro Arg Asp Ala
100 105 110
Pro Pro Ala Leu Arg Ala Val Leu Arg Ala Ala Gly Ala Gly Arg Ala
115 120 125
Asp Arg Gly Ala Leu Leu Val Arg Arg Arg Ala Leu Gln Arg Gly Asp
130 135 140
Gly Arg Arg Val Arg Gly Arg Gln Arg Gly Ala Asp Arg Arg Ala Asp
145 150 155 160
Pro Gly Asp Gly Asp Arg Arg Ala Gly Gly Gly His Ala Ala Gly Glu
165 170 175
Gly Gly Ala Gly Ala Gly Glu Ala Arg Arg Arg Gly Arg Val Gly Gly
180 185 190
Val Arg Ala Gly Ala Gly Ala Ala Arg Ala Gly Gly Ala Ala Gly Gly
195 200 205
Glu His Ala Ala Leu Asp Gly Asp Val Arg Gly Asp Gly Glu Gly Ala
210 215 220
Arg Gly Gly Pro Glu Ala Cys Ala Ala Gly Val Ser Gly Gly Thr Gly
225 230 235 240
Ala Ala Lys Val Val Ala Gly Glu Arg Ala Gly Gly Ala Gly Gly Gly
245 250 255
Ala Gly Ala Ala Gly Gly Gly Ala Arg Ala Gly Gln Arg Ala Ala Gly
260 265 270
Arg Ala Glu Pro Gly Gly Ala Ala Gly Gly Arg Ala Val Gln Ala Gly
275 280 285
Val Pro Glu Pro Glu Ala His Asp Asp Arg Val Ala Gly Gly Arg His
290 295 300
Gly Arg Ser Ala Val Gly Ala Asp Arg Ser Ala Gly Gly Ala Gly Gly
305 310 315 320
Ala Ala Asp Val Gly Gly Ala Val Ala Ala Gly Val Gly Gly Gly Glu
325 330 335
Leu Gln Gly Arg Ala Asp Pro Ala His Ala Gly Gly Pro Ala Gly Ala
340 345 350
Ala Arg Ile Gly Pro Val Arg Arg Ala Gly Asp Val Pro Arg Pro Gly
355 360 365
Gly Gly Gly Leu Gly Gly His Gly Gln Arg Val Leu Asp Ala Ala Val
370 375 380
Ala Glu Arg Gly Gly Glu Leu Asp Gln Gly Gly Ala Ala Pro Ala Gly
385 390 395 400
Gly Asp Leu Ala Gly Ala Val Gly Ala Gly Gly Ala Pro Ala Ala Gly
405 410 415
Gly Ala Val Asp Ala Arg Asp Gly Gly Asp Glu Gly Ala Trp Arg Pro
420 425 430
Pro Ala Arg Arg Arg Arg Ala Ala Ala Gly Ala Ala His Ala Gly Ala
435 440 445
Arg Ser Ala Gly Gly Arg Gly Arg Gly Arg Gly Phe Gly Ser Asp Ser
450 455 460
Gly Glu
465
<210> 29
<211> 1680
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(1680)
<400> 29
atg agt aac aag aac aac gat gag cta cag cgg cag gcc tcg gaa aac 48
Met Ser Asn Lys Asn Asn Asp Glu Leu Gln Arg Gln Ala Ser Glu Asn
1 5 10 15
acc atg ggg ctg aac ccg gtc atc ggc atc cgc cgc aag gac ctg ttg 96
Thr Met Gly Leu Asn Pro Val Ile Gly Ile Arg Arg Lys Asp Leu Leu
20 25 30
agc tcg gca cgc acc gtg ctg cgc cag gcc gtg cgc caa ccg ctg cac 144
Ser Ser Ala Arg Thr Val Leu Arg Gln Ala Val Arg Gln Pro Leu His
35 40 45
agc gcc aag cat gtg gct cac ttt ggc ctg gag ctg aag aac gtg ttg 192
Ser Ala Lys His Val Ala His Phe Gly Leu Glu Leu Lys Asn Val Leu
50 55 60
ctg ggc aaa tcc agc ctg gcc ccg gac agc gac gac cgt cgc ttc aat 240
Leu Gly Lys Ser Ser Leu Ala Pro Asp Ser Asp Asp Arg Arg Phe Asn
65 70 75 80
gac ccg gcc tgg agc aac aac ccg ctg tac cgc cgc tac ctg caa acc 288
Asp Pro Ala Trp Ser Asn Asn Pro Leu Tyr Arg Arg Tyr Leu Gln Thr
85 90 95
tac ctg gcc tgg cgc aag gag ctg cag gac tgg gtg agc agc agc gac 336
Tyr Leu Ala Trp Arg Lys Glu Leu Gln Asp Trp Val Ser Ser Ser Asp
100 105 110
ctg tcc ccc cag gac atc agc cgc ggc cag ttc gtc atc aac ctg atg 384
Leu Ser Pro Gln Asp Ile Ser Arg Gly Gln Phe Val Ile Asn Leu Met
115 120 125
acc gag gcc atg gcg ccg acc aat acc ctg tcc aac ccg gct gcg gtc 432
Thr Glu Ala Met Ala Pro Thr Asn Thr Leu Ser Asn Pro Ala Ala Val
130 135 140
aaa cgc ttc ttc gaa acc ggc ggc aag agc ctg ctc gat ggc ctg tcc 480
Lys Arg Phe Phe Glu Thr Gly Gly Lys Ser Leu Leu Asp Gly Leu Ser
145 150 155 160
aac ctg gcc aag gac atg gtc aac aac ggc ggc atg ccc agc cag gtg 528
Asn Leu Ala Lys Asp Met Val Asn Asn Gly Gly Met Pro Ser Gln Val
165 170 175
aac atg gat gcc ttc gaa gtg ggc aag aac ctg ggc acc agc gaa ggc 576
Asn Met Asp Ala Phe Glu Val Gly Lys Asn Leu Gly Thr Ser Glu Gly
180 185 190
gcg gtg gtg tac cgc aac gat gtg ctg gaa ctg atc cag tac agc ccc 624
Ala Val Val Tyr Arg Asn Asp Val Leu Glu Leu Ile Gln Tyr Ser Pro
195 200 205
atc acc gag cag gtg cat gcc cgt ccg ctg ctg gtg gtg cca ccg cag 672
Ile Thr Glu Gln Val His Ala Arg Pro Leu Leu Val Val Pro Pro Gln
210 215 220
atc aac aag ttc tac gtg ttc gac ctc agc ccg gaa aag agc ctg gcg 720
Ile Asn Lys Phe Tyr Val Phe Asp Leu Ser Pro Glu Lys Ser Leu Ala
225 230 235 240
cgc ttc tgc ctg cgc tcg cag cag cag acc ttc atc atc agc tgg cgc 768
Arg Phe Cys Leu Arg Ser Gln Gln Gln Thr Phe Ile Ile Ser Trp Arg
245 250 255
aac ccg acc aag gcc cag cgt gaa tgg ggc ctg tcc acc tac atc gat 816
Asn Pro Thr Lys Ala Gln Arg Glu Trp Gly Leu Ser Thr Tyr Ile Asp
260 265 270
gcg ctg aaa gaa gcc gtc gac gcg gtg ctg tcg att acc ggc agc aag 864
Ala Leu Lys Glu Ala Val Asp Ala Val Leu Ser Ile Thr Gly Ser Lys
275 280 285
gac ctg aac atg ctc ggc gcc tgc tcc ggt ggc atc act tgt acc gca 912
Asp Leu Asn Met Leu Gly Ala Cys Ser Gly Gly Ile Thr Cys Thr Ala
290 295 300
ctg gtc ggg cac tat gcc gca ttg ggc gag aac aag gtc aac gcc ctg 960
Leu Val Gly His Tyr Ala Ala Leu Gly Glu Asn Lys Val Asn Ala Leu
305 310 315 320
acc gtg ctg gtc agc gtg ctg gac acc acc atg gac aac cag gtt gct 1008
Thr Val Leu Val Ser Val Leu Asp Thr Thr Met Asp Asn Gln Val Ala
325 330 335
ttg ttt gtc gac gag cag acc ttg gag gcc gcc aag cgc cac tcc tat 1056
Leu Phe Val Asp Glu Gln Thr Leu Glu Ala Ala Lys Arg His Ser Tyr
340 345 350
cag gcg ggc gtg ctg gaa ggc agc gaa atg gcc aag gtg ttc gcc tgg 1104
Gln Ala Gly Val Leu Glu Gly Ser Glu Met Ala Lys Val Phe Ala Trp
355 360 365
atg cgc ccc aac gac ctg atc tgg aac tac tgg gta aac aac tac ctg 1152
Met Arg Pro Asn Asp Leu Ile Trp Asn Tyr Trp Val Asn Asn Tyr Leu
370 375 380
ctc ggc aat gag ccc ccc gtg ttc gac atc ctg ttc tgg aac aac gac 1200
Leu Gly Asn Glu Pro Pro Val Phe Asp Ile Leu Phe Trp Asn Asn Asp
385 390 395 400
acc acg cgc ctg ccg gcc gcc ttc cac ggc gac ctg atc gaa atg ttc 1248
Thr Thr Arg Leu Pro Ala Ala Phe His Gly Asp Leu Ile Glu Met Phe
405 410 415
aag agc aac ccg ctg acc cgc ccc gac gcc ctg aaa gtg tgc ggc acc 1296
Lys Ser Asn Pro Leu Thr Arg Pro Asp Ala Leu Lys Val Cys Gly Thr
420 425 430
gcg atc gac ctg aaa cag gtc aaa tgc gac atc tac agc ctc gcc ggc 1344
Ala Ile Asp Leu Lys Gln Val Lys Cys Asp Ile Tyr Ser Leu Ala Gly
435 440 445
acc aac gac cac atc acc ccc tgg ccg tca tgc tac cgc tcg gca cat 1392
Thr Asn Asp His Ile Thr Pro Trp Pro Ser Cys Tyr Arg Ser Ala His
450 455 460
ctg ttc ggc ggc aag atc gaa ttc gta ctg tcc aac agc ggg cat atc 1440
Leu Phe Gly Gly Lys Ile Glu Phe Val Leu Ser Asn Ser Gly His Ile
465 470 475 480
cag agc atc ctc aac ccg ccg ggc aac ccg aag gca cgt ttc atg acc 1488
Gln Ser Ile Leu Asn Pro Pro Gly Asn Pro Lys Ala Arg Phe Met Thr
485 490 495
ggt gcc gat cgc ccg ggt gac ccg gtg gcc tgg cag gaa aat gcc atc 1536
Gly Ala Asp Arg Pro Gly Asp Pro Val Ala Trp Gln Glu Asn Ala Ile
500 505 510
aag cat gca gac tcc tgg tgg ctg cac tgg cag agt tgg ctg ggc gag 1584
Lys His Ala Asp Ser Trp Trp Leu His Trp Gln Ser Trp Leu Gly Glu
515 520 525
cgt gcc ggc gcg ctg aaa aag gca ccg acc cgc ctg ggc aac cgt acc 1632
Arg Ala Gly Ala Leu Lys Lys Ala Pro Thr Arg Leu Gly Asn Arg Thr
530 535 540
tat gcc gcc ggc gaa gcc tcc cca ggc acc tac gtt cac gag cgt tga 1680
Tyr Ala Ala Gly Glu Ala Ser Pro Gly Thr Tyr Val His Glu Arg
545 550 555
<210> 30
<211> 559
<212> PRT
<213> Pseudomonas putida
<400> 30
Met Ser Asn Lys Asn Asn Asp Glu Leu Gln Arg Gln Ala Ser Glu Asn
1 5 10 15
Thr Met Gly Leu Asn Pro Val Ile Gly Ile Arg Arg Lys Asp Leu Leu
20 25 30
Ser Ser Ala Arg Thr Val Leu Arg Gln Ala Val Arg Gln Pro Leu His
35 40 45
Ser Ala Lys His Val Ala His Phe Gly Leu Glu Leu Lys Asn Val Leu
50 55 60
Leu Gly Lys Ser Ser Leu Ala Pro Asp Ser Asp Asp Arg Arg Phe Asn
65 70 75 80
Asp Pro Ala Trp Ser Asn Asn Pro Leu Tyr Arg Arg Tyr Leu Gln Thr
85 90 95
Tyr Leu Ala Trp Arg Lys Glu Leu Gln Asp Trp Val Ser Ser Ser Asp
100 105 110
Leu Ser Pro Gln Asp Ile Ser Arg Gly Gln Phe Val Ile Asn Leu Met
115 120 125
Thr Glu Ala Met Ala Pro Thr Asn Thr Leu Ser Asn Pro Ala Ala Val
130 135 140
Lys Arg Phe Phe Glu Thr Gly Gly Lys Ser Leu Leu Asp Gly Leu Ser
145 150 155 160
Asn Leu Ala Lys Asp Met Val Asn Asn Gly Gly Met Pro Ser Gln Val
165 170 175
Asn Met Asp Ala Phe Glu Val Gly Lys Asn Leu Gly Thr Ser Glu Gly
180 185 190
Ala Val Val Tyr Arg Asn Asp Val Leu Glu Leu Ile Gln Tyr Ser Pro
195 200 205
Ile Thr Glu Gln Val His Ala Arg Pro Leu Leu Val Val Pro Pro Gln
210 215 220
Ile Asn Lys Phe Tyr Val Phe Asp Leu Ser Pro Glu Lys Ser Leu Ala
225 230 235 240
Arg Phe Cys Leu Arg Ser Gln Gln Gln Thr Phe Ile Ile Ser Trp Arg
245 250 255
Asn Pro Thr Lys Ala Gln Arg Glu Trp Gly Leu Ser Thr Tyr Ile Asp
260 265 270
Ala Leu Lys Glu Ala Val Asp Ala Val Leu Ser Ile Thr Gly Ser Lys
275 280 285
Asp Leu Asn Met Leu Gly Ala Cys Ser Gly Gly Ile Thr Cys Thr Ala
290 295 300
Leu Val Gly His Tyr Ala Ala Leu Gly Glu Asn Lys Val Asn Ala Leu
305 310 315 320
Thr Val Leu Val Ser Val Leu Asp Thr Thr Met Asp Asn Gln Val Ala
325 330 335
Leu Phe Val Asp Glu Gln Thr Leu Glu Ala Ala Lys Arg His Ser Tyr
340 345 350
Gln Ala Gly Val Leu Glu Gly Ser Glu Met Ala Lys Val Phe Ala Trp
355 360 365
Met Arg Pro Asn Asp Leu Ile Trp Asn Tyr Trp Val Asn Asn Tyr Leu
370 375 380
Leu Gly Asn Glu Pro Pro Val Phe Asp Ile Leu Phe Trp Asn Asn Asp
385 390 395 400
Thr Thr Arg Leu Pro Ala Ala Phe His Gly Asp Leu Ile Glu Met Phe
405 410 415
Lys Ser Asn Pro Leu Thr Arg Pro Asp Ala Leu Lys Val Cys Gly Thr
420 425 430
Ala Ile Asp Leu Lys Gln Val Lys Cys Asp Ile Tyr Ser Leu Ala Gly
435 440 445
Thr Asn Asp His Ile Thr Pro Trp Pro Ser Cys Tyr Arg Ser Ala His
450 455 460
Leu Phe Gly Gly Lys Ile Glu Phe Val Leu Ser Asn Ser Gly His Ile
465 470 475 480
Gln Ser Ile Leu Asn Pro Pro Gly Asn Pro Lys Ala Arg Phe Met Thr
485 490 495
Gly Ala Asp Arg Pro Gly Asp Pro Val Ala Trp Gln Glu Asn Ala Ile
500 505 510
Lys His Ala Asp Ser Trp Trp Leu His Trp Gln Ser Trp Leu Gly Glu
515 520 525
Arg Ala Gly Ala Leu Lys Lys Ala Pro Thr Arg Leu Gly Asn Arg Thr
530 535 540
Tyr Ala Ala Gly Glu Ala Ser Pro Gly Thr Tyr Val His Glu Arg
545 550 555
<210> 31
<211> 1683
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(1683)
<400> 31
atg aca gac aaa ccg gcc aaa gga tcg aca acg ctc ccc gcc acc cgc 48
Met Thr Asp Lys Pro Ala Lys Gly Ser Thr Thr Leu Pro Ala Thr Arg
1 5 10 15
atg aac gtg cag aac gcc atc ctc ggc ctg cgc ggc cgc gac ctg ctt 96
Met Asn Val Gln Asn Ala Ile Leu Gly Leu Arg Gly Arg Asp Leu Leu
20 25 30
tcc acg ctg cgc aac gtc ggc cgc cac ggc ctg cgc cac ccg ctg cat 144
Ser Thr Leu Arg Asn Val Gly Arg His Gly Leu Arg His Pro Leu His
35 40 45
acc gcg cat cat ctg ctg gcg ctt ggc ggg cag ttg ggg cgg gtg atg 192
Thr Ala His His Leu Leu Ala Leu Gly Gly Gln Leu Gly Arg Val Met
50 55 60
ctg ggg gac acg ccc tac cag ccg aac ccg cgt gac gca cgc ttc agt 240
Leu Gly Asp Thr Pro Tyr Gln Pro Asn Pro Arg Asp Ala Arg Phe Ser
65 70 75 80
gac ccg acc tgg agc cag aac ccg ttc tac cgc cgc ggc ctg caa gcc 288
Asp Pro Thr Trp Ser Gln Asn Pro Phe Tyr Arg Arg Gly Leu Gln Ala
85 90 95
tat ctg gcc tgg cag aag cag aca cgc cag tgg atc gat gaa agc cat 336
Tyr Leu Ala Trp Gln Lys Gln Thr Arg Gln Trp Ile Asp Glu Ser His
100 105 110
ttg aac gac gat gat cga gcc cgc gcc cac ttc ctg ttc aac ctg atc 384
Leu Asn Asp Asp Asp Arg Ala Arg Ala His Phe Leu Phe Asn Leu Ile
115 120 125
aac gat gcg ctg gcg ccc agc aac tca ctg ctc aat ccg cag gcg gtc 432
Asn Asp Ala Leu Ala Pro Ser Asn Ser Leu Leu Asn Pro Gln Ala Val
130 135 140
aag ggg ctg ttc aac acc ggc ggc cag agc ctg gtg cgc ggc gtg gct 480
Lys Gly Leu Phe Asn Thr Gly Gly Gln Ser Leu Val Arg Gly Val Ala
145 150 155 160
cac ctg ctc gac gac ctg cgt cac aac gat ggg ctg cct cgt cag gtg 528
His Leu Leu Asp Asp Leu Arg His Asn Asp Gly Leu Pro Arg Gln Val
165 170 175
gac gag cgc gcc ttc gaa gtg ggc gtt aac ctg gcc gca acc cct ggc 576
Asp Glu Arg Ala Phe Glu Val Gly Val Asn Leu Ala Ala Thr Pro Gly
180 185 190
gca gtg gta ttt cgc aac gag ctg ctg gag ctg atc cag tac tcg ccg 624
Ala Val Val Phe Arg Asn Glu Leu Leu Glu Leu Ile Gln Tyr Ser Pro
195 200 205
atg agc gaa aag cag cac gca cgc cca ctg ctg gtc gtg ccg cct cag 672
Met Ser Glu Lys Gln His Ala Arg Pro Leu Leu Val Val Pro Pro Gln
210 215 220
atc aac agg ttc tac atc ttc gac ctc agc gcg acc aac agc ttc gtc 720
Ile Asn Arg Phe Tyr Ile Phe Asp Leu Ser Ala Thr Asn Ser Phe Val
225 230 235 240
cag tac atg ctc aaa agc ggc ttg cag gtg ttc atg gtc agc tgg agc 768
Gln Tyr Met Leu Lys Ser Gly Leu Gln Val Phe Met Val Ser Trp Ser
245 250 255
aac ccc gac cca cgc cac cgt gaa tgg ggc ctt tcc agc tat gtg caa 816
Asn Pro Asp Pro Arg His Arg Glu Trp Gly Leu Ser Ser Tyr Val Gln
260 265 270
gcc ctg gag gaa gcg ctc aat gcc tgc cgc agt atc agc ggc aac cgc 864
Ala Leu Glu Glu Ala Leu Asn Ala Cys Arg Ser Ile Ser Gly Asn Arg
275 280 285
gac ccc aac ctg atg ggt gcc tgt gcc ggc ggc ctg acc atg gcc gca 912
Asp Pro Asn Leu Met Gly Ala Cys Ala Gly Gly Leu Thr Met Ala Ala
290 295 300
ctg caa ggc cat ctg caa gcc aag aag caa ttg cgc cgg gtg cgc agt 960
Leu Gln Gly His Leu Gln Ala Lys Lys Gln Leu Arg Arg Val Arg Ser
305 310 315 320
gcc acg tat ctg gtc agc ttg ctg gac agc aag ttc gaa agc ccg gcc 1008
Ala Thr Tyr Leu Val Ser Leu Leu Asp Ser Lys Phe Glu Ser Pro Ala
325 330 335
agc ctg ttc gcc gat gag cag acc atc gaa gcg gcc aag cga cgc tcc 1056
Ser Leu Phe Ala Asp Glu Gln Thr Ile Glu Ala Ala Lys Arg Arg Ser
340 345 350
tat cag cgt ggc gtg ctg gac ggt ggt gaa gtg gcg cgg atc ttc gcc 1104
Tyr Gln Arg Gly Val Leu Asp Gly Gly Glu Val Ala Arg Ile Phe Ala
355 360 365
tgg atg cgg ccc aac gac ctg atc tgg aac tac tgg gta aac aac tac 1152
Trp Met Arg Pro Asn Asp Leu Ile Trp Asn Tyr Trp Val Asn Asn Tyr
370 375 380
ctg ctc ggc aag aca ccg cct gcg ttc gac atc ctg tac tgg aat gcc 1200
Leu Leu Gly Lys Thr Pro Pro Ala Phe Asp Ile Leu Tyr Trp Asn Ala
385 390 395 400
gac agc acg cgc ctg ccc gcc gcg ctg cat ggc gac ctg ctg gag ttt 1248
Asp Ser Thr Arg Leu Pro Ala Ala Leu His Gly Asp Leu Leu Glu Phe
405 410 415
ttc aag ctc aac ccc ttg acc tac gcg tcc ggg ctg gag gtg tgc ggt 1296
Phe Lys Leu Asn Pro Leu Thr Tyr Ala Ser Gly Leu Glu Val Cys Gly
420 425 430
acg ccg atc gac ctg cag cag gtc aat atc gac agc ttt acc gtg gcc 1344
Thr Pro Ile Asp Leu Gln Gln Val Asn Ile Asp Ser Phe Thr Val Ala
435 440 445
ggc agc aac gac cac atc aca cca tgg gat gcg gtg tac cgc tcg gcc 1392
Gly Ser Asn Asp His Ile Thr Pro Trp Asp Ala Val Tyr Arg Ser Ala
450 455 460
ttg ctg ctg ggt ggc gag cgg cgc ttc gtg ctg gcc aac agc ggg cat 1440
Leu Leu Leu Gly Gly Glu Arg Arg Phe Val Leu Ala Asn Ser Gly His
465 470 475 480
atc cag agc atc atc aac ccg cca ggc aac ccc aag gcc tac tac ctg 1488
Ile Gln Ser Ile Ile Asn Pro Pro Gly Asn Pro Lys Ala Tyr Tyr Leu
485 490 495
gcc aac ccc aag ctg agc agc gac cca cgc gcc tgg ttc cac gac gcc 1536
Ala Asn Pro Lys Leu Ser Ser Asp Pro Arg Ala Trp Phe His Asp Ala
500 505 510
aag cgc agt gaa ggc agc tgg tgg ccg ttg tgg ctg gag tgg atc acc 1584
Lys Arg Ser Glu Gly Ser Trp Trp Pro Leu Trp Leu Glu Trp Ile Thr
515 520 525
gca cgc tcc ggc ctg ctc aag gca ccg cgt act gaa ctg ggc aac gcc 1632
Ala Arg Ser Gly Leu Leu Lys Ala Pro Arg Thr Glu Leu Gly Asn Ala
530 535 540
act tac cca ctg cta ggc ccc gcg cca ggc acc tac gtg ctg acc cga 1680
Thr Tyr Pro Leu Leu Gly Pro Ala Pro Gly Thr Tyr Val Leu Thr Arg
545 550 555 560
tga 1683
<210> 32
<211> 560
<212> PRT
<213> Pseudomonas putida
<400> 32
Met Thr Asp Lys Pro Ala Lys Gly Ser Thr Thr Leu Pro Ala Thr Arg
1 5 10 15
Met Asn Val Gln Asn Ala Ile Leu Gly Leu Arg Gly Arg Asp Leu Leu
20 25 30
Ser Thr Leu Arg Asn Val Gly Arg His Gly Leu Arg His Pro Leu His
35 40 45
Thr Ala His His Leu Leu Ala Leu Gly Gly Gln Leu Gly Arg Val Met
50 55 60
Leu Gly Asp Thr Pro Tyr Gln Pro Asn Pro Arg Asp Ala Arg Phe Ser
65 70 75 80
Asp Pro Thr Trp Ser Gln Asn Pro Phe Tyr Arg Arg Gly Leu Gln Ala
85 90 95
Tyr Leu Ala Trp Gln Lys Gln Thr Arg Gln Trp Ile Asp Glu Ser His
100 105 110
Leu Asn Asp Asp Asp Arg Ala Arg Ala His Phe Leu Phe Asn Leu Ile
115 120 125
Asn Asp Ala Leu Ala Pro Ser Asn Ser Leu Leu Asn Pro Gln Ala Val
130 135 140
Lys Gly Leu Phe Asn Thr Gly Gly Gln Ser Leu Val Arg Gly Val Ala
145 150 155 160
His Leu Leu Asp Asp Leu Arg His Asn Asp Gly Leu Pro Arg Gln Val
165 170 175
Asp Glu Arg Ala Phe Glu Val Gly Val Asn Leu Ala Ala Thr Pro Gly
180 185 190
Ala Val Val Phe Arg Asn Glu Leu Leu Glu Leu Ile Gln Tyr Ser Pro
195 200 205
Met Ser Glu Lys Gln His Ala Arg Pro Leu Leu Val Val Pro Pro Gln
210 215 220
Ile Asn Arg Phe Tyr Ile Phe Asp Leu Ser Ala Thr Asn Ser Phe Val
225 230 235 240
Gln Tyr Met Leu Lys Ser Gly Leu Gln Val Phe Met Val Ser Trp Ser
245 250 255
Asn Pro Asp Pro Arg His Arg Glu Trp Gly Leu Ser Ser Tyr Val Gln
260 265 270
Ala Leu Glu Glu Ala Leu Asn Ala Cys Arg Ser Ile Ser Gly Asn Arg
275 280 285
Asp Pro Asn Leu Met Gly Ala Cys Ala Gly Gly Leu Thr Met Ala Ala
290 295 300
Leu Gln Gly His Leu Gln Ala Lys Lys Gln Leu Arg Arg Val Arg Ser
305 310 315 320
Ala Thr Tyr Leu Val Ser Leu Leu Asp Ser Lys Phe Glu Ser Pro Ala
325 330 335
Ser Leu Phe Ala Asp Glu Gln Thr Ile Glu Ala Ala Lys Arg Arg Ser
340 345 350
Tyr Gln Arg Gly Val Leu Asp Gly Gly Glu Val Ala Arg Ile Phe Ala
355 360 365
Trp Met Arg Pro Asn Asp Leu Ile Trp Asn Tyr Trp Val Asn Asn Tyr
370 375 380
Leu Leu Gly Lys Thr Pro Pro Ala Phe Asp Ile Leu Tyr Trp Asn Ala
385 390 395 400
Asp Ser Thr Arg Leu Pro Ala Ala Leu His Gly Asp Leu Leu Glu Phe
405 410 415
Phe Lys Leu Asn Pro Leu Thr Tyr Ala Ser Gly Leu Glu Val Cys Gly
420 425 430
Thr Pro Ile Asp Leu Gln Gln Val Asn Ile Asp Ser Phe Thr Val Ala
435 440 445
Gly Ser Asn Asp His Ile Thr Pro Trp Asp Ala Val Tyr Arg Ser Ala
450 455 460
Leu Leu Leu Gly Gly Glu Arg Arg Phe Val Leu Ala Asn Ser Gly His
465 470 475 480
Ile Gln Ser Ile Ile Asn Pro Pro Gly Asn Pro Lys Ala Tyr Tyr Leu
485 490 495
Ala Asn Pro Lys Leu Ser Ser Asp Pro Arg Ala Trp Phe His Asp Ala
500 505 510
Lys Arg Ser Glu Gly Ser Trp Trp Pro Leu Trp Leu Glu Trp Ile Thr
515 520 525
Ala Arg Ser Gly Leu Leu Lys Ala Pro Arg Thr Glu Leu Gly Asn Ala
530 535 540
Thr Tyr Pro Leu Leu Gly Pro Ala Pro Gly Thr Tyr Val Leu Thr Arg
545 550 555 560
<210> 33
<211> 888
<212> DNA
<213> Pseudomonas putida
<220>
<221> CDS
<222> (1)..(888)
<400> 33
atg agg cca gaa atc gct gta ctt gat atc caa ggt cag tat cgg gtt 48
Met Arg Pro Glu Ile Ala Val Leu Asp Ile Gln Gly Gln Tyr Arg Val
1 5 10 15
tac acg gag ttc tat cgc gcg gat gcg gcc gaa aac acg atc atc ctg 96
Tyr Thr Glu Phe Tyr Arg Ala Asp Ala Ala Glu Asn Thr Ile Ile Leu
20 25 30
atc aac ggc tcg ctg gcc acc acg gcc tcg ttc gcc cag acg gta cgt 144
Ile Asn Gly Ser Leu Ala Thr Thr Ala Ser Phe Ala Gln Thr Val Arg
35 40 45
aac ctg cac cca cag ttc aac gtg gtt ctg ttc gac cag ccg tat tca 192
Asn Leu His Pro Gln Phe Asn Val Val Leu Phe Asp Gln Pro Tyr Ser
50 55 60
ggc aag tcc aag ccg cac aac cgt cag gaa cgg ctg atc agc aag gag 240
Gly Lys Ser Lys Pro His Asn Arg Gln Glu Arg Leu Ile Ser Lys Glu
65 70 75 80
acc gag gcg cat atc ctc ctt gag ctg atc gag cac ttc cag gca gac 288
Thr Glu Ala His Ile Leu Leu Glu Leu Ile Glu His Phe Gln Ala Asp
85 90 95
cac gtg atg tct ttt tcg tgg ggt ggc gca agc acg ctg ctg gcg ctg 336
His Val Met Ser Phe Ser Trp Gly Gly Ala Ser Thr Leu Leu Ala Leu
100 105 110
gcg cac cag ccg cgg tac gtg aag aag gca gtg gtg agt tcg ttc tcg 384
Ala His Gln Pro Arg Tyr Val Lys Lys Ala Val Val Ser Ser Phe Ser
115 120 125
cca gtg atc aac gag cca atg cgc gac tat ctg gac cgt ggc tgc cag 432
Pro Val Ile Asn Glu Pro Met Arg Asp Tyr Leu Asp Arg Gly Cys Gln
130 135 140
tac ctg gcc gcc tgc gac cgt tat cag gtc ggc aac ctg gtc aat gac 480
Tyr Leu Ala Ala Cys Asp Arg Tyr Gln Val Gly Asn Leu Val Asn Asp
145 150 155 160
acc atc ggc aag cac ttg ccg tcg ctg ctc aaa cgc ttc aac tac cgc 528
Thr Ile Gly Lys His Leu Pro Ser Leu Leu Lys Arg Phe Asn Tyr Arg
165 170 175
cat gtg agc agc ctg gac agc cac gag tac gca cag atg cac ttc cac 576
His Val Ser Ser Leu Asp Ser His Glu Tyr Ala Gln Met His Phe His
180 185 190
atc aac caa gtg ctg gag cac gac ctg gaa cgt gcg ctg caa ggc gcg 624
Ile Asn Gln Val Leu Glu His Asp Leu Glu Arg Ala Leu Gln Gly Ala
195 200 205
cgc aat atc aac atc ccg gtg ttg ttc atc aac ggc gaa cgc gac gag 672
Arg Asn Ile Asn Ile Pro Val Leu Phe Ile Asn Gly Glu Arg Asp Glu
210 215 220
tac acc acg gtc gaa gat gcg cgg cag ttc agc aag cat gtg ggc aga 720
Tyr Thr Thr Val Glu Asp Ala Arg Gln Phe Ser Lys His Val Gly Arg
225 230 235 240
agc cag ttc agc gtg atc cgc gat gcg ggc cac ttc ctg gac atg gag 768
Ser Gln Phe Ser Val Ile Arg Asp Ala Gly His Phe Leu Asp Met Glu
245 250 255
aac aag acc gcc tgc gag aac acc cgc agt gtc atg ctg ggg ttc ctc 816
Asn Lys Thr Ala Cys Glu Asn Thr Arg Ser Val Met Leu Gly Phe Leu
260 265 270
aag cca acc gtg cgt gaa ccc cgc caa cgt tac caa ccc gtg caa cag 864
Lys Pro Thr Val Arg Glu Pro Arg Gln Arg Tyr Gln Pro Val Gln Gln
275 280 285
ggg cag cat gca ttg gcc atc tga 888
Gly Gln His Ala Leu Ala Ile
290 295
<210> 34
<211> 295
<212> PRT
<213> Pseudomonas putida
<400> 34
Met Arg Pro Glu Ile Ala Val Leu Asp Ile Gln Gly Gln Tyr Arg Val
1 5 10 15
Tyr Thr Glu Phe Tyr Arg Ala Asp Ala Ala Glu Asn Thr Ile Ile Leu
20 25 30
Ile Asn Gly Ser Leu Ala Thr Thr Ala Ser Phe Ala Gln Thr Val Arg
35 40 45
Asn Leu His Pro Gln Phe Asn Val Val Leu Phe Asp Gln Pro Tyr Ser
50 55 60
Gly Lys Ser Lys Pro His Asn Arg Gln Glu Arg Leu Ile Ser Lys Glu
65 70 75 80
Thr Glu Ala His Ile Leu Leu Glu Leu Ile Glu His Phe Gln Ala Asp
85 90 95
His Val Met Ser Phe Ser Trp Gly Gly Ala Ser Thr Leu Leu Ala Leu
100 105 110
Ala His Gln Pro Arg Tyr Val Lys Lys Ala Val Val Ser Ser Phe Ser
115 120 125
Pro Val Ile Asn Glu Pro Met Arg Asp Tyr Leu Asp Arg Gly Cys Gln
130 135 140
Tyr Leu Ala Ala Cys Asp Arg Tyr Gln Val Gly Asn Leu Val Asn Asp
145 150 155 160
Thr Ile Gly Lys His Leu Pro Ser Leu Leu Lys Arg Phe Asn Tyr Arg
165 170 175
His Val Ser Ser Leu Asp Ser His Glu Tyr Ala Gln Met His Phe His
180 185 190
Ile Asn Gln Val Leu Glu His Asp Leu Glu Arg Ala Leu Gln Gly Ala
195 200 205
Arg Asn Ile Asn Ile Pro Val Leu Phe Ile Asn Gly Glu Arg Asp Glu
210 215 220
Tyr Thr Thr Val Glu Asp Ala Arg Gln Phe Ser Lys His Val Gly Arg
225 230 235 240
Ser Gln Phe Ser Val Ile Arg Asp Ala Gly His Phe Leu Asp Met Glu
245 250 255
Asn Lys Thr Ala Cys Glu Asn Thr Arg Ser Val Met Leu Gly Phe Leu
260 265 270
Lys Pro Thr Val Arg Glu Pro Arg Gln Arg Tyr Gln Pro Val Gln Gln
275 280 285
Gly Gln His Ala Leu Ala Ile
290 295
<210> 35
<211> 903
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(903)
<400> 35
atg agg ccg gaa aca gcc atc atc gag atc cac ggg caa tac agg att 48
Met Arg Pro Glu Thr Ala Ile Ile Glu Ile His Gly Gln Tyr Arg Ile
1 5 10 15
cac acc gag ttc tac ggc aac ccc gcg gcg cag caa acc atc atc ctg 96
His Thr Glu Phe Tyr Gly Asn Pro Ala Ala Gln Gln Thr Ile Ile Leu
20 25 30
gtc aac ggc tcg ctg tcg acc aca gcg tcc ttc gcc cag acc gtg aag 144
Val Asn Gly Ser Leu Ser Thr Thr Ala Ser Phe Ala Gln Thr Val Lys
35 40 45
tac ctg cag ccg cat tac aac gtg gtg ctc tac gac cag ccg tat gcc 192
Tyr Leu Gln Pro His Tyr Asn Val Val Leu Tyr Asp Gln Pro Tyr Ala
50 55 60
ggc cag tcc aaa ccc cat aac gaa aac cac acg ccg atc agc aag gaa 240
Gly Gln Ser Lys Pro His Asn Glu Asn His Thr Pro Ile Ser Lys Glu
65 70 75 80
tgc gag gcc agg atc ctg ctg gaa ctg atc gaa cgc ttc cgt gcc gag 288
Cys Glu Ala Arg Ile Leu Leu Glu Leu Ile Glu Arg Phe Arg Ala Glu
85 90 95
gta gtg atg tcg ttc tcg tgg ggc ggc gtc gcc acc ctg ctg gcc ctg 336
Val Val Met Ser Phe Ser Trp Gly Gly Val Ala Thr Leu Leu Ala Leu
100 105 110
gcg caa cgt ccc gga cgg atc cgc agg gcg gtg gtc aac tca ttc tcg 384
Ala Gln Arg Pro Gly Arg Ile Arg Arg Ala Val Val Asn Ser Phe Ser
115 120 125
cct cag ctc aac ccg gcc atg ctc gac tac ctg cat cgc ggc ctc gac 432
Pro Gln Leu Asn Pro Ala Met Leu Asp Tyr Leu His Arg Gly Leu Asp
130 135 140
tac ctc gcc gcc tgc gat cgc acc cag atc ggc aac ctg gtc aac gaa 480
Tyr Leu Ala Ala Cys Asp Arg Thr Gln Ile Gly Asn Leu Val Asn Glu
145 150 155 160
acc atc ggc cgc tac ctg cca cag ttg ttc aag cgc tac aac ttc cgc 528
Thr Ile Gly Arg Tyr Leu Pro Gln Leu Phe Lys Arg Tyr Asn Phe Arg
165 170 175
cac gtc agc agc ctg gac gag cac gaa tac cac cag atg cac ttc cat 576
His Val Ser Ser Leu Asp Glu His Glu Tyr His Gln Met His Phe His
180 185 190
atc cgc gaa gtg ctg cgc ctg aac gcc gat agc tat acc gag agc ttc 624
Ile Arg Glu Val Leu Arg Leu Asn Ala Asp Ser Tyr Thr Glu Ser Phe
195 200 205
gcc ggc atc gag atc ccg atg ctg ttc atg aac ggc gag ctg gac atc 672
Ala Gly Ile Glu Ile Pro Met Leu Phe Met Asn Gly Glu Leu Asp Ile
210 215 220
tac acc acg ccc cac gaa gcc cgc cag ttc ggc caa ctg atc cgc ggc 720
Tyr Thr Thr Pro His Glu Ala Arg Gln Phe Gly Gln Leu Ile Arg Gly
225 230 235 240
gcg gaa ttc cac acc atc cgc aat gcc ggc cac ttc atc gac gtc gag 768
Ala Glu Phe His Thr Ile Arg Asn Ala Gly His Phe Ile Asp Val Glu
245 250 255
cac aag gcc gcc tgg cag cag acc cag gac gcc ctg ctg gcc ttc ctc 816
His Lys Ala Ala Trp Gln Gln Thr Gln Asp Ala Leu Leu Ala Phe Leu
260 265 270
cgc ccg cag cgc acg cag ccg ctc aac ccg atc tac cgc ccc cag ccc 864
Arg Pro Gln Arg Thr Gln Pro Leu Asn Pro Ile Tyr Arg Pro Gln Pro
275 280 285
aac ggc gcc agc gtc ccc ctc gcc gcc ctc gcc agc taa 903
Asn Gly Ala Ser Val Pro Leu Ala Ala Leu Ala Ser
290 295 300
<210> 36
<211> 300
<212> PRT
<213> Pseudomonas aeruginosa
<400> 36
Met Arg Pro Glu Thr Ala Ile Ile Glu Ile His Gly Gln Tyr Arg Ile
1 5 10 15
His Thr Glu Phe Tyr Gly Asn Pro Ala Ala Gln Gln Thr Ile Ile Leu
20 25 30
Val Asn Gly Ser Leu Ser Thr Thr Ala Ser Phe Ala Gln Thr Val Lys
35 40 45
Tyr Leu Gln Pro His Tyr Asn Val Val Leu Tyr Asp Gln Pro Tyr Ala
50 55 60
Gly Gln Ser Lys Pro His Asn Glu Asn His Thr Pro Ile Ser Lys Glu
65 70 75 80
Cys Glu Ala Arg Ile Leu Leu Glu Leu Ile Glu Arg Phe Arg Ala Glu
85 90 95
Val Val Met Ser Phe Ser Trp Gly Gly Val Ala Thr Leu Leu Ala Leu
100 105 110
Ala Gln Arg Pro Gly Arg Ile Arg Arg Ala Val Val Asn Ser Phe Ser
115 120 125
Pro Gln Leu Asn Pro Ala Met Leu Asp Tyr Leu His Arg Gly Leu Asp
130 135 140
Tyr Leu Ala Ala Cys Asp Arg Thr Gln Ile Gly Asn Leu Val Asn Glu
145 150 155 160
Thr Ile Gly Arg Tyr Leu Pro Gln Leu Phe Lys Arg Tyr Asn Phe Arg
165 170 175
His Val Ser Ser Leu Asp Glu His Glu Tyr His Gln Met His Phe His
180 185 190
Ile Arg Glu Val Leu Arg Leu Asn Ala Asp Ser Tyr Thr Glu Ser Phe
195 200 205
Ala Gly Ile Glu Ile Pro Met Leu Phe Met Asn Gly Glu Leu Asp Ile
210 215 220
Tyr Thr Thr Pro His Glu Ala Arg Gln Phe Gly Gln Leu Ile Arg Gly
225 230 235 240
Ala Glu Phe His Thr Ile Arg Asn Ala Gly His Phe Ile Asp Val Glu
245 250 255
His Lys Ala Ala Trp Gln Gln Thr Gln Asp Ala Leu Leu Ala Phe Leu
260 265 270
Arg Pro Gln Arg Thr Gln Pro Leu Asn Pro Ile Tyr Arg Pro Gln Pro
275 280 285
Asn Gly Ala Ser Val Pro Leu Ala Ala Leu Ala Ser
290 295 300
<210> 37
<211> 2310
<212> DNA
<213> Pseudomonas aeruginosa
<400> 37
aggtaccaga tctggcattt ttgggaggtg tgaaatgcgg cgcgaaagtc tgttggtatc 60
ggtttgcaag ggcctgcggg tacatgtcga gcgcgttggg caggatcccg ggcgcagcac 120
ggtgatgctg gtcaacggcg cgatggcgac caccgcctcg ttcgcccgga cctgcaagtg 180
cctggccgaa catttcaacg tggtgctgtt cgacctgccc ttcgccgggc agtcgcgtca 240
gcacaacccg cagcgggggt tgatcaccaa ggacgacgag gtggaaatcc tcctggcgct 300
gatcgagcgc ttcgaggtca atcacctggt ctccgcgtcc tggggcggta tctccacgct 360
gctggcgctg tcgcgcaatc cgcgcggcat ccgcagctcg gtggtgatgg cattcgcccc 420
tggactgaac caggcgatgc tcgactacgt cgggcgggcg caggcgctga tcgagctgga 480
cgacaagtcg gcgatcggcc atctgctcaa cgagaccgtc ggcaaatacc tgccgccgcg 540
cctgaaagcc agcaaccatc agcacatggc ttcgctggcc accggcgaat acgagcaggc 600
gcgctttcac atcgaccagg tgctggcgct caacgatcgg ggctacctgg cttgcctgga 660
gcggatccag agccacgtgc atttcatcaa cggcagctgg gacgaataca ccaccgccga 720
ggacgcccgc cagttccgcg actacctgcc gcactgcagt ttctcgcggg tggagggcac 780
cgggcatttc ctcgacctgg agtccaagct ggccgcggta cgcgtgcacc gcgccctgct 840
cgagcacctg ctgaagcaac cggagccgca gcgggcggaa cgcgcggcgg gattccacga 900
gatggccatc ggctacgcct gaacccttga cctgcgaaga cccggcctgg ccgggctttg 960
cggttgcata acgcacggag tagcaccatg cacgccatcc tcatcgccat cggctcggcc 1020
ggcgacgtat ttcccttcat cggcctggcc cggaccctga aattgcgcgg gcaccgcgtg 1080
agcctctgca ccatcccggt gtttcgcgac gcggtggagc agcacggcat cgcgttcgtc 1140
ccgctgagcg acgaactgac ctaccgccgg accatgggcg atccgcgcct gtgggacccc 1200
aagacgtcct tcggcgtgct ctggcaaacc atcgccggga tgatcgagcc ggtctacgag 1260
tacgtctcgg cgcagcgcca tgacgacatc gtggtggtcg gctcgctctg ggcgctgggc 1320
gcacgcatcg ctcacgagaa gtacgggatt ccctacctgt ccgcgcaggt ctcgccatcg 1380
accttgttgt cggcgcacct gccgccggta caccccaagt tcaacgtgcc cgagcagatg 1440
ccgctggcga tgcgcaagct gctctggcgc tgcatcgagc gcttcaagct ggatcgcacc 1500
tgcgcgccgg atatcaacgc ggtgcggcgc aaggtcggcc tggagacgcc ggtgaagcgc 1560
atcttcaccc aatggatgca ttcgccgcag ggcgtggtct gcctgttccc ggcctggttc 1620
gcgccgcccc agcaggattg gccgcaaccc ctgcacatga ccggcttccc gctgttcgac 1680
ggcagtatcc cggggacccc gctcgacgac gaactgcaac gctttctcga tcagggcagc 1740
cggccgctgg tgttcaccca gggctcgacc gaacacctgc agggcgactt ctacgccatg 1800
gccctgcgcg cgctggaacg cctcggcgcg cgtgggatct tcctcaccgg cgccggccag 1860
gaaccgctgc gcggcttgcc gaaccacgtg ctgcagcgcg cctacgcgcc actgggagcc 1920
ttgctgccat cgtgcgccgg gctggtccat ccgggcggta tcggcgccat gagcctggcc 1980
ttggcggcgg gggtgccgca ggtgctgctg ccctgcgccc acgaccagtt cgacaatgcc 2040
gaacggctgg tccggctcgg ctgcgggatg cgcctgggcg tgccattgcg cgagcaggag 2100
ttgcgcgggg cgctgtggcg cttgctcgag gacccggcca tggcggcggc ctgtcggcgt 2160
ttcatggaat tgtcacaacc gcacagtatc gcttgcggta aagcggccca ggtggtcgaa 2220
cgttgtcata gggaggggga tgcgcgatgg ctgaaggctg cgtcctgaac ggtgctggca 2280
taacagatag ggttgcctct agagagctca 2310
<210> 38
<211> 7422
<212> DNA
<213> Artificial
<220>
<223> Vector
<400> 38
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760
ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820
tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880
gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940
agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000
aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060
atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120
attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180
gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3240
tagaggcaac cctatctgtt atgccagcac cgttcaggac gcagccttca gccatcgcgc 3300
atccccctcc ctatgacaac gttcgaccac ctgggccgct ttaccgcaag cgatactgtg 3360
cggttgtgac aattccatga aacgccgaca ggccgccgcc atggccgggt cctcgagcaa 3420
gcgccacagc gccccgcgca actcctgctc gcgcaatggc acgcccaggc gcatcccgca 3480
gccgagccgg accagccgtt cggcattgtc gaactggtcg tgggcgcagg gcagcagcac 3540
ctgcggcacc cccgccgcca aggccaggct catggcgccg ataccgcccg gatggaccag 3600
cccggcgcac gatggcagca aggctcccag tggcgcgtag gcgcgctgca gcacgtggtt 3660
cggcaagccg cgcagcggtt cctggccggc gccggtgagg aagatcccac gcgcgccgag 3720
gcgttccagc gcgcgcaggg ccatggcgta gaagtcgccc tgcaggtgtt cggtcgagcc 3780
ctgggtgaac accagcggcc ggctgccctg atcgagaaag cgttgcagtt cgtcgtcgag 3840
cggggtcccc gggatactgc cgtcgaacag cgggaagccg gtcatgtgca ggggttgcgg 3900
ccaatcctgc tggggcggcg cgaaccaggc cgggaacagg cagaccacgc cctgcggcga 3960
atgcatccat tgggtgaaga tgcgcttcac cggcgtctcc aggccgacct tgcgccgcac 4020
cgcgttgata tccggcgcgc aggtgcgatc cagcttgaag cgctcgatgc agcgccagag 4080
cagcttgcgc atcgccagcg gcatctgctc gggcacgttg aacttggggt gtaccggcgg 4140
caggtgcgcc gacaacaagg tcgatggcga gacctgcgcg gacaggtagg gaatcccgta 4200
cttctcgtga gcgatgcgtg cgcccagcgc ccagagcgag ccgaccacca cgatgtcgtc 4260
atggcgctgc gccgagacgt actcgtagac cggctcgatc atcccggcga tggtttgcca 4320
gagcacgccg aaggacgtct tggggtccca caggcgcgga tcgcccatgg tccggcggta 4380
ggtcagttcg tcgctcagcg ggacgaacgc gatgccgtgc tgctccaccg cgtcgcgaaa 4440
caccgggatg gtgcagaggc tcacgcggtg cccgcgcaat ttcagggtcc gggccaggcc 4500
gatgaaggga aatacgtcgc cggccgagcc gatggcgatg aggatggcgt gcatggtgct 4560
actccgtgcg ttatgcaacc gcaaagcccg gccaggccgg gtcttcgcag gtcaagggtt 4620
caggcgtagc cgatggccat ctcgtggaat cccgccgcgc gttccgcccg ctgcggctcc 4680
ggttgcttca gcaggtgctc gagcagggcg cggtgcacgc gtaccgcggc cagcttggac 4740
tccaggtcga ggaaatgccc ggtgccctcc acccgcgaga aactgcagtg cggcaggtag 4800
tcgcggaact ggcgggcgtc ctcggcggtg gtgtattcgt cccagctgcc gttgatgaaa 4860
tgcacgtggc tctggatccg ctccaggcaa gccaggtagc cccgatcgtt gagcgccagc 4920
acctggtcga tgtgaaagcg cgcctgctcg tattcgccgg tggccagcga agccatgtgc 4980
tgatggttgc tggctttcag gcgcggcggc aggtatttgc cgacggtctc gttgagcaga 5040
tggccgatcg ccgacttgtc gtccagctcg atcagcgcct gcgcccgccc gacgtagtcg 5100
agcatcgcct ggttcagtcc aggggcgaat gccatcacca ccgagctgcg gatgccgcgc 5160
ggattgcgcg acagcgccag cagcgtggag ataccgcccc aggacgcgga gaccaggtga 5220
ttgacctcga agcgctcgat cagcgccagg aggatttcca cctcgtcgtc cttggtgatc 5280
aacccccgct gcgggttgtg ctgacgcgac tgcccggcga agggcaggtc gaacagcacc 5340
acgttgaaat gttcggccag gcacttgcag gtccgggcga acgaggcggt ggtcgccatc 5400
gcgccgttga ccagcatcac cgtgctgcgc ccgggatcct gcccaacgcg ctcgacatgt 5460
acccgcaggc ccttgcaaac cgataccaac agactttcgc gccgcatttc acacctccca 5520
aaaatgccag atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc 5580
tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg 5640
gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5700
aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5760
acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 5820
cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg catgcataaa 5880
aactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga 5940
acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatgggg 6000
gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg 6060
gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga 6120
tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac 6180
tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc 6240
acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac 6300
gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag 6360
cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc 6420
tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga 6480
tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc 6540
tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc 6600
cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg 6660
agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg 6720
tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg 6780
tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc 6840
tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca 6900
tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca 6960
atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag 7020
atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag 7080
ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat 7140
cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc 7200
cagatagccc agtagctgac attcatccca ggtggcactt ttcggggaaa tgtgcgcgcc 7260
cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt tccgtcagca 7320
gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc cgattcaacg 7380
gccccagggc gtccagaacg ggcttcaggc gctcccgaag gt 7422
<210> 39
<211> 3294
<212> DNA
<213> Pseudomonas aeruginosa
<400> 39
aggtaccaga tctggcattt ttgggaggtg tgaaatgcgg cgcgaaagtc tgttggtatc 60
ggtttgcaag ggcctgcggg tacatgtcga gcgcgttggg caggatcccg ggcgcagcac 120
ggtgatgctg gtcaacggcg cgatggcgac caccgcctcg ttcgcccgga cctgcaagtg 180
cctggccgaa catttcaacg tggtgctgtt cgacctgccc ttcgccgggc agtcgcgtca 240
gcacaacccg cagcgggggt tgatcaccaa ggacgacgag gtggaaatcc tcctggcgct 300
gatcgagcgc ttcgaggtca atcacctggt ctccgcgtcc tggggcggta tctccacgct 360
gctggcgctg tcgcgcaatc cgcgcggcat ccgcagctcg gtggtgatgg cattcgcccc 420
tggactgaac caggcgatgc tcgactacgt cgggcgggcg caggcgctga tcgagctgga 480
cgacaagtcg gcgatcggcc atctgctcaa cgagaccgtc ggcaaatacc tgccgccgcg 540
cctgaaagcc agcaaccatc agcacatggc ttcgctggcc accggcgaat acgagcaggc 600
gcgctttcac atcgaccagg tgctggcgct caacgatcgg ggctacctgg cttgcctgga 660
gcggatccag agccacgtgc atttcatcaa cggcagctgg gacgaataca ccaccgccga 720
ggacgcccgc cagttccgcg actacctgcc gcactgcagt ttctcgcggg tggagggcac 780
cgggcatttc ctcgacctgg agtccaagct ggccgcggta cgcgtgcacc gcgccctgct 840
cgagcacctg ctgaagcaac cggagccgca gcgggcggaa cgcgcggcgg gattccacga 900
gatggccatc ggctacgcct gaacccttga cctgcgaaga cccggcctgg ccgggctttg 960
cggttgcata acgcacggag tagcaccatg cacgccatcc tcatcgccat cggctcggcc 1020
ggcgacgtat ttcccttcat cggcctggcc cggaccctga aattgcgcgg gcaccgcgtg 1080
agcctctgca ccatcccggt gtttcgcgac gcggtggagc agcacggcat cgcgttcgtc 1140
ccgctgagcg acgaactgac ctaccgccgg accatgggcg atccgcgcct gtgggacccc 1200
aagacgtcct tcggcgtgct ctggcaaacc atcgccggga tgatcgagcc ggtctacgag 1260
tacgtctcgg cgcagcgcca tgacgacatc gtggtggtcg gctcgctctg ggcgctgggc 1320
gcacgcatcg ctcacgagaa gtacgggatt ccctacctgt ccgcgcaggt ctcgccatcg 1380
accttgttgt cggcgcacct gccgccggta caccccaagt tcaacgtgcc cgagcagatg 1440
ccgctggcga tgcgcaagct gctctggcgc tgcatcgagc gcttcaagct ggatcgcacc 1500
tgcgcgccgg atatcaacgc ggtgcggcgc aaggtcggcc tggagacgcc ggtgaagcgc 1560
atcttcaccc aatggatgca ttcgccgcag ggcgtggtct gcctgttccc ggcctggttc 1620
gcgccgcccc agcaggattg gccgcaaccc ctgcacatga ccggcttccc gctgttcgac 1680
ggcagtatcc cggggacccc gctcgacgac gaactgcaac gctttctcga tcagggcagc 1740
cggccgctgg tgttcaccca gggctcgacc gaacacctgc agggcgactt ctacgccatg 1800
gccctgcgcg cgctggaacg cctcggcgcg cgtgggatct tcctcaccgg cgccggccag 1860
gaaccgctgc gcggcttgcc gaaccacgtg ctgcagcgcg cctacgcgcc actgggagcc 1920
ttgctgccat cgtgcgccgg gctggtccat ccgggcggta tcggcgccat gagcctggcc 1980
ttggcggcgg gggtgccgca ggtgctgctg ccctgcgccc acgaccagtt cgacaatgcc 2040
gaacggctgg tccggctcgg ctgcgggatg cgcctgggcg tgccattgcg cgagcaggag 2100
ttgcgcgggg cgctgtggcg cttgctcgag gacccggcca tggcggcggc ctgtcggcgt 2160
ttcatggaat tgtcacaacc gcacagtatc gcttgcggta aagcggccca ggtggtcgaa 2220
cgttgtcata gggaggggga tgcgcgatgg ctgaaggctg cgtcctgacc tacgggagaa 2280
gaacgatcat ggaccggata gacatgggcg tgctggtggt actgttcaat cctggcgacg 2340
acgacctgga acaccttggc gaactggcgg cggcgtttcc gcaactgcgc ttccttgccg 2400
tcgacaactc accgcacagc gatccgcagc gcaatgcccg gctgcgcggg caaggcatcg 2460
ccgtgctgca ccacggcaac cggcagggca tcgccggcgc cttcaaccag ggactcgacg 2520
cgctattccg gcgtggcgtg cagggtgtgc tgctgctcga ccaggactcc cgtcccggcg 2580
gcgccttcct cgccgcccag tggcgcaacc tgcaggcgcg caacggtcag gcctgcctgc 2640
tcggcccacg gatcttcgac cggggtgacc ggcgcttcct gccggccatc catctcgacg 2700
gactgacgct caggcaattg tctctggacg gcctgacgac cccgcagcgc acctcgttcc 2760
tgatctcctc cggctgcctg ctgacccgcg aggcctacca gcgcctcggc cacttcgacg 2820
aggaactgtt catcgaccac gtggacaccg aatacagcct gcgcgcccag gcgctggacg 2880
tgcccctgta cgtcgacccg cggctggtcc tcgagcaccg catcggcacg cgcaagaccc 2940
gccgcctcgg cggtctcagc ctcagcgcga tgaaccacgc cccgctgcgc cgctactacc 3000
tggcgcgcaa cggcctgctg gtcctgcgcc gctacgcccg gtcctcgccg ctggccctgc 3060
tggcgaacct gccgaccctg acccagggcc tcgcggtgct cctgctcgaa cgcgacaagc 3120
tgctcaagct gcgctgcctg ggctggggcc tgtgggacgg cctgcgggga cgcggcggcg 3180
cgctggagac caaccgcccg cgcctgctga agcgcctcgc cggcccggcc gtggcgtccg 3240
tagcttccgg caaggccaag gcctagtcgg cgaaacgcat tccctctaga gagc 3294
<210> 40
<211> 8409
<212> DNA
<213> Artificial
<220>
<223> Vector
<400> 40
accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc 60
aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg 120
aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg 180
aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag 240
cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt 300
tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa 360
gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc 420
aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 480
cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 540
aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 600
tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 660
gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 720
aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 780
tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 840
ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 900
ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 960
cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 1020
tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 1080
ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 1140
tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 1200
tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 1260
ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 1320
accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 1380
atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta 1440
tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga 1500
tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat 1560
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 1620
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 1680
ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 1740
ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 1800
tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa 1860
gcttgatatc gaattcctgc agcccggggg atctggcatt tttgggaggt gtgaaatgcg 1920
gcgcgaaagt ctgttggtat cggtttgcaa gggcctgcgg gtacatgtcg agcgcgttgg 1980
gcaggatccc gggcgcagca cggtgatgct ggtcaacggc gcgatggcga ccaccgcctc 2040
gttcgcccgg acctgcaagt gcctggccga acatttcaac gtggtgctgt tcgacctgcc 2100
cttcgccggg cagtcgcgtc agcacaaccc gcagcggggg ttgatcacca aggacgacga 2160
ggtggaaatc ctcctggcgc tgatcgagcg cttcgaggtc aatcacctgg tctccgcgtc 2220
ctggggcggt atctccacgc tgctggcgct gtcgcgcaat ccgcgcggca tccgcagctc 2280
ggtggtgatg gcattcgccc ctggactgaa ccaggcgatg ctcgactacg tcgggcgggc 2340
gcaggcgctg atcgagctgg acgacaagtc ggcgatcggc catctgctca acgagaccgt 2400
cggcaaatac ctgccgccgc gcctgaaagc cagcaaccat cagcacatgg cttcgctggc 2460
caccggcgaa tacgagcagg cgcgctttca catcgaccag gtgctggcgc tcaacgatcg 2520
gggctacctg gcttgcctgg agcggatcca gagccacgtg catttcatca acggcagctg 2580
ggacgaatac accaccgccg aggacgcccg ccagttccgc gactacctgc cgcactgcag 2640
tttctcgcgg gtggagggca ccgggcattt cctcgacctg gagtccaagc tggccgcggt 2700
acgcgtgcac cgcgccctgc tcgagcacct gctgaagcaa ccggagccgc agcgggcgga 2760
acgcgcggcg ggattccacg agatggccat cggctacgcc tgaacccttg acctgcgaag 2820
acccggcctg gccgggcttt gcggttgcat aacgcacgga gtagcaccat gcacgccatc 2880
ctcatcgcca tcggctcggc cggcgacgta tttcccttca tcggcctggc ccggaccctg 2940
aaattgcgcg ggcaccgcgt gagcctctgc accatcccgg tgtttcgcga cgcggtggag 3000
cagcacggca tcgcgttcgt cccgctgagc gacgaactga cctaccgccg gaccatgggc 3060
gatccgcgcc tgtgggaccc caagacgtcc ttcggcgtgc tctggcaaac catcgccggg 3120
atgatcgagc cggtctacga gtacgtctcg gcgcagcgcc atgacgacat cgtggtggtc 3180
ggctcgctct gggcgctggg cgcacgcatc gctcacgaga agtacgggat tccctacctg 3240
tccgcgcagg tctcgccatc gaccttgttg tcggcgcacc tgccgccggt acaccccaag 3300
ttcaacgtgc ccgagcagat gccgctggcg atgcgcaagc tgctctggcg ctgcatcgag 3360
cgcttcaagc tggatcgcac ctgcgcgccg gatatcaacg cggtgcggcg caaggtcggc 3420
ctggagacgc cggtgaagcg catcttcacc caatggatgc attcgccgca gggcgtggtc 3480
tgcctgttcc cggcctggtt cgcgccgccc cagcaggatt ggccgcaacc cctgcacatg 3540
accggcttcc cgctgttcga cggcagtatc ccggggaccc cgctcgacga cgaactgcaa 3600
cgctttctcg atcagggcag ccggccgctg gtgttcaccc agggctcgac cgaacacctg 3660
cagggcgact tctacgccat ggccctgcgc gcgctggaac gcctcggcgc gcgtgggatc 3720
ttcctcaccg gcgccggcca ggaaccgctg cgcggcttgc cgaaccacgt gctgcagcgc 3780
gcctacgcgc cactgggagc cttgctgcca tcgtgcgccg ggctggtcca tccgggcggt 3840
atcggcgcca tgagcctggc cttggcggcg ggggtgccgc aggtgctgct gccctgcgcc 3900
cacgaccagt tcgacaatgc cgaacggctg gtccggctcg gctgcgggat gcgcctgggc 3960
gtgccattgc gcgagcagga gttgcgcggg gcgctgtggc gcttgctcga ggacccggcc 4020
atggcggcgg cctgtcggcg tttcatggaa ttgtcacaac cgcacagtat cgcttgcggt 4080
aaagcggccc aggtggtcga acgttgtcat agggaggggg atgcgcgatg gctgaaggct 4140
gcgtcctgac ctacgggaga agaacgatca tggaccggat agacatgggc gtgctggtgg 4200
tactgttcaa tcctggcgac gacgacctgg aacaccttgg cgaactggcg gcggcgtttc 4260
cgcaactgcg cttccttgcc gtcgacaact caccgcacag cgatccgcag cgcaatgccc 4320
ggctgcgcgg gcaaggcatc gccgtgctgc accacggcaa ccggcagggc atcgccggcg 4380
ccttcaacca gggactcgac gcgctattcc ggcgtggcgt gcagggtgtg ctgctgctcg 4440
accaggactc ccgtcccggc ggcgccttcc tcgccgccca gtggcgcaac ctgcaggcgc 4500
gcaacggtca ggcctgcctg ctcggcccac ggatcttcga ccggggtgac cggcgcttcc 4560
tgccggccat ccatctcgac ggactgacgc tcaggcaatt gtctctggac ggcctgacga 4620
ccccgcagcg cacctcgttc ctgatctcct ccggctgcct gctgacccgc gaggcctacc 4680
agcgcctcgg ccacttcgac gaggaactgt tcatcgacca cgtggacacc gaatacagcc 4740
tgcgcgccca ggcgctggac gtgcccctgt acgtcgaccc gcggctggtc ctcgagcacc 4800
gcatcggcac gcgcaagacc cgccgcctcg gcggtctcag cctcagcgcg atgaaccacg 4860
ccccgctgcg ccgctactac ctggcgcgca acggcctgct ggtcctgcgc cgctacgccc 4920
ggtcctcgcc gctggccctg ctggcgaacc tgccgaccct gacccagggc ctcgcggtgc 4980
tcctgctcga acgcgacaag ctgctcaagc tgcgctgcct gggctggggc ctgtgggacg 5040
gcctgcgggg acgcggcggc gcgctggaga ccaaccgccc gcgcctgctg aagcgcctcg 5100
ccggcccggc cgtggcgtcc gtagcttccg gcaaggccaa ggcctagtcg gcgaaacgca 5160
ttccctctag agcggccgcc accgcggtgg agctccaatt cgccctatag tgagtcgtat 5220
tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 5280
caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 5340
cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa attgtaagcg 5400
ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 5460
aggccgactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc 5520
cttaaacgcc tggtgctacg cctgaataag tgataataag cggatgaatg gcagaaattc 5580
gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc 5640
tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat 5700
gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcatt 5760
tattctgcct cccagagcct gataaaaacg gtgaatccgt tagcgaggtg ccgccggctt 5820
ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa 5880
ggtatagggc ggcgaggcgg ctacagccga tagtctggaa cagcgcactt acgggttgct 5940
gcgcaaccca agtgctaccg gcgcggcagc gtgacccgtg tcggcggctc caacggctcg 6000
ccatcgtcca gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc 6060
cattcctccg tttcggtcaa ggctggcagg tctggttcca tgcccggaat gccgggctgg 6120
ctgggcggct cctcgccggg gccggtcggt agttgctgct cgcccggata cagggtcggg 6180
atgcggcgca ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc 6240
acggcggcac tgaacaccga caggcgcaac tggtcgcggg gctggcccca cgccacgcgg 6300
tcattgacca cgtaggccga cacggtgccg gggccgttga gcttcacgac ggagatccag 6360
cgctcggcca ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc 6420
ttggaaagtg tcttctggct gaccaccacg gcgttctggt ggcccatctg cgccacgagg 6480
tgatgcagca gcattgccgc cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc 6540
gctttgcgtt ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat gccgatttct 6600
ctggactgcg tggccatgct tatctccatg cggtagggtg ccgcacggtt gcggcaccat 6660
gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt atgcgttgcg taaaagtggc 6720
agtcaattac agattttctt taacctacgc aatgagctat tgcggggggt gccgcaatga 6780
gctgttgcgt accccccttt tttaagttgt tgatttttaa gtctttcgca tttcgcccta 6840
tatctagttc tttggtgccc aaagaagggc acccctgcgg ggttccccca cgccttcggc 6900
gcggctcccc ctccggcaaa aagtggcccc tccggggctt gttgatcgac tgcgcggcct 6960
tcggccttgc ccaaggtggc gctgccccct tggaaccccc gcactcgccg ccgtgaggct 7020
cggggggcag gcgggcgggc ttcgccttcg actgccccca ctcgcatagg cttgggtcgt 7080
tccaggcgcg tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga cccgccttcc 7140
acttggtgtc caaccggcaa gcgaagcgcg caggccgcag gccggaggct tttccccaga 7200
gaaaattaaa aaaattgatg gggcaaggcc gcaggccgcg cagttggagc cggtgggtat 7260
gtggtcgaag gctgggtagc cggtgggcaa tccctgtggt caagctcgtg ggcaggcgca 7320
gcctgtccat cagcttgtcc agcagggttg tccacgggcc gagcgaagcg agccagccgg 7380
tggccgctcg cggccatcgt ccacatatcc acgggctggc aagggagcgc agcgaccgcg 7440
cagggcgaag cccggagagc aagcccgtag ggcgccgcag ccgccgtagg cggtcacgac 7500
tttgcgaagc aaagtctagt gagtatactc aagcattgag tggcccgccg gaggcaccgc 7560
cttgcgctgc ccccgtcgag ccggttggac accaaaaggg aggggcaggc atggcggcat 7620
acgcgatcat gcgatgcaag aagctggcga aaatgggcaa cgtggcggcc agtctcaagc 7680
acgcctaccg cgagcgcgag acgcccaacg ctgacgccag caggacgcca gagaacgagc 7740
actgggcggc cagcagcacc gatgaagcga tgggccgact gcgcgagttg ctgccagaga 7800
agcggcgcaa ggacgctgtg ttggcggtcg agtacgtcat gacggccagc ccggaatggt 7860
ggaagtcggc cagccaagaa cagcaggcgg cgttcttcga gaaggcgcac aagtggctgg 7920
cggacaagta cggggcggat cgcatcgtga cggccagcat ccaccgtgac gaaaccagcc 7980
cgcacatgac cgcgttcgtg gtgccgctga cgcaggacgg caggctgtcg gccaaggagt 8040
tcatcggcaa caaagcgcag atgacccgcg accagaccac gtttgcggcc gctgtggccg 8100
atctagggct gcaacggggc atcgagggca gcaaggcacg tcacacgcgc attcaggcgt 8160
tctacgaggc cctggagcgg ccaccagtgg gccacgtcac catcagcccg caagcggtcg 8220
agccacgcgc ctatgcaccg cagggattgg ccgaaaagct gggaatctca aagcgcgttg 8280
agacgccgga agccgtggcc gaccggctga caaaagcggt tcggcagggg tatgagcctg 8340
ccctacaggc cgccgcagga gcgcgtgaga tgcgcaagaa ggccgatcaa gcccaagaga 8400
cggcccgag 8409
<210> 41
<211> 3590
<212> DNA
<213> Pseudomonas aeruginosa
<400> 41
aggtaccaga tctggcattt ttgggaggtg tgaaatgcgg cgcgaaagtc tgttggtatc 60
ggtttgcaag ggcctgcggg tacatgtcga gcgcgttggg caggatcccg ggcgcagcac 120
ggtgatgctg gtcaacggcg cgatggcgac caccgcctcg ttcgcccgga cctgcaagtg 180
cctggccgaa catttcaacg tggtgctgtt cgacctgccc ttcgccgggc agtcgcgtca 240
gcacaacccg cagcgggggt tgatcaccaa ggacgacgag gtggaaatcc tcctggcgct 300
gatcgagcgc ttcgaggtca atcacctggt ctccgcgtcc tggggcggta tctccacgct 360
gctggcgctg tcgcgcaatc cgcgcggcat ccgcagctcg gtggtgatgg cattcgcccc 420
tggactgaac caggcgatgc tcgactacgt cgggcgggcg caggcgctga tcgagctgga 480
cgacaagtcg gcgatcggcc atctgctcaa cgagaccgtc ggcaaatacc tgccgccgcg 540
cctgaaagcc agcaaccatc agcacatggc ttcgctggcc accggcgaat acgagcaggc 600
gcgctttcac atcgaccagg tgctggcgct caacgatcgg ggctacctgg cttgcctgga 660
gcggatccag agccacgtgc atttcatcaa cggcagctgg gacgaataca ccaccgccga 720
ggacgcccgc cagttccgcg actacctgcc gcactgcagt ttctcgcggg tggagggcac 780
cgggcatttc ctcgacctgg agtccaagct ggccgcggta cgcgtgcacc gcgccctgct 840
cgagcacctg ctgaagcaac cggagccgca gcgggcggaa cgcgcggcgg gattccacga 900
gatggccatc ggctacgcct gaacccttga cctgcgaaga cccggcctgg ccgggctttg 960
cggttgcata acgcacggag tagcaccatg cacgccatcc tcatcgccat cggctcggcc 1020
ggcgacgtat ttcccttcat cggcctggcc cggaccctga aattgcgcgg gcaccgcgtg 1080
agcctctgca ccatcccggt gtttcgcgac gcggtggagc agcacggcat cgcgttcgtc 1140
ccgctgagcg acgaactgac ctaccgccgg accatgggcg atccgcgcct gtgggacccc 1200
aagacgtcct tcggcgtgct ctggcaaacc atcgccggga tgatcgagcc ggtctacgag 1260
tacgtctcgg cgcagcgcca tgacgacatc gtggtggtcg gctcgctctg ggcgctgggc 1320
gcacgcatcg ctcacgagaa gtacgggatt ccctacctgt ccgcgcaggt ctcgccatcg 1380
accttgttgt cggcgcacct gccgccggta caccccaagt tcaacgtgcc cgagcagatg 1440
ccgctggcga tgcgcaagct gctctggcgc tgcatcgagc gcttcaagct ggatcgcacc 1500
tgcgcgccgg atatcaacgc ggtgcggcgc aaggtcggcc tggagacgcc ggtgaagcgc 1560
atcttcaccc aatggatgca ttcgccgcag ggcgtggtct gcctgttccc ggcctggttc 1620
gcgccgcccc agcaggattg gccgcaaccc ctgcacatga ccggcttccc gctgttcgac 1680
ggcagtatcc cggggacccc gctcgacgac gaactgcaac gctttctcga tcagggcagc 1740
cggccgctgg tgttcaccca gggctcgacc gaacacctgc agggcgactt ctacgccatg 1800
gccctgcgcg cgctggaacg cctcggcgcg cgtgggatct tcctcaccgg cgccggccag 1860
gaaccgctgc gcggcttgcc gaaccacgtg ctgcagcgcg cctacgcgcc actgggagcc 1920
ttgctgccat cgtgcgccgg gctggtccat ccgggcggta tcggcgccat gagcctggcc 1980
ttggcggcgg gggtgccgca ggtgctgctg ccctgcgccc acgaccagtt cgacaatgcc 2040
gaacggctgg tccggctcgg ctgcgggatg cgcctgggcg tgccattgcg cgagcaggag 2100
ttgcgcgggg cgctgtggcg cttgctcgag gacccggcca tggcggcggc ctgtcggcgt 2160
ttcatggaat tgtcacaacc gcacagtatc gcttgcggta aagcggccca ggtggtcgaa 2220
cgttgtcata gggaggggga tgcgcgatgg ctgaaggctg cgtcctgacg ccgggaggat 2280
cctggcgtgt ccacgaccag cctctgcccc tccgccacgc gggaacacgg tcccggcgcg 2340
aaacgcgtcc tgcctctgct gttcctcacc tgcctgctgg atgccgctgg cgtcggcctg 2400
atcgtgcccc tgctgccgac gctgatcggc agcgtggcgc cgctggcggt ccgcgacgcg 2460
gccacctggg gcgccgccct ggtgatgacc ttcgcgctgc tgcaattgtt cttttcgccg 2520
gtcctcggca gcctcagcga ccgcttcgga cgccgccccg tcctggtcct ggcgatgctc 2580
ggcttcgccc tcagctatct gctgctggcg ctggccgaca gcctctggat gctgttcctc 2640
ggtcgcgcgc tggccgggct caccggcgcc agcgtggcca ccgcgatggc ctgcgcggct 2700
gacctcggca cgcacgggca gcgcacccgg cacttcggct ggctgtacgc cggcctcgcc 2760
ctgggcatga tcctcggccc cgccctcggt gggctgctgg cggtgcacgg cacgacgctg 2820
ccgctgttgc tggccgccgg cctgtgcctg ctcaacgccc tgctcgccgg cctgttcctc 2880
gaggaaaccc tgcccccgac gcgacgccgc cgcctggacc cgaggcggat gaatgccttg 2940
cgctcgatca gcggcctggc tcggcaaccg ggggtcggac gcctgctggc ggtgcttgcc 3000
ctggtattcc tcggcttgca ggcggtgatg gtggtctggc cgttcttcgt gatcgagaag 3060
tttcactgga gcagcgcctg gatcggctac tcgctggccc tctacggcgt gctcgcggtg 3120
ctcgcccaga ccctcggcgt gaacctctgc aagcggcgcc tggacgacgc ccgcctgctg 3180
cgcctgggcc tcgccctgca aggctgcggc ctgctgctgt tcgccctggt cgactcgtca 3240
ttctggctgg tctgcgcgct gctgcccttc gcgctcggca gcctcgccac cccggccatg 3300
caggggctgc tctcggcccg cgtgccggtc gaccgccagg gcgagttgca gggcgtgctg 3360
agcagcctga tgagcctcgc cgcgatcgtc ggtccgccgc tgatgagcgg cctgttccac 3420
tggggcagcg gtccgctcgc gccgctgccc ctggccggcg cgccattcct cgccggcgcc 3480
cttctcgttc tggccgggct ggtcctggcc tggcaacttc gacctacggg agaagaacga 3540
tcatggaccg gatagacatg ggcgtgctgg tggtacttct agagagctca 3590
<210> 42
<211> 8702
<212> DNA
<213> Artificial
<220>
<223> Vector
<400> 42
accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc 60
aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg 120
aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg 180
aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag 240
cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt 300
tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa 360
gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc 420
aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 480
cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 540
aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 600
tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 660
gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 720
aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 780
tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 840
ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 900
ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 960
cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 1020
tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 1080
ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 1140
tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 1200
tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 1260
ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 1320
accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 1380
atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta 1440
tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga 1500
tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat 1560
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 1620
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 1680
ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 1740
ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 1800
tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa 1860
gcttgatatc gaattcctgc agcccggggg atctggcatt tttgggaggt gtgaaatgcg 1920
gcgcgaaagt ctgttggtat cggtttgcaa gggcctgcgg gtacatgtcg agcgcgttgg 1980
gcaggatccc gggcgcagca cggtgatgct ggtcaacggc gcgatggcga ccaccgcctc 2040
gttcgcccgg acctgcaagt gcctggccga acatttcaac gtggtgctgt tcgacctgcc 2100
cttcgccggg cagtcgcgtc agcacaaccc gcagcggggg ttgatcacca aggacgacga 2160
ggtggaaatc ctcctggcgc tgatcgagcg cttcgaggtc aatcacctgg tctccgcgtc 2220
ctggggcggt atctccacgc tgctggcgct gtcgcgcaat ccgcgcggca tccgcagctc 2280
ggtggtgatg gcattcgccc ctggactgaa ccaggcgatg ctcgactacg tcgggcgggc 2340
gcaggcgctg atcgagctgg acgacaagtc ggcgatcggc catctgctca acgagaccgt 2400
cggcaaatac ctgccgccgc gcctgaaagc cagcaaccat cagcacatgg cttcgctggc 2460
caccggcgaa tacgagcagg cgcgctttca catcgaccag gtgctggcgc tcaacgatcg 2520
gggctacctg gcttgcctgg agcggatcca gagccacgtg catttcatca acggcagctg 2580
ggacgaatac accaccgccg aggacgcccg ccagttccgc gactacctgc cgcactgcag 2640
tttctcgcgg gtggagggca ccgggcattt cctcgacctg gagtccaagc tggccgcggt 2700
acgcgtgcac cgcgccctgc tcgagcacct gctgaagcaa ccggagccgc agcgggcgga 2760
acgcgcggcg ggattccacg agatggccat cggctacgcc tgaacccttg acctgcgaag 2820
acccggcctg gccgggcttt gcggttgcat aacgcacgga gtagcaccat gcacgccatc 2880
ctcatcgcca tcggctcggc cggcgacgta tttcccttca tcggcctggc ccggaccctg 2940
aaattgcgcg ggcaccgcgt gagcctctgc accatcccgg tgtttcgcga cgcggtggag 3000
cagcacggca tcgcgttcgt cccgctgagc gacgaactga cctaccgccg gaccatgggc 3060
gatccgcgcc tgtgggaccc caagacgtcc ttcggcgtgc tctggcaaac catcgccggg 3120
atgatcgagc cggtctacga gtacgtctcg gcgcagcgcc atgacgacat cgtggtggtc 3180
ggctcgctct gggcgctggg cgcacgcatc gctcacgaga agtacgggat tccctacctg 3240
tccgcgcagg tctcgccatc gaccttgttg tcggcgcacc tgccgccggt acaccccaag 3300
ttcaacgtgc ccgagcagat gccgctggcg atgcgcaagc tgctctggcg ctgcatcgag 3360
cgcttcaagc tggatcgcac ctgcgcgccg gatatcaacg cggtgcggcg caaggtcggc 3420
ctggagacgc cggtgaagcg catcttcacc caatggatgc attcgccgca gggcgtggtc 3480
tgcctgttcc cggcctggtt cgcgccgccc cagcaggatt ggccgcaacc cctgcacatg 3540
accggcttcc cgctgttcga cggcagtatc ccggggaccc cgctcgacga cgaactgcaa 3600
cgctttctcg atcagggcag ccggccgctg gtgttcaccc agggctcgac cgaacacctg 3660
cagggcgact tctacgccat ggccctgcgc gcgctggaac gcctcggcgc gcgtgggatc 3720
ttcctcaccg gcgccggcca ggaaccgctg cgcggcttgc cgaaccacgt gctgcagcgc 3780
gcctacgcgc cactgggagc cttgctgcca tcgtgcgccg ggctggtcca tccgggcggt 3840
atcggcgcca tgagcctggc cttggcggcg ggggtgccgc aggtgctgct gccctgcgcc 3900
cacgaccagt tcgacaatgc cgaacggctg gtccggctcg gctgcgggat gcgcctgggc 3960
gtgccattgc gcgagcagga gttgcgcggg gcgctgtggc gcttgctcga ggacccggcc 4020
atggcggcgg cctgtcggcg tttcatggaa ttgtcacaac cgcacagtat cgcttgcggt 4080
aaagcggccc aggtggtcga acgttgtcat agggaggggg atgcgcgatg gctgaaggct 4140
gcgtcctgac gccgggagga tcctggcgtg tccacgacca gcctctgccc ctccgccacg 4200
cgggaacacg gtcccggcgc gaaacgcgtc ctgcctctgc tgttcctcac ctgcctgctg 4260
gatgccgctg gcgtcggcct gatcgtgccc ctgctgccga cgctgatcgg cagcgtggcg 4320
ccgctggcgg tccgcgacgc ggccacctgg ggcgccgccc tggtgatgac cttcgcgctg 4380
ctgcaattgt tcttttcgcc ggtcctcggc agcctcagcg accgcttcgg acgccgcccc 4440
gtcctggtcc tggcgatgct cggcttcgcc ctcagctatc tgctgctggc gctggccgac 4500
agcctctgga tgctgttcct cggtcgcgcg ctggccgggc tcaccggcgc cagcgtggcc 4560
accgcgatgg cctgcgcggc tgacctcggc acgcacgggc agcgcacccg gcacttcggc 4620
tggctgtacg ccggcctcgc cctgggcatg atcctcggcc ccgccctcgg tgggctgctg 4680
gcggtgcacg gcacgacgct gccgctgttg ctggccgccg gcctgtgcct gctcaacgcc 4740
ctgctcgccg gcctgttcct cgaggaaacc ctgcccccga cgcgacgccg ccgcctggac 4800
ccgaggcgga tgaatgcctt gcgctcgatc agcggcctgg ctcggcaacc gggggtcgga 4860
cgcctgctgg cggtgcttgc cctggtattc ctcggcttgc aggcggtgat ggtggtctgg 4920
ccgttcttcg tgatcgagaa gtttcactgg agcagcgcct ggatcggcta ctcgctggcc 4980
ctctacggcg tgctcgcggt gctcgcccag accctcggcg tgaacctctg caagcggcgc 5040
ctggacgacg cccgcctgct gcgcctgggc ctcgccctgc aaggctgcgg cctgctgctg 5100
ttcgccctgg tcgactcgtc attctggctg gtctgcgcgc tgctgccctt cgcgctcggc 5160
agcctcgcca ccccggccat gcaggggctg ctctcggccc gcgtgccggt cgaccgccag 5220
ggcgagttgc agggcgtgct gagcagcctg atgagcctcg ccgcgatcgt cggtccgccg 5280
ctgatgagcg gcctgttcca ctggggcagc ggtccgctcg cgccgctgcc cctggccggc 5340
gcgccattcc tcgccggcgc ccttctcgtt ctggccgggc tggtcctggc ctggcaactt 5400
cgacctacgg gagaagaacg atcatggacc ggatagacat gggcgtgctg gtggtacttc 5460
tagagcggcc gccaccgcgg tggagctcca attcgcccta tagtgagtcg tattacgcgc 5520
gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta 5580
atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg 5640
atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat 5700
tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 5760
ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac 5820
gcctggtgct acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgaaagca 5880
aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct 5940
ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca aatgcctgag 6000
gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc atttattctg 6060
cctcccagag cctgataaaa acggtgaatc cgttagcgag gtgccgccgg cttccattca 6120
ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag 6180
ggcggcgagg cggctacagc cgatagtctg gaacagcgca cttacgggtt gctgcgcaac 6240
ccaagtgcta ccggcgcggc agcgtgaccc gtgtcggcgg ctccaacggc tcgccatcgt 6300
ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg cccgcgccgt tcccattcct 6360
ccgtttcggt caaggctggc aggtctggtt ccatgcccgg aatgccgggc tggctgggcg 6420
gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg atacagggtc gggatgcggc 6480
gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc gtgatcaacc accacggcgg 6540
cactgaacac cgacaggcgc aactggtcgc ggggctggcc ccacgccacg cggtcattga 6600
ccacgtaggc cgacacggtg ccggggccgt tgagcttcac gacggagatc cagcgctcgg 6660
ccaccaagtc cttgactgcg tattggaccg tccgcaaaga acgtccgatg agcttggaaa 6720
gtgtcttctg gctgaccacc acggcgttct ggtggcccat ctgcgccacg aggtgatgca 6780
gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc 6840
gttccgtttg cacccagtga ccgggcttgt tcttggcttg aatgccgatt tctctggact 6900
gcgtggccat gcttatctcc atgcggtagg gtgccgcacg gttgcggcac catgcgcaat 6960
cagctgcaac ttttcggcag cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat 7020
tacagatttt ctttaaccta cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg 7080
cgtacccccc ttttttaagt tgttgatttt taagtctttc gcatttcgcc ctatatctag 7140
ttctttggtg cccaaagaag ggcacccctg cggggttccc ccacgccttc ggcgcggctc 7200
cccctccggc aaaaagtggc ccctccgggg cttgttgatc gactgcgcgg ccttcggcct 7260
tgcccaaggt ggcgctgccc ccttggaacc cccgcactcg ccgccgtgag gctcgggggg 7320
caggcgggcg ggcttcgcct tcgactgccc ccactcgcat aggcttgggt cgttccaggc 7380
gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct tgacccgcct tccacttggt 7440
gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcttttcccc agagaaaatt 7500
aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg agccggtggg tatgtggtcg 7560
aaggctgggt agccggtggg caatccctgt ggtcaagctc gtgggcaggc gcagcctgtc 7620
catcagcttg tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc cggtggccgc 7680
tcgcggccat cgtccacata tccacgggct ggcaagggag cgcagcgacc gcgcagggcg 7740
aagcccggag agcaagcccg tagggcgccg cagccgccgt aggcggtcac gactttgcga 7800
agcaaagtct agtgagtata ctcaagcatt gagtggcccg ccggaggcac cgccttgcgc 7860
tgcccccgtc gagccggttg gacaccaaaa gggaggggca ggcatggcgg catacgcgat 7920
catgcgatgc aagaagctgg cgaaaatggg caacgtggcg gccagtctca agcacgccta 7980
ccgcgagcgc gagacgccca acgctgacgc cagcaggacg ccagagaacg agcactgggc 8040
ggccagcagc accgatgaag cgatgggccg actgcgcgag ttgctgccag agaagcggcg 8100
caaggacgct gtgttggcgg tcgagtacgt catgacggcc agcccggaat ggtggaagtc 8160
ggccagccaa gaacagcagg cggcgttctt cgagaaggcg cacaagtggc tggcggacaa 8220
gtacggggcg gatcgcatcg tgacggccag catccaccgt gacgaaacca gcccgcacat 8280
gaccgcgttc gtggtgccgc tgacgcagga cggcaggctg tcggccaagg agttcatcgg 8340
caacaaagcg cagatgaccc gcgaccagac cacgtttgcg gccgctgtgg ccgatctagg 8400
gctgcaacgg ggcatcgagg gcagcaaggc acgtcacacg cgcattcagg cgttctacga 8460
ggccctggag cggccaccag tgggccacgt caccatcagc ccgcaagcgg tcgagccacg 8520
cgcctatgca ccgcagggat tggccgaaaa gctgggaatc tcaaagcgcg ttgagacgcc 8580
ggaagccgtg gccgaccggc tgacaaaagc ggttcggcag gggtatgagc ctgccctaca 8640
ggccgccgca ggagcgcgtg agatgcgcaa gaaggccgat caagcccaag agacggcccg 8700
ag 8702
<210> 43
<211> 36
<212> DNA
<213> Artificial
<220>
<223> Primer
<400> 43
tatatataga attcggctgc gctaccgcag cccttc 36
<210> 44
<211> 34
<212> DNA
<213> Artificial
<220>
<223> Primer
<400> 44
tatatatatc tagaattaat gcagctggca cgac 34
<210> 45
<211> 8672
<212> DNA
<213> Artificial
<220>
<223> Vector
<400> 45
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760
ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820
tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880
gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940
agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000
aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060
atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120
attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180
gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3240
tagaactagt ggatcccccg ggctgcagga attcggctgc gctaccgcag cccttctccg 3300
ctaaaaccgt tagtcgaaca gttcggcgtc agccaatgcg accccaagct ggtccttgcc 3360
agacaagctt ggtacggagt cgatgggcca ttcgataccc actgccggat cattccaggc 3420
aatgcagcgc tcgcactgcg gcgagtagaa gtcggtggtc ttgtagagga actctgcggt 3480
ttcactcaac gtgacgaacc cgtgtgcgaa ccctggcggg atccacagct ggttcttgtt 3540
ctcggccgac aacaccgcac ctacccattt accgaaggtt gtggacgagc gacggatatc 3600
caccgcaaca tcgaagactt cgccttgcac cacacgcacc agcttgccct gggcgtgagg 3660
tgccagctga tagtgcaggc cacggagcac gccttttacc gagcgcgagt ggttgtcttg 3720
tacgaagtcg ggctgcaggc cggtcacttc gctgaaaaca cgggcgttga agctctcgta 3780
gaagaaacca cgttcgtcgc caaaaacctt gggggtaaac agcacgactt cggggatatc 3840
cagcggaatg gcttgcatca gaacaccttc tctttcagca agttctgcag atacttgcca 3900
taaccgtttt tcagcagtgg ttgagccagg cactcgagtt gctcagcgtt gatccagcca 3960
gcgcggtagc aaatttcctc agggcaggcg actttcaagc cctgacggcg ctccatggtt 4020
gcgatgtact ggctagcctc cagcagactg tcgtgcgtgc cggtgtcgag ccacgcatag 4080
ccacggccca tgatttcgac ctgcaactgc tgctgctgca agtaaaggtt gttgaggtcg 4140
gtgatttcca gctcgccacg tggggaaggc ttcagctcgc gagccagatt gactacctga 4200
ttgtcataga aatacaggcc ggtgaccgca tagctagact ttggaactgc cggtttttct 4260
tccagcgaca atacgcgacc gctatcgtca aactccgcta cgccatagcg ttctgggtca 4320
tgaacatgat aagcgaatac tgaagcaccg gattcacgtt tatctgcgtt caatagcagt 4380
gcctggaagt catggccgta gaaaatattg tcaccgagaa ccaacgcaga agggtcgtta 4440
ccgatgaagt cagcgccgat ggtgaacgct tgcgccaagc catccgggct tggttgtatt 4500
gcgtatgaca ggttcaggcc ccactggctg ccatcgccca gcagctgttc gaagcgcggg 4560
gtgtcctgcg gggtggaaat gatcaggatg tcccggatac cagcgagcag cagggtgctc 4620
agcgggtagt agatcatcgg tttgtcatac accggcagca gctgcttcga aaccgaaagt 4680
gtggccggat gcaggcgtgt acccgaaccg ccggccagaa taattccttt acgagccatg 4740
agagtcccta ttactggatt tcgtccagca tacgttgcac gccttgctcc caaagcggca 4800
ttttgaaatt gaacgtgttt tccagtttgc ccagtgccag gcgcgagttg cgcggacgtg 4860
gtgcaggtac tggataagct tcggtgctga ttgcggcaac cttatcagct gtcactttca 4920
gcgctacgcc agtgcgttga gcatgcgcca gcacgaactg agcaaaacca tgccaagagg 4980
tttcaccgga cgcagccaag tggtaaatcc ctgccaggtg acggttgtct tgcccattga 5040
agatttgccg caggatgtgt gcagtaacgt cggcgatcag gtcagcgccc gtgggtgcgc 5100
caaattggtc tgctaccacg ctcaacgtct cacgctccgc cgccaggcgc agcatggtct 5160
tggcaaaatt gtgcccgcgc gcagcataca cccagctggt gcgcagtacc acggccttgg 5220
cgccgctggc gagaatggca tgctcgcctt ccagcttggt ccggccgtag accgaaaggg 5280
ggccggtagg cgcagtttcc tcccagcgct gactgccgct gccgtcaaat acataatcgg 5340
tggaatagtg aatcaaccag gcgcccaaag ctgctgtttc acgtgctaat acagcaggag 5400
ccgcggcatt gatcattgca gccagtgcct gatcgctctc agctttatcc actgcagtgt 5460
aggcagcagc gttgacgatc acgtccggcg ccagctgacg aatcgtagcg gccaagccgt 5520
ccaggttgga caagtcgcca cataagccct cggccccctg acgatccagc gcaatgacct 5580
cacccagcgg cgccaaggcg cgctgtagct cccagcctac ttgcccgttt ttccccaaca 5640
gcaggatttt cacgctttat ttgccccgta ttgttgtgcc acccagtcac ggtagctgcc 5700
gtccatgaca ccttttaccc atttctggtt ggccaagtac caagcgactg tctttcgaat 5760
gcccgtctcg aaggtttcgg cagtttccag ccgagctccc gctcgatctt gcgtgcatcg 5820
atggcataac ggcggtcatg gcctgggcgg tcggttacgt aggcgatgag ttctgcatac 5880
tgttcgacag gctcgccggt cttctgattg attacctggc gcgatgccgc aggtgccatc 5940
tcgtcgagaa ggctgcagag tgtacgcaca atgtcaatgt tggctttttc attccagccg 6000
ccaatattgt acgtctcgcc gaacgcaccg gcttccagta cgcgacggat gcccgagcag 6060
tgatcttcga catacagcca gtcgcggatt tgctggccgt cgccatagac aggcagcgcc 6120
ttaccggcga gtgcgttgac gatcatcagc gggatcagtt tttccgggaa gtggagcggc 6180
ccgtaattgt tggagcagtt ggtagtgagt accggcatgc cgtaggtatg gaaatacgag 6240
cgtaccagat ggtcgctggc tgccttgctg gcggagtatg ggctgttcgg cgcgtacggc 6300
gtggtttcgg tgaacgccgg gtcgtttggc cctagtgtgc cgtagacttc gtcggtagag 6360
acatggagga aacggaaggc ctccttctct gcaccttcca aactattcca atgcgcccgg 6420
gcggcttcaa gcaagcgaaa cgtgcccatc acgttggttt cgacaaacgc ttcggggccg 6480
gtgattgagc ggtctacatg ggattccgcc gcgaagtgaa ccacggcgcg cgggcggtgc 6540
tctgcgaaca gcttggtcag aagcgcagca tcgcaaatat tgccttgcac aaagcgatgc 6600
tgagggttgc cttccagcgg ctgcaggttg gccaggttgc ctgcgtaggt cagggcgtcg 6660
aggttgagga cgggttcctc attgtgcgca caccattgca gtacgaaatt tgagccgatg 6720
aagccggctc cgcctgttac tagaatcata atttggctct aattggacaa aaggtgttgt 6780
cgtagacaga tgacgcgaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg 6840
gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat 6900
ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6960
ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 7020
cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 7080
tcggccaacg cgcggggaga ggcggtttgc gtattgggcg catgcataaa aactgttgta 7140
attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg 7200
ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatgggg gtgggcgaag 7260
aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt 7320
ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg 7380
ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa 7440
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 7500
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 7560
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 7620
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 7680
gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 7740
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 7800
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 7860
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 7920
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 7980
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt 8040
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 8100
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 8160
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 8220
acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg 8280
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag 8340
ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa 8400
gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc 8460
agtagctgac attcatccca ggtggcactt ttcggggaaa tgtgcgcgcc cgcgttcctg 8520
ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt tccgtcagca gcttttcgcc 8580
cacggccttg atgatcgcgg cggccttggc ctgcatatcc cgattcaacg gccccagggc 8640
gtccagaacg ggcttcaggc gctcccgaag gt 8672
<210> 46
<211> 21
<212> DNA
<213> Artificial
<220>
<223> Primer
<400> 46
ggccgctcta gaactagtgg a 21
<210> 47
<211> 12249
<212> DNA
<213> Artificial
<220>
<223> Vector
<400> 47
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760
ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820
tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880
gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940
agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000
aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060
atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120
attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180
gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3240
tagaactagt ggatcccccg ggctgcagga attcggctgc gctaccgcag cccttctccg 3300
ctaaaaccgt tagtcgaaca gttcggcgtc agccaatgcg accccaagct ggtccttgcc 3360
agacaagctt ggtacggagt cgatgggcca ttcgataccc actgccggat cattccaggc 3420
aatgcagcgc tcgcactgcg gcgagtagaa gtcggtggtc ttgtagagga actctgcggt 3480
ttcactcaac gtgacgaacc cgtgtgcgaa ccctggcggg atccacagct ggttcttgtt 3540
ctcggccgac aacaccgcac ctacccattt accgaaggtt gtggacgagc gacggatatc 3600
caccgcaaca tcgaagactt cgccttgcac cacacgcacc agcttgccct gggcgtgagg 3660
tgccagctga tagtgcaggc cacggagcac gccttttacc gagcgcgagt ggttgtcttg 3720
tacgaagtcg ggctgcaggc cggtcacttc gctgaaaaca cgggcgttga agctctcgta 3780
gaagaaacca cgttcgtcgc caaaaacctt gggggtaaac agcacgactt cggggatatc 3840
cagcggaatg gcttgcatca gaacaccttc tctttcagca agttctgcag atacttgcca 3900
taaccgtttt tcagcagtgg ttgagccagg cactcgagtt gctcagcgtt gatccagcca 3960
gcgcggtagc aaatttcctc agggcaggcg actttcaagc cctgacggcg ctccatggtt 4020
gcgatgtact ggctagcctc cagcagactg tcgtgcgtgc cggtgtcgag ccacgcatag 4080
ccacggccca tgatttcgac ctgcaactgc tgctgctgca agtaaaggtt gttgaggtcg 4140
gtgatttcca gctcgccacg tggggaaggc ttcagctcgc gagccagatt gactacctga 4200
ttgtcataga aatacaggcc ggtgaccgca tagctagact ttggaactgc cggtttttct 4260
tccagcgaca atacgcgacc gctatcgtca aactccgcta cgccatagcg ttctgggtca 4320
tgaacatgat aagcgaatac tgaagcaccg gattcacgtt tatctgcgtt caatagcagt 4380
gcctggaagt catggccgta gaaaatattg tcaccgagaa ccaacgcaga agggtcgtta 4440
ccgatgaagt cagcgccgat ggtgaacgct tgcgccaagc catccgggct tggttgtatt 4500
gcgtatgaca ggttcaggcc ccactggctg ccatcgccca gcagctgttc gaagcgcggg 4560
gtgtcctgcg gggtggaaat gatcaggatg tcccggatac cagcgagcag cagggtgctc 4620
agcgggtagt agatcatcgg tttgtcatac accggcagca gctgcttcga aaccgaaagt 4680
gtggccggat gcaggcgtgt acccgaaccg ccggccagaa taattccttt acgagccatg 4740
agagtcccta ttactggatt tcgtccagca tacgttgcac gccttgctcc caaagcggca 4800
ttttgaaatt gaacgtgttt tccagtttgc ccagtgccag gcgcgagttg cgcggacgtg 4860
gtgcaggtac tggataagct tcggtgctga ttgcggcaac cttatcagct gtcactttca 4920
gcgctacgcc agtgcgttga gcatgcgcca gcacgaactg agcaaaacca tgccaagagg 4980
tttcaccgga cgcagccaag tggtaaatcc ctgccaggtg acggttgtct tgcccattga 5040
agatttgccg caggatgtgt gcagtaacgt cggcgatcag gtcagcgccc gtgggtgcgc 5100
caaattggtc tgctaccacg ctcaacgtct cacgctccgc cgccaggcgc agcatggtct 5160
tggcaaaatt gtgcccgcgc gcagcataca cccagctggt gcgcagtacc acggccttgg 5220
cgccgctggc gagaatggca tgctcgcctt ccagcttggt ccggccgtag accgaaaggg 5280
ggccggtagg cgcagtttcc tcccagcgct gactgccgct gccgtcaaat acataatcgg 5340
tggaatagtg aatcaaccag gcgcccaaag ctgctgtttc acgtgctaat acagcaggag 5400
ccgcggcatt gatcattgca gccagtgcct gatcgctctc agctttatcc actgcagtgt 5460
aggcagcagc gttgacgatc acgtccggcg ccagctgacg aatcgtagcg gccaagccgt 5520
ccaggttgga caagtcgcca cataagccct cggccccctg acgatccagc gcaatgacct 5580
cacccagcgg cgccaaggcg cgctgtagct cccagcctac ttgcccgttt ttccccaaca 5640
gcaggatttt cacgctttat ttgccccgta ttgttgtgcc acccagtcac ggtagctgcc 5700
gtccatgaca ccttttaccc atttctggtt ggccaagtac caagcgactg tctttcgaat 5760
gcccgtctcg aaggtttcgg caggtttcca gccgagctcc cgctcgatct tgcgtgcatc 5820
gatggcataa cggcggtcat ggcctgggcg gtcggttacg taggcgatga gttctgcata 5880
ctgttcgaca ggctcgccgg tcttctgatt gattacctgg cgcgatgccg caggtgccat 5940
ctcgtcgaga aggctgcaga gtgtacgcac aatgtcaatg ttggcttttt cattccagcc 6000
gccaatattg tacgtctcgc cgaacgcacc ggcttccagt acgcgacgga tgcccgagca 6060
gtgatcttcg acatacagcc agtcgcggat ttgctggccg tcgccataga caggcagcgc 6120
cttaccggcg agtgcgttga cgatcatcag cgggatcagt ttttccggga agtggagcgg 6180
cccgtaattg ttggagcagt tggtagtgag taccggcatg ccgtaggtat ggaaatacga 6240
gcgtaccaga tggtcgctgg ctgccttgct ggcggagtat gggctgttcg gcgcgtacgg 6300
cgtggtttcg gtgaacgccg ggtcgtttgg ccctagtgtg ccgtagactt cgtcggtaga 6360
gacatggagg aaacggaagg cctccttctc tgcaccttcc aaactattcc aatgcgcccg 6420
ggcggcttca agcaagcgaa acgtgcccat cacgttggtt tcgacaaacg cttcggggcc 6480
ggtgattgag cggtctacat gggattccgc cgcgaagtga accacggcgc gcgggcggtg 6540
ctctgcgaac agcttggtca gaagcgcagc atcgcaaata ttgccttgca caaagcgatg 6600
ctgagggttg ccttccagcg gctgcaggtt ggccaggttg cctgcgtagg tcagggcgtc 6660
gaggttgagg acgggttcct cattgtgcgc acaccattgc agtacgaaat ttgagccgat 6720
gaagccggct ccgcctgtta ctagaatcat aatttggctc taattggaca aaaggtgttg 6780
tcgtagacag atgacgcgaa ttcgatatca agcttatcga taccgtcgac ctcgaggggg 6840
ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca 6900
tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6960
gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 7020
gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaattc 7080
tagagggaat gcgtttcgcc gactaggcct tggccttgcc ggaagctacg gacgccacgg 7140
ccgggccggc gaggcgcttc agcaggcgcg ggcggttggt ctccagcgcg ccgccgcgtc 7200
cccgcaggcc gtcccacagg ccccagccca ggcagcgcag cttgagcagc ttgtcgcgtt 7260
cgagcaggag caccgcgagg ccctgggtca gggtcggcag gttcgccagc agggccagcg 7320
gcgaggaccg ggcgtagcgg cgcaggacca gcaggccgtt gcgcgccagg tagtagcggc 7380
gcagcggggc gtggttcatc gcgctgaggc tgagaccgcc gaggcggcgg gtcttgcgcg 7440
tgccgatgcg gtgctcgagg accagccgcg ggtcgacgta caggggcacg tccagcgcct 7500
gggcgcgcag gctgtattcg gtgtccacgt ggtcgatgaa cagttcctcg tcgaagtggc 7560
cgaggcgctg gtaggcctcg cgggtcagca ggcagccgga ggagatcagg aacgaggtgc 7620
gctgcggggt cgtcaggccg tccagagaca attgcctgag cgtcagtccg tcgagatgga 7680
tggccggcag gaagcgccgg tcaccccggt cgaagatccg tgggccgagc aggcaggcct 7740
gaccgttgcg cgcctgcagg ttgcgccact gggcggcgag gaaggcgccg ccgggacggg 7800
agtcctggtc gagcagcagc acaccctgca cgccacgccg gaatagcgcg tcgagtccct 7860
ggttgaaggc gccggcgatg ccctgccggt tgccgtggtg cagcacggcg atgccttgcc 7920
cgcgcagccg ggcattgcgc tgcggatcgc tgtgcggtga gttgtcgacg gcaaggaagc 7980
gcagttgcgg aaacgccgcc gccagttcgc caaggtgttc caggtcgtcg tcgccaggat 8040
tgaacagtac caccagcacg cccatgtcta tccggtccat gatcgttctt ctcccgtagg 8100
tcaggacgca gccttcagcc atcgcgcatc cccctcccta tgacaacgtt cgaccacctg 8160
ggccgcttta ccgcaagcga tactgtgcgg ttgtgacaat tccatgaaac gccgacaggc 8220
cgccgccatg gccgggtcct cgagcaagcg ccacagcgcc ccgcgcaact cctgctcgcg 8280
caatggcacg cccaggcgca tcccgcagcc gagccggacc agccgttcgg cattgtcgaa 8340
ctggtcgtgg gcgcagggca gcagcacctg cggcaccccc gccgccaagg ccaggctcat 8400
ggcgccgata ccgcccggat ggaccagccc ggcgcacgat ggcagcaagg ctcccagtgg 8460
cgcgtaggcg cgctgcagca cgtggttcgg caagccgcgc agcggttcct ggccggcgcc 8520
ggtgaggaag atcccacgcg cgccgaggcg ttccagcgcg cgcagggcca tggcgtagaa 8580
gtcgccctgc aggtgttcgg tcgagccctg ggtgaacacc agcggccggc tgccctgatc 8640
gagaaagcgt tgcagttcgt cgtcgagcgg ggtccccggg atactgccgt cgaacagcgg 8700
gaagccggtc atgtgcaggg gttgcggcca atcctgctgg ggcggcgcga accaggccgg 8760
gaacaggcag accacgccct gcggcgaatg catccattgg gtgaagatgc gcttcaccgg 8820
cgtctccagg ccgaccttgc gccgcaccgc gttgatatcc ggcgcgcagg tgcgatccag 8880
cttgaagcgc tcgatgcagc gccagagcag cttgcgcatc gccagcggca tctgctcggg 8940
cacgttgaac ttggggtgta ccggcggcag gtgcgccgac aacaaggtcg atggcgagac 9000
ctgcgcggac aggtagggaa tcccgtactt ctcgtgagcg atgcgtgcgc ccagcgccca 9060
gagcgagccg accaccacga tgtcgtcatg gcgctgcgcc gagacgtact cgtagaccgg 9120
ctcgatcatc ccggcgatgg tttgccagag cacgccgaag gacgtcttgg ggtcccacag 9180
gcgcggatcg cccatggtcc ggcggtaggt cagttcgtcg ctcagcggga cgaacgcgat 9240
gccgtgctgc tccaccgcgt cgcgaaacac cgggatggtg cagaggctca cgcggtgccc 9300
gcgcaatttc agggtccggg ccaggccgat gaagggaaat acgtcgccgg ccgagccgat 9360
ggcgatgagg atggcgtgca tggtgctact ccgtgcgtta tgcaaccgca aagcccggcc 9420
aggccgggtc ttcgcaggtc aagggttcag gcgtagccga tggccatctc gtggaatccc 9480
gccgcgcgtt ccgcccgctg cggctccggt tgcttcagca ggtgctcgag cagggcgcgg 9540
tgcacgcgta ccgcggccag cttggactcc aggtcgagga aatgcccggt gccctccacc 9600
cgcgagaaac tgcagtgcgg caggtagtcg cggaactggc gggcgtcctc ggcggtggtg 9660
tattcgtccc agctgccgtt gatgaaatgc acgtggctct ggatccgctc caggcaagcc 9720
aggtagcccc gatcgttgag cgccagcacc tggtcgatgt gaaagcgcgc ctgctcgtat 9780
tcgccggtgg ccagcgaagc catgtgctga tggttgctgg ctttcaggcg cggcggcagg 9840
tatttgccga cggtctcgtt gagcagatgg ccgatcgccg acttgtcgtc cagctcgatc 9900
agcgcctgcg cccgcccgac gtagtcgagc atcgcctggt tcagtccagg ggcgaatgcc 9960
atcaccaccg agctgcggat gccgcgcgga ttgcgcgaca gcgccagcag cgtggagata 10020
ccgccccagg acgcggagac caggtgattg acctcgaagc gctcgatcag cgccaggagg 10080
atttccacct cgtcgtcctt ggtgatcaac ccccgctgcg ggttgtgctg acgcgactgc 10140
ccggcgaagg gcaggtcgaa cagcaccacg ttgaaatgtt cggccaggca cttgcaggtc 10200
cgggcgaacg aggcggtggt cgccatcgcg ccgttgacca gcatcaccgt gctgcgcccg 10260
ggatcctgcc caacgcgctc gacatgtacc cgcaggccct tgcaaaccga taccaacaga 10320
ctttcgcgcc gcatttcaca cctcccaaaa atgccagatc ccccgggctg caggaattcg 10380
atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccag cttttgttcc 10440
ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga 10500
aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 10560
tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 10620
cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 10680
ggtttgcgta ttgggcgcat gcataaaaac tgttgtaatt cattaagcat tctgccgaca 10740
tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg 10800
ccttgcgtat aatatttgcc catgggggtg ggcgaagaac tccagcatga gatccccgcg 10860
ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa 10920
ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc 10980
gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 11040
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 11100
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 11160
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 11220
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg 11280
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 11340
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 11400
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 11460
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 11520
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 11580
gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg 11640
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 11700
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 11760
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 11820
tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact 11880
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 11940
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 12000
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catcccaggt 12060
ggcacttttc ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc 12120
gctggacttc ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg 12180
ccttggcctg catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct 12240
cccgaaggt 12249
<210> 48
<211> 37
<212> DNA
<213> Artificial
<220>
<223> Primer
<400> 48
tatatataga attcgcgtca tctgtctacg acaacac 37
<210> 49
<211> 5144
<212> DNA
<213> Artificial
<220>
<223> Vector
<400> 49
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760
ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820
tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880
gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940
agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000
aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060
atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120
attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180
gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3240
tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga 3300
cctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct 3360
tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 3420
acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 3480
tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 3540
tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgcatgcata 3600
aaaactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat 3660
gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg 3720
gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc 3780
cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt 3840
gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga 3900
actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa 3960
gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca 4020
acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa 4080
agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat 4140
cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct 4200
gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc 4260
gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca 4320
gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca 4380
ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa 4440
cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct 4500
cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc 4560
cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt 4620
catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt 4680
caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc 4740
agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag 4800
agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc cagtctagct 4860
atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg 4920
tccagatagc ccagtagctg acattcatcc caggtggcac ttttcgggga aatgtgcgcg 4980
cccgcgttcc tgctggcgct gggcctgttt ctggcgctgg acttcccgct gttccgtcag 5040
cagcttttcg cccacggcct tgatgatcgc ggcggccttg gcctgcatat cccgattcaa 5100
cggccccagg gcgtccagaa cgggcttcag gcgctcccga aggt 5144
<210> 50
<211> 4549
<212> DNA
<213> Pseudomonas aeruginosa
<400> 50
ggtaccagat ctggcatttt tgggaggtgt gaaatgcggc gcgaaagtct gttggtatcg 60
gtttgcaagg gcctgcgggt acatgtcgag cgcgttgggc aggatcccgg gcgcagcacg 120
gtgatgctgg tcaacggcgc gatggcgacc accgcctcgt tcgcccggac ctgcaagtgc 180
ctggccgaac atttcaacgt ggtgctgttc gacctgccct tcgccgggca gtcgcgtcag 240
cacaacccgc agcgggggtt gatcaccaag gacgacgagg tggaaatcct cctggcgctg 300
atcgagcgct tcgaggtcaa tcacctggtc tccgcgtcct ggggcggtat ctccacgctg 360
ctggcgctgt cgcgcaatcc gcgcggcatc cgcagctcgg tggtgatggc attcgcccct 420
ggactgaacc aggcgatgct cgactacgtc gggcgggcgc aggcgctgat cgagctggac 480
gacaagtcgg cgatcggcca tctgctcaac gagaccgtcg gcaaatacct gccgccgcgc 540
ctgaaagcca gcaaccatca gcacatggct tcgctggcca ccggcgaata cgagcaggcg 600
cgctttcaca tcgaccaggt gctggcgctc aacgatcggg gctacctggc ttgcctggag 660
cggatccaga gccacgtgca tttcatcaac ggcagctggg acgaatacac caccgccgag 720
gacgcccgcc agttccgcga ctacctgccg cactgcagtt tctcgcgggt ggagggcacc 780
gggcatttcc tcgacctgga gtccaagctg gccgcggtac gcgtgcaccg cgccctgctc 840
gagcacctgc tgaagcaacc ggagccgcag cgggcggaac gcgcggcggg attccacgag 900
atggccatcg gctacgcctg aacccttgac ctgcgaagac ccggcctggc cgggctttgc 960
ggttgcataa cgcacggagt agcaccatgc acgccatcct catcgccatc ggctcggccg 1020
gcgacgtatt tcccttcatc ggcctggccc ggaccctgaa attgcgcggg caccgcgtga 1080
gcctctgcac catcccggtg tttcgcgacg cggtggagca gcacggcatc gcgttcgtcc 1140
cgctgagcga cgaactgacc taccgccgga ccatgggcga tccgcgcctg tgggacccca 1200
agacgtcctt cggcgtgctc tggcaaacca tcgccgggat gatcgagccg gtctacgagt 1260
acgtctcggc gcagcgccat gacgacatcg tggtggtcgg ctcgctctgg gcgctgggcg 1320
cacgcatcgc tcacgagaag tacgggattc cctacctgtc cgcgcaggtc tcgccatcga 1380
ccttgttgtc ggcgcacctg ccgccggtac accccaagtt caacgtgccc gagcagatgc 1440
cgctggcgat gcgcaagctg ctctggcgct gcatcgagcg cttcaagctg gatcgcacct 1500
gcgcgccgga tatcaacgcg gtgcggcgca aggtcggcct ggagacgccg gtgaagcgca 1560
tcttcaccca atggatgcat tcgccgcagg gcgtggtctg cctgttcccg gcctggttcg 1620
cgccgcccca gcaggattgg ccgcaacccc tgcacatgac cggcttcccg ctgttcgacg 1680
gcagtatccc ggggaccccg ctcgacgacg aactgcaacg ctttctcgat cagggcagcc 1740
ggccgctggt gttcacccag ggctcgaccg aacacctgca gggcgacttc tacgccatgg 1800
ccctgcgcgc gctggaacgc ctcggcgcgc gtgggatctt cctcaccggc gccggccagg 1860
aaccgctgcg cggcttgccg aaccacgtgc tgcagcgcgc ctacgcgcca ctgggagcct 1920
tgctgccatc gtgcgccggg ctggtccatc cgggcggtat cggcgccatg agcctggcct 1980
tggcggcggg ggtgccgcag gtgctgctgc cctgcgccca cgaccagttc gacaatgccg 2040
aacggctggt ccggctcggc tgcgggatgc gcctgggcgt gccattgcgc gagcaggagt 2100
tgcgcggggc gctgtggcgc ttgctcgagg acccggccat ggcggcggcc tgtcggcgtt 2160
tcatggaatt gtcacaaccg cacagtatcg cttgcggtaa agcggcccag gtggtcgaac 2220
gttgtcatag ggagggggat gcgcgatggc tgaaggctgc gtcctgacgc cgggaggatc 2280
ctggcgtgtc cacgaccagc ctctgcccct ccgccacgcg ggaacacggt cccggcgcga 2340
aacgcgtcct gcctctgctg ttcctcacct gcctgctgga tgccgctggc gtcggcctga 2400
tcgtgcccct gctgccgacg ctgatcggca gcgtggcgcc gctggcggtc cgcgacgcgg 2460
ccacctgggg cgccgccctg gtgatgacct tcgcgctgct gcaattgttc ttttcgccgg 2520
tcctcggcag cctcagcgac cgcttcggac gccgccccgt cctggtcctg gcgatgctcg 2580
gcttcgccct cagctatctg ctgctggcgc tggccgacag cctctggatg ctgttcctcg 2640
gtcgcgcgct ggccgggctc accggcgcca gcgtggccac cgcgatggcc tgcgcggctg 2700
acctcggcac gcacgggcag cgcacccggc acttcggctg gctgtacgcc ggcctcgccc 2760
tgggcatgat cctcggcccc gccctcggtg ggctgctggc ggtgcacggc acgacgctgc 2820
cgctgttgct ggccgccggc ctgtgcctgc tcaacgccct gctcgccggc ctgttcctcg 2880
aggaaaccct gcccccgacg cgacgccgcc gcctggaccc gaggcggatg aatgccttgc 2940
gctcgatcag cggcctggct cggcaaccgg gggtcggacg cctgctggcg gtgcttgccc 3000
tggtattcct cggcttgcag gcggtgatgg tggtctggcc gttcttcgtg atcgagaagt 3060
ttcactggag cagcgcctgg atcggctact cgctggccct ctacggcgtg ctcgcggtgc 3120
tcgcccagac cctcggcgtg aacctctgca agcggcgcct ggacgacgcc cgcctgctgc 3180
gcctgggcct cgccctgcaa ggctgcggcc tgctgctgtt cgccctggtc gactcgtcat 3240
tctggctggt ctgcgcgctg ctgcccttcg cgctcggcag cctcgccacc ccggccatgc 3300
aggggctgct ctcggcccgc gtgccggtcg accgccaggg cgagttgcag ggcgtgctga 3360
gcagcctgat gagcctcgcc gcgatcgtcg gtccgccgct gatgagcggc ctgttccact 3420
ggggcagcgg tccgctcgcg ccgctgcccc tggccggcgc gccattcctc gccggcgccc 3480
ttctcgttct ggccgggctg gtcctggcct ggcaacttcg acctacggga gaagaacgat 3540
catggaccgg atagacatgg gcgtgctggt ggtactgttc aatcctggcg acgacgacct 3600
ggaacacctt ggcgaactgg cggcggcgtt tccgcaactg cgcttccttg ccgtcgacaa 3660
ctcaccgcac agcgatccgc agcgcaatgc ccggctgcgc gggcaaggca tcgccgtgct 3720
gcaccacggc aaccggcagg gcatcgccgg cgccttcaac cagggactcg acgcgctatt 3780
ccggcgtggc gtgcagggtg tgctgctgct cgaccaggac tcccgtcccg gcggcgcctt 3840
cctcgccgcc cagtggcgca acctgcaggc gcgcaacggt caggcctgcc tgctcggccc 3900
acggatcttc gaccggggtg accggcgctt cctgccggcc atccatctcg acggactgac 3960
gctcaggcaa ttgtctctgg acggcctgac gaccccgcag cgcacctcgt tcctgatctc 4020
ctccggctgc ctgctgaccc gcgaggccta ccagcgcctc ggccacttcg acgaggaact 4080
gttcatcgac cacgtggaca ccgaatacag cctgcgcgcc caggcgctgg acgtgcccct 4140
gtacgtcgac ccgcggctgg tcctcgagca ccgcatcggc acgcgcaaga cccgccgcct 4200
cggcggtctc agcctcagcg cgatgaacca cgccccgctg cgccgctact acctggcgcg 4260
caacggcctg ctggtcctgc gccgctacgc ccggtcctcg ccgctggccc tgctggcgaa 4320
cctgccgacc ctgacccagg gcctcgcggt gctcctgctc gaacgcgaca agctgctcaa 4380
gctgcgctgc ctgggctggg gcctgtggga cggcctgcgg ggacgcggcg gcgcgctgga 4440
gaccaaccgc ccgcgcctgc tgaagcgcct cgccggcccg gccgtggcgt ccgtagcttc 4500
cggcaaggcc aaggcctagt cggcgaaacg cattccctct agagagctc 4549
<210> 51
<211> 9663
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 51
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760
ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820
tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880
gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940
agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000
aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060
atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120
attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180
gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3240
tagagggaat gcgtttcgcc gactaggcct tggccttgcc ggaagctacg gacgccacgg 3300
ccgggccggc gaggcgcttc agcaggcgcg ggcggttggt ctccagcgcg ccgccgcgtc 3360
cccgcaggcc gtcccacagg ccccagccca ggcagcgcag cttgagcagc ttgtcgcgtt 3420
cgagcaggag caccgcgagg ccctgggtca gggtcggcag gttcgccagc agggccagcg 3480
gcgaggaccg ggcgtagcgg cgcaggacca gcaggccgtt gcgcgccagg tagtagcggc 3540
gcagcggggc gtggttcatc gcgctgaggc tgagaccgcc gaggcggcgg gtcttgcgcg 3600
tgccgatgcg gtgctcgagg accagccgcg ggtcgacgta caggggcacg tccagcgcct 3660
gggcgcgcag gctgtattcg gtgtccacgt ggtcgatgaa cagttcctcg tcgaagtggc 3720
cgaggcgctg gtaggcctcg cgggtcagca ggcagccgga ggagatcagg aacgaggtgc 3780
gctgcggggt cgtcaggccg tccagagaca attgcctgag cgtcagtccg tcgagatgga 3840
tggccggcag gaagcgccgg tcaccccggt cgaagatccg tgggccgagc aggcaggcct 3900
gaccgttgcg cgcctgcagg ttgcgccact gggcggcgag gaaggcgccg ccgggacggg 3960
agtcctggtc gagcagcagc acaccctgca cgccacgccg gaatagcgcg tcgagtccct 4020
ggttgaaggc gccggcgatg ccctgccggt tgccgtggtg cagcacggcg atgccttgcc 4080
cgcgcagccg ggcattgcgc tgcggatcgc tgtgcggtga gttgtcgacg gcaaggaagc 4140
gcagttgcgg aaacgccgcc gccagttcgc caaggtgttc caggtcgtcg tcgccaggat 4200
tgaacagtac caccagcacg cccatgtcta tccggtccat gatcgttctt ctcccgtagg 4260
tcgaagttgc caggccagga ccagcccggc cagaacgaga agggcgccgg cgaggaatgg 4320
cgcgccggcc aggggcagcg gcgcgagcgg accgctgccc cagtggaaca ggccgctcat 4380
cagcggcgga ccgacgatcg cggcgaggct catcaggctg ctcagcacgc cctgcaactc 4440
gccctggcgg tcgaccggca cgcgggccga gagcagcccc tgcatggccg gggtggcgag 4500
gctgccgagc gcgaagggca gcagcgcgca gaccagccag aatgacgagt cgaccagggc 4560
gaacagcagc aggccgcagc cttgcagggc gaggcccagg cgcagcaggc gggcgtcgtc 4620
caggcgccgc ttgcagaggt tcacgccgag ggtctgggcg agcaccgcga gcacgccgta 4680
gagggccagc gagtagccga tccaggcgct gctccagtga aacttctcga tcacgaagaa 4740
cggccagacc accatcaccg cctgcaagcc gaggaatacc agggcaagca ccgccagcag 4800
gcgtccgacc cccggttgcc gagccaggcc gctgatcgag cgcaaggcat tcatccgcct 4860
cgggtccagg cggcggcgtc gcgtcggggg cagggtttcc tcgaggaaca ggccggcgag 4920
cagggcgttg agcaggcaca ggccggcggc cagcaacagc ggcagcgtcg tgccgtgcac 4980
cgccagcagc ccaccgaggg cggggccgag gatcatgccc agggcgaggc cggcgtacag 5040
ccagccgaag tgccgggtgc gctgcccgtg cgtgccgagg tcagccgcgc aggccatcgc 5100
ggtggccacg ctggcgccgg tgagcccggc cagcgcgcga ccgaggaaca gcatccagag 5160
gctgtcggcc agcgccagca gcagatagct gagggcgaag ccgagcatcg ccaggaccag 5220
gacggggcgg cgtccgaagc ggtcgctgag gctgccgagg accggcgaaa agaacaattg 5280
cagcagcgcg aaggtcatca ccagggcggc gccccaggtg gccgcgtcgc ggaccgccag 5340
cggcgccacg ctgccgatca gcgtcggcag caggggcacg atcaggccga cgccagcggc 5400
atccagcagg caggtgagga acagcagagg caggacgcgt ttcgcgccgg gaccgtgttc 5460
ccgcgtggcg gaggggcaga ggctggtcgt ggacacgcca ggatcctccc ggcgtcagga 5520
cgcagccttc agccatcgcg catccccctc cctatgacaa cgttcgacca cctgggccgc 5580
tttaccgcaa gcgatactgt gcggttgtga caattccatg aaacgccgac aggccgccgc 5640
catggccggg tcctcgagca agcgccacag cgccccgcgc aactcctgct cgcgcaatgg 5700
cacgcccagg cgcatcccgc agccgagccg gaccagccgt tcggcattgt cgaactggtc 5760
gtgggcgcag ggcagcagca cctgcggcac ccccgccgcc aaggccaggc tcatggcgcc 5820
gataccgccc ggatggacca gcccggcgca cgatggcagc aaggctccca gtggcgcgta 5880
ggcgcgctgc agcacgtggt tcggcaagcc gcgcagcggt tcctggccgg cgccggtgag 5940
gaagatccca cgcgcgccga ggcgttccag cgcgcgcagg gccatggcgt agaagtcgcc 6000
ctgcaggtgt tcggtcgagc cctgggtgaa caccagcggc cggctgccct gatcgagaaa 6060
gcgttgcagt tcgtcgtcga gcggggtccc cgggatactg ccgtcgaaca gcgggaagcc 6120
ggtcatgtgc aggggttgcg gccaatcctg ctggggcggc gcgaaccagg ccgggaacag 6180
gcagaccacg ccctgcggcg aatgcatcca ttgggtgaag atgcgcttca ccggcgtctc 6240
caggccgacc ttgcgccgca ccgcgttgat atccggcgcg caggtgcgat ccagcttgaa 6300
gcgctcgatg cagcgccaga gcagcttgcg catcgccagc ggcatctgct cgggcacgtt 6360
gaacttgggg tgtaccggcg gcaggtgcgc cgacaacaag gtcgatggcg agacctgcgc 6420
ggacaggtag ggaatcccgt acttctcgtg agcgatgcgt gcgcccagcg cccagagcga 6480
gccgaccacc acgatgtcgt catggcgctg cgccgagacg tactcgtaga ccggctcgat 6540
catcccggcg atggtttgcc agagcacgcc gaaggacgtc ttggggtccc acaggcgcgg 6600
atcgcccatg gtccggcggt aggtcagttc gtcgctcagc gggacgaacg cgatgccgtg 6660
ctgctccacc gcgtcgcgaa acaccgggat ggtgcagagg ctcacgcggt gcccgcgcaa 6720
tttcagggtc cgggccaggc cgatgaaggg aaatacgtcg ccggccgagc cgatggcgat 6780
gaggatggcg tgcatggtgc tactccgtgc gttatgcaac cgcaaagccc ggccaggccg 6840
ggtcttcgca ggtcaagggt tcaggcgtag ccgatggcca tctcgtggaa tcccgccgcg 6900
cgttccgccc gctgcggctc cggttgcttc agcaggtgct cgagcagggc gcggtgcacg 6960
cgtaccgcgg ccagcttgga ctccaggtcg aggaaatgcc cggtgccctc cacccgcgag 7020
aaactgcagt gcggcaggta gtcgcggaac tggcgggcgt cctcggcggt ggtgtattcg 7080
tcccagctgc cgttgatgaa atgcacgtgg ctctggatcc gctccaggca agccaggtag 7140
ccccgatcgt tgagcgccag cacctggtcg atgtgaaagc gcgcctgctc gtattcgccg 7200
gtggccagcg aagccatgtg ctgatggttg ctggctttca ggcgcggcgg caggtatttg 7260
ccgacggtct cgttgagcag atggccgatc gccgacttgt cgtccagctc gatcagcgcc 7320
tgcgcccgcc cgacgtagtc gagcatcgcc tggttcagtc caggggcgaa tgccatcacc 7380
accgagctgc ggatgccgcg cggattgcgc gacagcgcca gcagcgtgga gataccgccc 7440
caggacgcgg agaccaggtg attgacctcg aagcgctcga tcagcgccag gaggatttcc 7500
acctcgtcgt ccttggtgat caacccccgc tgcgggttgt gctgacgcga ctgcccggcg 7560
aagggcaggt cgaacagcac cacgttgaaa tgttcggcca ggcacttgca ggtccgggcg 7620
aacgaggcgg tggtcgccat cgcgccgttg accagcatca ccgtgctgcg cccgggatcc 7680
tgcccaacgc gctcgacatg tacccgcagg cccttgcaaa ccgataccaa cagactttcg 7740
cgccgcattt cacacctccc aaaaatgcca gatcccccgg gctgcaggaa ttcgatatca 7800
agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg ttccctttag 7860
tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 7920
tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 7980
gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 8040
ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 8100
cgtattgggc gcatgcataa aaactgttgt aattcattaa gcattctgcc gacatggaag 8160
ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc 8220
gtataatatt tgcccatggg ggtgggcgaa gaactccagc atgagatccc cgcgctggag 8280
gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc 8340
ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc 8400
cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg 8460
ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca 8520
gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca 8580
cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg 8640
ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt 8700
tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct 8760
tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta 8820
gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca 8880
ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc 8940
cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc 9000
cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg 9060
acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg 9120
attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct 9180
gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat 9240
cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag 9300
ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat 9360
aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt 9420
gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccc aggtggcact 9480
tttcggggaa atgtgcgcgc ccgcgttcct gctggcgctg ggcctgtttc tggcgctgga 9540
cttcccgctg ttccgtcagc agcttttcgc ccacggcctt gatgatcgcg gcggccttgg 9600
cctgcatatc ccgattcaac ggccccaggg cgtccagaac gggcttcagg cgctcccgaa 9660
ggt 9663
<210> 52
<211> 9793
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 52
gatctggcat ttttgggagg tgtgaaatgc ggcgcgaaag tctgttggta tcggtttgca 60
agggcctgcg ggtacatgtc gagcgcgttg ggcaggatcc cgggcgcagc acggtgatgc 120
tggtcaacgg cgcgatggcg accaccgcct cgttcgcccg gacctgcaag tgcctggccg 180
aacatttcaa cgtggtgctg ttcgacctgc ccttcgccgg gcagtcgcgt cagcacaacc 240
cgcagcgggg gttgatcacc aaggacgacg aggtggaaat cctcctggcg ctgatcgagc 300
gcttcgaggt caatcacctg gtctccgcgt cctggggcgg tatctccacg ctgctggcgc 360
tgtcgcgcaa tccgcgcggc atccgcagct cggtggtgat ggcattcgcc cctggactga 420
accaggcgat gctcgactac gtcgggcggg cgcaggcgct gatcgagctg gacgacaagt 480
cggcgatcgg ccatctgctc aacgagaccg tcggcaaata cctgccgccg cgcctgaaag 540
ccagcaacca tcagcacatg gcttcgctgg ccaccggcga atacgagcag gcgcgctttc 600
acatcgacca ggtgctggcg ctcaacgatc ggggctacct ggcttgcctg gagcggatcc 660
agagccacgt gcatttcatc aacggcagct gggacgaata caccaccgcc gaggacgccc 720
gccagttccg cgactacctg ccgcactgca gtttctcgcg ggtggagggc accgggcatt 780
tcctcgacct ggagtccaag ctggccgcgg tacgcgtgca ccgcgccctg ctcgagcacc 840
tgctgaagca accggagccg cagcgggcgg aacgcgcggc gggattccac gagatggcca 900
tcggctacgc ctgaaccctt gacctgcgaa gacccggcct ggccgggctt tgcggttgca 960
taacgcacgg agtagcacca tgcacgccat cctcatcgcc atcggctcgg ccggcgacgt 1020
atttcccttc atcggcctgg cccggaccct gaaattgcgc gggcaccgcg tgagcctctg 1080
caccatcccg gtgtttcgcg acgcggtgga gcagcacggc atcgcgttcg tcccgctgag 1140
cgacgaactg acctaccgcc ggaccatggg cgatccgcgc ctgtgggacc ccaagacgtc 1200
cttcggcgtg ctctggcaaa ccatcgccgg gatgatcgag ccggtctacg agtacgtctc 1260
ggcgcagcgc catgacgaca tcgtggtggt cggctcgctc tgggcgctgg gcgcacgcat 1320
cgctcacgag aagtacggga ttccctacct gtccgcgcag gtctcgccat cgaccttgtt 1380
gtcggcgcac ctgccgccgg tacaccccaa gttcaacgtg cccgagcaga tgccgctggc 1440
gatgcgcaag ctgctctggc gctgcatcga gcgcttcaag ctggatcgca cctgcgcgcc 1500
ggatatcaac gcggtgcggc gcaaggtcgg cctggagacg ccggtgaagc gcatcttcac 1560
ccaatggatg cattcgccgc agggcgtggt ctgcctgttc ccggcctggt tcgcgccgcc 1620
ccagcaggat tggccgcaac ccctgcacat gaccggcttc ccgctgttcg acggcagtat 1680
cccggggacc ccgctcgacg acgaactgca acgctttctc gatcagggca gccggccgct 1740
ggtgttcacc cagggctcga ccgaacacct gcagggcgac ttctacgcca tggccctgcg 1800
cgcgctggaa cgcctcggcg cgcgtgggat cttcctcacc ggcgccggcc aggaaccgct 1860
gcgcggcttg ccgaaccacg tgctgcagcg cgcctacgcg ccactgggag ccttgctgcc 1920
atcgtgcgcc gggctggtcc atccgggcgg tatcggcgcc atgagcctgg ccttggcggc 1980
gggggtgccg caggtgctgc tgccctgcgc ccacgaccag ttcgacaatg ccgaacggct 2040
ggtccggctc ggctgcggga tgcgcctggg cgtgccattg cgcgagcagg agttgcgcgg 2100
ggcgctgtgg cgcttgctcg aggacccggc catggcggcg gcctgtcggc gtttcatgga 2160
attgtcacaa ccgcacagta tcgcttgcgg taaagcggcc caggtggtcg aacgttgtca 2220
tagggagggg gatgcgcgat ggctgaaggc tgcgtcctga acggtgctgg cataacagat 2280
agggttgcct ctagagtcga cctgcaggca tgcaagcttg gctgttttgg cggatgagag 2340
aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat 2400
ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa 2460
cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca 2520
tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc 2580
ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca 2640
acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca 2700
gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttttg tttatttttc 2760
taaatacatt caaatatgta tccgctcatg ctccttcgtc ggtgtcgtcg ccggatggtc 2820
tgcggtggtg ctcagcgtgg agacgcgcac cgtcacggac ccccatcaat cctgcctatt 2880
tgccacgttt aacaaggtag ttaagcgttc atttacgaag aaaacacgat aagctgcaca 2940
aatacctgaa aaagttgaac gccccgtgag cgggaactca cagggcgtcg gctaaccccc 3000
agtcatcagc tgggagaaag cactcaagac atgactctag ccgatccgca ggacacagtc 3060
acagctagcg cgtggaaatt gtccgccgat ctgttcgaca cccaccccga agctatgcgc 3120
tgcggctcac gcggctggac ggcagaagat cgccgcgaac tgctcgctca cctgggacgc 3180
gaaagcttcc agggcagcaa gacaagagat ttcgcgagcg cctggattaa aaacccggat 3240
accggcgaaa cccaaccaaa gctctaccgg gctggctcaa aagcgctgac gcggtgccag 3300
tacgttgcgc tgacgcacgc gcaacatgcc gcggtgatcg tgcttgacat cgatgtgccc 3360
agccaccagg ccggcgggaa gattgagcac gtaaacccgc aggtctacgc gattttagag 3420
aaatgggcac gcctagaaaa agcgccggct tggatcggcg tgaatccgct gagcgggaaa 3480
tgccagctca tctggctcat tgacccggtg tatgccgcag caggtaaaac cagcccaaat 3540
atgcgcctgc tggctgcaac gacggaagaa atgactcgtg ttttcggcgc tgaccaggct 3600
ttttcgcata ggctgagccg gtggccgctg cacgtctcag acgatccgac agcctataaa 3660
tggcactgcc agcatgatcg tgtggatcgg ctggccgacc taatggagat tgctcgaacg 3720
atgaccggat cacagaagcc gaaaaagtac attgagcagg acttttccag cggacgcgcc 3780
cgcattgaag cggcacaacg cgccaccgca gaagccaagg cgctagcgat tttggacgcg 3840
agcctgccga gcgccctgga cgcgtccggc gacctgatcg acggcgtgcg agtgctctgg 3900
acaaatccag agcgagcgcg cgacgagacc gcgtttcgcc acgcgttgac cgtgggatac 3960
cagctcaaag ctgctggtga gcgcctaaaa gatgccaaga tcatcgacgc gtatgaagtg 4020
gcgtacaacg ttgcccaggc ggtcggtgca gacggccggg agccggatct tcccgccatg 4080
cgtgatcgcc tgacgatggc gcgtcgtgtg cgcggctacg tggctaaagg ccagccagtc 4140
gtccctgctc gtcgggtgga aacgcagagc agccgagggc ggaaagctct agcgacgatg 4200
gggcgacggg gcgcagctac atcgaatgca cgcagatggg ctgacccaga aagtaagtat 4260
gcgcaggaga cgcgacagcg attagcggaa gcaaacaaac gccgagaaat gacaggcgag 4320
ttgctcgaac ttcgcgtcaa aactgcgatc ctggatgccc gttctcaatc ggttgctgat 4380
ccctcgactc gtgagcttgc aggcgaacta ggtgtcagtg aaaggcgcat ccaacaagtc 4440
agaaaggcac ttggaatgga agctaaacgc ggccgtccac gggctgaaaa ctaataaacg 4500
aaacaccgtc agcagaaaac ggttcccccc tttaggggtc ccgtccttgc tctggctctc 4560
acttgccctc accctccgct atccacgggc tgaaaactaa taaacgaaac accgtcagca 4620
gaaaacggtt cccccccttt agggtgtctc gctcctagct ctgatccctc cccggttcct 4680
ccccggcctg atttttaagg ggggctcacg ctgtcggcag agaacggttc cccgccttct 4740
gctctggctc ttcctcgact ccctccccct caaaaatctc ctcgagatcc tggagacctt 4800
tttggagcta gcgcgttgct gcttcgcacc aacttgctca tgatgatttt catttttgct 4860
tgtgtgcttt tttgggttga accctccaaa gaggggaaac caggggcaca cctcatgcac 4920
taaagtgccg cttcgctggt cagggtgaaa tcacctggaa aaaaagtgcg gtaaccgctg 4980
cgcttggcgt tttttctggg caagaagtct cgcaggtttt cgcaggagtg ccggaagaaa 5040
ttatcagaat tggggctaga atttttaacg aacgttcgtt ataatggtgt catgaccttc 5100
acgacgaagt accaaaactg gcctgaagca tcagcggtgg atctctccga tgtcgcgctg 5160
gagtccgacg cactcgatgc cgccgtcgat ttaaaaacgg tgatcggatt tttccgcgcc 5220
ctcgatacga cagacgcgcc agcatcacgc gactgggcaa gtgccgcgag cgacctagaa 5280
acgcttgtgg ccgaccttga agagctggcc gacgagctgc gtgctcggca gcgccaggag 5340
gacgcgcagt agtggaggat cgcatcagct gcgcctactg cggtggcctg atcccacccc 5400
ggcctgaccc acgaggacgg cgcgcaaaat actgctcaga cgcgtgtcgt gccgcagcca 5460
gccgcgagcg cgccaacaag cgccacgccc aggaggtcga agccgcacgt cgaccgcgtg 5520
tagtgcgtgg cggaaacttc ttgcgtttcg caagagaaat gcgtcccatt tctcgtcgga 5580
ctcggggaag gaagcgtgat gctctcggtc aagcacgtcg ctcgccagcg ctgcgaggag 5640
ttcggccttc gtgcggaagt gccagtagag gccgggctgc tgtacctgta agtgagccgc 5700
cagcgcgcga gtggtgaagc catcgagccc agtctcgtcg agcacctgcc gggccccgag 5760
caacacggac gtgcggtcga gacgcttccg gtggtgagtc atagttgcac tttatcatcg 5820
ataactttat cttagataaa gtgactgctc gctactctca tctgactgct cgctactctc 5880
atcgtggaat cctgacagcc gtgctcatca cggcgaccct cgatgctgca gggctgggcc 5940
tcgtgatgcc gatcttgcct acccttctcg accaggtcgg tgcccccgac gacatgatcc 6000
cactgcacgt cggactactg acagcgctct atgcgatcat gcagtttctt tgcgccccga 6060
tccttggccg actctctgac cgtttcggac gccgccgcgt gcttgtcgcc tccctcgcag 6120
gcgcgacgat cgactacctc gtgctcgcac tgacggacac gctgtgggtc ttttacctcg 6180
cccgcgcggt tgcaggcatt accggcgcca cgaacgccgt caccgcgacg gtgatcgccg 6240
acattactcc gccggatcag cgcgcaaaac gctacgggtg gctcggcgca tgctacggcg 6300
gtggcatgat cgcgggtccc gccattggcg gtcttttcgg cggggtctca ccgcatctgc 6360
cattcctcgt cgccgccgcg ctcgccggaa tcaccctcgt actcagcgcg agtcttctgc 6420
gtgagacgcg gccaccgggc agcaacggct cgcacgcaca gcaacccggt acggcgaagc 6480
gaaccgcagt gccggggatg cttatccttc tcgcagtctt cggcatcgtg cagttcatcg 6540
gccaagcacc aggctccacc tgggtgctct tcacgcagca gcgcctcgac tggaaccccg 6600
tcgaagtcgg cgtttcgcta tccatcttcg gaatggtgca agtattcgtg caggcggcac 6660
tgaccggacg catcgtgtcc cggatcggcg agacccgggc gatcctcgtc ggtatcgccg 6720
cagacgccat tgggctcatc ggccttgccc tcatcgccag cacatgggcg atgctaccga 6780
tcctcgcagc gctcggactc ggcagcatca cgttgcccgc actgcagacg ctgctctcga 6840
gacgcgcgcc cgagcagcag cagggacgcc tgcagggaac acttgcaagc ctgaacagcc 6900
tcacctcgat catcggcccg gtcaccttca ccggcatttt cgcactcacc cgaacgaatg 6960
cagacggcac cctctggatc tgcgccgcag cgctctacgt tctctgcgcc ctcctgatga 7020
tccgtgagac atgcgcctca cggcgatctc gataaccgcg ctaaggtgcc atcccgatgc 7080
gacgggatcg ctctgccacc agtcaagtct cccgtagccg gtatgagcat gaccaaaatc 7140
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 7200
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 7260
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 7320
ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 7380
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 7440
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 7500
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 7560
acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 7620
gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 7680
gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 7740
cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 7800
aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 7860
gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 7920
cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg 7980
atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc 8040
agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg 8100
actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 8160
gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 8220
agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg 8280
cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc gcggtatggc 8340
atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata 8400
cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc 8460
cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta 8520
cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc 8580
cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc 8640
cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg 8700
taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc 8760
gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt 8820
tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac 8880
gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg 8940
cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg 9000
caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca 9060
acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga 9120
tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga 9180
tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgtcaac 9240
caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact 9300
ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa 9360
aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 9420
gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg 9480
tgagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg tgcaccaatg 9540
cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg 9600
cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc 9660
ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc cggctcgtat 9720
aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaccat ggaattcgag 9780
ctcggtaccc ggg 9793
<210> 53
<211> 10780
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 53
gatctggcat ttttgggagg tgtgaaatgc ggcgcgaaag tctgttggta tcggtttgca 60
agggcctgcg ggtacatgtc gagcgcgttg ggcaggatcc cgggcgcagc acggtgatgc 120
tggtcaacgg cgcgatggcg accaccgcct cgttcgcccg gacctgcaag tgcctggccg 180
aacatttcaa cgtggtgctg ttcgacctgc ccttcgccgg gcagtcgcgt cagcacaacc 240
cgcagcgggg gttgatcacc aaggacgacg aggtggaaat cctcctggcg ctgatcgagc 300
gcttcgaggt caatcacctg gtctccgcgt cctggggcgg tatctccacg ctgctggcgc 360
tgtcgcgcaa tccgcgcggc atccgcagct cggtggtgat ggcattcgcc cctggactga 420
accaggcgat gctcgactac gtcgggcggg cgcaggcgct gatcgagctg gacgacaagt 480
cggcgatcgg ccatctgctc aacgagaccg tcggcaaata cctgccgccg cgcctgaaag 540
ccagcaacca tcagcacatg gcttcgctgg ccaccggcga atacgagcag gcgcgctttc 600
acatcgacca ggtgctggcg ctcaacgatc ggggctacct ggcttgcctg gagcggatcc 660
agagccacgt gcatttcatc aacggcagct gggacgaata caccaccgcc gaggacgccc 720
gccagttccg cgactacctg ccgcactgca gtttctcgcg ggtggagggc accgggcatt 780
tcctcgacct ggagtccaag ctggccgcgg tacgcgtgca ccgcgccctg ctcgagcacc 840
tgctgaagca accggagccg cagcgggcgg aacgcgcggc gggattccac gagatggcca 900
tcggctacgc ctgaaccctt gacctgcgaa gacccggcct ggccgggctt tgcggttgca 960
taacgcacgg agtagcacca tgcacgccat cctcatcgcc atcggctcgg ccggcgacgt 1020
atttcccttc atcggcctgg cccggaccct gaaattgcgc gggcaccgcg tgagcctctg 1080
caccatcccg gtgtttcgcg acgcggtgga gcagcacggc atcgcgttcg tcccgctgag 1140
cgacgaactg acctaccgcc ggaccatggg cgatccgcgc ctgtgggacc ccaagacgtc 1200
cttcggcgtg ctctggcaaa ccatcgccgg gatgatcgag ccggtctacg agtacgtctc 1260
ggcgcagcgc catgacgaca tcgtggtggt cggctcgctc tgggcgctgg gcgcacgcat 1320
cgctcacgag aagtacggga ttccctacct gtccgcgcag gtctcgccat cgaccttgtt 1380
gtcggcgcac ctgccgccgg tacaccccaa gttcaacgtg cccgagcaga tgccgctggc 1440
gatgcgcaag ctgctctggc gctgcatcga gcgcttcaag ctggatcgca cctgcgcgcc 1500
ggatatcaac gcggtgcggc gcaaggtcgg cctggagacg ccggtgaagc gcatcttcac 1560
ccaatggatg cattcgccgc agggcgtggt ctgcctgttc ccggcctggt tcgcgccgcc 1620
ccagcaggat tggccgcaac ccctgcacat gaccggcttc ccgctgttcg acggcagtat 1680
cccggggacc ccgctcgacg acgaactgca acgctttctc gatcagggca gccggccgct 1740
ggtgttcacc cagggctcga ccgaacacct gcagggcgac ttctacgcca tggccctgcg 1800
cgcgctggaa cgcctcggcg cgcgtgggat cttcctcacc ggcgccggcc aggaaccgct 1860
gcgcggcttg ccgaaccacg tgctgcagcg cgcctacgcg ccactgggag ccttgctgcc 1920
atcgtgcgcc gggctggtcc atccgggcgg tatcggcgcc atgagcctgg ccttggcggc 1980
gggggtgccg caggtgctgc tgccctgcgc ccacgaccag ttcgacaatg ccgaacggct 2040
ggtccggctc ggctgcggga tgcgcctggg cgtgccattg cgcgagcagg agttgcgcgg 2100
ggcgctgtgg cgcttgctcg aggacccggc catggcggcg gcctgtcggc gtttcatgga 2160
attgtcacaa ccgcacagta tcgcttgcgg taaagcggcc caggtggtcg aacgttgtca 2220
tagggagggg gatgcgcgat ggctgaaggc tgcgtcctga cctacgggag aagaacgatc 2280
atggaccgga tagacatggg cgtgctggtg gtactgttca atcctggcga cgacgacctg 2340
gaacaccttg gcgaactggc ggcggcgttt ccgcaactgc gcttccttgc cgtcgacaac 2400
tcaccgcaca gcgatccgca gcgcaatgcc cggctgcgcg ggcaaggcat cgccgtgctg 2460
caccacggca accggcaggg catcgccggc gccttcaacc agggactcga cgcgctattc 2520
cggcgtggcg tgcagggtgt gctgctgctc gaccaggact cccgtcccgg cggcgccttc 2580
ctcgccgccc agtggcgcaa cctgcaggcg cgcaacggtc aggcctgcct gctcggccca 2640
cggatcttcg accggggtga ccggcgcttc ctgccggcca tccatctcga cggactgacg 2700
ctcaggcaat tgtctctgga cggcctgacg accccgcagc gcacctcgtt cctgatctcc 2760
tccggctgcc tgctgacccg cgaggcctac cagcgcctcg gccacttcga cgaggaactg 2820
ttcatcgacc acgtggacac cgaatacagc ctgcgcgccc aggcgctgga cgtgcccctg 2880
tacgtcgacc cgcggctggt cctcgagcac cgcatcggca cgcgcaagac ccgccgcctc 2940
ggcggtctca gcctcagcgc gatgaaccac gccccgctgc gccgctacta cctggcgcgc 3000
aacggcctgc tggtcctgcg ccgctacgcc cggtcctcgc cgctggccct gctggcgaac 3060
ctgccgaccc tgacccaggg cctcgcggtg ctcctgctcg aacgcgacaa gctgctcaag 3120
ctgcgctgcc tgggctgggg cctgtgggac ggcctgcggg gacgcggcgg cgcgctggag 3180
accaaccgcc cgcgcctgct gaagcgcctc gccggcccgg ccgtggcgtc cgtagcttcc 3240
ggcaaggcca aggcctagtc ggcgaaacgc attccctcta gagtcgacct gcaggcatgc 3300
aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa 3360
cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct 3420
gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc 3480
catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg 3540
ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc 3600
gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc 3660
ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt 3720
tctacaaact ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgctc 3780
cttcgtcggt gtcgtcgccg gatggtctgc ggtggtgctc agcgtggaga cgcgcaccgt 3840
cacggacccc catcaatcct gcctatttgc cacgtttaac aaggtagtta agcgttcatt 3900
tacgaagaaa acacgataag ctgcacaaat acctgaaaaa gttgaacgcc ccgtgagcgg 3960
gaactcacag ggcgtcggct aacccccagt catcagctgg gagaaagcac tcaagacatg 4020
actctagccg atccgcagga cacagtcaca gctagcgcgt ggaaattgtc cgccgatctg 4080
ttcgacaccc accccgaagc tatgcgctgc ggctcacgcg gctggacggc agaagatcgc 4140
cgcgaactgc tcgctcacct gggacgcgaa agcttccagg gcagcaagac aagagatttc 4200
gcgagcgcct ggattaaaaa cccggatacc ggcgaaaccc aaccaaagct ctaccgggct 4260
ggctcaaaag cgctgacgcg gtgccagtac gttgcgctga cgcacgcgca acatgccgcg 4320
gtgatcgtgc ttgacatcga tgtgcccagc caccaggccg gcgggaagat tgagcacgta 4380
aacccgcagg tctacgcgat tttagagaaa tgggcacgcc tagaaaaagc gccggcttgg 4440
atcggcgtga atccgctgag cgggaaatgc cagctcatct ggctcattga cccggtgtat 4500
gccgcagcag gtaaaaccag cccaaatatg cgcctgctgg ctgcaacgac ggaagaaatg 4560
actcgtgttt tcggcgctga ccaggctttt tcgcataggc tgagccggtg gccgctgcac 4620
gtctcagacg atccgacagc ctataaatgg cactgccagc atgatcgtgt ggatcggctg 4680
gccgacctaa tggagattgc tcgaacgatg accggatcac agaagccgaa aaagtacatt 4740
gagcaggact tttccagcgg acgcgcccgc attgaagcgg cacaacgcgc caccgcagaa 4800
gccaaggcgc tagcgatttt ggacgcgagc ctgccgagcg ccctggacgc gtccggcgac 4860
ctgatcgacg gcgtgcgagt gctctggaca aatccagagc gagcgcgcga cgagaccgcg 4920
tttcgccacg cgttgaccgt gggataccag ctcaaagctg ctggtgagcg cctaaaagat 4980
gccaagatca tcgacgcgta tgaagtggcg tacaacgttg cccaggcggt cggtgcagac 5040
ggccgggagc cggatcttcc cgccatgcgt gatcgcctga cgatggcgcg tcgtgtgcgc 5100
ggctacgtgg ctaaaggcca gccagtcgtc cctgctcgtc gggtggaaac gcagagcagc 5160
cgagggcgga aagctctagc gacgatgggg cgacggggcg cagctacatc gaatgcacgc 5220
agatgggctg acccagaaag taagtatgcg caggagacgc gacagcgatt agcggaagca 5280
aacaaacgcc gagaaatgac aggcgagttg ctcgaacttc gcgtcaaaac tgcgatcctg 5340
gatgcccgtt ctcaatcggt tgctgatccc tcgactcgtg agcttgcagg cgaactaggt 5400
gtcagtgaaa ggcgcatcca acaagtcaga aaggcacttg gaatggaagc taaacgcggc 5460
cgtccacggg ctgaaaacta ataaacgaaa caccgtcagc agaaaacggt tccccccttt 5520
aggggtcccg tccttgctct ggctctcact tgccctcacc ctccgctatc cacgggctga 5580
aaactaataa acgaaacacc gtcagcagaa aacggttccc cccctttagg gtgtctcgct 5640
cctagctctg atccctcccc ggttcctccc cggcctgatt tttaaggggg gctcacgctg 5700
tcggcagaga acggttcccc gccttctgct ctggctcttc ctcgactccc tccccctcaa 5760
aaatctcctc gagatcctgg agaccttttt ggagctagcg cgttgctgct tcgcaccaac 5820
ttgctcatga tgattttcat ttttgcttgt gtgctttttt gggttgaacc ctccaaagag 5880
gggaaaccag gggcacacct catgcactaa agtgccgctt cgctggtcag ggtgaaatca 5940
cctggaaaaa aagtgcggta accgctgcgc ttggcgtttt ttctgggcaa gaagtctcgc 6000
aggttttcgc aggagtgccg gaagaaatta tcagaattgg ggctagaatt tttaacgaac 6060
gttcgttata atggtgtcat gaccttcacg acgaagtacc aaaactggcc tgaagcatca 6120
gcggtggatc tctccgatgt cgcgctggag tccgacgcac tcgatgccgc cgtcgattta 6180
aaaacggtga tcggattttt ccgcgccctc gatacgacag acgcgccagc atcacgcgac 6240
tgggcaagtg ccgcgagcga cctagaaacg cttgtggccg accttgaaga gctggccgac 6300
gagctgcgtg ctcggcagcg ccaggaggac gcgcagtagt ggaggatcgc atcagctgcg 6360
cctactgcgg tggcctgatc ccaccccggc ctgacccacg aggacggcgc gcaaaatact 6420
gctcagacgc gtgtcgtgcc gcagccagcc gcgagcgcgc caacaagcgc cacgcccagg 6480
aggtcgaagc cgcacgtcga ccgcgtgtag tgcgtggcgg aaacttcttg cgtttcgcaa 6540
gagaaatgcg tcccatttct cgtcggactc ggggaaggaa gcgtgatgct ctcggtcaag 6600
cacgtcgctc gccagcgctg cgaggagttc ggccttcgtg cggaagtgcc agtagaggcc 6660
gggctgctgt acctgtaagt gagccgccag cgcgcgagtg gtgaagccat cgagcccagt 6720
ctcgtcgagc acctgccggg ccccgagcaa cacggacgtg cggtcgagac gcttccggtg 6780
gtgagtcata gttgcacttt atcatcgata actttatctt agataaagtg actgctcgct 6840
actctcatct gactgctcgc tactctcatc gtggaatcct gacagccgtg ctcatcacgg 6900
cgaccctcga tgctgcaggg ctgggcctcg tgatgccgat cttgcctacc cttctcgacc 6960
aggtcggtgc ccccgacgac atgatcccac tgcacgtcgg actactgaca gcgctctatg 7020
cgatcatgca gtttctttgc gccccgatcc ttggccgact ctctgaccgt ttcggacgcc 7080
gccgcgtgct tgtcgcctcc ctcgcaggcg cgacgatcga ctacctcgtg ctcgcactga 7140
cggacacgct gtgggtcttt tacctcgccc gcgcggttgc aggcattacc ggcgccacga 7200
acgccgtcac cgcgacggtg atcgccgaca ttactccgcc ggatcagcgc gcaaaacgct 7260
acgggtggct cggcgcatgc tacggcggtg gcatgatcgc gggtcccgcc attggcggtc 7320
ttttcggcgg ggtctcaccg catctgccat tcctcgtcgc cgccgcgctc gccggaatca 7380
ccctcgtact cagcgcgagt cttctgcgtg agacgcggcc accgggcagc aacggctcgc 7440
acgcacagca acccggtacg gcgaagcgaa ccgcagtgcc ggggatgctt atccttctcg 7500
cagtcttcgg catcgtgcag ttcatcggcc aagcaccagg ctccacctgg gtgctcttca 7560
cgcagcagcg cctcgactgg aaccccgtcg aagtcggcgt ttcgctatcc atcttcggaa 7620
tggtgcaagt attcgtgcag gcggcactga ccggacgcat cgtgtcccgg atcggcgaga 7680
cccgggcgat cctcgtcggt atcgccgcag acgccattgg gctcatcggc cttgccctca 7740
tcgccagcac atgggcgatg ctaccgatcc tcgcagcgct cggactcggc agcatcacgt 7800
tgcccgcact gcagacgctg ctctcgagac gcgcgcccga gcagcagcag ggacgcctgc 7860
agggaacact tgcaagcctg aacagcctca cctcgatcat cggcccggtc accttcaccg 7920
gcattttcgc actcacccga acgaatgcag acggcaccct ctggatctgc gccgcagcgc 7980
tctacgttct ctgcgccctc ctgatgatcc gtgagacatg cgcctcacgg cgatctcgat 8040
aaccgcgcta aggtgccatc ccgatgcgac gggatcgctc tgccaccagt caagtctccc 8100
gtagccggta tgagcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 8160
gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 8220
tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 8280
ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 8340
ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 8400
gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 8460
ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 8520
tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 8580
ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 8640
agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 8700
agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 8760
gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 8820
tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 8880
accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 8940
gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt 9000
atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 9060
cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa 9120
cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 9180
tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 9240
ggcagcagat caattcgcgc gcgaaggcga agcggcatgc atttacgttg acaccatcga 9300
atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt 9360
ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca 9420
gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt 9480
ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg 9540
caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca 9600
aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat 9660
ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg 9720
cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc 9780
tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag 9840
tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg 9900
tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct 9960
ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg 10020
cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt 10080
tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac 10140
cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga 10200
cagctcatgt tatatcccgc cgtcaaccac catcaaacag gattttcgcc tgctggggca 10260
aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct 10320
gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc 10380
tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 10440
cgggcagtga gcgcaacgca attaatgtga gttagcgcga attgatctgg tttgacagct 10500
tatcatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg 10560
tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt 10620
ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct 10680
gttgacaatt aatcatccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca 10740
cacaggaaac agaccatgga attcgagctc ggtacccggg 10780
<210> 54
<211> 11073
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 54
gatctggcat ttttgggagg tgtgaaatgc ggcgcgaaag tctgttggta tcggtttgca 60
agggcctgcg ggtacatgtc gagcgcgttg ggcaggatcc cgggcgcagc acggtgatgc 120
tggtcaacgg cgcgatggcg accaccgcct cgttcgcccg gacctgcaag tgcctggccg 180
aacatttcaa cgtggtgctg ttcgacctgc ccttcgccgg gcagtcgcgt cagcacaacc 240
cgcagcgggg gttgatcacc aaggacgacg aggtggaaat cctcctggcg ctgatcgagc 300
gcttcgaggt caatcacctg gtctccgcgt cctggggcgg tatctccacg ctgctggcgc 360
tgtcgcgcaa tccgcgcggc atccgcagct cggtggtgat ggcattcgcc cctggactga 420
accaggcgat gctcgactac gtcgggcggg cgcaggcgct gatcgagctg gacgacaagt 480
cggcgatcgg ccatctgctc aacgagaccg tcggcaaata cctgccgccg cgcctgaaag 540
ccagcaacca tcagcacatg gcttcgctgg ccaccggcga atacgagcag gcgcgctttc 600
acatcgacca ggtgctggcg ctcaacgatc ggggctacct ggcttgcctg gagcggatcc 660
agagccacgt gcatttcatc aacggcagct gggacgaata caccaccgcc gaggacgccc 720
gccagttccg cgactacctg ccgcactgca gtttctcgcg ggtggagggc accgggcatt 780
tcctcgacct ggagtccaag ctggccgcgg tacgcgtgca ccgcgccctg ctcgagcacc 840
tgctgaagca accggagccg cagcgggcgg aacgcgcggc gggattccac gagatggcca 900
tcggctacgc ctgaaccctt gacctgcgaa gacccggcct ggccgggctt tgcggttgca 960
taacgcacgg agtagcacca tgcacgccat cctcatcgcc atcggctcgg ccggcgacgt 1020
atttcccttc atcggcctgg cccggaccct gaaattgcgc gggcaccgcg tgagcctctg 1080
caccatcccg gtgtttcgcg acgcggtgga gcagcacggc atcgcgttcg tcccgctgag 1140
cgacgaactg acctaccgcc ggaccatggg cgatccgcgc ctgtgggacc ccaagacgtc 1200
cttcggcgtg ctctggcaaa ccatcgccgg gatgatcgag ccggtctacg agtacgtctc 1260
ggcgcagcgc catgacgaca tcgtggtggt cggctcgctc tgggcgctgg gcgcacgcat 1320
cgctcacgag aagtacggga ttccctacct gtccgcgcag gtctcgccat cgaccttgtt 1380
gtcggcgcac ctgccgccgg tacaccccaa gttcaacgtg cccgagcaga tgccgctggc 1440
gatgcgcaag ctgctctggc gctgcatcga gcgcttcaag ctggatcgca cctgcgcgcc 1500
ggatatcaac gcggtgcggc gcaaggtcgg cctggagacg ccggtgaagc gcatcttcac 1560
ccaatggatg cattcgccgc agggcgtggt ctgcctgttc ccggcctggt tcgcgccgcc 1620
ccagcaggat tggccgcaac ccctgcacat gaccggcttc ccgctgttcg acggcagtat 1680
cccggggacc ccgctcgacg acgaactgca acgctttctc gatcagggca gccggccgct 1740
ggtgttcacc cagggctcga ccgaacacct gcagggcgac ttctacgcca tggccctgcg 1800
cgcgctggaa cgcctcggcg cgcgtgggat cttcctcacc ggcgccggcc aggaaccgct 1860
gcgcggcttg ccgaaccacg tgctgcagcg cgcctacgcg ccactgggag ccttgctgcc 1920
atcgtgcgcc gggctggtcc atccgggcgg tatcggcgcc atgagcctgg ccttggcggc 1980
gggggtgccg caggtgctgc tgccctgcgc ccacgaccag ttcgacaatg ccgaacggct 2040
ggtccggctc ggctgcggga tgcgcctggg cgtgccattg cgcgagcagg agttgcgcgg 2100
ggcgctgtgg cgcttgctcg aggacccggc catggcggcg gcctgtcggc gtttcatgga 2160
attgtcacaa ccgcacagta tcgcttgcgg taaagcggcc caggtggtcg aacgttgtca 2220
tagggagggg gatgcgcgat ggctgaaggc tgcgtcctga cgccgggagg atcctggcgt 2280
gtccacgacc agcctctgcc cctccgccac gcgggaacac ggtcccggcg cgaaacgcgt 2340
cctgcctctg ctgttcctca cctgcctgct ggatgccgct ggcgtcggcc tgatcgtgcc 2400
cctgctgccg acgctgatcg gcagcgtggc gccgctggcg gtccgcgacg cggccacctg 2460
gggcgccgcc ctggtgatga ccttcgcgct gctgcaattg ttcttttcgc cggtcctcgg 2520
cagcctcagc gaccgcttcg gacgccgccc cgtcctggtc ctggcgatgc tcggcttcgc 2580
cctcagctat ctgctgctgg cgctggccga cagcctctgg atgctgttcc tcggtcgcgc 2640
gctggccggg ctcaccggcg ccagcgtggc caccgcgatg gcctgcgcgg ctgacctcgg 2700
cacgcacggg cagcgcaccc ggcacttcgg ctggctgtac gccggcctcg ccctgggcat 2760
gatcctcggc cccgccctcg gtgggctgct ggcggtgcac ggcacgacgc tgccgctgtt 2820
gctggccgcc ggcctgtgcc tgctcaacgc cctgctcgcc ggcctgttcc tcgaggaaac 2880
cctgcccccg acgcgacgcc gccgcctgga cccgaggcgg atgaatgcct tgcgctcgat 2940
cagcggcctg gctcggcaac cgggggtcgg acgcctgctg gcggtgcttg ccctggtatt 3000
cctcggcttg caggcggtga tggtggtctg gccgttcttc gtgatcgaga agtttcactg 3060
gagcagcgcc tggatcggct actcgctggc cctctacggc gtgctcgcgg tgctcgccca 3120
gaccctcggc gtgaacctct gcaagcggcg cctggacgac gcccgcctgc tgcgcctggg 3180
cctcgccctg caaggctgcg gcctgctgct gttcgccctg gtcgactcgt cattctggct 3240
ggtctgcgcg ctgctgccct tcgcgctcgg cagcctcgcc accccggcca tgcaggggct 3300
gctctcggcc cgcgtgccgg tcgaccgcca gggcgagttg cagggcgtgc tgagcagcct 3360
gatgagcctc gccgcgatcg tcggtccgcc gctgatgagc ggcctgttcc actggggcag 3420
cggtccgctc gcgccgctgc ccctggccgg cgcgccattc ctcgccggcg cccttctcgt 3480
tctggccggg ctggtcctgg cctggcaact tcgacctacg ggagaagaac gatcatggac 3540
cggatagaca tgggcgtgct ggtggtactt ctagagtcga cctgcaggca tgcaagcttg 3600
gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa 3660
gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca 3720
tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga 3780
gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt 3840
cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg 3900
gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact 3960
gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa 4020
actctttttg tttatttttc taaatacatt caaatatgta tccgctcatg ctccttcgtc 4080
ggtgtcgtcg ccggatggtc tgcggtggtg ctcagcgtgg agacgcgcac cgtcacggac 4140
ccccatcaat cctgcctatt tgccacgttt aacaaggtag ttaagcgttc atttacgaag 4200
aaaacacgat aagctgcaca aatacctgaa aaagttgaac gccccgtgag cgggaactca 4260
cagggcgtcg gctaaccccc agtcatcagc tgggagaaag cactcaagac atgactctag 4320
ccgatccgca ggacacagtc acagctagcg cgtggaaatt gtccgccgat ctgttcgaca 4380
cccaccccga agctatgcgc tgcggctcac gcggctggac ggcagaagat cgccgcgaac 4440
tgctcgctca cctgggacgc gaaagcttcc agggcagcaa gacaagagat ttcgcgagcg 4500
cctggattaa aaacccggat accggcgaaa cccaaccaaa gctctaccgg gctggctcaa 4560
aagcgctgac gcggtgccag tacgttgcgc tgacgcacgc gcaacatgcc gcggtgatcg 4620
tgcttgacat cgatgtgccc agccaccagg ccggcgggaa gattgagcac gtaaacccgc 4680
aggtctacgc gattttagag aaatgggcac gcctagaaaa agcgccggct tggatcggcg 4740
tgaatccgct gagcgggaaa tgccagctca tctggctcat tgacccggtg tatgccgcag 4800
caggtaaaac cagcccaaat atgcgcctgc tggctgcaac gacggaagaa atgactcgtg 4860
ttttcggcgc tgaccaggct ttttcgcata ggctgagccg gtggccgctg cacgtctcag 4920
acgatccgac agcctataaa tggcactgcc agcatgatcg tgtggatcgg ctggccgacc 4980
taatggagat tgctcgaacg atgaccggat cacagaagcc gaaaaagtac attgagcagg 5040
acttttccag cggacgcgcc cgcattgaag cggcacaacg cgccaccgca gaagccaagg 5100
cgctagcgat tttggacgcg agcctgccga gcgccctgga cgcgtccggc gacctgatcg 5160
acggcgtgcg agtgctctgg acaaatccag agcgagcgcg cgacgagacc gcgtttcgcc 5220
acgcgttgac cgtgggatac cagctcaaag ctgctggtga gcgcctaaaa gatgccaaga 5280
tcatcgacgc gtatgaagtg gcgtacaacg ttgcccaggc ggtcggtgca gacggccggg 5340
agccggatct tcccgccatg cgtgatcgcc tgacgatggc gcgtcgtgtg cgcggctacg 5400
tggctaaagg ccagccagtc gtccctgctc gtcgggtgga aacgcagagc agccgagggc 5460
ggaaagctct agcgacgatg gggcgacggg gcgcagctac atcgaatgca cgcagatggg 5520
ctgacccaga aagtaagtat gcgcaggaga cgcgacagcg attagcggaa gcaaacaaac 5580
gccgagaaat gacaggcgag ttgctcgaac ttcgcgtcaa aactgcgatc ctggatgccc 5640
gttctcaatc ggttgctgat ccctcgactc gtgagcttgc aggcgaacta ggtgtcagtg 5700
aaaggcgcat ccaacaagtc agaaaggcac ttggaatgga agctaaacgc ggccgtccac 5760
gggctgaaaa ctaataaacg aaacaccgtc agcagaaaac ggttcccccc tttaggggtc 5820
ccgtccttgc tctggctctc acttgccctc accctccgct atccacgggc tgaaaactaa 5880
taaacgaaac accgtcagca gaaaacggtt cccccccttt agggtgtctc gctcctagct 5940
ctgatccctc cccggttcct ccccggcctg atttttaagg ggggctcacg ctgtcggcag 6000
agaacggttc cccgccttct gctctggctc ttcctcgact ccctccccct caaaaatctc 6060
ctcgagatcc tggagacctt tttggagcta gcgcgttgct gcttcgcacc aacttgctca 6120
tgatgatttt catttttgct tgtgtgcttt tttgggttga accctccaaa gaggggaaac 6180
caggggcaca cctcatgcac taaagtgccg cttcgctggt cagggtgaaa tcacctggaa 6240
aaaaagtgcg gtaaccgctg cgcttggcgt tttttctggg caagaagtct cgcaggtttt 6300
cgcaggagtg ccggaagaaa ttatcagaat tggggctaga atttttaacg aacgttcgtt 6360
ataatggtgt catgaccttc acgacgaagt accaaaactg gcctgaagca tcagcggtgg 6420
atctctccga tgtcgcgctg gagtccgacg cactcgatgc cgccgtcgat ttaaaaacgg 6480
tgatcggatt tttccgcgcc ctcgatacga cagacgcgcc agcatcacgc gactgggcaa 6540
gtgccgcgag cgacctagaa acgcttgtgg ccgaccttga agagctggcc gacgagctgc 6600
gtgctcggca gcgccaggag gacgcgcagt agtggaggat cgcatcagct gcgcctactg 6660
cggtggcctg atcccacccc ggcctgaccc acgaggacgg cgcgcaaaat actgctcaga 6720
cgcgtgtcgt gccgcagcca gccgcgagcg cgccaacaag cgccacgccc aggaggtcga 6780
agccgcacgt cgaccgcgtg tagtgcgtgg cggaaacttc ttgcgtttcg caagagaaat 6840
gcgtcccatt tctcgtcgga ctcggggaag gaagcgtgat gctctcggtc aagcacgtcg 6900
ctcgccagcg ctgcgaggag ttcggccttc gtgcggaagt gccagtagag gccgggctgc 6960
tgtacctgta agtgagccgc cagcgcgcga gtggtgaagc catcgagccc agtctcgtcg 7020
agcacctgcc gggccccgag caacacggac gtgcggtcga gacgcttccg gtggtgagtc 7080
atagttgcac tttatcatcg ataactttat cttagataaa gtgactgctc gctactctca 7140
tctgactgct cgctactctc atcgtggaat cctgacagcc gtgctcatca cggcgaccct 7200
cgatgctgca gggctgggcc tcgtgatgcc gatcttgcct acccttctcg accaggtcgg 7260
tgcccccgac gacatgatcc cactgcacgt cggactactg acagcgctct atgcgatcat 7320
gcagtttctt tgcgccccga tccttggccg actctctgac cgtttcggac gccgccgcgt 7380
gcttgtcgcc tccctcgcag gcgcgacgat cgactacctc gtgctcgcac tgacggacac 7440
gctgtgggtc ttttacctcg cccgcgcggt tgcaggcatt accggcgcca cgaacgccgt 7500
caccgcgacg gtgatcgccg acattactcc gccggatcag cgcgcaaaac gctacgggtg 7560
gctcggcgca tgctacggcg gtggcatgat cgcgggtccc gccattggcg gtcttttcgg 7620
cggggtctca ccgcatctgc cattcctcgt cgccgccgcg ctcgccggaa tcaccctcgt 7680
actcagcgcg agtcttctgc gtgagacgcg gccaccgggc agcaacggct cgcacgcaca 7740
gcaacccggt acggcgaagc gaaccgcagt gccggggatg cttatccttc tcgcagtctt 7800
cggcatcgtg cagttcatcg gccaagcacc aggctccacc tgggtgctct tcacgcagca 7860
gcgcctcgac tggaaccccg tcgaagtcgg cgtttcgcta tccatcttcg gaatggtgca 7920
agtattcgtg caggcggcac tgaccggacg catcgtgtcc cggatcggcg agacccgggc 7980
gatcctcgtc ggtatcgccg cagacgccat tgggctcatc ggccttgccc tcatcgccag 8040
cacatgggcg atgctaccga tcctcgcagc gctcggactc ggcagcatca cgttgcccgc 8100
actgcagacg ctgctctcga gacgcgcgcc cgagcagcag cagggacgcc tgcagggaac 8160
acttgcaagc ctgaacagcc tcacctcgat catcggcccg gtcaccttca ccggcatttt 8220
cgcactcacc cgaacgaatg cagacggcac cctctggatc tgcgccgcag cgctctacgt 8280
tctctgcgcc ctcctgatga tccgtgagac atgcgcctca cggcgatctc gataaccgcg 8340
ctaaggtgcc atcccgatgc gacgggatcg ctctgccacc agtcaagtct cccgtagccg 8400
gtatgagcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 8460
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 8520
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 8580
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 8640
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 8700
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 8760
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 8820
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 8880
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 8940
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 9000
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 9060
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 9120
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 9180
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 9240
aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 9300
accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata 9360
cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc 9420
tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 9480
ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca 9540
gatcaattcg cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggtgc 9600
aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat 9660
gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt 9720
tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg 9780
gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag 9840
tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc 9900
gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa 9960
cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt 10020
gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc 10080
actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt 10140
ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag 10200
caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc 10260
tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg 10320
agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact 10380
gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc 10440
gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca 10500
tgttatatcc cgccgtcaac caccatcaaa caggattttc gcctgctggg gcaaaccagc 10560
gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc 10620
gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc 10680
gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 10740
tgagcgcaac gcaattaatg tgagttagcg cgaattgatc tggtttgaca gcttatcatc 10800
gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct 10860
gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata 10920
atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca 10980
attaatcatc cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga 11040
aacagaccat ggaattcgag ctcggtaccc ggg 11073
<210> 55
<211> 12263
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 55
gatctggcat ttttgggagg tgtgaaatgc ggcgcgaaag tctgttggta tcggtttgca 60
agggcctgcg ggtacatgtc gagcgcgttg ggcaggatcc cgggcgcagc acggtgatgc 120
tggtcaacgg cgcgatggcg accaccgcct cgttcgcccg gacctgcaag tgcctggccg 180
aacatttcaa cgtggtgctg ttcgacctgc ccttcgccgg gcagtcgcgt cagcacaacc 240
cgcagcgggg gttgatcacc aaggacgacg aggtggaaat cctcctggcg ctgatcgagc 300
gcttcgaggt caatcacctg gtctccgcgt cctggggcgg tatctccacg ctgctggcgc 360
tgtcgcgcaa tccgcgcggc atccgcagct cggtggtgat ggcattcgcc cctggactga 420
accaggcgat gctcgactac gtcgggcggg cgcaggcgct gatcgagctg gacgacaagt 480
cggcgatcgg ccatctgctc aacgagaccg tcggcaaata cctgccgccg cgcctgaaag 540
ccagcaacca tcagcacatg gcttcgctgg ccaccggcga atacgagcag gcgcgctttc 600
acatcgacca ggtgctggcg ctcaacgatc ggggctacct ggcttgcctg gagcggatcc 660
agagccacgt gcatttcatc aacggcagct gggacgaata caccaccgcc gaggacgccc 720
gccagttccg cgactacctg ccgcactgca gtttctcgcg ggtggagggc accgggcatt 780
tcctcgacct ggagtccaag ctggccgcgg tacgcgtgca ccgcgccctg ctcgagcacc 840
tgctgaagca accggagccg cagcgggcgg aacgcgcggc gggattccac gagatggcca 900
tcggctacgc ctgaaccctt gacctgcgaa gacccggcct ggccgggctt tgcggttgca 960
taacgcacgg agtagcacca tgcacgccat cctcatcgcc atcggctcgg ccggcgacgt 1020
atttcccttc atcggcctgg cccggaccct gaaattgcgc gggcaccgcg tgagcctctg 1080
caccatcccg gtgtttcgcg acgcggtgga gcagcacggc atcgcgttcg tcccgctgag 1140
cgacgaactg acctaccgcc ggaccatggg cgatccgcgc ctgtgggacc ccaagacgtc 1200
cttcggcgtg ctctggcaaa ccatcgccgg gatgatcgag ccggtctacg agtacgtctc 1260
ggcgcagcgc catgacgaca tcgtggtggt cggctcgctc tgggcgctgg gcgcacgcat 1320
cgctcacgag aagtacggga ttccctacct gtccgcgcag gtctcgccat cgaccttgtt 1380
gtcggcgcac ctgccgccgg tacaccccaa gttcaacgtg cccgagcaga tgccgctggc 1440
gatgcgcaag ctgctctggc gctgcatcga gcgcttcaag ctggatcgca cctgcgcgcc 1500
ggatatcaac gcggtgcggc gcaaggtcgg cctggagacg ccggtgaagc gcatcttcac 1560
ccaatggatg cattcgccgc agggcgtggt ctgcctgttc ccggcctggt tcgcgccgcc 1620
ccagcaggat tggccgcaac ccctgcacat gaccggcttc ccgctgttcg acggcagtat 1680
cccggggacc ccgctcgacg acgaactgca acgctttctc gatcagggca gccggccgct 1740
ggtgttcacc cagggctcga ccgaacacct gcagggcgac ttctacgcca tggccctgcg 1800
cgcgctggaa cgcctcggcg cgcgtgggat cttcctcacc ggcgccggcc aggaaccgct 1860
gcgcggcttg ccgaaccacg tgctgcagcg cgcctacgcg ccactgggag ccttgctgcc 1920
atcgtgcgcc gggctggtcc atccgggcgg tatcggcgcc atgagcctgg ccttggcggc 1980
gggggtgccg caggtgctgc tgccctgcgc ccacgaccag ttcgacaatg ccgaacggct 2040
ggtccggctc ggctgcggga tgcgcctggg cgtgccattg cgcgagcagg agttgcgcgg 2100
ggcgctgtgg cgcttgctcg aggacccggc catggcggcg gcctgtcggc gtttcatgga 2160
attgtcacaa ccgcacagta tcgcttgcgg taaagcggcc caggtggtcg aacgttgtca 2220
tagggagggg gatgcgcgat ggctgaaggc tgcgtcctga cctacgggag aagaacgatc 2280
atggaccgga tagacatggg cgtgctggtg gtactgttca atcctggcga cgacgacctg 2340
gaacaccttg gcgaactggc ggcggcgttt ccgcaactgc gcttccttgc cgtcgacaac 2400
tcaccgcaca gcgatccgca gcgcaatgcc cggctgcgcg ggcaaggcat cgccgtgctg 2460
caccacggca accggcaggg catcgccggc gccttcaacc agggactcga cgcgctattc 2520
cggcgtggcg tgcagggtgt gctgctgctc gaccaggact cccgtcccgg cggcgccttc 2580
ctcgccgccc agtggcgcaa cctgcaggcg cgcaacggtc aggcctgcct gctcggccca 2640
cggatcttcg accggggtga ccggcgcttc ctgccggcca tccatctcga cggactgacg 2700
ctcaggcaat tgtctctgga cggcctgacg accccgcagc gcacctcgtt cctgatctcc 2760
tccggctgcc tgctgacccg cgaggcctac cagcgcctcg gccacttcga cgaggaactg 2820
ttcatcgacc acgtggacac cgaatacagc ctgcgcgccc aggcgctgga cgtgcccctg 2880
tacgtcgacc cgcggctggt cctcgagcac cgcatcggca cgcgcaagac ccgccgcctc 2940
ggcggtctca gcctcagcgc gatgaaccac gccccgctgc gccgctacta cctggcgcgc 3000
aacggcctgc tggtcctgcg ccgctacgcc cggtcctcgc cgctggccct gctggcgaac 3060
ctgccgaccc tgacccaggg cctcgcggtg ctcctgctcg aacgcgacaa gctgctcaag 3120
ctgcgctgcc tgggctgggg cctgtgggac ggcctgcggg gacgcggcgg cgcgctggag 3180
accaaccgcc cgcgcctgct gaagcgcctc gccggcccgg ccgtggcgtc cgtagcttcc 3240
ggcaaggcca aggcctagtc ggcgaaacgc attccctcta gatgagaggc cggcaaggat 3300
acccgactgg cgcacgggtc gcatcattat gacatcacgc cgcccgccgg cgttgccgcg 3360
accgttcgtc gaacctgtga attccggtag tttcccttgc cctcgctggc gtcccaagat 3420
caggatttcc tgtgttcgcc gggaggatcc tggcgtgtcc acgaccagcc tctgcccctc 3480
cgccacgcgg gaacacggtc ccggcgcgaa acgcgtcctg cctctgctgt tcctcacctg 3540
cctgctggat gccgctggcg tcggcctgat cgtgcccctg ctgccgacgc tgatcggcag 3600
cgtggcgccg ctggcggtcc gcgacgcggc cacctggggc gccgccctgg tgatgacctt 3660
cgcgctgctg caattgttct tttcgccggt cctcggcagc ctcagcgacc gcttcggacg 3720
ccgccccgtc ctggtcctgg cgatgctcgg cttcgccctc agctatctgc tgctggcgct 3780
ggccgacagc ctctggatgc tgttcctcgg tcgcgcgctg gccgggctca ccggcgccag 3840
cgtggccacc gcgatggcct gcgcggctga cctcggcacg cacgggcagc gcacccggca 3900
cttcggctgg ctgtacgccg gcctcgccct gggcatgatc ctcggccccg ccctcggtgg 3960
gctgctggcg gtgcacggca cgacgctgcc gctgttgctg gccgccggcc tgtgcctgct 4020
caacgccctg ctcgccggcc tgttcctcga ggaaaccctg cccccgacgc gacgccgccg 4080
cctggacccg aggcggatga atgccttgcg ctcgatcagc ggcctggctc ggcaaccggg 4140
ggtcggacgc ctgctggcgg tgcttgccct ggtattcctc ggcttgcagg cggtgatggt 4200
ggtctggccg ttcttcgtga tcgagaagtt tcactggagc agcgcctgga tcggctactc 4260
gctggccctc tacggcgtgc tcgcggtgct cgcccagacc ctcggcgtga acctctgcaa 4320
gcggcgcctg gacgacgccc gcctgctgcg cctgggcctc gccctgcaag gctgcggcct 4380
gctgctgttc gccctggtcg actcgtcatt ctggctggtc tgcgcgctgc tgcccttcgc 4440
gctcggcagc ctcgccaccc cggccatgca ggggctgctc tcggcccgcg tgccggtcga 4500
ccgccagggc gagttgcagg gcgtgctgag cagcctgatg agcctcgccg cgatcgtcgg 4560
tccgccgctg atgagcggcc tgttccactg gggcagcggt ccgctcgcgc cgctgcccct 4620
ggccggcgcg ccattcctcg ccggcgccct tctcgttctg gccgggctgg tcctggcctg 4680
gcaacttcga cctacgggag aagaacgatc atggaccgga tagacatggg cgtgctggtg 4740
gtactgttca atcctggcgt ctagagtcga cctgcaggca tgcaagcttg gctgttttgg 4800
cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat 4860
aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc 4920
agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa 4980
ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct 5040
gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg 5100
ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc 5160
aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttttg 5220
tttatttttc taaatacatt caaatatgta tccgctcatg ctccttcgtc ggtgtcgtcg 5280
ccggatggtc tgcggtggtg ctcagcgtgg agacgcgcac cgtcacggac ccccatcaat 5340
cctgcctatt tgccacgttt aacaaggtag ttaagcgttc atttacgaag aaaacacgat 5400
aagctgcaca aatacctgaa aaagttgaac gccccgtgag cgggaactca cagggcgtcg 5460
gctaaccccc agtcatcagc tgggagaaag cactcaagac atgactctag ccgatccgca 5520
ggacacagtc acagctagcg cgtggaaatt gtccgccgat ctgttcgaca cccaccccga 5580
agctatgcgc tgcggctcac gcggctggac ggcagaagat cgccgcgaac tgctcgctca 5640
cctgggacgc gaaagcttcc agggcagcaa gacaagagat ttcgcgagcg cctggattaa 5700
aaacccggat accggcgaaa cccaaccaaa gctctaccgg gctggctcaa aagcgctgac 5760
gcggtgccag tacgttgcgc tgacgcacgc gcaacatgcc gcggtgatcg tgcttgacat 5820
cgatgtgccc agccaccagg ccggcgggaa gattgagcac gtaaacccgc aggtctacgc 5880
gattttagag aaatgggcac gcctagaaaa agcgccggct tggatcggcg tgaatccgct 5940
gagcgggaaa tgccagctca tctggctcat tgacccggtg tatgccgcag caggtaaaac 6000
cagcccaaat atgcgcctgc tggctgcaac gacggaagaa atgactcgtg ttttcggcgc 6060
tgaccaggct ttttcgcata ggctgagccg gtggccgctg cacgtctcag acgatccgac 6120
agcctataaa tggcactgcc agcatgatcg tgtggatcgg ctggccgacc taatggagat 6180
tgctcgaacg atgaccggat cacagaagcc gaaaaagtac attgagcagg acttttccag 6240
cggacgcgcc cgcattgaag cggcacaacg cgccaccgca gaagccaagg cgctagcgat 6300
tttggacgcg agcctgccga gcgccctgga cgcgtccggc gacctgatcg acggcgtgcg 6360
agtgctctgg acaaatccag agcgagcgcg cgacgagacc gcgtttcgcc acgcgttgac 6420
cgtgggatac cagctcaaag ctgctggtga gcgcctaaaa gatgccaaga tcatcgacgc 6480
gtatgaagtg gcgtacaacg ttgcccaggc ggtcggtgca gacggccggg agccggatct 6540
tcccgccatg cgtgatcgcc tgacgatggc gcgtcgtgtg cgcggctacg tggctaaagg 6600
ccagccagtc gtccctgctc gtcgggtgga aacgcagagc agccgagggc ggaaagctct 6660
agcgacgatg gggcgacggg gcgcagctac atcgaatgca cgcagatggg ctgacccaga 6720
aagtaagtat gcgcaggaga cgcgacagcg attagcggaa gcaaacaaac gccgagaaat 6780
gacaggcgag ttgctcgaac ttcgcgtcaa aactgcgatc ctggatgccc gttctcaatc 6840
ggttgctgat ccctcgactc gtgagcttgc aggcgaacta ggtgtcagtg aaaggcgcat 6900
ccaacaagtc agaaaggcac ttggaatgga agctaaacgc ggccgtccac gggctgaaaa 6960
ctaataaacg aaacaccgtc agcagaaaac ggttcccccc tttaggggtc ccgtccttgc 7020
tctggctctc acttgccctc accctccgct atccacgggc tgaaaactaa taaacgaaac 7080
accgtcagca gaaaacggtt cccccccttt agggtgtctc gctcctagct ctgatccctc 7140
cccggttcct ccccggcctg atttttaagg ggggctcacg ctgtcggcag agaacggttc 7200
cccgccttct gctctggctc ttcctcgact ccctccccct caaaaatctc ctcgagatcc 7260
tggagacctt tttggagcta gcgcgttgct gcttcgcacc aacttgctca tgatgatttt 7320
catttttgct tgtgtgcttt tttgggttga accctccaaa gaggggaaac caggggcaca 7380
cctcatgcac taaagtgccg cttcgctggt cagggtgaaa tcacctggaa aaaaagtgcg 7440
gtaaccgctg cgcttggcgt tttttctggg caagaagtct cgcaggtttt cgcaggagtg 7500
ccggaagaaa ttatcagaat tggggctaga atttttaacg aacgttcgtt ataatggtgt 7560
catgaccttc acgacgaagt accaaaactg gcctgaagca tcagcggtgg atctctccga 7620
tgtcgcgctg gagtccgacg cactcgatgc cgccgtcgat ttaaaaacgg tgatcggatt 7680
tttccgcgcc ctcgatacga cagacgcgcc agcatcacgc gactgggcaa gtgccgcgag 7740
cgacctagaa acgcttgtgg ccgaccttga agagctggcc gacgagctgc gtgctcggca 7800
gcgccaggag gacgcgcagt agtggaggat cgcatcagct gcgcctactg cggtggcctg 7860
atcccacccc ggcctgaccc acgaggacgg cgcgcaaaat actgctcaga cgcgtgtcgt 7920
gccgcagcca gccgcgagcg cgccaacaag cgccacgccc aggaggtcga agccgcacgt 7980
cgaccgcgtg tagtgcgtgg cggaaacttc ttgcgtttcg caagagaaat gcgtcccatt 8040
tctcgtcgga ctcggggaag gaagcgtgat gctctcggtc aagcacgtcg ctcgccagcg 8100
ctgcgaggag ttcggccttc gtgcggaagt gccagtagag gccgggctgc tgtacctgta 8160
agtgagccgc cagcgcgcga gtggtgaagc catcgagccc agtctcgtcg agcacctgcc 8220
gggccccgag caacacggac gtgcggtcga gacgcttccg gtggtgagtc atagttgcac 8280
tttatcatcg ataactttat cttagataaa gtgactgctc gctactctca tctgactgct 8340
cgctactctc atcgtggaat cctgacagcc gtgctcatca cggcgaccct cgatgctgca 8400
gggctgggcc tcgtgatgcc gatcttgcct acccttctcg accaggtcgg tgcccccgac 8460
gacatgatcc cactgcacgt cggactactg acagcgctct atgcgatcat gcagtttctt 8520
tgcgccccga tccttggccg actctctgac cgtttcggac gccgccgcgt gcttgtcgcc 8580
tccctcgcag gcgcgacgat cgactacctc gtgctcgcac tgacggacac gctgtgggtc 8640
ttttacctcg cccgcgcggt tgcaggcatt accggcgcca cgaacgccgt caccgcgacg 8700
gtgatcgccg acattactcc gccggatcag cgcgcaaaac gctacgggtg gctcggcgca 8760
tgctacggcg gtggcatgat cgcgggtccc gccattggcg gtcttttcgg cggggtctca 8820
ccgcatctgc cattcctcgt cgccgccgcg ctcgccggaa tcaccctcgt actcagcgcg 8880
agtcttctgc gtgagacgcg gccaccgggc agcaacggct cgcacgcaca gcaacccggt 8940
acggcgaagc gaaccgcagt gccggggatg cttatccttc tcgcagtctt cggcatcgtg 9000
cagttcatcg gccaagcacc aggctccacc tgggtgctct tcacgcagca gcgcctcgac 9060
tggaaccccg tcgaagtcgg cgtttcgcta tccatcttcg gaatggtgca agtattcgtg 9120
caggcggcac tgaccggacg catcgtgtcc cggatcggcg agacccgggc gatcctcgtc 9180
ggtatcgccg cagacgccat tgggctcatc ggccttgccc tcatcgccag cacatgggcg 9240
atgctaccga tcctcgcagc gctcggactc ggcagcatca cgttgcccgc actgcagacg 9300
ctgctctcga gacgcgcgcc cgagcagcag cagggacgcc tgcagggaac acttgcaagc 9360
ctgaacagcc tcacctcgat catcggcccg gtcaccttca ccggcatttt cgcactcacc 9420
cgaacgaatg cagacggcac cctctggatc tgcgccgcag cgctctacgt tctctgcgcc 9480
ctcctgatga tccgtgagac atgcgcctca cggcgatctc gataaccgcg ctaaggtgcc 9540
atcccgatgc gacgggatcg ctctgccacc agtcaagtct cccgtagccg gtatgagcat 9600
gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 9660
caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 9720
accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 9780
ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt 9840
aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 9900
accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 9960
gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 10020
ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 10080
gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 10140
gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 10200
ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 10260
aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 10320
gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 10380
tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 10440
agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg 10500
gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta 10560
tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc 10620
tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc 10680
tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg 10740
cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc 10800
gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag 10860
taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg 10920
tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg 10980
agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga 11040
ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta 11100
aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg 11160
tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca 11220
ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc 11280
cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg 11340
aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc 11400
tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat 11460
atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt 11520
ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg 11580
ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg 11640
ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc 11700
cgccgtcaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct 11760
tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg 11820
tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 11880
attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 11940
gcaattaatg tgagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg 12000
tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg 12060
taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg 12120
cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc 12180
cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaccat 12240
ggaattcgag ctcggtaccc ggg 12263
<210> 56
<211> 8471
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 56
aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattcact 60
ggccgtcgtt ttacagccaa gcttggctgt tttggcggat gagagaagat tttcagcctg 120
atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt 180
agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat 240
ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa 300
ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct 360
gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg 420
gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac 480
ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata 540
tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 600
gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 660
ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 720
cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 780
ccgaagaacg ttttccaatg atgagcactt ttgatccccc tgcggcgtcg ctgatcgccc 840
tcgcgacgtt gtgcgggtgg cttgtccctg agggcgctgc gacagatagc taaaaatctg 900
cgtcaggatc gccgtagagc gcgcgtcgcg tcgattggag gcttcccctt tggttgacgg 960
tcttcaatcg ctctacggcg atcctgacgc ttttttgttg cgtaccgtcg atcgttttat 1020
ttctgtcgat cccgaaaaag tttttgcctt ttgtaaaaaa cttctcggtc gccccgcaaa 1080
ttttcgattc cagatttttt aaaaaccaag ccagaaatac gacacaccgt ttgcagataa 1140
tctgtctttc ggaaaaatca agtgcgatac aaaattttta gcacccctga gctgcgcaaa 1200
gtcccgcttc gtgaaaattt tcgtgccgcg tgattttccg ccaaaaactt taacgaacgt 1260
tcgttataat ggtgtcatga ccttcacgac gaagtaccaa aattggcccg aatcatcagc 1320
tatggatctc tctgatgtcg cgctggagtc cgacgcgctc gatgctgccg tcgatttaaa 1380
aacggtgatc ggatttttcc gagctctcga tacgacggac gcgccagcat cacgagactg 1440
ggccagtgcc gcgagcgacc tagaaactct cgtggcggat cttgaggagc tggctgacga 1500
gctgcgtgct cggcagcgcc aggaggacgc acagtagtgg aggatcgaat cagttgcgcc 1560
tactgcggtg gcctgattcc tccccggcct gacccgcgag gacggcgcgc aaaatattgc 1620
tcagatgcgt gtcgtgccgc agccagccgc gagcgcgcca acaaacgcca cgccgaggag 1680
ctggaggcgg ctaggtcgca aatggcgctg gaagtgcgtc ccccgagcga aattttggcc 1740
atggtcgtca cagagctgga agcggcagcg agaattatcc gcgatcgtgg cgcggtgccc 1800
gcaggcatga caaacatcgt aaatgccgcg tttcgtgtgg ccgtggccgc ccaggacgtg 1860
tcagcgccgc caccacctgc accgaatcgg cagcagcgtc gcgcgtcgaa aaagcgcaca 1920
ggcggcaaga agcgataagc tgcacgaata cctgaaaaat gttgaacgcc ccgtgagcgg 1980
taactcacag ggcgtcggct aacccccagt ccaaacctgg gagaaagcgc tcaaaaatga 2040
ctctagcgga ttcacgagac attgacacac cggcctggaa attttccgct gatctgttcg 2100
acacccatcc cgagctcgcg ctgcgatcac gtggctggac gagcgaagac cgccgcgaat 2160
tcctcgctca cctgggcaga gaaaatttcc agggcagcaa gacccgcgac ttcgccagcg 2220
cttggatcaa agacccggac acgggagaaa cacagccgaa gttataccga gttggttcaa 2280
aatcgcttgc ccggtgccag tatgttgctc tgacgcacgc gcagcacgca gccgtgcttg 2340
tcctggacat tgatgtgccg agccaccagg ccggcgggaa aatcgagcac gtaaaccccg 2400
aggtctacgc gattttggag cgctgggcac gcctggaaaa agcgccagct tggatcggcg 2460
tgaatccact gagcgggaaa tgccagctca tctggctcat tgatccggtg tatgccgcag 2520
caggcatgag cagcccgaat atgcgcctgc tggctgcaac gaccgaggaa atgacccgcg 2580
ttttcggcgc tgaccaggct ttttcacata ggctgagccg gtggccactg cacgtctccg 2640
acgatcccac cgcgtaccgc tggcatgccc agcacaatcg cgtggatcgc ctagctgatc 2700
ttatggaggt tgctcgcatg atctcaggca cagaaaaacc taaaaaacgc tatgagcagg 2760
agttttctag cggacgggca cgtatcgaag cggcaagaaa agccactgcg gaagcaaaag 2820
cacttgccac gcttgaagca agcctgccga gcgccgctga agcgtctgga gagctgatcg 2880
acggcgtccg tgtcctctgg actgctccag ggcgtgccgc ccgtgatgag acggcttttc 2940
gccacgcttt gactgtggga taccagttaa aagcggctgg tgagcgccta aaagacacca 3000
agatcatcga cgcctacgag cgtgcctaca ccgtcgctca ggcggtcgga gcagacggcc 3060
gtgagcctga tctgccgccg atgcgtgacc gccagacgat ggcgcgacgt gtgcgcggct 3120
acgtcgctaa aggccagcca gtcgtccctg ctcgtcagac agagacgcag agcagccgag 3180
ggcgaaaagc tctggccact atgggaagac gtggcggtaa aaaggccgca gaacgctgga 3240
aagacccaaa cagtgagtac gcccgagcac agcgagaaaa actagctaag tccagtcaac 3300
gacaagctag gaaagctaaa ggaaatcgct tgaccattgc aggttggttt atgactgttg 3360
agggagagac tggctcgtgg ccgacaatca atgaagctat gtctgaattt agcgtgtcac 3420
gtcagaccgt gaatagagca cttaagtctg cgggcattga acttccacga ggacgccgta 3480
aagcttccca gtaaatgtgc catctcgtag gcagaaaacg gttccccccg taggggtctc 3540
tctcttggcc tcctttctag gtcgggctga ttgctcttga agctctctag gggggctcac 3600
accataggca gataacggtt ccccaccggc tcacctcgta agcgcacaag gactgctccc 3660
aaagatcttc aaagccactg ccgcgactcc gcttcgcgaa gccttgcccc gcggaaattt 3720
cctccaccga gttcgtgcac acccctatgc caagcttctt tcaccctaaa ttcgagagat 3780
tggattctta ccgtggaaat tcttcgcaaa aatcgtcccc tgatcgccct tgcgacgttg 3840
ctcgcggcgg tgccgctggt tgcgcttggc ttgaccgact tgatcctccg gcgttcagcc 3900
tgtgccacag ccgacaggat ggtgaccacc atttgcccca tatcaccgtc ggtactgatc 3960
ccgtcgtcaa taaaccgaac cgctacaccc tgagcatcaa actcttttat cagttggatc 4020
atgtcggcgg tgtcgcggcc aagacggtcg agcttcttca ccagaatgac atcaccttcc 4080
tccaccttca tcctcagcaa atccagccct tcccgatctg ttgaactgcc ggatgccttg 4140
tcggtaaaga tgcggttagc ttttacccct gcatctttga gcgctgaggt ctgcctcgtg 4200
aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga 4260
gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt 4320
gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag 4380
caaaagttcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca 4440
gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg 4500
caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga 4560
aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat 4620
tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc 4680
aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat 4740
ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc 4800
aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt 4860
aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc 4920
aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg 4980
gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg 5040
aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc 5100
aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg 5160
atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc 5220
agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct 5280
cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat 5340
atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat 5400
aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg 5460
ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat 5520
caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc 5580
agcaacacct tcttcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat 5640
gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac 5700
cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc 5760
tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc 5820
tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt 5880
ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg 5940
aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc 6000
tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca 6060
ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg 6120
aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6180
cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta 6240
gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct 6300
cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg 6360
ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct 6420
caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc acgttaaggg 6480
attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6540
agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6600
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6660
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6720
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6780
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6840
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6900
gccgatgata agctgtcaaa catggcctgt cgcttgcggt attcggaatc ttgcacgccc 6960
tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta 7020
tcgccggcat ggcggccgac gcgcggggag aggcggtttg cgtattgggc gccagggtgg 7080
tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag 7140
agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg 7200
tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga 7260
tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct 7320
gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt 7380
gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat 7440
tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg 7500
ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc 7560
gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa 7620
gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca 7680
gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt 7740
tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat 7800
cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg 7860
tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa 7920
tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc 7980
tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat 8040
cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc 8100
atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc aacgtaaatg catgccgctt 8160
cgccttcgcg cgcgaattgc aagctgatcc gggcttatcg actgcacggt gcaccaatgc 8220
ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc 8280
ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca 8340
taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa 8400
tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagaattaaa agatatgacc 8460
atgattacgc c 8471
<210> 57
<211> 12311
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 57
aagcttgcat gcctgcaggt cgactctaga attaatgcag ctggcacgac aggtttcccg 60
actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac 120
cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac 180
aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact 240
aaagggaaca aaagctgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg 300
atatcgaatt cgcgtcatct gtctacgaca acaccttttg tccaattaga gccaaattat 360
gattctagta acaggcggag ccggcttcat cggctcaaat ttcgtactgc aatggtgtgc 420
gcacaatgag gaacccgtcc tcaacctcga cgccctgacc tacgcaggca acctggccaa 480
cctgcagccg ctggaaggca accctcagca tcgctttgtg caaggcaata tttgcgatgc 540
tgcgcttctg accaagctgt tcgcagagca ccgcccgcgc gccgtggttc acttcgcggc 600
ggaatcccat gtagaccgct caatcaccgg ccccgaagcg tttgtcgaaa ccaacgtgat 660
gggcacgttt cgcttgcttg aagccgcccg ggcgcattgg aatagtttgg aaggtgcaga 720
gaaggaggcc ttccgtttcc tccatgtctc taccgacgaa gtctacggca cactagggcc 780
aaacgacccg gcgttcaccg aaaccacgcc gtacgcgccg aacagcccat actccgccag 840
caaggcagcc agcgaccatc tggtacgctc gtatttccat acctacggca tgccggtact 900
cactaccaac tgctccaaca attacgggcc gctccacttc ccggaaaaac tgatcccgct 960
gatgatcgtc aacgcactcg ccggtaaggc gctgcctgtc tatggcgacg gccagcaaat 1020
ccgcgactgg ctgtatgtcg aagatcactg ctcgggcatc cgtcgcgtac tggaagccgg 1080
tgcgttcggc gagacgtaca atattggcgg ctggaatgaa aaagccaaca ttgacattgt 1140
gcgtacactc tgcagccttc tcgacgagat ggcacctgcg gcatcgcgcc aggtaatcaa 1200
tcagaagacc ggcgagcctg tcgaacagta tgcagaactc atcgcctacg taaccgaccg 1260
cccaggccat gaccgccgtt atgccatcga tgcacgcaag atcgagcggg agctcggctg 1320
gaaacctgcc gaaaccttcg agacgggcat tcgaaagaca gtcgcttggt acttggccaa 1380
ccagaaatgg gtaaaaggtg tcatggacgg cagctaccgt gactgggtgg cacaacaata 1440
cggggcaaat aaagcgtgaa aatcctgctg ttggggaaaa acgggcaagt aggctgggag 1500
ctacagcgcg ccttggcgcc gctgggtgag gtcattgcgc tggatcgtca gggggccgag 1560
ggcttatgtg gcgacttgtc caacctggac ggcttggccg ctacgattcg tcagctggcg 1620
ccggacgtga tcgtcaacgc tgctgcctac actgcagtgg ataaagctga gagcgatcag 1680
gcactggctg caatgatcaa tgccgcggct cctgctgtat tagcacgtga aacagcagct 1740
ttgggcgcct ggttgattca ctattccacc gattatgtat ttgacggcag cggcagtcag 1800
cgctgggagg aaactgcgcc taccggcccc ctttcggtct acggccggac caagctggaa 1860
ggcgagcatg ccattctcgc cagcggcgcc aaggccgtgg tactgcgcac cagctgggtg 1920
tatgctgcgc gcgggcacaa ttttgccaag accatgctgc gcctggcggc ggagcgtgag 1980
acgttgagcg tggtagcaga ccaatttggc gcacccacgg gcgctgacct gatcgccgac 2040
gttactgcac acatcctgcg gcaaatcttc aatgggcaag acaaccgtca cctggcaggg 2100
atttaccact tggctgcgtc cggtgaaacc tcttggcatg gttttgctca gttcgtgctg 2160
gcgcatgctc aacgcactgg cgtagcgctg aaagtgacag ctgataaggt tgccgcaatc 2220
agcaccgaag cttatccagt acctgcacca cgtccgcgca actcgcgcct ggcactgggc 2280
aaactggaaa acacgttcaa tttcaaaatg ccgctttggg agcaaggcgt gcaacgtatg 2340
ctggacgaaa tccagtaata gggactctca tggctcgtaa aggaattatt ctggccggcg 2400
gttcgggtac acgcctgcat ccggccacac tttcggtttc gaagcagctg ctgccggtgt 2460
atgacaaacc gatgatctac tacccgctga gcaccctgct gctcgctggt atccgggaca 2520
tcctgatcat ttccaccccg caggacaccc cgcgcttcga acagctgctg ggcgatggca 2580
gccagtgggg cctgaacctg tcatacgcaa tacaaccaag cccggatggc ttggcgcaag 2640
cgttcaccat cggcgctgac ttcatcggta acgacccttc tgcgttggtt ctcggtgaca 2700
atattttcta cggccatgac ttccaggcac tgctattgaa cgcagataaa cgtgaatccg 2760
gtgcttcagt attcgcttat catgttcatg acccagaacg ctatggcgta gcggagtttg 2820
acgatagcgg tcgcgtattg tcgctggaag aaaaaccggc agttccaaag tctagctatg 2880
cggtcaccgg cctgtatttc tatgacaatc aggtagtcaa tctggctcgc gagctgaagc 2940
cttccccacg tggcgagctg gaaatcaccg acctcaacaa cctttacttg cagcagcagc 3000
agttgcaggt cgaaatcatg ggccgtggct atgcgtggct cgacaccggc acgcacgaca 3060
gtctgctgga ggctagccag tacatcgcaa ccatggagcg ccgtcagggc ttgaaagtcg 3120
cctgccctga ggaaatttgc taccgcgctg gctggatcaa cgctgagcaa ctcgagtgcc 3180
tggctcaacc actgctgaaa aacggttatg gcaagtatct gcagaacttg ctgaaagaga 3240
aggtgttctg atgcaagcca ttccgctgga tatccccgaa gtcgtgctgt ttacccccaa 3300
ggtttttggc gacgaacgtg gtttcttcta cgagagcttc aacgcccgtg ttttcagcga 3360
agtgaccggc ctgcagcccg acttcgtaca agacaaccac tcgcgctcgg taaaaggcgt 3420
gctccgtggc ctgcactatc agctggcacc tcacgcccag ggcaagctgg tgcgtgtggt 3480
gcaaggcgaa gtcttcgatg ttgcggtgga tatccgtcgc tcgtccacaa ccttcggtaa 3540
atgggtaggt gcggtgttgt cggccgagaa caagaaccag ctgtggatcc cgccagggtt 3600
cgcacacggg ttcgtcacgt tgagtgaaac cgcagagttc ctctacaaga ccaccgactt 3660
ctactcgccg cagtgcgagc gctgcattgc ctggaatgat ccggcagtgg gtatcgaatg 3720
gcccatcgac tccgtaccaa gcttgtctgg caaggaccag cttggggtcg cattggctga 3780
cgccgaactg ttcgactaac ggttttagcg gagaagggct gcggtagcgc agccgaattc 3840
ctgcagcccg ggggatccac tagttctaga ggatccccgg gtaccgagct cgaattcact 3900
ggccgtcgtt ttacagccaa gcttggctgt tttggcggat gagagaagat tttcagcctg 3960
atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt 4020
agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat 4080
ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa 4140
ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct 4200
gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg 4260
gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac 4320
ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata 4380
tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 4440
gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 4500
ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 4560
cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 4620
ccgaagaacg ttttccaatg atgagcactt ttgatccccc tgcggcgtcg ctgatcgccc 4680
tcgcgacgtt gtgcgggtgg cttgtccctg agggcgctgc gacagatagc taaaaatctg 4740
cgtcaggatc gccgtagagc gcgcgtcgcg tcgattggag gcttcccctt tggttgacgg 4800
tcttcaatcg ctctacggcg atcctgacgc ttttttgttg cgtaccgtcg atcgttttat 4860
ttctgtcgat cccgaaaaag tttttgcctt ttgtaaaaaa cttctcggtc gccccgcaaa 4920
ttttcgattc cagatttttt aaaaaccaag ccagaaatac gacacaccgt ttgcagataa 4980
tctgtctttc ggaaaaatca agtgcgatac aaaattttta gcacccctga gctgcgcaaa 5040
gtcccgcttc gtgaaaattt tcgtgccgcg tgattttccg ccaaaaactt taacgaacgt 5100
tcgttataat ggtgtcatga ccttcacgac gaagtaccaa aattggcccg aatcatcagc 5160
tatggatctc tctgatgtcg cgctggagtc cgacgcgctc gatgctgccg tcgatttaaa 5220
aacggtgatc ggatttttcc gagctctcga tacgacggac gcgccagcat cacgagactg 5280
ggccagtgcc gcgagcgacc tagaaactct cgtggcggat cttgaggagc tggctgacga 5340
gctgcgtgct cggcagcgcc aggaggacgc acagtagtgg aggatcgaat cagttgcgcc 5400
tactgcggtg gcctgattcc tccccggcct gacccgcgag gacggcgcgc aaaatattgc 5460
tcagatgcgt gtcgtgccgc agccagccgc gagcgcgcca acaaacgcca cgccgaggag 5520
ctggaggcgg ctaggtcgca aatggcgctg gaagtgcgtc ccccgagcga aattttggcc 5580
atggtcgtca cagagctgga agcggcagcg agaattatcc gcgatcgtgg cgcggtgccc 5640
gcaggcatga caaacatcgt aaatgccgcg tttcgtgtgg ccgtggccgc ccaggacgtg 5700
tcagcgccgc caccacctgc accgaatcgg cagcagcgtc gcgcgtcgaa aaagcgcaca 5760
ggcggcaaga agcgataagc tgcacgaata cctgaaaaat gttgaacgcc ccgtgagcgg 5820
taactcacag ggcgtcggct aacccccagt ccaaacctgg gagaaagcgc tcaaaaatga 5880
ctctagcgga ttcacgagac attgacacac cggcctggaa attttccgct gatctgttcg 5940
acacccatcc cgagctcgcg ctgcgatcac gtggctggac gagcgaagac cgccgcgaat 6000
tcctcgctca cctgggcaga gaaaatttcc agggcagcaa gacccgcgac ttcgccagcg 6060
cttggatcaa agacccggac acgggagaaa cacagccgaa gttataccga gttggttcaa 6120
aatcgcttgc ccggtgccag tatgttgctc tgacgcacgc gcagcacgca gccgtgcttg 6180
tcctggacat tgatgtgccg agccaccagg ccggcgggaa aatcgagcac gtaaaccccg 6240
aggtctacgc gattttggag cgctgggcac gcctggaaaa agcgccagct tggatcggcg 6300
tgaatccact gagcgggaaa tgccagctca tctggctcat tgatccggtg tatgccgcag 6360
caggcatgag cagcccgaat atgcgcctgc tggctgcaac gaccgaggaa atgacccgcg 6420
ttttcggcgc tgaccaggct ttttcacata ggctgagccg gtggccactg cacgtctccg 6480
acgatcccac cgcgtaccgc tggcatgccc agcacaatcg cgtggatcgc ctagctgatc 6540
ttatggaggt tgctcgcatg atctcaggca cagaaaaacc taaaaaacgc tatgagcagg 6600
agttttctag cggacgggca cgtatcgaag cggcaagaaa agccactgcg gaagcaaaag 6660
cacttgccac gcttgaagca agcctgccga gcgccgctga agcgtctgga gagctgatcg 6720
acggcgtccg tgtcctctgg actgctccag ggcgtgccgc ccgtgatgag acggcttttc 6780
gccacgcttt gactgtggga taccagttaa aagcggctgg tgagcgccta aaagacacca 6840
agatcatcga cgcctacgag cgtgcctaca ccgtcgctca ggcggtcgga gcagacggcc 6900
gtgagcctga tctgccgccg atgcgtgacc gccagacgat ggcgcgacgt gtgcgcggct 6960
acgtcgctaa aggccagcca gtcgtccctg ctcgtcagac agagacgcag agcagccgag 7020
ggcgaaaagc tctggccact atgggaagac gtggcggtaa aaaggccgca gaacgctgga 7080
aagacccaaa cagtgagtac gcccgagcac agcgagaaaa actagctaag tccagtcaac 7140
gacaagctag gaaagctaaa ggaaatcgct tgaccattgc aggttggttt atgactgttg 7200
agggagagac tggctcgtgg ccgacaatca atgaagctat gtctgaattt agcgtgtcac 7260
gtcagaccgt gaatagagca cttaagtctg cgggcattga acttccacga ggacgccgta 7320
aagcttccca gtaaatgtgc catctcgtag gcagaaaacg gttccccccg taggggtctc 7380
tctcttggcc tcctttctag gtcgggctga ttgctcttga agctctctag gggggctcac 7440
accataggca gataacggtt ccccaccggc tcacctcgta agcgcacaag gactgctccc 7500
aaagatcttc aaagccactg ccgcgactcc gcttcgcgaa gccttgcccc gcggaaattt 7560
cctccaccga gttcgtgcac acccctatgc caagcttctt tcaccctaaa ttcgagagat 7620
tggattctta ccgtggaaat tcttcgcaaa aatcgtcccc tgatcgccct tgcgacgttg 7680
ctcgcggcgg tgccgctggt tgcgcttggc ttgaccgact tgatcctccg gcgttcagcc 7740
tgtgccacag ccgacaggat ggtgaccacc atttgcccca tatcaccgtc ggtactgatc 7800
ccgtcgtcaa taaaccgaac cgctacaccc tgagcatcaa actcttttat cagttggatc 7860
atgtcggcgg tgtcgcggcc aagacggtcg agcttcttca ccagaatgac atcaccttcc 7920
tccaccttca tcctcagcaa atccagccct tcccgatctg ttgaactgcc ggatgccttg 7980
tcggtaaaga tgcggttagc ttttacccct gcatctttga gcgctgaggt ctgcctcgtg 8040
aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga 8100
gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt 8160
gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag 8220
caaaagttcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca 8280
gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg 8340
caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga 8400
aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat 8460
tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc 8520
aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat 8580
ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc 8640
aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt 8700
aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc 8760
aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg 8820
gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg 8880
aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc 8940
aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg 9000
atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc 9060
agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct 9120
cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat 9180
atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat 9240
aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg 9300
ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat 9360
caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc 9420
agcaacacct tcttcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat 9480
gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac 9540
cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc 9600
tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc 9660
tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt 9720
ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg 9780
aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc 9840
tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca 9900
ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg 9960
aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 10020
cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta 10080
gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct 10140
cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg 10200
ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct 10260
caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc acgttaaggg 10320
attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 10380
agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 10440
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 10500
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 10560
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 10620
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 10680
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 10740
gccgatgata agctgtcaaa catggcctgt cgcttgcggt attcggaatc ttgcacgccc 10800
tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta 10860
tcgccggcat ggcggccgac gcgcggggag aggcggtttg cgtattgggc gccagggtgg 10920
tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag 10980
agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg 11040
tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga 11100
tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct 11160
gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt 11220
gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat 11280
tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg 11340
ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc 11400
gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa 11460
gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca 11520
gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt 11580
tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat 11640
cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg 11700
tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa 11760
tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc 11820
tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat 11880
cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc 11940
atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc aacgtaaatg catgccgctt 12000
cgccttcgcg cgcgaattgc aagctgatcc gggcttatcg actgcacggt gcaccaatgc 12060
ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc 12120
ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca 12180
taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa 12240
tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagaattaaa agatatgacc 12300
atgattacgc c 12311
<210> 58
<211> 9892
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 58
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760
ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820
tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880
gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940
agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000
aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060
atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120
attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180
gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3240
tagacgccag gattgaacag taccaccagc acgcccatgt ctatccggtc catgatcgtt 3300
cttctcccgt aggtcgaagt tgccaggcca ggaccagccc ggccagaacg agaagggcgc 3360
cggcgaggaa tggcgcgccg gccaggggca gcggcgcgag cggaccgctg ccccagtgga 3420
acaggccgct catcagcggc ggaccgacga tcgcggcgag gctcatcagg ctgctcagca 3480
cgccctgcaa ctcgccctgg cggtcgaccg gcacgcgggc cgagagcagc ccctgcatgg 3540
ccggggtggc gaggctgccg agcgcgaagg gcagcagcgc gcagaccagc cagaatgacg 3600
agtcgaccag ggcgaacagc agcaggccgc agccttgcag ggcgaggccc aggcgcagca 3660
ggcgggcgtc gtccaggcgc cgcttgcaga ggttcacgcc gagggtctgg gcgagcaccg 3720
cgagcacgcc gtagagggcc agcgagtagc cgatccaggc gctgctccag tgaaacttct 3780
cgatcacgaa gaacggccag accaccatca ccgcctgcaa gccgaggaat accagggcaa 3840
gcaccgccag caggcgtccg acccccggtt gccgagccag gccgctgatc gagcgcaagg 3900
cattcatccg cctcgggtcc aggcggcggc gtcgcgtcgg gggcagggtt tcctcgagga 3960
acaggccggc gagcagggcg ttgagcaggc acaggccggc ggccagcaac agcggcagcg 4020
tcgtgccgtg caccgccagc agcccaccga gggcggggcc gaggatcatg cccagggcga 4080
ggccggcgta cagccagccg aagtgccggg tgcgctgccc gtgcgtgccg aggtcagccg 4140
cgcaggccat cgcggtggcc acgctggcgc cggtgagccc ggccagcgcg cgaccgagga 4200
acagcatcca gaggctgtcg gccagcgcca gcagcagata gctgagggcg aagccgagca 4260
tcgccaggac caggacgggg cggcgtccga agcggtcgct gaggctgccg aggaccggcg 4320
aaaagaacaa ttgcagcagc gcgaaggtca tcaccagggc ggcgccccag gtggccgcgt 4380
cgcggaccgc cagcggcgcc acgctgccga tcagcgtcgg cagcaggggc acgatcaggc 4440
cgacgccagc ggcatccagc aggcaggtga ggaacagcag aggcaggacg cgtttcgcgc 4500
cgggaccgtg ttcccgcgtg gcggaggggc agaggctggt cgtggacacg ccaggatcct 4560
cccggcgaac acaggaaatc ctgatcttgg gacgccagcg agggcaaggg aaactaccgg 4620
aattcacagg ttcgacgaac ggtcgcggca acgccggcgg gcggcgtgat gtcataatga 4680
tgcgacccgt gcgccagtcg ggtatccttg ccggcctctc atctagaggg aatgcgtttc 4740
gccgactagg ccttggcctt gccggaagct acggacgcca cggccgggcc ggcgaggcgc 4800
ttcagcaggc gcgggcggtt ggtctccagc gcgccgccgc gtccccgcag gccgtcccac 4860
aggccccagc ccaggcagcg cagcttgagc agcttgtcgc gttcgagcag gagcaccgcg 4920
aggccctggg tcagggtcgg caggttcgcc agcagggcca gcggcgagga ccgggcgtag 4980
cggcgcagga ccagcaggcc gttgcgcgcc aggtagtagc ggcgcagcgg ggcgtggttc 5040
atcgcgctga ggctgagacc gccgaggcgg cgggtcttgc gcgtgccgat gcggtgctcg 5100
aggaccagcc gcgggtcgac gtacaggggc acgtccagcg cctgggcgcg caggctgtat 5160
tcggtgtcca cgtggtcgat gaacagttcc tcgtcgaagt ggccgaggcg ctggtaggcc 5220
tcgcgggtca gcaggcagcc ggaggagatc aggaacgagg tgcgctgcgg ggtcgtcagg 5280
ccgtccagag acaattgcct gagcgtcagt ccgtcgagat ggatggccgg caggaagcgc 5340
cggtcacccc ggtcgaagat ccgtgggccg agcaggcagg cctgaccgtt gcgcgcctgc 5400
aggttgcgcc actgggcggc gaggaaggcg ccgccgggac gggagtcctg gtcgagcagc 5460
agcacaccct gcacgccacg ccggaatagc gcgtcgagtc cctggttgaa ggcgccggcg 5520
atgccctgcc ggttgccgtg gtgcagcacg gcgatgcctt gcccgcgcag ccgggcattg 5580
cgctgcggat cgctgtgcgg tgagttgtcg acggcaagga agcgcagttg cggaaacgcc 5640
gccgccagtt cgccaaggtg ttccaggtcg tcgtcgccag gattgaacag taccaccagc 5700
acgcccatgt ctatccggtc catgatcgtt cttctcccgt aggtcaggac gcagccttca 5760
gccatcgcgc atccccctcc ctatgacaac gttcgaccac ctgggccgct ttaccgcaag 5820
cgatactgtg cggttgtgac aattccatga aacgccgaca ggccgccgcc atggccgggt 5880
cctcgagcaa gcgccacagc gccccgcgca actcctgctc gcgcaatggc acgcccaggc 5940
gcatcccgca gccgagccgg accagccgtt cggcattgtc gaactggtcg tgggcgcagg 6000
gcagcagcac ctgcggcacc cccgccgcca aggccaggct catggcgccg ataccgcccg 6060
gatggaccag cccggcgcac gatggcagca aggctcccag tggcgcgtag gcgcgctgca 6120
gcacgtggtt cggcaagccg cgcagcggtt cctggccggc gccggtgagg aagatcccac 6180
gcgcgccgag gcgttccagc gcgcgcaggg ccatggcgta gaagtcgccc tgcaggtgtt 6240
cggtcgagcc ctgggtgaac accagcggcc ggctgccctg atcgagaaag cgttgcagtt 6300
cgtcgtcgag cggggtcccc gggatactgc cgtcgaacag cgggaagccg gtcatgtgca 6360
ggggttgcgg ccaatcctgc tggggcggcg cgaaccaggc cgggaacagg cagaccacgc 6420
cctgcggcga atgcatccat tgggtgaaga tgcgcttcac cggcgtctcc aggccgacct 6480
tgcgccgcac cgcgttgata tccggcgcgc aggtgcgatc cagcttgaag cgctcgatgc 6540
agcgccagag cagcttgcgc atcgccagcg gcatctgctc gggcacgttg aacttggggt 6600
gtaccggcgg caggtgcgcc gacaacaagg tcgatggcga gacctgcgcg gacaggtagg 6660
gaatcccgta cttctcgtga gcgatgcgtg cgcccagcgc ccagagcgag ccgaccacca 6720
cgatgtcgtc atggcgctgc gccgagacgt actcgtagac cggctcgatc atcccggcga 6780
tggtttgcca gagcacgccg aaggacgtct tggggtccca caggcgcgga tcgcccatgg 6840
tccggcggta ggtcagttcg tcgctcagcg ggacgaacgc gatgccgtgc tgctccaccg 6900
cgtcgcgaaa caccgggatg gtgcagaggc tcacgcggtg cccgcgcaat ttcagggtcc 6960
gggccaggcc gatgaaggga aatacgtcgc cggccgagcc gatggcgatg aggatggcgt 7020
gcatggtgct actccgtgcg ttatgcaacc gcaaagcccg gccaggccgg gtcttcgcag 7080
gtcaagggtt caggcgtagc cgatggccat ctcgtggaat cccgccgcgc gttccgcccg 7140
ctgcggctcc ggttgcttca gcaggtgctc gagcagggcg cggtgcacgc gtaccgcggc 7200
cagcttggac tccaggtcga ggaaatgccc ggtgccctcc acccgcgaga aactgcagtg 7260
cggcaggtag tcgcggaact ggcgggcgtc ctcggcggtg gtgtattcgt cccagctgcc 7320
gttgatgaaa tgcacgtggc tctggatccg ctccaggcaa gccaggtagc cccgatcgtt 7380
gagcgccagc acctggtcga tgtgaaagcg cgcctgctcg tattcgccgg tggccagcga 7440
agccatgtgc tgatggttgc tggctttcag gcgcggcggc aggtatttgc cgacggtctc 7500
gttgagcaga tggccgatcg ccgacttgtc gtccagctcg atcagcgcct gcgcccgccc 7560
gacgtagtcg agcatcgcct ggttcagtcc aggggcgaat gccatcacca ccgagctgcg 7620
gatgccgcgc ggattgcgcg acagcgccag cagcgtggag ataccgcccc aggacgcgga 7680
gaccaggtga ttgacctcga agcgctcgat cagcgccagg aggatttcca cctcgtcgtc 7740
cttggtgatc aacccccgct gcgggttgtg ctgacgcgac tgcccggcga agggcaggtc 7800
gaacagcacc acgttgaaat gttcggccag gcacttgcag gtccgggcga acgaggcggt 7860
ggtcgccatc gcgccgttga ccagcatcac cgtgctgcgc ccgggatcct gcccaacgcg 7920
ctcgacatgt acccgcaggc ccttgcaaac cgataccaac agactttcgc gccgcatttc 7980
acacctccca aaaatgccag atcccccggg ctgcaggaat tcgatatcaa gcttatcgat 8040
accgtcgacc tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat 8100
tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 8160
aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 8220
gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 8280
gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 8340
catgcataaa aactgttgta attcattaag cattctgccg acatggaagc catcacaaac 8400
ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt 8460
gcccatgggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc 8520
cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga 8580
aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc 8640
tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat 8700
accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg 8760
ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa 8820
tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac 8880
gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc 8940
gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt 9000
acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag 9060
cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg 9120
agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc 9180
agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg 9240
cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac 9300
cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg 9360
tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc 9420
atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct 9480
gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac 9540
cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca 9600
gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt 9660
ttcccttgtc cagatagccc agtagctgac attcatccca ggtggcactt ttcggggaaa 9720
tgtgcgcgcc cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt 9780
tccgtcagca gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc 9840
cgattcaacg gccccagggc gtccagaacg ggcttcaggc gctcccgaag gt 9892
<210> 59
<211> 1477
<212> DNA
<213> Pseudomonas aeruginosa
<400> 59
tgagaggccg gcaaggatac ccgactggcg cacgggtcgc atcattatga catcacgccg 60
cccgccggcg ttgccgcgac cgttcgtcga acctgtgaat tccggtagtt tcccttgccc 120
tcgctggcgt cccaagatca ggatttcctg tgttcgccgg gaggatcctg gcgtgtccac 180
gaccagcctc tgcccctccg ccacgcggga acacggtccc ggcgcgaaac gcgtcctgcc 240
tctgctgttc ctcacctgcc tgctggatgc cgctggcgtc ggcctgatcg tgcccctgct 300
gccgacgctg atcggcagcg tggcgccgct ggcggtccgc gacgcggcca cctggggcgc 360
cgccctggtg atgaccttcg cgctgctgca attgttcttt tcgccggtcc tcggcagcct 420
cagcgaccgc ttcggacgcc gccccgtcct ggtcctggcg atgctcggct tcgccctcag 480
ctatctgctg ctggcgctgg ccgacagcct ctggatgctg ttcctcggtc gcgcgctggc 540
cgggctcacc ggcgccagcg tggccaccgc gatggcctgc gcggctgacc tcggcacgca 600
cgggcagcgc acccggcact tcggctggct gtacgccggc ctcgccctgg gcatgatcct 660
cggccccgcc ctcggtgggc tgctggcggt gcacggcacg acgctgccgc tgttgctggc 720
cgccggcctg tgcctgctca acgccctgct cgccggcctg ttcctcgagg aaaccctgcc 780
cccgacgcga cgccgccgcc tggacccgag gcggatgaat gccttgcgct cgatcagcgg 840
cctggctcgg caaccggggg tcggacgcct gctggcggtg cttgccctgg tattcctcgg 900
cttgcaggcg gtgatggtgg tctggccgtt cttcgtgatc gagaagtttc actggagcag 960
cgcctggatc ggctactcgc tggccctcta cggcgtgctc gcggtgctcg cccagaccct 1020
cggcgtgaac ctctgcaagc ggcgcctgga cgacgcccgc ctgctgcgcc tgggcctcgc 1080
cctgcaaggc tgcggcctgc tgctgttcgc cctggtcgac tcgtcattct ggctggtctg 1140
cgcgctgctg cccttcgcgc tcggcagcct cgccaccccg gccatgcagg ggctgctctc 1200
ggcccgcgtg ccggtcgacc gccagggcga gttgcagggc gtgctgagca gcctgatgag 1260
cctcgccgcg atcgtcggtc cgccgctgat gagcggcctg ttccactggg gcagcggtcc 1320
gctcgcgccg ctgcccctgg ccggcgcgcc attcctcgcc ggcgcccttc tcgttctggc 1380
cgggctggtc ctggcctggc aacttcgacc tacgggagaa gaacgatcat ggaccggata 1440
gacatgggcg tgctggtggt actgttcaat cctggcg 1477
<210> 60
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> primer
<400> 60
aggaaatcta gatgagaggc cggcaaggat ac 32
<210> 61
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> primer
<400> 61
ccaggttcta gacgccagga ttgaacagta cc 32
<210> 62
<211> 7332
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 62
cgaaaatttt tgggaggtgt gaaatgcggc gcgaaagtct gttggtatcg gtttgcaagg 60
gcctgcgggt acatgtcgag cgcgttgggc aggatcccgg gcgcagcacg gtgatgctgg 120
tcaacggcgc gatggcgacc accgcctcgt tcgcccggac ctgcaagtgc ctggccgaac 180
atttcaacgt ggtgctgttc gacctgccct tcgccgggca gtcgcgtcag cacaacccgc 240
agcgcgggtt gatcaccaag gacgacgagg tggaaatcct cctggcgctg atcgagcgct 300
tcgaggtcaa tcacctggtc tccgcgtcct ggggcggtat ctccacgctg ctggcgctgt 360
cgcgcaatcc gcgcggcatc cgcagctcgg tggtgatggc attcgcccct ggactgaacc 420
aggcgatgct cgactacgtc gggcgggcgc aggcgctgat cgagctggac gacaagtcgg 480
cgatcggcca tctgctcaac gagaccgtcg gcaaatacct gccgcagcgc ctgaaagcca 540
gcaaccatca gcacatggct tcgctggcca ccggcgaata cgagcaggcg cgctttcaca 600
tcgaccaggt gctggcgctc aacgatcggg gctacttggc ttgcctggag cggatccaga 660
gccacgtgca tttcatcaac ggcagctggg acgaatacac caccgccgag gacgcccgcc 720
agttccgcga ctacctgccg cactgcagtt tctcgcgggt ggagggcacc gggcatttcc 780
tcgacctgga gtccaagctg gcagcggtac gcgtgcaccg cgccctgctc gagcacctgc 840
tgaagcaacc ggagccgcag cgggcggaac gcgcggcggg attccacgag atggccatcg 900
gctacgcctg aacccttgac ctgcgaagac ccggcctggc cgggctttgc ggttgcataa 960
cgcacggagt agccccatgc acgccatcct catcgccatc ggctcggccg gcgacgtatt 1020
tcccttcatc ggcctggccc ggaccctgaa actgcgcggg caccgcgtga gcctctgcac 1080
catcccggtg tttcgcgacg cggtggagca gcacggcatc gcgttcgtcc cgctgagcga 1140
cgaactgacc taccgccgga ccatgggcga tccgcgcctg tgggacccca agacgtcctt 1200
cggcgtgctc tggcaagcca tcgccgggat gatcgagccg gtctacgagt acgtctcggc 1260
gcagcgccat gacgacatcg tggtggtcgg ctcgctatgg gcgctgggcg cacgcatcgc 1320
tcacgagaag tacgggattc cctacctgtc cgcgcaggtc tcgccatcga ccctgttgtc 1380
ggcgcacctg ccgccggtac accccaagtt caacgtgccc gagcagatgc cgctggcgat 1440
gcgcaagctg ctctggcgct gcatcgagcg cttcaagctg gatcgcacct gcgcgccgga 1500
gatcaacgcg gtgcgccgca aggtcggcct ggaaacgccg gtgaagcgca tcttcaccca 1560
atggatgcat tcgccgcagg gcgtggtctg cctgttcccg gcctggttcg cgccgcccca 1620
gcaggattgg ccgcaacccc tgcacatgac cggcttcccg ctgttcgacg gcagtatccc 1680
ggggaccccg ctcgacgacg aactgcaacg ctttctcgat cagggcagcc ggccgctggt 1740
gttcacccag ggctcgaccg aacacctgca gggcgacttc tacgccatgg ccctgcgcgc 1800
gctggaacgc ctcggcgcgc gtgggatctt cctcaccggc gccggccagg aaccgctgcg 1860
cggcttgccg aaccacgtgc tgcagcgcgc ctacgcgcca ctgggagcct tgctgccatc 1920
gtgcgccggg ctggtccatc cgggcggtat cggcgccatg agcctagcct tggcggcggg 1980
ggtgccgcag gtgctgctgc cctgtgccca cgaccagttc gacaatgccg aacggctggt 2040
ccggctcggc tgcgggatgc gcctgggcgt gccgttgcgc gagcaggagt tgcgcggggc 2100
gctgtggcgc ttgctcgagg acccggccat ggcggcggcc tgtcggcgtt tcatggaatt 2160
gtcacaaccg cacagtatcg cttgcggtaa agcggcccag gtggtcgaac gttgtcatag 2220
ggagggggat gctcgatggc tgaaggctgc gtcctgaacg gtctagagcg gccgccaccg 2280
cggtggagct ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt 2340
ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 2400
ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 2460
ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt 2520
taaatttttg ttaaatcagc tcatttttta accaataggc cgactgcgat gagtggcagg 2580
gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt gctacgcctg 2640
aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc 2700
ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg 2760
actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct 2820
ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca gagcctgata 2880
aaaacggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc 2940
tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg aggcggctac 3000
agccgatagt ctggaacagc gcacttacgg gttgctgcgc aacccaagtg ctaccggcgc 3060
ggcagcgtga cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa acacggctca 3120
tcgggcatcg gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct 3180
ggcaggtctg gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg 3240
gtcggtagtt gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc 3300
aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg 3360
cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg 3420
gtgccggggc cgttgagctt cacgacggag atccagcgct cggccaccaa gtccttgact 3480
gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagtgtctt ctggctgacc 3540
accacggcgt tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg 3600
ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag 3660
tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc 3720
tccatgcggt agggtgccgc acggttgcgg caccatgcgc aatcagctgc aacttttcgg 3780
cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc aattacagat tttctttaac 3840
ctacgcaatg agctattgcg gggggtgccg caatgagctg ttgcgtaccc ccctttttta 3900
agttgttgat ttttaagtct ttcgcatttc gccctatatc tagttctttg gtgcccaaag 3960
aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg ctccccctcc ggcaaaaagt 4020
ggcccctccg gggcttgttg atcgactgcg cggccttcgg ccttgcccaa ggtggcgctg 4080
cccccttgga acccccgcac tcgccgccgt gaggctcggg gggcaggcgg gcgggcttcg 4140
ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg 4200
ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac cggcaagcga 4260
agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa ttgatggggc 4320
aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg ggtagccggt 4380
gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc ttgtccagca 4440
gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc catcgtccac 4500
atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg gagagcaagc 4560
ccgtagggcg ccgcagccgc cgtaggcggt cacgactttg cgaagcaaag tctagtgagt 4620
atactcaagc attgagtggc ccgccggagg caccgccttg cgctgccccc gtcgagccgg 4680
ttggacacca aaagggaggg gcaggcatgg cggcatacgc gatcatgcga tgcaagaagc 4740
tggcgaaaat gggcaacgtg gcggccagtc tcaagcacgc ctaccgcgag cgcgagacgc 4800
ccaacgctga cgccagcagg acgccagaga acgagcactg ggcggccagc agcaccgatg 4860
aagcgatggg ccgactgcgc gagttgctgc cagagaagcg gcgcaaggac gctgtgttgg 4920
cggtcgagta cgtcatgacg gccagcccgg aatggtggaa gtcggccagc caagaacagc 4980
aggcggcgtt cttcgagaag gcgcacaagt ggctggcgga caagtacggg gcggatcgca 5040
tcgtgacggc cagcatccac cgtgacgaaa ccagcccgca catgaccgcg ttcgtggtgc 5100
cgctgacgca ggacggcagg ctgtcggcca aggagttcat cggcaacaaa gcgcagatga 5160
cccgcgacca gaccacgttt gcggccgctg tggccgatct agggctgcaa cggggcatcg 5220
agggcagcaa ggcacgtcac acgcgcattc aggcgttcta cgaggccctg gagcggccac 5280
cagtgggcca cgtcaccatc agcccgcaag cggtcgagcc acgcgcctat gcaccgcagg 5340
gattggccga aaagctggga atctcaaagc gcgttgagac gccggaagcc gtggccgacc 5400
ggctgacaaa agcggttcgg caggggtatg agcctgccct acaggccgcc gcaggagcgc 5460
gtgagatgcg caagaaggcc gatcaagccc aagagacggc ccgagacctt cgggagcgcc 5520
tgaagcccgt tctggacgcc ctggggccgt tgaatcggga tatgcaggcc aaggccgccg 5580
cgatcatcaa ggccgtgggc gaaaagctgc tgacggaaca gcgggaagtc cagcgccaga 5640
aacaggccca gcgccagcag gaacgcgggc gcgcacattt ccccgaaaag tgccacctgg 5700
gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca 5760
ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag 5820
cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa 5880
ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga 5940
gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 6000
cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 6060
tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 6120
gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 6180
gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 6240
attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 6300
atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 6360
cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 6420
cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 6480
gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 6540
gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 6600
tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 6660
cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 6720
catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 6780
accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 6840
gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 6900
gatctcatgc tggagttctt cgcccacccc catgggcaaa tattatacgc aaggcgacaa 6960
ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg 7020
cagaatgctt aatgaattac aacagttttt atgcatgcgc ccaatacgca aaccgcctct 7080
ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc 7140
gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt 7200
acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac 7260
aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca 7320
aaagctgggt ac 7332
<210> 63
<211> 7354
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 63
cgaattcaaa actttttggg aggtgtgaga tgcggcgcga aagtctgttg gtaacggtat 60
gcaagggcct gcgggtacat gtcgagcgcg tggggcagga tcccgggcgc gacacggtga 120
tgctggtcaa cggcgcgatg gcgaccaccg cctcgttcgc ccggacctgc aagtgcctgg 180
ccgaacattt caacgtggtg ctgttcgacc tgcccttcgc cgggcagtcg cggcagcaca 240
atccgcagcg cgggttgatc accaaggacg acgaggtgga gattctcctg gcgctgatcg 300
agcgcttcgc tgtcaaccac ctggtctcgg cctcctgggg cggcatctcc acgctgctgg 360
cgctgtcgcg caacccgcgc ggggtccgca gctcggtggt gatggcgttc gcgccggggc 420
tgaaccaggc gatgctcgat tatgtcgggc gggcccagga actgatcgaa ctggacgaca 480
agtcggcgat cggccacctg ctcaacgaga ccgtcggcaa gtacctgccg ccgcggctga 540
aggccagcaa ccatcagcac atggcctccc tggccactgg cgagtacgag caggcgcgtt 600
tccacatcga ccaggtgctg gcgctcaatg accgtggcta cctgagctgc ctggggcaga 660
tccagagtca cgtgcatttc atcaacggca gctgggacga gtacaccacc gccgaggacg 720
cccgccagtt ccgcgattac ctgccgcatt gcagtttttc gcgggtggaa ggcaccgggc 780
acttcctcga cctggagtcc aagctggcgg cggcgcgtgt gcaccgggcg ttgctcgagc 840
acctgctggc gcaaccggaa ccgtggcgct ccgagcaggc ggcgggattc cacgagatgg 900
ccatcggcta cgcctgaccc gtcgggatct gcgaaggccc ggcatggccg ggccttgccg 960
ttgcacaacg caaggagtag ccccatgcac gccattctca tcgccatcgg ttcggccggc 1020
gacgtgttcc ccttcatcgg cctggcccgc accctgaagt tgcgcggcca ccgcgtcagc 1080
ctgtgcacca ttccggtgtt tcgcgccgcg gtggagcagc acggcatcga gttcgtcccg 1140
ctcagcgacg aactgaccta ccgccggacc atgggcgacc cgcgcctgtg ggatccgaag 1200
acctcgttcg gagtgctctg gcaggccatc gccgggatga tcgagccggt ctacgagtac 1260
gtctgcgcac agcgccacga cgacatcgtg gtggtcggtt cgctgtgggc cctgggcgcg 1320
cggatcgccc atgagaaata cgggattccc tacctgtcgg tgcaggtctc gccgtcgacc 1380
ctgctgtcgg cgcacctgcc gccggtccac cccaggttca acgtgcccga gcaggtcccg 1440
ctggcgatgc gcaagttgct ctggcgctgc atcgaacgct tcaagctgga ccgcacctgc 1500
gccccggaga tcaacgcggt gcgccgcaag gtcggcctgg tcggcccggc gaagcgcatc 1560
ttcacccagt ggatgcattc gccacaggga gtgctctgcc tgttcccggc ctggttcgca 1620
ccgccccagc aggactggcc gcaaccgctg cacatgaccg gcttcccgct gttcgacggc 1680
agcgtcccgg ggacccgcct cgacgacgag ttgcagcgct tcctcgagca gggcagtcgg 1740
ccgctggtgt tcacccaggg ttcgaccgag cacctgcagg gagacttcta tgccatggcc 1800
ttgcgcgcgc tggagcgtct cggcgcccgc ggcatcttcc tcaccggcgc cggccaggag 1860
ccgctgcgtg gcttgccgag ccacgtgctg caacgctcgt acgtgccgtt gggggccttg 1920
ctgccggcgt gcgccgggct ggtccacccg gccggcatcg gcgccatgag cctggcgctg 1980
gcggcggggg tgccgcaggt gctgctgcct tgcgcccacg accagttcga caacgccgaa 2040
cgcctggtcc gcctcggctg cggtatccgc ctgggcctgc cgctacgcga gcaggcgctg 2100
cgcgagtcgc tctggcggct gctcgaggac ccggcgctgg cggcggcctg tcggcgtttc 2160
atggaattgt cacaaccgca cagtatcgct tgcggtaaag cggcccaagt ggtcgaacgt 2220
tgtcataggg agggggatgt gcgatggctg aaagccgcgt cctgagccgt gctggcagaa 2280
ttctctagag cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta 2340
cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca 2400
acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg 2460
caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt 2520
aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 2580
gccgactgcg atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct 2640
taaacgcctg gtgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga 2700
aagcaaattc gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta 2760
ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc 2820
ctgaggccag tttgctcagg ctctccccgt ggaggtaata attgacgata tgatcattta 2880
ttctgcctcc cagagcctga taaaaacggt gaatccgtta gcgaggtgcc gccggcttcc 2940
attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg 3000
tatagggcgg cgaggcggct acagccgata gtctggaaca gcgcacttac gggttgctgc 3060
gcaacccaag tgctaccggc gcggcagcgt gacccgtgtc ggcggctcca acggctcgcc 3120
atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc gccgttccca 3180
ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc cgggctggct 3240
gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca gggtcgggat 3300
gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac 3360
ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc 3420
attgaccacg taggccgaca cggtgccggg gccgttgagc ttcacgacgg agatccagcg 3480
ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt 3540
ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg ccacgaggtg 3600
atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg cctcatgcgc 3660
tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc cgatttctct 3720
ggactgcgtg gccatgctta tctccatgcg gtagggtgcc gcacggttgc ggcaccatgc 3780
gcaatcagct gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag 3840
tcaattacag attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc 3900
tgttgcgtac cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata 3960
tctagttctt tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc 4020
ggctccccct ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggccttc 4080
ggccttgccc aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg 4140
gggggcaggc gggcgggctt cgccttcgac tgcccccact cgcataggct tgggtcgttc 4200
caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc cgccttccac 4260
ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc cggaggcttt tccccagaga 4320
aaattaaaaa aattgatggg gcaaggccgc aggccgcgca gttggagccg gtgggtatgt 4380
ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca agctcgtggg caggcgcagc 4440
ctgtccatca gcttgtccag cagggttgtc cacgggccga gcgaagcgag ccagccggtg 4500
gccgctcgcg gccatcgtcc acatatccac gggctggcaa gggagcgcag cgaccgcgca 4560
gggcgaagcc cggagagcaa gcccgtaggg cgccgcagcc gccgtaggcg gtcacgactt 4620
tgcgaagcaa agtctagtga gtatactcaa gcattgagtg gcccgccgga ggcaccgcct 4680
tgcgctgccc ccgtcgagcc ggttggacac caaaagggag gggcaggcat ggcggcatac 4740
gcgatcatgc gatgcaagaa gctggcgaaa atgggcaacg tggcggccag tctcaagcac 4800
gcctaccgcg agcgcgagac gcccaacgct gacgccagca ggacgccaga gaacgagcac 4860
tgggcggcca gcagcaccga tgaagcgatg ggccgactgc gcgagttgct gccagagaag 4920
cggcgcaagg acgctgtgtt ggcggtcgag tacgtcatga cggccagccc ggaatggtgg 4980
aagtcggcca gccaagaaca gcaggcggcg ttcttcgaga aggcgcacaa gtggctggcg 5040
gacaagtacg gggcggatcg catcgtgacg gccagcatcc accgtgacga aaccagcccg 5100
cacatgaccg cgttcgtggt gccgctgacg caggacggca ggctgtcggc caaggagttc 5160
atcggcaaca aagcgcagat gacccgcgac cagaccacgt ttgcggccgc tgtggccgat 5220
ctagggctgc aacggggcat cgagggcagc aaggcacgtc acacgcgcat tcaggcgttc 5280
tacgaggccc tggagcggcc accagtgggc cacgtcacca tcagcccgca agcggtcgag 5340
ccacgcgcct atgcaccgca gggattggcc gaaaagctgg gaatctcaaa gcgcgttgag 5400
acgccggaag ccgtggccga ccggctgaca aaagcggttc ggcaggggta tgagcctgcc 5460
ctacaggccg ccgcaggagc gcgtgagatg cgcaagaagg ccgatcaagc ccaagagacg 5520
gcccgagacc ttcgggagcg cctgaagccc gttctggacg ccctggggcc gttgaatcgg 5580
gatatgcagg ccaaggccgc cgcgatcatc aaggccgtgg gcgaaaagct gctgacggaa 5640
cagcgggaag tccagcgcca gaaacaggcc cagcgccagc aggaacgcgg gcgcgcacat 5700
ttccccgaaa agtgccacct gggatgaatg tcagctactg ggctatctgg acaagggaaa 5760
acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact 5820
gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg 5880
ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca 5940
ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 6000
gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 6060
aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 6120
ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 6180
ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 6240
aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 6300
accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 6360
ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 6420
ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 6480
cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 6540
tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 6600
tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 6660
gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 6720
tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 6780
cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 6840
cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 6900
ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc cccatgggca 6960
aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg 7020
tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagttt ttatgcatgc 7080
gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 7140
acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca 7200
ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg 7260
tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa 7320
ttaaccctca ctaaagggaa caaaagctgg gtac 7354
<210> 64
<211> 2298
<212> DNA
<213> Artificial Sequence
<220>
<223> syn operon
<400> 64
aagcttgaat tcggtaccga aaatttttgg gaggtgtgaa atgcggcgcg aaagtctgtt 60
ggtatcggtt tgcaagggcc tgcgggtaca tgtcgagcgc gttgggcagg atcccgggcg 120
cagcacggtg atgctggtca acggcgcgat ggcgaccacc gcctcgttcg cccggacctg 180
caagtgcctg gccgaacatt tcaacgtggt gctgttcgac ctgcccttcg ccgggcagtc 240
gcgtcagcac aacccgcagc gcgggttgat caccaaggac gacgaggtgg aaatcctcct 300
ggcgctgatc gagcgcttcg aggtcaatca cctggtctcc gcgtcctggg gcggtatctc 360
cacgctgctg gcgctgtcgc gcaatccgcg cggcatccgc agctcggtgg tgatggcatt 420
cgcccctgga ctgaaccagg cgatgctcga ctacgtcggg cgggcgcagg cgctgatcga 480
gctggacgac aagtcggcga tcggccatct gctcaacgag accgtcggca aatacctgcc 540
gcagcgcctg aaagccagca accatcagca catggcttcg ctggccaccg gcgaatacga 600
gcaggcgcgc tttcacatcg accaggtgct ggcgctcaac gatcggggct acttggcttg 660
cctggagcgg atccagagcc acgtgcattt catcaacggc agctgggacg aatacaccac 720
cgccgaggac gcccgccagt tccgcgacta cctgccgcac tgcagtttct cgcgggtgga 780
gggcaccggg catttcctcg acctggagtc caagctggca gcggtacgcg tgcaccgcgc 840
cctgctcgag cacctgctga agcaaccgga gccgcagcgg gcggaacgcg cggcgggatt 900
ccacgagatg gccatcggct acgcctgaac ccttgacctg cgaagacccg gcctggccgg 960
gctttgcggt tgcataacgc acggagtagc cccatgcacg ccatcctcat cgccatcggc 1020
tcggccggcg acgtatttcc cttcatcggc ctggcccgga ccctgaaact gcgcgggcac 1080
cgcgtgagcc tctgcaccat cccggtgttt cgcgacgcgg tggagcagca cggcatcgcg 1140
ttcgtcccgc tgagcgacga actgacctac cgccggacca tgggcgatcc gcgcctgtgg 1200
gaccccaaga cgtccttcgg cgtgctctgg caagccatcg ccgggatgat cgagccggtc 1260
tacgagtacg tctcggcgca gcgccatgac gacatcgtgg tggtcggctc gctatgggcg 1320
ctgggcgcac gcatcgctca cgagaagtac gggattccct acctgtccgc gcaggtctcg 1380
ccatcgaccc tgttgtcggc gcacctgccg ccggtacacc ccaagttcaa cgtgcccgag 1440
cagatgccgc tggcgatgcg caagctgctc tggcgctgca tcgagcgctt caagctggat 1500
cgcacctgcg cgccggagat caacgcggtg cgccgcaagg tcggcctgga aacgccggtg 1560
aagcgcatct tcacccaatg gatgcattcg ccgcagggcg tggtctgcct gttcccggcc 1620
tggttcgcgc cgccccagca ggattggccg caacccctgc acatgaccgg cttcccgctg 1680
ttcgacggca gtatcccggg gaccccgctc gacgacgaac tgcaacgctt tctcgatcag 1740
ggcagccggc cgctggtgtt cacccagggc tcgaccgaac acctgcaggg cgacttctac 1800
gccatggccc tgcgcgcgct ggaacgcctc ggcgcgcgtg ggatcttcct caccggcgcc 1860
ggccaggaac cgctgcgcgg cttgccgaac cacgtgctgc agcgcgccta cgcgccactg 1920
ggagccttgc tgccatcgtg cgccgggctg gtccatccgg gcggtatcgg cgccatgagc 1980
ctagccttgg cggcgggggt gccgcaggtg ctgctgccct gtgcccacga ccagttcgac 2040
aatgccgaac ggctggtccg gctcggctgc gggatgcgcc tgggcgtgcc gttgcgcgag 2100
caggagttgc gcggggcgct gtggcgcttg ctcgaggacc cggccatggc ggcggcctgt 2160
cggcgtttca tggaattgtc acaaccgcac agtatcgctt gcggtaaagc ggcccaggtg 2220
gtcgaacgtt gtcataggga gggggatgct cgatggctga aggctgcgtc ctgaacggtc 2280
tagagaattc ggcgcgcc 2298
<210> 65
<211> 2302
<212> DNA
<213> Artificial Sequence
<220>
<223> syn operon
<400> 65
ggtaccgaat tcaaaacttt ttgggaggtg tgagatgcgg cgcgaaagtc tgttggtaac 60
ggtatgcaag ggcctgcggg tacatgtcga gcgcgtgggg caggatcccg ggcgcgacac 120
ggtgatgctg gtcaacggcg cgatggcgac caccgcctcg ttcgcccgga cctgcaagtg 180
cctggccgaa catttcaacg tggtgctgtt cgacctgccc ttcgccgggc agtcgcggca 240
gcacaatccg cagcgcgggt tgatcaccaa ggacgacgag gtggagattc tcctggcgct 300
gatcgagcgc ttcgctgtca accacctggt ctcggcctcc tggggcggca tctccacgct 360
gctggcgctg tcgcgcaacc cgcgcggggt ccgcagctcg gtggtgatgg cgttcgcgcc 420
ggggctgaac caggcgatgc tcgattatgt cgggcgggcc caggaactga tcgaactgga 480
cgacaagtcg gcgatcggcc acctgctcaa cgagaccgtc ggcaagtacc tgccgccgcg 540
gctgaaggcc agcaaccatc agcacatggc ctccctggcc actggcgagt acgagcaggc 600
gcgtttccac atcgaccagg tgctggcgct caatgaccgt ggctacctga gctgcctggg 660
gcagatccag agtcacgtgc atttcatcaa cggcagctgg gacgagtaca ccaccgccga 720
ggacgcccgc cagttccgcg attacctgcc gcattgcagt ttttcgcggg tggaaggcac 780
cgggcacttc ctcgacctgg agtccaagct ggcggcggcg cgtgtgcacc gggcgttgct 840
cgagcacctg ctggcgcaac cggaaccgtg gcgctccgag caggcggcgg gattccacga 900
gatggccatc ggctacgcct gacccgtcgg gatctgcgaa ggcccggcat ggccgggcct 960
tgccgttgca caacgcaagg agtagcccca tgcacgccat tctcatcgcc atcggttcgg 1020
ccggcgacgt gttccccttc atcggcctgg cccgcaccct gaagttgcgc ggccaccgcg 1080
tcagcctgtg caccattccg gtgtttcgcg ccgcggtgga gcagcacggc atcgagttcg 1140
tcccgctcag cgacgaactg acctaccgcc ggaccatggg cgacccgcgc ctgtgggatc 1200
cgaagacctc gttcggagtg ctctggcagg ccatcgccgg gatgatcgag ccggtctacg 1260
agtacgtctg cgcacagcgc cacgacgaca tcgtggtggt cggttcgctg tgggccctgg 1320
gcgcgcggat cgcccatgag aaatacggga ttccctacct gtcggtgcag gtctcgccgt 1380
cgaccctgct gtcggcgcac ctgccgccgg tccaccccag gttcaacgtg cccgagcagg 1440
tcccgctggc gatgcgcaag ttgctctggc gctgcatcga acgcttcaag ctggaccgca 1500
cctgcgcccc ggagatcaac gcggtgcgcc gcaaggtcgg cctggtcggc ccggcgaagc 1560
gcatcttcac ccagtggatg cattcgccac agggagtgct ctgcctgttc ccggcctggt 1620
tcgcaccgcc ccagcaggac tggccgcaac cgctgcacat gaccggcttc ccgctgttcg 1680
acggcagcgt cccggggacc cgcctcgacg acgagttgca gcgcttcctc gagcagggca 1740
gtcggccgct ggtgttcacc cagggttcga ccgagcacct gcagggagac ttctatgcca 1800
tggccttgcg cgcgctggag cgtctcggcg cccgcggcat cttcctcacc ggcgccggcc 1860
aggagccgct gcgtggcttg ccgagccacg tgctgcaacg ctcgtacgtg ccgttggggg 1920
ccttgctgcc ggcgtgcgcc gggctggtcc acccggccgg catcggcgcc atgagcctgg 1980
cgctggcggc gggggtgccg caggtgctgc tgccttgcgc ccacgaccag ttcgacaacg 2040
ccgaacgcct ggtccgcctc ggctgcggta tccgcctggg cctgccgcta cgcgagcagg 2100
cgctgcgcga gtcgctctgg cggctgctcg aggacccggc gctggcggcg gcctgtcggc 2160
gtttcatgga attgtcacaa ccgcacagta tcgcttgcgg taaagcggcc caagtggtcg 2220
aacgttgtca tagggagggg gatgtgcgat ggctgaaagc cgcgtcctga gccgtgctgg 2280
cagaattctc tagaggcgcg cc 2302
<210> 66
<211> 8325
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 66
ctagatacgg gagaagaacg atcatggacc ggatagacat gggcgtgctg gtggtgctgt 60
tcaatcctgg cgacgacgac ctggaacacc ttggcgaact ggcggcggcc tttccgcaac 120
tgcgcttcct cgccgtcgac aactcgccgc acagcgatcc gcagcgcaac gcccggctgc 180
gcgggcaagg catcgccgtg ctctaccacg gcaaccggca gggcatcgcc ggcgccttca 240
accaggggct cgacacgctg ttccggcgcg gcctgcaggg tgtgctgctg ctcgaccagg 300
actcccgtcc cggcggcgcc ttcctcgccg cccagtggcg caacctgcag gcatgcaacg 360
gccaggcctg cctgctcggc ccacggatct tcgaccgggg cgaccggcgc ttcctgccgg 420
ccatccacct cgacgggctg gcgctcaggc aactgtccct ggacggcctg acgaccccac 480
agcgcacctc gttcctgatc tcctccggct gcctgctgac ccgcgaggcc taccagcgcc 540
tcggccactt cgacgaggaa ctgttcatcg accacgtgga caccgagtac agcctgcgcg 600
cccaggcgct ggacgtgccc ctgtacgtcg acccgcggct ggtcctcgag caccgcatcg 660
gcacgcgcaa gacccgccgc ctcggcggtc tcagcctcag cgcgatgaac cacgccccac 720
tgcgccgcta ctacctggcg cgcaacggcc tgctggtcct gcgccgctac gcccggtcct 780
cgccgctggc cctgctggcg aacctgccga ccctgaccca gggcctcgcg gtgctcctgc 840
tcgaacgcga caagctgctc aagctgcgct gcctgggctg gggcctgtgg gacggcctgc 900
gggggcgcgg cggcgcgctg gagcgcaacc gcccgcgcct gctgaagcgc ctcgccggtc 960
cggcggtggc gcccacagtt cccggcaagg ccaaggccta gtcggcgaaa cgcattgagc 1020
tccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt 1080
cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc 1140
gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc 1200
ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 1260
gttaaatcag ctcatttttt aaccaatagg ccgactgcga tgagtggcag ggcggggcgt 1320
aattttttta aggcagttat tggtgccctt aaacgcctgg tgctacgcct gaataagtga 1380
taataagcgg atgaatggca gaaattcgaa agcaaattcg acccggtcgt cggttcaggg 1440
cagggtcgtt aaatagccgc ttatgtctat tgctggttta ccggtttatt gactaccgga 1500
agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg 1560
gaggtaataa ttgacgatat gatcatttat tctgcctccc agagcctgat aaaaacggtg 1620
aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac 1680
cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gaggcggcta cagccgatag 1740
tctggaacag cgcacttacg ggttgctgcg caacccaagt gctaccggcg cggcagcgtg 1800
acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc 1860
ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct 1920
ggttccatgc ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt 1980
tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat 2040
tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg 2100
tcgcggggct ggccccacgc cacgcggtca ttgaccacgt aggccgacac ggtgccgggg 2160
ccgttgagct tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg 2220
accgtccgca aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg 2280
ttctggtggc ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc 2340
gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc 2400
ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg 2460
tagggtgccg cacggttgcg gcaccatgcg caatcagctg caacttttcg gcagcgcgac 2520
aacaattatg cgttgcgtaa aagtggcagt caattacaga ttttctttaa cctacgcaat 2580
gagctattgc ggggggtgcc gcaatgagct gttgcgtacc cccctttttt aagttgttga 2640
tttttaagtc tttcgcattt cgccctatat ctagttcttt ggtgcccaaa gaagggcacc 2700
cctgcggggt tcccccacgc cttcggcgcg gctccccctc cggcaaaaag tggcccctcc 2760
ggggcttgtt gatcgactgc gcggccttcg gccttgccca aggtggcgct gcccccttgg 2820
aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc gccttcgact 2880
gcccccactc gcataggctt gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt 2940
cgctgcgcga gccttgaccc gccttccact tggtgtccaa ccggcaagcg aagcgcgcag 3000
gccgcaggcc ggaggctttt ccccagagaa aattaaaaaa attgatgggg caaggccgca 3060
ggccgcgcag ttggagccgg tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc 3120
ctgtggtcaa gctcgtgggc aggcgcagcc tgtccatcag cttgtccagc agggttgtcc 3180
acgggccgag cgaagcgagc cagccggtgg ccgctcgcgg ccatcgtcca catatccacg 3240
ggctggcaag ggagcgcagc gaccgcgcag ggcgaagccc ggagagcaag cccgtagggc 3300
gccgcagccg ccgtaggcgg tcacgacttt gcgaagcaaa gtctagtgag tatactcaag 3360
cattgagtgg cccgccggag gcaccgcctt gcgctgcccc cgtcgagccg gttggacacc 3420
aaaagggagg ggcaggcatg gcggcatacg cgatcatgcg atgcaagaag ctggcgaaaa 3480
tgggcaacgt ggcggccagt ctcaagcacg cctaccgcga gcgcgagacg cccaacgctg 3540
acgccagcag gacgccagag aacgagcact gggcggccag cagcaccgat gaagcgatgg 3600
gccgactgcg cgagttgctg ccagagaagc ggcgcaagga cgctgtgttg gcggtcgagt 3660
acgtcatgac ggccagcccg gaatggtgga agtcggccag ccaagaacag caggcggcgt 3720
tcttcgagaa ggcgcacaag tggctggcgg acaagtacgg ggcggatcgc atcgtgacgg 3780
ccagcatcca ccgtgacgaa accagcccgc acatgaccgc gttcgtggtg ccgctgacgc 3840
aggacggcag gctgtcggcc aaggagttca tcggcaacaa agcgcagatg acccgcgacc 3900
agaccacgtt tgcggccgct gtggccgatc tagggctgca acggggcatc gagggcagca 3960
aggcacgtca cacgcgcatt caggcgttct acgaggccct ggagcggcca ccagtgggcc 4020
acgtcaccat cagcccgcaa gcggtcgagc cacgcgccta tgcaccgcag ggattggccg 4080
aaaagctggg aatctcaaag cgcgttgaga cgccggaagc cgtggccgac cggctgacaa 4140
aagcggttcg gcaggggtat gagcctgccc tacaggccgc cgcaggagcg cgtgagatgc 4200
gcaagaaggc cgatcaagcc caagagacgg cccgagacct tcgggagcgc ctgaagcccg 4260
ttctggacgc cctggggccg ttgaatcggg atatgcaggc caaggccgcc gcgatcatca 4320
aggccgtggg cgaaaagctg ctgacggaac agcgggaagt ccagcgccag aaacaggccc 4380
agcgccagca ggaacgcggg cgcgcacatt tccccgaaaa gtgccacctg ggatgaatgt 4440
cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg 4500
cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga 4560
attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc 4620
tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg 4680
aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 4740
ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 4800
gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 4860
cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 4920
ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 4980
agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 5040
ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 5100
agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 5160
tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 5220
gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 5280
catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 5340
ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 5400
ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 5460
ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 5520
gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 5580
ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 5640
ctggagttct tcgcccaccc ccatgggcaa atattatacg caaggcgaca aggtgctgat 5700
gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct 5760
taatgaatta caacagtttt tatgcatgcg cccaatacgc aaaccgcctc tccccgcgcg 5820
ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 5880
gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat 5940
gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag 6000
ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctggg 6060
taccgaaaat ttttgggagg tgtgaaatgc ggcgcgaaag tctgttggta tcggtttgca 6120
agggcctgcg ggtacatgtc gagcgcgttg ggcaggatcc cgggcgcagc acggtgatgc 6180
tggtcaacgg cgcgatggcg accaccgcct cgttcgcccg gacctgcaag tgcctggccg 6240
aacatttcaa cgtggtgctg ttcgacctgc ccttcgccgg gcagtcgcgt cagcacaacc 6300
cgcagcgcgg gttgatcacc aaggacgacg aggtggaaat cctcctggcg ctgatcgagc 6360
gcttcgaggt caatcacctg gtctccgcgt cctggggcgg tatctccacg ctgctggcgc 6420
tgtcgcgcaa tccgcgcggc atccgcagct cggtggtgat ggcattcgcc cctggactga 6480
accaggcgat gctcgactac gtcgggcggg cgcaggcgct gatcgagctg gacgacaagt 6540
cggcgatcgg ccatctgctc aacgagaccg tcggcaaata cctgccgcag cgcctgaaag 6600
ccagcaacca tcagcacatg gcttcgctgg ccaccggcga atacgagcag gcgcgctttc 6660
acatcgacca ggtgctggcg ctcaacgatc ggggctactt ggcttgcctg gagcggatcc 6720
agagccacgt gcatttcatc aacggcagct gggacgaata caccaccgcc gaggacgccc 6780
gccagttccg cgactacctg ccgcactgca gtttctcgcg ggtggagggc accgggcatt 6840
tcctcgacct ggagtccaag ctggcagcgg tacgcgtgca ccgcgccctg ctcgagcacc 6900
tgctgaagca accggagccg cagcgggcgg aacgcgcggc gggattccac gagatggcca 6960
tcggctacgc ctgaaccctt gacctgcgaa gacccggcct ggccgggctt tgcggttgca 7020
taacgcacgg agtagcccca tgcacgccat cctcatcgcc atcggctcgg ccggcgacgt 7080
atttcccttc atcggcctgg cccggaccct gaaactgcgc gggcaccgcg tgagcctctg 7140
caccatcccg gtgtttcgcg acgcggtgga gcagcacggc atcgcgttcg tcccgctgag 7200
cgacgaactg acctaccgcc ggaccatggg cgatccgcgc ctgtgggacc ccaagacgtc 7260
cttcggcgtg ctctggcaag ccatcgccgg gatgatcgag ccggtctacg agtacgtctc 7320
ggcgcagcgc catgacgaca tcgtggtggt cggctcgcta tgggcgctgg gcgcacgcat 7380
cgctcacgag aagtacggga ttccctacct gtccgcgcag gtctcgccat cgaccctgtt 7440
gtcggcgcac ctgccgccgg tacaccccaa gttcaacgtg cccgagcaga tgccgctggc 7500
gatgcgcaag ctgctctggc gctgcatcga gcgcttcaag ctggatcgca cctgcgcgcc 7560
ggagatcaac gcggtgcgcc gcaaggtcgg cctggaaacg ccggtgaagc gcatcttcac 7620
ccaatggatg cattcgccgc agggcgtggt ctgcctgttc ccggcctggt tcgcgccgcc 7680
ccagcaggat tggccgcaac ccctgcacat gaccggcttc ccgctgttcg acggcagtat 7740
cccggggacc ccgctcgacg acgaactgca acgctttctc gatcagggca gccggccgct 7800
ggtgttcacc cagggctcga ccgaacacct gcagggcgac ttctacgcca tggccctgcg 7860
cgcgctggaa cgcctcggcg cgcgtgggat cttcctcacc ggcgccggcc aggaaccgct 7920
gcgcggcttg ccgaaccacg tgctgcagcg cgcctacgcg ccactgggag ccttgctgcc 7980
atcgtgcgcc gggctggtcc atccgggcgg tatcggcgcc atgagcctag ccttggcggc 8040
gggggtgccg caggtgctgc tgccctgtgc ccacgaccag ttcgacaatg ccgaacggct 8100
ggtccggctc ggctgcggga tgcgcctggg cgtgccgttg cgcgagcagg agttgcgcgg 8160
ggcgctgtgg cgcttgctcg aggacccggc catggcggcg gcctgtcggc gtttcatgga 8220
attgtcacaa ccgcacagta tcgcttgcgg taaagcggcc caggtggtcg aacgttgtca 8280
tagggagggg gatgctcgat ggctgaaggc tgcgtcctga acggt 8325
<210> 67
<211> 8335
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 67
ctagatacgg gagaagaacg atcatgacga tcctgggggc gctggtgatt ctgtacgacc 60
cgacggacga gcagttgtcg gggctggagg cgctcgcgcg cgacagcgac gcgctcgtgg 120
tcgtggacaa cacgccgcac gagcacgcgg cggcgcgcga gcgggtgcgt gcgctgtcgg 180
cgcggacgaa cacggtgtgg cgacaccacg gcaaccgggg cggggtcgcg ggcgggtaca 240
acgcggggct gtcggtgctg ttcgcgcagg gcgtcgaggc ggtcgcgctg ttcgaccagg 300
actcgacggt gccggccggg tacttcgagc ggatgcgcga ggcgtgcgcg caactgggtg 360
agcaaccggg cgcgcacgcg ggcgcgttca tcgcgggccc gcggatctac gacgcgaacg 420
agcagcgctt cctgccggag ctgatgacga gcggggtgac ggtgcgccgc gtgcgggtgg 480
agggcgagac ggcgccgcag cgctgcgcgt tcctgatctc gtcgggcagc gtgatttcgc 540
gggccgcgta cgcgcggctc ggtcgattcg acgaggcgct gttcatcgat cacgtcgaca 600
ccgagtattg cctgcgcgcg ctcgcgcaca acgtgccgct gtacgtggtg ccgccgctcg 660
tgctgacgca ccggatcggc gcgcggcgcc ggcacaaggt ggggccgttc gagctgacgg 720
cgatgcatca cgggtggttg cgccgatact acggcgcgcg caacgcgatg caactggggc 780
tgcagtacgg cttgcggttt ccggtggcgc tggtgccgaa tctgctgacg atatggcagg 840
tgatccaggt ggtgctgtgc gagcgggaga agggcgcgaa gctgcgcggg atcgcgctgg 900
gcgtgctcga cggcctgttc gggcggctgg gatcgttcga cgatgcgcgc gcgggcgcgg 960
cggcgcgcga gccggtgcgg caggaatgat cggcgaaacg cattgagctc caattcgccc 1020
tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa 1080
aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 1140
aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 1200
tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 1260
cattttttaa ccaataggcc gactgcgatg agtggcaggg cggggcgtaa tttttttaag 1320
gcagttattg gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat 1380
gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa 1440
atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc 1500
gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt 1560
gacgatatga tcatttattc tgcctcccag agcctgataa aaacggtgaa tccgttagcg 1620
aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg 1680
cggggaggca gacaaggtat agggcggcga ggcggctaca gccgatagtc tggaacagcg 1740
cacttacggg ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc 1800
ggctccaacg gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc 1860
tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc 1920
ggaatgccgg gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc 1980
ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg 2040
tcgtgatcaa ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg 2100
ccccacgcca cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc 2160
acgacggaga tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa 2220
gaacgtccga tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc 2280
atctgcgcca cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg 2340
gcccacgcct catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct 2400
tgaatgccga tttctctgga ctgcgtggcc atgcttatct ccatgcggta gggtgccgca 2460
cggttgcggc accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg 2520
ttgcgtaaaa gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg 2580
ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt 2640
tcgcatttcg ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc 2700
ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga 2760
tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact 2820
cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc 2880
ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc 2940
cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg 3000
aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt 3060
ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc 3120
tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg 3180
aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg 3240
agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc 3300
gtaggcggtc acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc 3360
cgccggaggc accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg 3420
caggcatggc ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg 3480
cggccagtct caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga 3540
cgccagagaa cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg 3600
agttgctgcc agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg 3660
ccagcccgga atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg 3720
cgcacaagtg gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc 3780
gtgacgaaac cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc 3840
tgtcggccaa ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg 3900
cggccgctgt ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca 3960
cgcgcattca ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca 4020
gcccgcaagc ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa 4080
tctcaaagcg cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc 4140
aggggtatga gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg 4200
atcaagccca agagacggcc cgagaccttc gggagcgcct gaagcccgtt ctggacgccc 4260
tggggccgtt gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg 4320
aaaagctgct gacggaacag cgggaagtcc agcgccagaa acaggcccag cgccagcagg 4380
aacgcgggcg cgcacatttc cccgaaaagt gccacctggg atgaatgtca gctactgggc 4440
tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac 4500
atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg 4560
ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc 4620
aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg 4680
catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 4740
cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 4800
agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 4860
gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 4920
gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 4980
ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 5040
gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 5100
catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 5160
agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 5220
cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 5280
tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 5340
catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 5400
cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 5460
tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac 5520
ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 5580
gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 5640
gcccaccccc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat 5700
tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca 5760
acagttttta tgcatgcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 5820
ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 5880
taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 5940
tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 6000
ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctgggta ccgaattcaa 6060
aactttttgg gaggtgtgag atgcggcgcg aaagtctgtt ggtaacggta tgcaagggcc 6120
tgcgggtaca tgtcgagcgc gtggggcagg atcccgggcg cgacacggtg atgctggtca 6180
acggcgcgat ggcgaccacc gcctcgttcg cccggacctg caagtgcctg gccgaacatt 6240
tcaacgtggt gctgttcgac ctgcccttcg ccgggcagtc gcggcagcac aatccgcagc 6300
gcgggttgat caccaaggac gacgaggtgg agattctcct ggcgctgatc gagcgcttcg 6360
ctgtcaacca cctggtctcg gcctcctggg gcggcatctc cacgctgctg gcgctgtcgc 6420
gcaacccgcg cggggtccgc agctcggtgg tgatggcgtt cgcgccgggg ctgaaccagg 6480
cgatgctcga ttatgtcggg cgggcccagg aactgatcga actggacgac aagtcggcga 6540
tcggccacct gctcaacgag accgtcggca agtacctgcc gccgcggctg aaggccagca 6600
accatcagca catggcctcc ctggccactg gcgagtacga gcaggcgcgt ttccacatcg 6660
accaggtgct ggcgctcaat gaccgtggct acctgagctg cctggggcag atccagagtc 6720
acgtgcattt catcaacggc agctgggacg agtacaccac cgccgaggac gcccgccagt 6780
tccgcgatta cctgccgcat tgcagttttt cgcgggtgga aggcaccggg cacttcctcg 6840
acctggagtc caagctggcg gcggcgcgtg tgcaccgggc gttgctcgag cacctgctgg 6900
cgcaaccgga accgtggcgc tccgagcagg cggcgggatt ccacgagatg gccatcggct 6960
acgcctgacc cgtcgggatc tgcgaaggcc cggcatggcc gggccttgcc gttgcacaac 7020
gcaaggagta gccccatgca cgccattctc atcgccatcg gttcggccgg cgacgtgttc 7080
cccttcatcg gcctggcccg caccctgaag ttgcgcggcc accgcgtcag cctgtgcacc 7140
attccggtgt ttcgcgccgc ggtggagcag cacggcatcg agttcgtccc gctcagcgac 7200
gaactgacct accgccggac catgggcgac ccgcgcctgt gggatccgaa gacctcgttc 7260
ggagtgctct ggcaggccat cgccgggatg atcgagccgg tctacgagta cgtctgcgca 7320
cagcgccacg acgacatcgt ggtggtcggt tcgctgtggg ccctgggcgc gcggatcgcc 7380
catgagaaat acgggattcc ctacctgtcg gtgcaggtct cgccgtcgac cctgctgtcg 7440
gcgcacctgc cgccggtcca ccccaggttc aacgtgcccg agcaggtccc gctggcgatg 7500
cgcaagttgc tctggcgctg catcgaacgc ttcaagctgg accgcacctg cgccccggag 7560
atcaacgcgg tgcgccgcaa ggtcggcctg gtcggcccgg cgaagcgcat cttcacccag 7620
tggatgcatt cgccacaggg agtgctctgc ctgttcccgg cctggttcgc accgccccag 7680
caggactggc cgcaaccgct gcacatgacc ggcttcccgc tgttcgacgg cagcgtcccg 7740
gggacccgcc tcgacgacga gttgcagcgc ttcctcgagc agggcagtcg gccgctggtg 7800
ttcacccagg gttcgaccga gcacctgcag ggagacttct atgccatggc cttgcgcgcg 7860
ctggagcgtc tcggcgcccg cggcatcttc ctcaccggcg ccggccagga gccgctgcgt 7920
ggcttgccga gccacgtgct gcaacgctcg tacgtgccgt tgggggcctt gctgccggcg 7980
tgcgccgggc tggtccaccc ggccggcatc ggcgccatga gcctggcgct ggcggcgggg 8040
gtgccgcagg tgctgctgcc ttgcgcccac gaccagttcg acaacgccga acgcctggtc 8100
cgcctcggct gcggtatccg cctgggcctg ccgctacgcg agcaggcgct gcgcgagtcg 8160
ctctggcggc tgctcgagga cccggcgctg gcggcggcct gtcggcgttt catggaattg 8220
tcacaaccgc acagtatcgc ttgcggtaaa gcggcccaag tggtcgaacg ttgtcatagg 8280
gagggggatg tgcgatggct gaaagccgcg tcctgagccg tgctggcaga attct 8335
<210> 68
<211> 1023
<212> DNA
<213> Pseudomonas aeruginosa
<400> 68
tctagatacg ggagaagaac gatcatggac cggatagaca tgggcgtgct ggtggtgctg 60
ttcaatcctg gcgacgacga cctggaacac cttggcgaac tggcggcggc ctttccgcaa 120
ctgcgcttcc tcgccgtcga caactcgccg cacagcgatc cgcagcgcaa cgcccggctg 180
cgcgggcaag gcatcgccgt gctctaccac ggcaaccggc agggcatcgc cggcgccttc 240
aaccaggggc tcgacacgct gttccggcgc ggcctgcagg gtgtgctgct gctcgaccag 300
gactcccgtc ccggcggcgc cttcctcgcc gcccagtggc gcaacctgca ggcatgcaac 360
ggccaggcct gcctgctcgg cccacggatc ttcgaccggg gcgaccggcg cttcctgccg 420
gccatccacc tcgacgggct ggcgctcagg caactgtccc tggacggcct gacgacccca 480
cagcgcacct cgttcctgat ctcctccggc tgcctgctga cccgcgaggc ctaccagcgc 540
ctcggccact tcgacgagga actgttcatc gaccacgtgg acaccgagta cagcctgcgc 600
gcccaggcgc tggacgtgcc cctgtacgtc gacccgcggc tggtcctcga gcaccgcatc 660
ggcacgcgca agacccgccg cctcggcggt ctcagcctca gcgcgatgaa ccacgcccca 720
ctgcgccgct actacctggc gcgcaacggc ctgctggtcc tgcgccgcta cgcccggtcc 780
tcgccgctgg ccctgctggc gaacctgccg accctgaccc agggcctcgc ggtgctcctg 840
ctcgaacgcg acaagctgct caagctgcgc tgcctgggct ggggcctgtg ggacggcctg 900
cgggggcgcg gcggcgcgct ggagcgcaac cgcccgcgcc tgctgaagcg cctcgccggt 960
ccggcggtgg cgcccacagt tcccggcaag gccaaggcct agtcggcgaa acgcattgag 1020
ctc 1023
<210> 69
<211> 13768
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 69
ctagaggtgg acggccgcac gtatgagcac gcggtgacgc aggtgctgca ggccacgggg 60
gtgcggggga ttctgctcgc gccggatgcg ccggatgcgc cggcggcatc ggacggggcg 120
gcgctgctca agcgccgcta cgtgccgctc gcggcgttgc tgccgcgctg ccgggcgctg 180
gtgcaccacg gggggatcgg gacggcgtcg ctcgcgtacg cggcgggggt gccgcaggtg 240
gtgacgccgt tcgcgcacga ccagttcgac aacgcgcagc gggtggcggc gagcggctgc 300
ggggtgcggc tggacgcgcc ggtgcgcggc gagccgctcg cgcgggcgct ggcgcaggtg 360
ctgggcgacg cggcgatggc ggcgcgctgc gcgcaggtgc gcgcgcggat ggcggcggag 420
ccgaacggct gcgacgcggc ggcgcgcttc atcgagcgct tcgcgccggg cgtcgcggcg 480
cggcgggcgc agccggcatg agcgcgcagg cgatgtcggc ggatcaggcg ggcgttgcgc 540
cgccggcggc cgccccgctg cgcggcgcga agctcgcgct gctgacgttc gcgctgtcgc 600
tcgcgacgtt catcgaagtg ctggattcga cggtggcgaa cgtggcggtg ccggcgatct 660
cgggcagcct cggggtgtcg aacagccagg gcacgtgggt gatcagctcg tactcggtgg 720
ccgcggcgat cgcggtgccg ctgacggggt ggcttgcgcg gcgcgtgggc gagctgaggc 780
tgttcgtggc gtcggtgatc ctgttcacgc tgacgtcgct gctgtgcggg ctcgcgcggg 840
acctggaggt gctggttgcg tgccgggcgc tgcaggggct gttctcgggg ccgatggtgc 900
cgctgtcgca gacgatcctg atgcgcgcgt tcccgccggc gcggcgcacg ctggcgctgg 960
cgctgtgggg gatgacggtg ctgctcgcgc cgatcttcgg gccggtggtg ggcggctggc 1020
tgatcgacaa cttctcgtgg ccgtggatct tcctgatcaa cctgccgatc gggctgttct 1080
cgttcgcggt gtgcacgctg atgctgcgcc cgcaggcgca gcgcggcgag gcgagcccga 1140
tcgacgcgcc ggggatcgtg ctgctggtga tcggggtggg ctcgctgcag gcgatgctgg 1200
acctggggca cgaccggggc tggttcgatt cgccgctgat cacggcgctg gcgatcgcgg 1260
cgggggtgtc gctcgtgtcg ctgctgatct gggagctggg cgaggcgcat ccggtggtgg 1320
atctgagcct gttccgggag cggaccttca cgttctgcgt ggtgatcatc tcgctgggga 1380
tgatgagctt ctcggtggtg ggggtggtgt ttccgctgtg gctgcaggcg gtgatgggat 1440
acacggcgta ccaggcgggg ctggcgacgg cgtcgatggg ggtgctggcg ctggtgttct 1500
cgatcctggt ggggctgtac gcgagccggg tggacgcgcg ggtgctggtg acgttcgggt 1560
tcggggtgtt tgcggcggtg atgtggtgga gcacgcactt cacgctgtcg atgacgttcg 1620
cgcaggtggt gacgccgcgg ctgattcagg ggatggggct gccgtgcttc ttcataccgc 1680
tgacggcggc gacgctgtcg cgggtgccgg acgagaagct ggcggcggcg tcgagcctgt 1740
cgaacttcct gcggacgctg tcggcggcgt tcggcacggc gctgagcgtg acgtggtggg 1800
acaaccgggc gacgtaccac tacgcggtgg tgtcgcaatc ggtgacgcgc gcctcggaga 1860
acacgcagcg gtacgtggac gcgctgcacg cgatggggct gcacggcgcg cgggagctga 1920
gctcgctgca ccaggtggtg cggcagcagg cgtacatgat ggcgacgaac gacatgttct 1980
acatggcgag cgcgacgtgc ctgctgctgg cggggctgat gtggctgacg cggccgaagc 2040
ggggcgcggc ggcggcgctc gggcactgag gcgaggcatg tcgcgccccg catgacgaag 2100
gcgaaggaga agggcgatgc gccgaagtcc tggggacgcg gcgcgtcgat gcggcaacga 2160
agcgggcatt tcggcattcc gaaccaccaa agggaagagc gatgacgatc ctgggggcgc 2220
tggtgttcgg gcggctggga tcgttcgacg atgcgcgcgc gggcgcggcg gcgcgcgagc 2280
cggtgcggca ggaatgaacg gaacgggccg cagcgggata ccggaaagca agaaggacgc 2340
atcatacgaa tgacgcagac agcaacgcaa gcagccactc gcgcgatgat cgcgacagga 2400
agccgcgcgg cgcgccggct cgcggcagcc gcgctcgcgt gggcgctcgc cggctgcgtg 2460
ccgtcgggct tcgagccggc gctcgcgccg cgcacgccgg gcgacgacgc gctcgcgcac 2520
acggcggggg gcgccgcgca cggcgcatgg ccgagccccg actgggtccg gcagctcggc 2580
gatccgcaac tcgacgcgct cgtcgacgag gcgctgcggc agaacccgac gctgcaggcc 2640
gcgcaggcgc gcatcggcgt cgcgcagtcg cagctgcagc agttcgaatc gctgacgggg 2700
ctcaccgcga cggcgggcgc gtcgctctcc aaggcgcacg tgccgcgctc gggcggcacc 2760
atcaatacga cgttcaacgg cttgccggtg tcggtgccgc tcgtcggcga atcggtggtg 2820
tcgtcgtcgt cgctgttcgt cgggctgaac tatcagctgg acctgtgggg caagaacgcg 2880
gcggccacgc gcgggctgct gtcgatgcgc gatgcggcgc gcgtggaggc cgagcaggcg 2940
cggctcgcgc tgtcggtggc gatcgtgacg ctgtacggcg agctggaccg cgcgtatgcg 3000
ctgcgcgagc tgctgcagca gaagcgccgc gcgagcgagc aggtggagac ggtgctgcgc 3060
gagcgcgcgg cgcgcgggat cgacaacggc tacgatgcgg acgacgcggc gctcaagcgg 3120
ggcaagctgc tcgagcagct cgcgctgacc gacgagcaga tccagttgca gaagctgcaa 3180
ctgggggtgc tgagcgggcg ggggccggag cgcgggctgt cgctcgcgcg gccgaagctc 3240
gcgccgctcg cggacgcgcc gctgccggcg cggctgccgg ccgggctgct ggggcggcgg 3300
ccggacatcg tcgcggcgcg gctgcgggtg gaggcggcgt acgcggcgat cgacggcacg 3360
cgcgcgtcgt tctacccgga cgtgaacctg gcggcgctgg gcgggctgtt cgcgctcacg 3420
ccggcgtcgc tgttcaagca cgatgcgctg gggggctcga tcggtccggc gctgtcgctg 3480
ccgatcttcg atcgcggccg gctgaaggcg aagctggggg gcgacgtggc gaacgcggac 3540
gtggcgctgg cgctgtacaa ccagacggtg gatgcggcgc tgggcgaggt ggcgcggcag 3600
ttgacgtcgc tgtcgacggt ggatgcgctg ctcgaggcgc agcagcaggc ggtgcgctcg 3660
gcgcagcgga tggtggcgct ggcgcaggac cggcaccggc gggggatggg gatgcgcaag 3720
gacgtgaacg tggcgaagct gacgctgctg gacgagcgtg cgcacgtgat cgagctgcag 3780
gcgcggcggc ggacgctgcg ggtggggctg atcggggcgc tgggcggcgg cttcgacgcg 3840
cggccggcgg gcggcgcgcc gctcgcgcag ggcaagccgt tcgcggcggc gagcgacagg 3900
ccgcccgatt gagcggcacg cacgcatgcg gcccgaagcc accgacaccc gaagacaccg 3960
acaccaacgc caccttcacc gtgtacacga gcgattcaac cgacaccgcc cccgagcatc 4020
gaagcccgtc gggccgatcc gcgacggctt gcgggccggc ccggccgttg ccggccggcg 4080
ccaccgacat cacgcacgcg aagaccttga acgataccgc caccgatacc ccgcgcgcga 4140
aggcgcccac cgatccggcc gccctcgacg gcgcgcacgc gcagcccgtg ccggcgcacg 4200
agcgcggatc gcctccgccg ccggaagccg cggcgacgct cgccgcgcgc cgcgcgacgc 4260
gccgccggcg cttcgcgctg ttcttcgggc tgctggcgct ggccgcgctg accgcggggc 4320
tctactggtt cgtcgccggg cgcttcagcg aggagacgga cgacgcgtac gtggccggca 4380
acgtggtgca gatcgccgcg cagatccagg ggacggtgac cgacgtgctg gtggcggaca 4440
cgcagcaggt gaaggcgggg caggcgctgg tgaagctcga cgacgcggac gcgtcggcgg 4500
cgttcgcgca ggcgcgggcg cagctcgcgc aggcggtgcg gcaggtggcg aacacgcggc 4560
tctcgatggg gatgtacgag gagacggtga aggcgcgcga ggcggacctg aagcttgcgc 4620
agcaggcgta tccggaggaa ctggcgcggc gaaagtcgtc gctggcgaac gcgcaggcgg 4680
cgctggcggg ggcgcaggcg cagctggagg cggcgcgcgc gctgggcagc gagcggccgg 4740
tcgagcagaa cccggcggtg cagcaggcgg ccgcgcagtt caagctggcg taccggaacc 4800
tgaggcgcac gacgatcgtg tcgccggtgg acggcacggt cggtcagcgg tcggtgcaga 4860
tcggtcagca ggtggggccg ggggtgccgc tgatgtcggt ggtgcagttg cggcaggtgt 4920
gggtggaggc gaacttcaag gaagggcaga tccggcacat gcgggtgggc cagccggtgc 4980
ggctcgaatc ggacctgtac ggcgcgcggg tgacgtacca cggccgggtg gagggggtct 5040
cggcgggcac gggcagcgcg ttctcgatgc tgccgtcgca gaacgcggcg gggaactgga 5100
tcaaggtggt gcagcgcctg ccggtggtga tctcgctgga gccgtcggag ctggcggcgc 5160
acccgctgcg ggtggggctg tcgatgcgcg cgacggtgga gacgaaggtg cgtggcggcc 5220
gcctgctcga cggcgacgcg ccgctgccgg ggctgcgcac gcgggtgcac gaagcgcagg 5280
cgggcgaggc cgaggccgcg gcttcggcag tgattcggga gaatgacggc cgcaggtgac 5340
gggcggttgc gggatcgctc tagagcggcc gccaccgcgg tggagctcca attcgcccta 5400
tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa 5460
ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 5520
tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 5580
gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca 5640
ttttttaacc aataggccga ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc 5700
agttattggt gcccttaaac gcctggtgct acgcctgaat aagtgataat aagcggatga 5760
atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat 5820
agccgcttat gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt 5880
gtgcttctca aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga 5940
cgatatgatc atttattctg cctcccagag cctgataaaa acggtgaatc cgttagcgag 6000
gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg 6060
gggaggcaga caaggtatag ggcggcgagg cggctacagc cgatagtctg gaacagcgca 6120
cttacgggtt gctgcgcaac ccaagtgcta ccggcgcggc agcgtgaccc gtgtcggcgg 6180
ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg 6240
cccgcgccgt tcccattcct ccgtttcggt caaggctggc aggtctggtt ccatgcccgg 6300
aatgccgggc tggctgggcg gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg 6360
atacagggtc gggatgcggc gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc 6420
gtgatcaacc accacggcgg cactgaacac cgacaggcgc aactggtcgc ggggctggcc 6480
ccacgccacg cggtcattga ccacgtaggc cgacacggtg ccggggccgt tgagcttcac 6540
gacggagatc cagcgctcgg ccaccaagtc cttgactgcg tattggaccg tccgcaaaga 6600
acgtccgatg agcttggaaa gtgtcttctg gctgaccacc acggcgttct ggtggcccat 6660
ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc 6720
ccacgcctca tgcgctttgc gttccgtttg cacccagtga ccgggcttgt tcttggcttg 6780
aatgccgatt tctctggact gcgtggccat gcttatctcc atgcggtagg gtgccgcacg 6840
gttgcggcac catgcgcaat cagctgcaac ttttcggcag cgcgacaaca attatgcgtt 6900
gcgtaaaagt ggcagtcaat tacagatttt ctttaaccta cgcaatgagc tattgcgggg 6960
ggtgccgcaa tgagctgttg cgtacccccc ttttttaagt tgttgatttt taagtctttc 7020
gcatttcgcc ctatatctag ttctttggtg cccaaagaag ggcacccctg cggggttccc 7080
ccacgccttc ggcgcggctc cccctccggc aaaaagtggc ccctccgggg cttgttgatc 7140
gactgcgcgg ccttcggcct tgcccaaggt ggcgctgccc ccttggaacc cccgcactcg 7200
ccgccgtgag gctcgggggg caggcgggcg ggcttcgcct tcgactgccc ccactcgcat 7260
aggcttgggt cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct 7320
tgacccgcct tccacttggt gtccaaccgg caagcgaagc gcgcaggccg caggccggag 7380
gcttttcccc agagaaaatt aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg 7440
agccggtggg tatgtggtcg aaggctgggt agccggtggg caatccctgt ggtcaagctc 7500
gtgggcaggc gcagcctgtc catcagcttg tccagcaggg ttgtccacgg gccgagcgaa 7560
gcgagccagc cggtggccgc tcgcggccat cgtccacata tccacgggct ggcaagggag 7620
cgcagcgacc gcgcagggcg aagcccggag agcaagcccg tagggcgccg cagccgccgt 7680
aggcggtcac gactttgcga agcaaagtct agtgagtata ctcaagcatt gagtggcccg 7740
ccggaggcac cgccttgcgc tgcccccgtc gagccggttg gacaccaaaa gggaggggca 7800
ggcatggcgg catacgcgat catgcgatgc aagaagctgg cgaaaatggg caacgtggcg 7860
gccagtctca agcacgccta ccgcgagcgc gagacgccca acgctgacgc cagcaggacg 7920
ccagagaacg agcactgggc ggccagcagc accgatgaag cgatgggccg actgcgcgag 7980
ttgctgccag agaagcggcg caaggacgct gtgttggcgg tcgagtacgt catgacggcc 8040
agcccggaat ggtggaagtc ggccagccaa gaacagcagg cggcgttctt cgagaaggcg 8100
cacaagtggc tggcggacaa gtacggggcg gatcgcatcg tgacggccag catccaccgt 8160
gacgaaacca gcccgcacat gaccgcgttc gtggtgccgc tgacgcagga cggcaggctg 8220
tcggccaagg agttcatcgg caacaaagcg cagatgaccc gcgaccagac cacgtttgcg 8280
gccgctgtgg ccgatctagg gctgcaacgg ggcatcgagg gcagcaaggc acgtcacacg 8340
cgcattcagg cgttctacga ggccctggag cggccaccag tgggccacgt caccatcagc 8400
ccgcaagcgg tcgagccacg cgcctatgca ccgcagggat tggccgaaaa gctgggaatc 8460
tcaaagcgcg ttgagacgcc ggaagccgtg gccgaccggc tgacaaaagc ggttcggcag 8520
gggtatgagc ctgccctaca ggccgccgca ggagcgcgtg agatgcgcaa gaaggccgat 8580
caagcccaag agacggcccg agaccttcgg gagcgcctga agcccgttct ggacgccctg 8640
gggccgttga atcgggatat gcaggccaag gccgccgcga tcatcaaggc cgtgggcgaa 8700
aagctgctga cggaacagcg ggaagtccag cgccagaaac aggcccagcg ccagcaggaa 8760
cgcgggcgcg cacatttccc cgaaaagtgc cacctgggat gaatgtcagc tactgggcta 8820
tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat 8880
ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg 8940
cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa 9000
ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga tcgtttcgca 9060
tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 9120
gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 9180
cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 9240
aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 9300
tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 9360
atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 9420
ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 9480
tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 9540
agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 9600
gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 9660
gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 9720
tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 9780
tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 9840
acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 9900
gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 9960
tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 10020
ccacccccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc 10080
aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac 10140
agtttttatg catgcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 10200
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 10260
atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 10320
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 10380
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctgggtacc gggccccccc 10440
tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatctggca 10500
tttttgggag gtgtgaaatg cggcgcgaaa gtctgttggt atcggtttgc aagggcctgc 10560
gggtacatgt cgagcgcgtt gggcaggatc ccgggcgcag cacggtgatg ctggtcaacg 10620
gcgcgatggc gaccaccgcc tcgttcgccc ggacctgcaa gtgcctggcc gaacatttca 10680
acgtggtgct gttcgacctg cccttcgccg ggcagtcgcg tcagcacaac ccgcagcggg 10740
ggttgatcac caaggacgac gaggtggaaa tcctcctggc gctgatcgag cgcttcgagg 10800
tcaatcacct ggtctccgcg tcctggggcg gtatctccac gctgctggcg ctgtcgcgca 10860
atccgcgcgg catccgcagc tcggtggtga tggcattcgc ccctggactg aaccaggcga 10920
tgctcgacta cgtcgggcgg gcgcaggcgc tgatcgagct ggacgacaag tcggcgatcg 10980
gccatctgct caacgagacc gtcggcaaat acctgccgcc gcgcctgaaa gccagcaacc 11040
atcagcacat ggcttcgctg gccaccggcg aatacgagca ggcgcgcttt cacatcgacc 11100
aggtgctggc gctcaacgat cggggctacc tggcttgcct ggagcggatc cagagccacg 11160
tgcatttcat caacggcagc tgggacgaat acaccaccgc cgaggacgcc cgccagttcc 11220
gcgactacct gccgcactgc agtttctcgc gggtggaggg caccgggcat ttcctcgacc 11280
tggagtccaa gctggccgcg gtacgcgtgc accgcgccct gctcgagcac ctgctgaagc 11340
aaccggagcc gcagcgggcg gaacgcgcgg cgggattcca cgagatggcc atcggctacg 11400
cctgaaccct tgacctgcga agacccggcc tggccgggct ttgcggttgc ataacgcacg 11460
gagtagcacc atgcacgcca tcctcatcgc catcggctcg gccggcgacg tatttccctt 11520
catcggcctg gcccggaccc tgaaattgcg cgggcaccgc gtgagcctct gcaccatccc 11580
ggtgtttcgc gacgcggtgg agcagcacgg catcgcgttc gtcccgctga gcgacgaact 11640
gacctaccgc cggaccatgg gcgatccgcg cctgtgggac cccaagacgt ccttcggcgt 11700
gctctggcaa accatcgccg ggatgatcga gccggtctac gagtacgtct cggcgcagcg 11760
ccatgacgac atcgtggtgg tcggctcgct ctgggcgctg ggcgcacgca tcgctcacga 11820
gaagtacggg attccctacc tgtccgcgca ggtctcgcca tcgaccttgt tgtcggcgca 11880
cctgccgccg gtacacccca agttcaacgt gcccgagcag atgccgctgg cgatgcgcaa 11940
gctgctctgg cgctgcatcg agcgcttcaa gctggatcgc acctgcgcgc cggatatcaa 12000
cgcggtgcgg cgcaaggtcg gcctggagac gccggtgaag cgcatcttca cccaatggat 12060
gcattcgccg cagggcgtgg tctgcctgtt cccggcctgg ttcgcgccgc cccagcagga 12120
ttggccgcaa cccctgcaca tgaccggctt cccgctgttc gacggcagta tcccggggac 12180
cccgctcgac gacgaactgc aacgctttct cgatcagggc agccggccgc tggtgttcac 12240
ccagggctcg accgaacacc tgcagggcga cttctacgcc atggccctgc gcgcgctgga 12300
acgcctcggc gcgcgtggga tcttcctcac cggcgccggc caggaaccgc tgcgcggctt 12360
gccgaaccac gtgctgcagc gcgcctacgc gccactggga gccttgctgc catcgtgcgc 12420
cgggctggtc catccgggcg gtatcggcgc catgagcctg gccttggcgg cgggggtgcc 12480
gcaggtgctg ctgccctgcg cccacgacca gttcgacaat gccgaacggc tggtccggct 12540
cggctgcggg atgcgcctgg gcgtgccatt gcgcgagcag gagttgcgcg gggcgctgtg 12600
gcgcttgctc gaggacccgg ccatggcggc ggcctgtcgg cgtttcatgg aattgtcaca 12660
accgcacagt atcgcttgcg gtaaagcggc ccaggtggtc gaacgttgtc atagggaggg 12720
ggatgcgcga tggctgaagg ctgcgtcctg acctacggga gaagaacgat catggaccgg 12780
atagacatgg gcgtgctggt ggtactgttc aatcctggcg acgacgacct ggaacacctt 12840
ggcgaactgg cggcggcgtt tccgcaactg cgcttccttg ccgtcgacaa ctcaccgcac 12900
agcgatccgc agcgcaatgc ccggctgcgc gggcaaggca tcgccgtgct gcaccacggc 12960
aaccggcagg gcatcgccgg cgccttcaac cagggactcg acgcgctatt ccggcgtggc 13020
gtgcagggtg tgctgctgct cgaccaggac tcccgtcccg gcggcgcctt cctcgccgcc 13080
cagtggcgca acctgcaggc gcgcaacggt caggcctgcc tgctcggccc acggatcttc 13140
gaccggggtg accggcgctt cctgccggcc atccatctcg acggactgac gctcaggcaa 13200
ttgtctctgg acggcctgac gaccccgcag cgcacctcgt tcctgatctc ctccggctgc 13260
ctgctgaccc gcgaggccta ccagcgcctc ggccacttcg acgaggaact gttcatcgac 13320
cacgtggaca ccgaatacag cctgcgcgcc caggcgctgg acgtgcccct gtacgtcgac 13380
ccgcggctgg tcctcgagca ccgcatcggc acgcgcaaga cccgccgcct cggcggtctc 13440
agcctcagcg cgatgaacca cgccccgctg cgccgctact acctggcgcg caacggcctg 13500
ctggtcctgc gccgctacgc ccggtcctcg ccgctggccc tgctggcgaa cctgccgacc 13560
ctgacccagg gcctcgcggt gctcctgctc gaacgcgaca agctgctcaa gctgcgctgc 13620
ctgggctggg gcctgtggga cggcctgcgg ggacgcggcg gcgcgctgga gaccaaccgc 13680
ccgcgcctgc tgaagcgcct cgccggcccg gccgtggcgt ccgtagcttc cggcaaggcc 13740
aaggcctagt cggcgaaacg cattccct 13768
<210> 70
<211> 5365
<212> DNA
<213> Artificial Sequence
<220>
<223> synth operon
<400> 70
tctagaggtg gacggccgca cgtatgagca cgcggtgacg caggtgctgc aggccacggg 60
ggtgcggggg attctgctcg cgccggatgc gccggatgcg ccggcggcat cggacggggc 120
ggcgctgctc aagcgccgct acgtgccgct cgcggcgttg ctgccgcgct gccgggcgct 180
ggtgcaccac ggggggatcg ggacggcgtc gctcgcgtac gcggcggggg tgccgcaggt 240
ggtgacgccg ttcgcgcacg accagttcga caacgcgcag cgggtggcgg cgagcggctg 300
cggggtgcgg ctggacgcgc cggtgcgcgg cgagccgctc gcgcgggcgc tggcgcaggt 360
gctgggcgac gcggcgatgg cggcgcgctg cgcgcaggtg cgcgcgcgga tggcggcgga 420
gccgaacggc tgcgacgcgg cggcgcgctt catcgagcgc ttcgcgccgg gcgtcgcggc 480
gcggcgggcg cagccggcat gagcgcgcag gcgatgtcgg cggatcaggc gggcgttgcg 540
ccgccggcgg ccgccccgct gcgcggcgcg aagctcgcgc tgctgacgtt cgcgctgtcg 600
ctcgcgacgt tcatcgaagt gctggattcg acggtggcga acgtggcggt gccggcgatc 660
tcgggcagcc tcggggtgtc gaacagccag ggcacgtggg tgatcagctc gtactcggtg 720
gccgcggcga tcgcggtgcc gctgacgggg tggcttgcgc ggcgcgtggg cgagctgagg 780
ctgttcgtgg cgtcggtgat cctgttcacg ctgacgtcgc tgctgtgcgg gctcgcgcgg 840
gacctggagg tgctggttgc gtgccgggcg ctgcaggggc tgttctcggg gccgatggtg 900
ccgctgtcgc agacgatcct gatgcgcgcg ttcccgccgg cgcggcgcac gctggcgctg 960
gcgctgtggg ggatgacggt gctgctcgcg ccgatcttcg ggccggtggt gggcggctgg 1020
ctgatcgaca acttctcgtg gccgtggatc ttcctgatca acctgccgat cgggctgttc 1080
tcgttcgcgg tgtgcacgct gatgctgcgc ccgcaggcgc agcgcggcga ggcgagcccg 1140
atcgacgcgc cggggatcgt gctgctggtg atcggggtgg gctcgctgca ggcgatgctg 1200
gacctggggc acgaccgggg ctggttcgat tcgccgctga tcacggcgct ggcgatcgcg 1260
gcgggggtgt cgctcgtgtc gctgctgatc tgggagctgg gcgaggcgca tccggtggtg 1320
gatctgagcc tgttccggga gcggaccttc acgttctgcg tggtgatcat ctcgctgggg 1380
atgatgagct tctcggtggt gggggtggtg tttccgctgt ggctgcaggc ggtgatggga 1440
tacacggcgt accaggcggg gctggcgacg gcgtcgatgg gggtgctggc gctggtgttc 1500
tcgatcctgg tggggctgta cgcgagccgg gtggacgcgc gggtgctggt gacgttcggg 1560
ttcggggtgt ttgcggcggt gatgtggtgg agcacgcact tcacgctgtc gatgacgttc 1620
gcgcaggtgg tgacgccgcg gctgattcag gggatggggc tgccgtgctt cttcataccg 1680
ctgacggcgg cgacgctgtc gcgggtgccg gacgagaagc tggcggcggc gtcgagcctg 1740
tcgaacttcc tgcggacgct gtcggcggcg ttcggcacgg cgctgagcgt gacgtggtgg 1800
gacaaccggg cgacgtacca ctacgcggtg gtgtcgcaat cggtgacgcg cgcctcggag 1860
aacacgcagc ggtacgtgga cgcgctgcac gcgatggggc tgcacggcgc gcgggagctg 1920
agctcgctgc accaggtggt gcggcagcag gcgtacatga tggcgacgaa cgacatgttc 1980
tacatggcga gcgcgacgtg cctgctgctg gcggggctga tgtggctgac gcggccgaag 2040
cggggcgcgg cggcggcgct cgggcactga ggcgaggcat gtcgcgcccc gcatgacgaa 2100
ggcgaaggag aagggcgatg cgccgaagtc ctggggacgc ggcgcgtcga tgcggcaacg 2160
aagcgggcat ttcggcattc cgaaccacca aagggaagag cgatgacgat cctgggggcg 2220
ctggtgttcg ggcggctggg atcgttcgac gatgcgcgcg cgggcgcggc ggcgcgcgag 2280
ccggtgcggc aggaatgaac ggaacgggcc gcagcgggat accggaaagc aagaaggacg 2340
catcatacga atgacgcaga cagcaacgca agcagccact cgcgcgatga tcgcgacagg 2400
aagccgcgcg gcgcgccggc tcgcggcagc cgcgctcgcg tgggcgctcg ccggctgcgt 2460
gccgtcgggc ttcgagccgg cgctcgcgcc gcgcacgccg ggcgacgacg cgctcgcgca 2520
cacggcgggg ggcgccgcgc acggcgcatg gccgagcccc gactgggtcc ggcagctcgg 2580
cgatccgcaa ctcgacgcgc tcgtcgacga ggcgctgcgg cagaacccga cgctgcaggc 2640
cgcgcaggcg cgcatcggcg tcgcgcagtc gcagctgcag cagttcgaat cgctgacggg 2700
gctcaccgcg acggcgggcg cgtcgctctc caaggcgcac gtgccgcgct cgggcggcac 2760
catcaatacg acgttcaacg gcttgccggt gtcggtgccg ctcgtcggcg aatcggtggt 2820
gtcgtcgtcg tcgctgttcg tcgggctgaa ctatcagctg gacctgtggg gcaagaacgc 2880
ggcggccacg cgcgggctgc tgtcgatgcg cgatgcggcg cgcgtggagg ccgagcaggc 2940
gcggctcgcg ctgtcggtgg cgatcgtgac gctgtacggc gagctggacc gcgcgtatgc 3000
gctgcgcgag ctgctgcagc agaagcgccg cgcgagcgag caggtggaga cggtgctgcg 3060
cgagcgcgcg gcgcgcggga tcgacaacgg ctacgatgcg gacgacgcgg cgctcaagcg 3120
gggcaagctg ctcgagcagc tcgcgctgac cgacgagcag atccagttgc agaagctgca 3180
actgggggtg ctgagcgggc gggggccgga gcgcgggctg tcgctcgcgc ggccgaagct 3240
cgcgccgctc gcggacgcgc cgctgccggc gcggctgccg gccgggctgc tggggcggcg 3300
gccggacatc gtcgcggcgc ggctgcgggt ggaggcggcg tacgcggcga tcgacggcac 3360
gcgcgcgtcg ttctacccgg acgtgaacct ggcggcgctg ggcgggctgt tcgcgctcac 3420
gccggcgtcg ctgttcaagc acgatgcgct ggggggctcg atcggtccgg cgctgtcgct 3480
gccgatcttc gatcgcggcc ggctgaaggc gaagctgggg ggcgacgtgg cgaacgcgga 3540
cgtggcgctg gcgctgtaca accagacggt ggatgcggcg ctgggcgagg tggcgcggca 3600
gttgacgtcg ctgtcgacgg tggatgcgct gctcgaggcg cagcagcagg cggtgcgctc 3660
ggcgcagcgg atggtggcgc tggcgcagga ccggcaccgg cgggggatgg ggatgcgcaa 3720
ggacgtgaac gtggcgaagc tgacgctgct ggacgagcgt gcgcacgtga tcgagctgca 3780
ggcgcggcgg cggacgctgc gggtggggct gatcggggcg ctgggcggcg gcttcgacgc 3840
gcggccggcg ggcggcgcgc cgctcgcgca gggcaagccg ttcgcggcgg cgagcgacag 3900
gccgcccgat tgagcggcac gcacgcatgc ggcccgaagc caccgacacc cgaagacacc 3960
gacaccaacg ccaccttcac cgtgtacacg agcgattcaa ccgacaccgc ccccgagcat 4020
cgaagcccgt cgggccgatc cgcgacggct tgcgggccgg cccggccgtt gccggccggc 4080
gccaccgaca tcacgcacgc gaagaccttg aacgataccg ccaccgatac cccgcgcgcg 4140
aaggcgccca ccgatccggc cgccctcgac ggcgcgcacg cgcagcccgt gccggcgcac 4200
gagcgcggat cgcctccgcc gccggaagcc gcggcgacgc tcgccgcgcg ccgcgcgacg 4260
cgccgccggc gcttcgcgct gttcttcggg ctgctggcgc tggccgcgct gaccgcgggg 4320
ctctactggt tcgtcgccgg gcgcttcagc gaggagacgg acgacgcgta cgtggccggc 4380
aacgtggtgc agatcgccgc gcagatccag gggacggtga ccgacgtgct ggtggcggac 4440
acgcagcagg tgaaggcggg gcaggcgctg gtgaagctcg acgacgcgga cgcgtcggcg 4500
gcgttcgcgc aggcgcgggc gcagctcgcg caggcggtgc ggcaggtggc gaacacgcgg 4560
ctctcgatgg ggatgtacga ggagacggtg aaggcgcgcg aggcggacct gaagcttgcg 4620
cagcaggcgt atccggagga actggcgcgg cgaaagtcgt cgctggcgaa cgcgcaggcg 4680
gcgctggcgg gggcgcaggc gcagctggag gcggcgcgcg cgctgggcag cgagcggccg 4740
gtcgagcaga acccggcggt gcagcaggcg gccgcgcagt tcaagctggc gtaccggaac 4800
ctgaggcgca cgacgatcgt gtcgccggtg gacggcacgg tcggtcagcg gtcggtgcag 4860
atcggtcagc aggtggggcc gggggtgccg ctgatgtcgg tggtgcagtt gcggcaggtg 4920
tgggtggagg cgaacttcaa ggaagggcag atccggcaca tgcgggtggg ccagccggtg 4980
cggctcgaat cggacctgta cggcgcgcgg gtgacgtacc acggccgggt ggagggggtc 5040
tcggcgggca cgggcagcgc gttctcgatg ctgccgtcgc agaacgcggc ggggaactgg 5100
atcaaggtgg tgcagcgcct gccggtggtg atctcgctgg agccgtcgga gctggcggcg 5160
cacccgctgc gggtggggct gtcgatgcgc gcgacggtgg agacgaaggt gcgtggcggc 5220
cgcctgctcg acggcgacgc gccgctgccg gggctgcgca cgcgggtgca cgaagcgcag 5280
gcgggcgagg ccgaggccgc ggcttcggca gtgattcggg agaatgacgg ccgcaggtga 5340
cgggcggttg cgggatcgct ctaga 5365
<210> 71
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> primer
<400> 71
tatatataac cggtattaat gcagctggca cgac 34
<210> 72
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> primer
<400> 72
ggccgaccgg tactagtgga 20
<210> 73
<211> 11960
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 73
accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc 60
aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg 120
aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg 180
aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag 240
cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt 300
tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa 360
gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc 420
aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 480
cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 540
aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 600
tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 660
gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 720
aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 780
tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 840
ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 900
ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 960
cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 1020
tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 1080
ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 1140
tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 1200
tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 1260
ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 1320
accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 1380
atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta 1440
tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga 1500
tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat 1560
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 1620
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 1680
ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 1740
ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 1800
tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa 1860
gctttgggag gtgtgaaatg cggcgcgaaa gtctgttggt atcggtttgc aagggcctgc 1920
gggtacatgt cgagcgcgtt gggcaggatc ccgggcgcag cacggtgatg ctggtcaacg 1980
gcgcgatggc gaccaccgcc tcgttcgccc ggacctgcaa gtgcctggcc gaacatttca 2040
acgtggtgct gttcgacctg cccttcgccg ggcagtcgcg tcagcacaac ccgcagcgcg 2100
ggttgatcac caaggacgac gaggtggaaa tcctcctggc gctgatcgag cgcttcgagg 2160
tcaatcacct ggtctccgcg tcctggggcg gtatctccac gctgctggcg ctgtcgcgca 2220
atccgcgcgg catccgcagc tcggtggtga tggcattcgc ccctggactg aaccaggcga 2280
tgctcgacta cgtcgggcgg gcgcaggcgc tgatcgagct ggacgacaag tcggcgatcg 2340
gccatctgct caacgagacc gtcggcaaat acctgccgca gcgcctgaaa gccagcaacc 2400
atcagcacat ggcttcgctg gccaccggcg aatacgagca ggcgcgcttt cacatcgacc 2460
aggtgctggc gctcaacgat cggggctact tggcttgcct ggagcggatc cagagccacg 2520
tgcatttcat caacggcagc tgggacgaat acaccaccgc cgaggacgcc cgccagttcc 2580
gcgactacct gccgcactgc agtttctcgc gggtggaggg caccgggcat ttcctcgacc 2640
tggagtccaa gctggcagcg gtacgcgtgc accgcgccct gctcgagcac ctgctgaagc 2700
aaccggagcc gcagcgggcg gaacgcgcgg cgggattcca cgagatggcc atcggctacg 2760
cctgaaccct tgacctgcga agacccggcc tggccgggct ttgcggttgc ataacgcacg 2820
gagtagcccc atgcacgcca tcctcatcgc catcggctcg gccggcgacg tatttccctt 2880
catcggcctg gcccggaccc tgaaactgcg cgggcaccgc gtgagcctct gcaccatccc 2940
ggtgtttcgc gacgcggtgg agcagcacgg catcgcgttc gtcccgctga gcgacgaact 3000
gacctaccgc cggaccatgg gcgatccgcg cctgtgggac cccaagacgt ccttcggcgt 3060
gctctggcaa gccatcgccg ggatgatcga gccggtctac gagtacgtct cggcgcagcg 3120
ccatgacgac atcgtggtgg tcggctcgct atgggcgctg ggcgcacgca tcgctcacga 3180
gaagtacggg attccctacc tgtccgcgca ggtctcgcca tcgaccctgt tgtcggcgca 3240
cctgccgccg gtacacccca agttcaacgt gcccgagcag atgccgctgg cgatgcgcaa 3300
gctgctctgg cgctgcatcg agcgcttcaa gctggatcgc acctgcgcgc cggagatcaa 3360
cgcggtgcgc cgcaaggtcg gcctggaaac gccggtgaag cgcatcttca cccaatggat 3420
gcattcgccg cagggcgtgg tctgcctgtt cccggcctgg ttcgcgccgc cccagcagga 3480
ttggccgcaa cccctgcaca tgaccggctt cccgctgttc gacggcagta tcccggggac 3540
cccgctcgac gacgaactgc aacgctttct cgatcagggc agccggccgc tggtgttcac 3600
ccagggctcg accgaacacc tgcagggcga cttctacgcc atggccctgc gcgcgctgga 3660
acgcctcggc gcgcgtggga tcttcctcac cggcgccggc caggaaccgc tgcgcggctt 3720
gccgaaccac gtgctgcagc gcgcctacgc gccactggga gccttgctgc catcgtgcgc 3780
cgggctggtc catccgggcg gtatcggcgc catgagccta gccttggcgg cgggggtgcc 3840
gcaggtgctg ctgccctgtg cccacgacca gttcgacaat gccgaacggc tggtccggct 3900
cggctgcggg atgcgcctgg gcgtgccgtt gcgcgagcag gagttgcgcg gggcgctgtg 3960
gcgcttgctc gaggacccgg ccatggcggc ggcctgtcgg cgtttcatgg aattgtcaca 4020
accgcacagt atcgcttgcg gtaaagcggc ccaggtggtc gaacgttgtc atagggaggg 4080
ggatgctcga tggctgaagg ctgcgtcctg aacggtgctg gcataacagt ctagagcggc 4140
cgccaccgcg gtggagctcc aattcgccct atagtgagtc gtattacgcg cgctcactgg 4200
ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg 4260
cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt 4320
cccaacagtt gcgcagcctg aatggcgaat ggaaattgta agcgttaata ttttgttaaa 4380
attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg actgcgatga 4440
gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggtgc 4500
tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc 4560
cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg 4620
gtaggtcaac tttcgcaaca tccggcttga ccataacggc gctgtcggcc gcattgagaa 4680
ctgcttcagc gataagctcc gcatcaccac tttcgtgcga agccgaggca taacggcctt 4740
cttgatgacc gaacgcatgc caatgctgtt cgagcgcatc gtccgtgagc gaagataggt 4800
ctgggtatcg attccgataa taagccagat ccagcacaaa tcgccttgag ccagtctttt 4860
tctgggggac gggcactgaa tttcccttaa catgcatgct cggctgcaca tctacgctcg 4920
gcaagccttc cttaacagac atttttcact ttcctatgaa tattcaagag cgccagaccg 4980
ctgaaacatg aagaatgacg tctaacctgc cgagccccga ggctattata atttttgttg 5040
ggttgttcaa catgaatgca agaactgcca tcactctaat tcctatttca ggaatactga 5100
caaataaaaa tgacactttc caacccccgg aaacgaaccc gacttccaga tgcgcacccc 5160
gcgctgccat gcccagcgaa ctagagtccc aggatagcgg tagcttaagg ccctctagac 5220
ttgtcagcac ccagatttgt ctccgtgagt tgcatcgctc aaacaactgt ttcttgcatg 5280
gacacctgac gactccctgt tgtgtctagg caaccatgag gtcaccttcc accgctgcaa 5340
cggctcctcc tcttccagcg tgagcagatc ggcacggtca gtaccggcac ccagcagtgt 5400
atttttcgac atcagggtgg tctaattccg gcagcgctag cagctcgcct tgcgttgccg 5460
gggcgaagtt tctcagcgtc atctgtctac gacaacacct tttgtccaat tagagccaaa 5520
ttatgattct agtaacaggc ggagccggct tcatcggctc aaatttcgta ctgcaatggt 5580
gtgcgcacaa tgaggaaccc gtcctcaacc tcgacgccct gacctacgca ggcaacctgg 5640
ccaacctgca gccgctggaa ggcaaccctc agcatcgctt tgtgcaaggc aatatttgcg 5700
atgctgcgct tctgaccaag ctgttcgcag agcaccgccc gcgcgccgtg gttcacttcg 5760
cggcggaatc ccatgtagac cgctcaatca ccggccccga agcgtttgtc gaaaccaacg 5820
tgatgggcac gtttcgcttg cttgaagccg cccgggcgca ttggaatagt ttggaaggtg 5880
cagagaagga ggccttccgt ttcctccatg tctctaccga cgaagtctac ggcacactag 5940
ggccaaacga cccggcgttc accgaaacca cgccgtacgc gccgaacagc ccatactccg 6000
ccagcaaggc agccagcgac catctggtac gctcgtattt ccatacctac ggcatgccgg 6060
tactcactac caactgctcc aacaattacg ggccgctcca cttcccggaa aaactgatcc 6120
cgctgatgat cgtcaacgca ctcgccggta aggcgctgcc tgtctatggc gacggccagc 6180
aaatccgcga ctggctgtat gtcgaagatc actgctcggg catccgtcgc gtactggaag 6240
ccggtgcgtt cggcgagacg tacaatattg gcggctggaa tgaaaaagcc aacattgaca 6300
ttgtgcgtac actctgcagc cttctcgacg agatggcacc tgcggcatcg cgccaggtaa 6360
tcaatcagaa gaccggcgag cctgtcgaac agtatgcaga actcatcgcc tacgtaaccg 6420
accgcccagg ccatgaccgc cgttatgcca tcgatgcacg caagatcgag cgggagctcg 6480
gctggaaacc tgccgaaacc ttcgagacgg gcattcgaaa gacagtcgct tggtacttgg 6540
ccaaccagaa atgggtaaaa ggtgtcatgg acggcagcta ccgtgactgg gtggcacaac 6600
aatacggggc aaataaagcg tgaaaatcct gctgttgggg aaaaacgggc aagtaggctg 6660
ggagctacag cgcgccttgg cgccgctggg tgaggtcatt gcgctggatc gtcagggggc 6720
cgagggctta tgtggcgact tgtccaacct ggacggcttg gccgctacga ttcgtcagct 6780
ggcgccggac gtgatcgtca acgctgctgc ctacactgca gtggataaag ctgagagcga 6840
tcaggcactg gctgcaatga tcaatgccgc ggctcctgct gtattagcac gtgaaacagc 6900
agctttgggc gcctggttga ttcactattc caccgattat gtatttgacg gcagcggcag 6960
tcagcgctgg gaggaaactg cgcctaccgg ccccctttcg gtctacggcc ggaccaagct 7020
ggaaggcgag catgccattc tcgccagcgg cgccaaggcc gtggtactgc gcaccagctg 7080
ggtgtatgct gcgcgcgggc acaattttgc caagaccatg ctgcgcctgg cggcggagcg 7140
tgagacgttg agcgtggtag cagaccaatt tggcgcaccc acgggcgctg acctgatcgc 7200
cgacgttact gcacacatcc tgcggcaaat cttcaatggg caagacaacc gtcacctggc 7260
agggatttac cacttggctg cgtccggtga aacctcttgg catggttttg ctcagttcgt 7320
gctggcgcat gctcaacgca ctggcgtagc gctgaaagtg acagctgata aggttgccgc 7380
aatcagcacc gaagcttatc cagtacctgc accacgtccg cgcaactcgc gcctggcact 7440
gggcaaactg gaaaacacgt tcaatttcaa aatgccgctt tgggagcaag gcgtgcaacg 7500
tatgctggac gaaatccagt aatagggact ctcatggctc gtaaaggaat tattctggcc 7560
ggcggttcgg gtacacgcct gcatccggcc acactttcgg tttcgaagca gctgctgccg 7620
gtgtatgaca aaccgatgat ctactacccg ctgagcaccc tgctgctcgc tggtatccgg 7680
gacatcctga tcatttccac cccgcaggac accccgcgct tcgaacagct gctgggcgat 7740
ggcagccagt ggggcctgaa cctgtcatac gcaatacaac caagcccgga tggcttggcg 7800
caagcgttca ccatcggcgc tgacttcatc ggtaacgacc cttctgcgtt ggttctcggt 7860
gacaatattt tctacggcca tgacttccag gcactgctat tgaacgcaga taaacgtgaa 7920
tccggtgctt cagtattcgc ttatcatgtt catgacccag aacgctatgg cgtagcggag 7980
tttgacgata gcggtcgcgt attgtcgctg gaagaaaaac cggcagttcc aaagtctagc 8040
tatgcggtca ccggcctgta tttctatgac aatcaggtag tcaatctggc tcgcgagctg 8100
aagccttccc cacgtggcga gctggaaatc accgacctca acaaccttta cttgcagcag 8160
cagcagttgc aggtcgaaat catgggccgt ggctatgcgt ggctcgacac cggcacgcac 8220
gacagtctgc tggaggctag ccagtacatc gcaaccatgg agcgccgtca gggcttgaaa 8280
gtcgcctgcc ctgaggaaat ttgctaccgc gctggctgga tcaacgctga gcaactcgag 8340
tgcctggctc aaccactgct gaaaaacggt tatggcaagt atctgcagaa cttgctgaaa 8400
gagaaggtgt tctgatgcaa gccattccgc tggatatccc cgaagtcgtg ctgtttaccc 8460
ccaaggtttt tggcgacgaa cgtggtttct tctacgagag cttcaacgcc cgtgttttca 8520
gcgaagtgac cggcctgcag cccgacttcg tacaagacaa ccactcgcgc tcggtaaaag 8580
gcgtgctccg tggcctgcac tatcagctgg cacctcacgc ccagggcaag ctggtgcgtg 8640
tggtgcaagg cgaagtcttc gatgttgcgg tggatatccg tcgctcgtcc acaaccttcg 8700
gtaaatgggt aggtgcggtg ttgtcggccg agaacaagaa ccagctgtgg atcccgccag 8760
ggttcgcaca cgggttcgtc acgttgagtg aaaccgcaga gttcctctac aagaccaccg 8820
acttctactc gccgcagtgc gagcgctgca ttgcctggaa tgatccggca gtgggtatcg 8880
aatggcccat cgactccgta ccaagcttgt ctggcaagga ccagcttggg gtcgcattgg 8940
ctgacgccga actgttcgac taacggtttt agcggagaag ggctgcggta gcgcagcctt 9000
gtctctgaac acatgccata ccgggtcttg ccgatagtgg cgtttttcac acgccactaa 9060
gaagcaaccg ctgcatggcc tggcaaataa tcagaatttg ccccttcctt gtaggccatt 9120
tcccaaagat acccctgcgc ctgttttcca ttgcacacgt taaacgtgag acttagtctc 9180
gacccgtcgc tgccaaatca gtgaccggtt tattgactac cggaagcagt gtgaccgtgt 9240
gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgacg 9300
atatgatcat ttattctgcc tcccagagcc tgataaaaac ggtgaatccg ttagcgaggt 9360
gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg 9420
gaggcagaca aggtataggg cggcgaggcg gctacagccg atagtctgga acagcgcact 9480
tacgggttgc tgcgcaaccc aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct 9540
ccaacggctc gccatcgtcc agaaaacacg gctcatcggg catcggcagg cgctgctgcc 9600
cgcgccgttc ccattcctcc gtttcggtca aggctggcag gtctggttcc atgcccggaa 9660
tgccgggctg gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat 9720
acagggtcgg gatgcggcgc aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt 9780
gatcaaccac cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc 9840
acgccacgcg gtcattgacc acgtaggccg acacggtgcc ggggccgttg agcttcacga 9900
cggagatcca gcgctcggcc accaagtcct tgactgcgta ttggaccgtc cgcaaagaac 9960
gtccgatgag cttggaaagt gtcttctggc tgaccaccac ggcgttctgg tggcccatct 10020
gcgccacgag gtgatgcagc agcattgccg ccgtgggttt cctcgcaata agcccggccc 10080
acgcctcatg cgctttgcgt tccgtttgca cccagtgacc gggcttgttc ttggcttgaa 10140
tgccgatttc tctggactgc gtggccatgc ttatctccat gcggtagggt gccgcacggt 10200
tgcggcacca tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc 10260
gtaaaagtgg cagtcaatta cagattttct ttaacctacg caatgagcta ttgcgggggg 10320
tgccgcaatg agctgttgcg tacccccctt ttttaagttg ttgattttta agtctttcgc 10380
atttcgccct atatctagtt ctttggtgcc caaagaaggg cacccctgcg gggttccccc 10440
acgccttcgg cgcggctccc cctccggcaa aaagtggccc ctccggggct tgttgatcga 10500
ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc 10560
gccgtgaggc tcggggggca ggcgggcggg cttcgccttc gactgccccc actcgcatag 10620
gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc gcgagccttg 10680
acccgccttc cacttggtgt ccaaccggca agcgaagcgc gcaggccgca ggccggaggc 10740
ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc gcagttggag 10800
ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg tcaagctcgt 10860
gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc 10920
gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg caagggagcg 10980
cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggcgccgca gccgccgtag 11040
gcggtcacga ctttgcgaag caaagtctag tgagtatact caagcattga gtggcccgcc 11100
ggaggcaccg ccttgcgctg cccccgtcga gccggttgga caccaaaagg gaggggcagg 11160
catggcggca tacgcgatca tgcgatgcaa gaagctggcg aaaatgggca acgtggcggc 11220
cagtctcaag cacgcctacc gcgagcgcga gacgcccaac gctgacgcca gcaggacgcc 11280
agagaacgag cactgggcgg ccagcagcac cgatgaagcg atgggccgac tgcgcgagtt 11340
gctgccagag aagcggcgca aggacgctgt gttggcggtc gagtacgtca tgacggccag 11400
cccggaatgg tggaagtcgg ccagccaaga acagcaggcg gcgttcttcg agaaggcgca 11460
caagtggctg gcggacaagt acggggcgga tcgcatcgtg acggccagca tccaccgtga 11520
cgaaaccagc ccgcacatga ccgcgttcgt ggtgccgctg acgcaggacg gcaggctgtc 11580
ggccaaggag ttcatcggca acaaagcgca gatgacccgc gaccagacca cgtttgcggc 11640
cgctgtggcc gatctagggc tgcaacgggg catcgagggc agcaaggcac gtcacacgcg 11700
cattcaggcg ttctacgagg ccctggagcg gccaccagtg ggccacgtca ccatcagccc 11760
gcaagcggtc gagccacgcg cctatgcacc gcagggattg gccgaaaagc tgggaatctc 11820
aaagcgcgtt gagacgccgg aagccgtggc cgaccggctg acaaaagcgg ttcggcaggg 11880
gtatgagcct gccctacagg ccgccgcagg agcgcgtgag atgcgcaaga aggccgatca 11940
agcccaagag acggcccgag 11960
<210> 74
<211> 13289
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 74
ccggtcactg atttggcagc gacgggtcga gactaagtct cacgtttaac gtgtgcaatg 60
gaaaacaggc gcaggggtat ctttgggaaa tggcctacaa ggaaggggca aattctgatt 120
atttgccagg ccatgcagcg gttgcttctt agtggcgtgt gaaaaacgcc actatcggca 180
agacccggta tggcatgtgt tcagagacaa ggctgcgcta ccgcagccct tctccgctaa 240
aaccgttagt cgaacagttc ggcgtcagcc aatgcgaccc caagctggtc cttgccagac 300
aagcttggta cggagtcgat gggccattcg atacccactg ccggatcatt ccaggcaatg 360
cagcgctcgc actgcggcga gtagaagtcg gtggtcttgt agaggaactc tgcggtttca 420
ctcaacgtga cgaacccgtg tgcgaaccct ggcgggatcc acagctggtt cttgttctcg 480
gccgacaaca ccgcacctac ccatttaccg aaggttgtgg acgagcgacg gatatccacc 540
gcaacatcga agacttcgcc ttgcaccaca cgcaccagct tgccctgggc gtgaggtgcc 600
agctgatagt gcaggccacg gagcacgcct tttaccgagc gcgagtggtt gtcttgtacg 660
aagtcgggct gcaggccggt cacttcgctg aaaacacggg cgttgaagct ctcgtagaag 720
aaaccacgtt cgtcgccaaa aaccttgggg gtaaacagca cgacttcggg gatatccagc 780
ggaatggctt gcatcagaac accttctctt tcagcaagtt ctgcagatac ttgccataac 840
cgtttttcag cagtggttga gccaggcact cgagttgctc agcgttgatc cagccagcgc 900
ggtagcaaat ttcctcaggg caggcgactt tcaagccctg acggcgctcc atggttgcga 960
tgtactggct agcctccagc agactgtcgt gcgtgccggt gtcgagccac gcatagccac 1020
ggcccatgat ttcgacctgc aactgctgct gctgcaagta aaggttgttg aggtcggtga 1080
tttccagctc gccacgtggg gaaggcttca gctcgcgagc cagattgact acctgattgt 1140
catagaaata caggccggtg accgcatagc tagactttgg aactgccggt ttttcttcca 1200
gcgacaatac gcgaccgcta tcgtcaaact ccgctacgcc atagcgttct gggtcatgaa 1260
catgataagc gaatactgaa gcaccggatt cacgtttatc tgcgttcaat agcagtgcct 1320
ggaagtcatg gccgtagaaa atattgtcac cgagaaccaa cgcagaaggg tcgttaccga 1380
tgaagtcagc gccgatggtg aacgcttgcg ccaagccatc cgggcttggt tgtattgcgt 1440
atgacaggtt caggccccac tggctgccat cgcccagcag ctgttcgaag cgcggggtgt 1500
cctgcggggt ggaaatgatc aggatgtccc ggataccagc gagcagcagg gtgctcagcg 1560
ggtagtagat catcggtttg tcatacaccg gcagcagctg cttcgaaacc gaaagtgtgg 1620
ccggatgcag gcgtgtaccc gaaccgccgg ccagaataat tcctttacga gccatgagag 1680
tccctattac tggatttcgt ccagcatacg ttgcacgcct tgctcccaaa gcggcatttt 1740
gaaattgaac gtgttttcca gtttgcccag tgccaggcgc gagttgcgcg gacgtggtgc 1800
aggtactgga taagcttcgg tgctgattgc ggcaacctta tcagctgtca ctttcagcgc 1860
tacgccagtg cgttgagcat gcgccagcac gaactgagca aaaccatgcc aagaggtttc 1920
accggacgca gccaagtggt aaatccctgc caggtgacgg ttgtcttgcc cattgaagat 1980
ttgccgcagg atgtgtgcag taacgtcggc gatcaggtca gcgcccgtgg gtgcgccaaa 2040
ttggtctgct accacgctca acgtctcacg ctccgccgcc aggcgcagca tggtcttggc 2100
aaaattgtgc ccgcgcgcag catacaccca gctggtgcgc agtaccacgg ccttggcgcc 2160
gctggcgaga atggcatgct cgccttccag cttggtccgg ccgtagaccg aaagggggcc 2220
ggtaggcgca gtttcctccc agcgctgact gccgctgccg tcaaatacat aatcggtgga 2280
atagtgaatc aaccaggcgc ccaaagctgc tgtttcacgt gctaatacag caggagccgc 2340
ggcattgatc attgcagcca gtgcctgatc gctctcagct ttatccactg cagtgtaggc 2400
agcagcgttg acgatcacgt ccggcgccag ctgacgaatc gtagcggcca agccgtccag 2460
gttggacaag tcgccacata agccctcggc cccctgacga tccagcgcaa tgacctcacc 2520
cagcggcgcc aaggcgcgct gtagctccca gcctacttgc ccgtttttcc ccaacagcag 2580
gattttcacg ctttatttgc cccgtattgt tgtgccaccc agtcacggta gctgccgtcc 2640
atgacacctt ttacccattt ctggttggcc aagtaccaag cgactgtctt tcgaatgccc 2700
gtctcgaagg tttcggcagg tttccagccg agctcccgct cgatcttgcg tgcatcgatg 2760
gcataacggc ggtcatggcc tgggcggtcg gttacgtagg cgatgagttc tgcatactgt 2820
tcgacaggct cgccggtctt ctgattgatt acctggcgcg atgccgcagg tgccatctcg 2880
tcgagaaggc tgcagagtgt acgcacaatg tcaatgttgg ctttttcatt ccagccgcca 2940
atattgtacg tctcgccgaa cgcaccggct tccagtacgc gacggatgcc cgagcagtga 3000
tcttcgacat acagccagtc gcggatttgc tggccgtcgc catagacagg cagcgcctta 3060
ccggcgagtg cgttgacgat catcagcggg atcagttttt ccgggaagtg gagcggcccg 3120
taattgttgg agcagttggt agtgagtacc ggcatgccgt aggtatggaa atacgagcgt 3180
accagatggt cgctggctgc cttgctggcg gagtatgggc tgttcggcgc gtacggcgtg 3240
gtttcggtga acgccgggtc gtttggccct agtgtgccgt agacttcgtc ggtagagaca 3300
tggaggaaac ggaaggcctc cttctctgca ccttccaaac tattccaatg cgcccgggcg 3360
gcttcaagca agcgaaacgt gcccatcacg ttggtttcga caaacgcttc ggggccggtg 3420
attgagcggt ctacatggga ttccgccgcg aagtgaacca cggcgcgcgg gcggtgctct 3480
gcgaacagct tggtcagaag cgcagcatcg caaatattgc cttgcacaaa gcgatgctga 3540
gggttgcctt ccagcggctg caggttggcc aggttgcctg cgtaggtcag ggcgtcgagg 3600
ttgaggacgg gttcctcatt gtgcgcacac cattgcagta cgaaatttga gccgatgaag 3660
ccggctccgc ctgttactag aatcataatt tggctctaat tggacaaaag gtgttgtcgt 3720
agacagatga cgctgagaaa cttcgccccg gcaacgcaag gcgagctgct agcgctgccg 3780
gaattagacc accctgatgt cgaaaaatac actgctgggt gccggtactg accgtgccga 3840
tctgctcacg ctggaagagg aggagccgtt gcagcggtgg aaggtgacct catggttgcc 3900
tagacacaac agggagtcgt caggtgtcca tgcaagaaac agttgtttga gcgatgcaac 3960
tcacggagac aaatctgggt gctgacaagt ctagagggcc ttaagctacc gctatcctgg 4020
gactctagtt cgctgggcat ggcagcgcgg ggtgcgcatc tggaagtcgg gttcgtttcc 4080
gggggttgga aagtgtcatt tttatttgtc agtattcctg aaataggaat tagagtgatg 4140
gcagttcttg cattcatgtt gaacaaccca acaaaaatta taatagcctc ggggctcggc 4200
aggttagacg tcattcttca tgtttcagcg gtctggcgct cttgaatatt cataggaaag 4260
tgaaaaatgt ctgttaagga aggcttgccg agcgtagatg tgcagccgag catgcatgtt 4320
aagggaaatt cagtgcccgt cccccagaaa aagactggct caaggcgatt tgtgctggat 4380
ctggcttatt atcggaatcg atacccagac ctatcttcgc tcacggacga tgcgctcgaa 4440
cagcattggc atgcgttcgg tcatcaagaa ggccgttatg cctcggcttc gcacgaaagt 4500
ggtgatgcgg agcttatcgc tgaagcagtt ctcaatgcgg ccgacagcgc cgttatggtc 4560
aagccggatg ttgcgaaagt tgacctaccg gtaaaccagc aatagacata agcggctatt 4620
taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca 4680
tccgcttatt atcacttatt caggcgtagc accaggcgtt taagggcacc aataactgcc 4740
ttaaaaaaat tacgccccgc cctgccactc atcgcagtcg gcctattggt taaaaaatga 4800
gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat 4860
tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 4920
cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 4980
tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag 5040
ggcgaattgg agctccaccg cggtggcggc cgctctagaa gtaccaccag cacgcccatg 5100
tctatccggt ccatgatcgt tcttctcccg taggtcgaag ttgccaggcc aggaccagcc 5160
cggccagaac gagaagggcg ccggcgagga atggcgcgcc ggccaggggc agcggcgcga 5220
gcggaccgct gccccagtgg aacaggccgc tcatcagcgg cggaccgacg atcgcggcga 5280
ggctcatcag gctgctcagc acgccctgca actcgccctg gcggtcgacc ggcacgcggg 5340
ccgagagcag cccctgcatg gccggggtgg cgaggctgcc gagcgcgaag ggcagcagcg 5400
cgcagaccag ccagaatgac gagtcgacca gggcgaacag cagcaggccg cagccttgca 5460
gggcgaggcc caggcgcagc aggcgggcgt cgtccaggcg ccgcttgcag aggttcacgc 5520
cgagggtctg ggcgagcacc gcgagcacgc cgtagagggc cagcgagtag ccgatccagg 5580
cgctgctcca gtgaaacttc tcgatcacga agaacggcca gaccaccatc accgcctgca 5640
agccgaggaa taccagggca agcaccgcca gcaggcgtcc gacccccggt tgccgagcca 5700
ggccgctgat cgagcgcaag gcattcatcc gcctcgggtc caggcggcgg cgtcgcgtcg 5760
ggggcagggt ttcctcgagg aacaggccgg cgagcagggc gttgagcagg cacaggccgg 5820
cggccagcaa cagcggcagc gtcgtgccgt gcaccgccag cagcccaccg agggcggggc 5880
cgaggatcat gcccagggcg aggccggcgt acagccagcc gaagtgccgg gtgcgctgcc 5940
cgtgcgtgcc gaggtcagcc gcgcaggcca tcgcggtggc cacgctggcg ccggtgagcc 6000
cggccagcgc gcgaccgagg aacagcatcc agaggctgtc ggccagcgcc agcagcagat 6060
agctgagggc gaagccgagc atcgccagga ccaggacggg gcggcgtccg aagcggtcgc 6120
tgaggctgcc gaggaccggc gaaaagaaca attgcagcag cgcgaaggtc atcaccaggg 6180
cggcgcccca ggtggccgcg tcgcggaccg ccagcggcgc cacgctgccg atcagcgtcg 6240
gcagcagggg cacgatcagg ccgacgccag cggcatccag caggcaggtg aggaacagca 6300
gaggcaggac gcgtttcgcg ccgggaccgt gttcccgcgt ggcggagggg cagaggctgg 6360
tcgtggacac gccaggatcc tcccggcgtc aggacgcagc cttcagccat cgcgcatccc 6420
cctccctatg acaacgttcg accacctggg ccgctttacc gcaagcgata ctgtgcggtt 6480
gtgacaattc catgaaacgc cgacaggccg ccgccatggc cgggtcctcg agcaagcgcc 6540
acagcgcccc gcgcaactcc tgctcgcgca atggcacgcc caggcgcatc ccgcagccga 6600
gccggaccag ccgttcggca ttgtcgaact ggtcgtgggc gcagggcagc agcacctgcg 6660
gcacccccgc cgccaaggcc aggctcatgg cgccgatacc gcccggatgg accagcccgg 6720
cgcacgatgg cagcaaggct cccagtggcg cgtaggcgcg ctgcagcacg tggttcggca 6780
agccgcgcag cggttcctgg ccggcgccgg tgaggaagat cccacgcgcg ccgaggcgtt 6840
ccagcgcgcg cagggccatg gcgtagaagt cgccctgcag gtgttcggtc gagccctggg 6900
tgaacaccag cggccggctg ccctgatcga gaaagcgttg cagttcgtcg tcgagcgggg 6960
tccccgggat actgccgtcg aacagcggga agccggtcat gtgcaggggt tgcggccaat 7020
cctgctgggg cggcgcgaac caggccggga acaggcagac cacgccctgc ggcgaatgca 7080
tccattgggt gaagatgcgc ttcaccggcg tctccaggcc gaccttgcgc cgcaccgcgt 7140
tgatatccgg cgcgcaggtg cgatccagct tgaagcgctc gatgcagcgc cagagcagct 7200
tgcgcatcgc cagcggcatc tgctcgggca cgttgaactt ggggtgtacc ggcggcaggt 7260
gcgccgacaa caaggtcgat ggcgagacct gcgcggacag gtagggaatc ccgtacttct 7320
cgtgagcgat gcgtgcgccc agcgcccaga gcgagccgac caccacgatg tcgtcatggc 7380
gctgcgccga gacgtactcg tagaccggct cgatcatccc ggcgatggtt tgccagagca 7440
cgccgaagga cgtcttgggg tcccacaggc gcggatcgcc catggtccgg cggtaggtca 7500
gttcgtcgct cagcgggacg aacgcgatgc cgtgctgctc caccgcgtcg cgaaacaccg 7560
ggatggtgca gaggctcacg cggtgcccgc gcaatttcag ggtccgggcc aggccgatga 7620
agggaaatac gtcgccggcc gagccgatgg cgatgaggat ggcgtgcatg gtgctactcc 7680
gtgcgttatg caaccgcaaa gcccggccag gccgggtctt cgcaggtcaa gggttcaggc 7740
gtagccgatg gccatctcgt ggaatcccgc cgcgcgttcc gcccgctgcg gctccggttg 7800
cttcagcagg tgctcgagca gggcgcggtg cacgcgtacc gcggccagct tggactccag 7860
gtcgaggaaa tgcccggtgc cctccacccg cgagaaactg cagtgcggca ggtagtcgcg 7920
gaactggcgg gcgtcctcgg cggtggtgta ttcgtcccag ctgccgttga tgaaatgcac 7980
gtggctctgg atccgctcca ggcaagccag gtagccccga tcgttgagcg ccagcacctg 8040
gtcgatgtga aagcgcgcct gctcgtattc gccggtggcc agcgaagcca tgtgctgatg 8100
gttgctggct ttcaggcgcg gcggcaggta tttgccgacg gtctcgttga gcagatggcc 8160
gatcgccgac ttgtcgtcca gctcgatcag cgcctgcgcc cgcccgacgt agtcgagcat 8220
cgcctggttc agtccagggg cgaatgccat caccaccgag ctgcggatgc cgcgcggatt 8280
gcgcgacagc gccagcagcg tggagatacc gccccaggac gcggagacca ggtgattgac 8340
ctcgaagcgc tcgatcagcg ccaggaggat ttccacctcg tcgtccttgg tgatcaaccc 8400
ccgctgcggg ttgtgctgac gcgactgccc ggcgaagggc aggtcgaaca gcaccacgtt 8460
gaaatgttcg gccaggcact tgcaggtccg ggcgaacgag gcggtggtcg ccatcgcgcc 8520
gttgaccagc atcaccgtgc tgcgcccggg atcctgccca acgcgctcga catgtacccg 8580
caggcccttg caaaccgata ccaacagact ttcgcgccgc atttcacacc tcccaaaaat 8640
gccagatccc ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag 8700
ggggggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta 8760
atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 8820
acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 8880
aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 8940
atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcatgc ataaaaactg 9000
ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg 9060
aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tgggggtggg 9120
cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa 9180
cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca 9240
ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc 9300
aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag 9360
gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat 9420
gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc 9480
attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc 9540
gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc 9600
ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat 9660
gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg 9720
cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc 9780
ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag 9840
cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg 9900
cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc 9960
tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc 10020
gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat 10080
gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct 10140
tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc 10200
cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca 10260
tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat 10320
agcccagtag ctgacattca tcccaggtgg cacttttcgg ggaaatgtgc gcgcccgcgt 10380
tcctgctggc gctgggcctg tttctggcgc tggacttccc gctgttccgt cagcagcttt 10440
tcgcccacgg ccttgatgat cgcggcggcc ttggcctgca tatcccgatt caacggcccc 10500
agggcgtcca gaacgggctt caggcgctcc cgaaggtctc gggccgtctc ttgggcttga 10560
tcggccttct tgcgcatctc acgcgctcct gcggcggcct gtagggcagg ctcatacccc 10620
tgccgaaccg cttttgtcag ccggtcggcc acggcttccg gcgtctcaac gcgctttgag 10680
attcccagct tttcggccaa tccctgcggt gcataggcgc gtggctcgac cgcttgcggg 10740
ctgatggtga cgtggcccac tggtggccgc tccagggcct cgtagaacgc ctgaatgcgc 10800
gtgtgacgtg ccttgctgcc ctcgatgccc cgttgcagcc ctagatcggc cacagcggcc 10860
gcaaacgtgg tctggtcgcg ggtcatctgc gctttgttgc cgatgaactc cttggccgac 10920
agcctgccgt cctgcgtcag cggcaccacg aacgcggtca tgtgcgggct ggtttcgtca 10980
cggtggatgc tggccgtcac gatgcgatcc gccccgtact tgtccgccag ccacttgtgc 11040
gccttctcga agaacgccgc ctgctgttct tggctggccg acttccacca ttccgggctg 11100
gccgtcatga cgtactcgac cgccaacaca gcgtccttgc gccgcttctc tggcagcaac 11160
tcgcgcagtc ggcccatcgc ttcatcggtg ctgctggccg cccagtgctc gttctctggc 11220
gtcctgctgg cgtcagcgtt gggcgtctcg cgctcgcggt aggcgtgctt gagactggcc 11280
gccacgttgc ccattttcgc cagcttcttg catcgcatga tcgcgtatgc cgccatgcct 11340
gcccctccct tttggtgtcc aaccggctcg acgggggcag cgcaaggcgg tgcctccggc 11400
gggccactca atgcttgagt atactcacta gactttgctt cgcaaagtcg tgaccgccta 11460
cggcggctgc ggcgccctac gggcttgctc tccgggcttc gccctgcgcg gtcgctgcgc 11520
tcccttgcca gcccgtggat atgtggacga tggccgcgag cggccaccgg ctggctcgct 11580
tcgctcggcc cgtggacaac cctgctggac aagctgatgg acaggctgcg cctgcccacg 11640
agcttgacca cagggattgc ccaccggcta cccagccttc gaccacatac ccaccggctc 11700
caactgcgcg gcctgcggcc ttgccccatc aattttttta attttctctg gggaaaagcc 11760
tccggcctgc ggcctgcgcg cttcgcttgc cggttggaca ccaagtggaa ggcgggtcaa 11820
ggctcgcgca gcgaccgcgc agcggcttgg ccttgacgcg cctggaacga cccaagccta 11880
tgcgagtggg ggcagtcgaa ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc 11940
gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg 12000
atcaacaagc cccggagggg ccactttttg ccggaggggg agccgcgccg aaggcgtggg 12060
ggaaccccgc aggggtgccc ttctttgggc accaaagaac tagatatagg gcgaaatgcg 12120
aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc aacagctcat tgcggcaccc 12180
cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca cttttacgca 12240
acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga ttgcgcatgg tgccgcaacc 12300
gtgcggcacc ctaccgcatg gagataagca tggccacgca gtccagagaa atcggcattc 12360
aagccaagaa caagcccggt cactgggtgc aaacggaacg caaagcgcat gaggcgtggg 12420
ccgggcttat tgcgaggaaa cccacggcgg caatgctgct gcatcacctc gtggcgcaga 12480
tgggccacca gaacgccgtg gtggtcagcc agaagacact ttccaagctc atcggacgtt 12540
ctttgcggac ggtccaatac gcagtcaagg acttggtggc cgagcgctgg atctccgtcg 12600
tgaagctcaa cggccccggc accgtgtcgg cctacgtggt caatgaccgc gtggcgtggg 12660
gccagccccg cgaccagttg cgcctgtcgg tgttcagtgc cgccgtggtg gttgatcacg 12720
acgaccagga cgaatcgctg ttggggcatg gcgacctgcg ccgcatcccg accctgtatc 12780
cgggcgagca gcaactaccg accggccccg gcgaggagcc gcccagccag cccggcattc 12840
cgggcatgga accagacctg ccagccttga ccgaaacgga ggaatgggaa cggcgcgggc 12900
agcagcgcct gccgatgccc gatgagccgt gttttctgga cgatggcgag ccgttggagc 12960
cgccgacacg ggtcacgctg ccgcgccggt agcacttggg ttgcgcagca acccgtaagt 13020
gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc tataccttgt ctgcctcccc 13080
gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc 13140
tcgctaacgg attcaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt 13200
caattattac ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca 13260
cggtcacact gcttccggta gtcaataaa 13289
<210> 75
<211> 14250
<212> DNA
<213> Artificial Sequence
<220>
<223> vector
<400> 75
ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60
cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120
ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180
cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240
cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300
gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360
tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420
tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480
acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540
ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600
tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660
ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720
ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780
tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840
cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900
cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960
ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020
gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080
tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140
ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200
ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260
acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320
gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380
tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440
caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500
gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560
aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620
cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680
taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740
tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800
gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860
acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920
gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980
actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040
ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100
ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160
tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220
gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280
gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340
ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400
ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460
gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520
ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580
cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640
ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700
gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtcac 2760
tgatttggca gcgacgggtc gagactaagt ctcacgttta acgtgtgcaa tggaaaacag 2820
gcgcaggggt atctttggga aatggcctac aaggaagggg caaattctga ttatttgcca 2880
ggccatgcag cggttgcttc ttagtggcgt gtgaaaaacg ccactatcgg caagacccgg 2940
tatggcatgt gttcagagac aaggctgcgc taccgcagcc cttctccgct aaaaccgtta 3000
gtcgaacagt tcggcgtcag ccaatgcgac cccaagctgg tccttgccag acaagcttgg 3060
tacggagtcg atgggccatt cgatacccac tgccggatca ttccaggcaa tgcagcgctc 3120
gcactgcggc gagtagaagt cggtggtctt gtagaggaac tctgcggttt cactcaacgt 3180
gacgaacccg tgtgcgaacc ctggcgggat ccacagctgg ttcttgttct cggccgacaa 3240
caccgcacct acccatttac cgaaggttgt ggacgagcga cggatatcca ccgcaacatc 3300
gaagacttcg ccttgcacca cacgcaccag cttgccctgg gcgtgaggtg ccagctgata 3360
gtgcaggcca cggagcacgc cttttaccga gcgcgagtgg ttgtcttgta cgaagtcggg 3420
ctgcaggccg gtcacttcgc tgaaaacacg ggcgttgaag ctctcgtaga agaaaccacg 3480
ttcgtcgcca aaaaccttgg gggtaaacag cacgacttcg gggatatcca gcggaatggc 3540
ttgcatcaga acaccttctc tttcagcaag ttctgcagat acttgccata accgtttttc 3600
agcagtggtt gagccaggca ctcgagttgc tcagcgttga tccagccagc gcggtagcaa 3660
atttcctcag ggcaggcgac tttcaagccc tgacggcgct ccatggttgc gatgtactgg 3720
ctagcctcca gcagactgtc gtgcgtgccg gtgtcgagcc acgcatagcc acggcccatg 3780
atttcgacct gcaactgctg ctgctgcaag taaaggttgt tgaggtcggt gatttccagc 3840
tcgccacgtg gggaaggctt cagctcgcga gccagattga ctacctgatt gtcatagaaa 3900
tacaggccgg tgaccgcata gctagacttt ggaactgccg gtttttcttc cagcgacaat 3960
acgcgaccgc tatcgtcaaa ctccgctacg ccatagcgtt ctgggtcatg aacatgataa 4020
gcgaatactg aagcaccgga ttcacgttta tctgcgttca atagcagtgc ctggaagtca 4080
tggccgtaga aaatattgtc accgagaacc aacgcagaag ggtcgttacc gatgaagtca 4140
gcgccgatgg tgaacgcttg cgccaagcca tccgggcttg gttgtattgc gtatgacagg 4200
ttcaggcccc actggctgcc atcgcccagc agctgttcga agcgcggggt gtcctgcggg 4260
gtggaaatga tcaggatgtc ccggatacca gcgagcagca gggtgctcag cgggtagtag 4320
atcatcggtt tgtcatacac cggcagcagc tgcttcgaaa ccgaaagtgt ggccggatgc 4380
aggcgtgtac ccgaaccgcc ggccagaata attcctttac gagccatgag agtccctatt 4440
actggatttc gtccagcata cgttgcacgc cttgctccca aagcggcatt ttgaaattga 4500
acgtgttttc cagtttgccc agtgccaggc gcgagttgcg cggacgtggt gcaggtactg 4560
gataagcttc ggtgctgatt gcggcaacct tatcagctgt cactttcagc gctacgccag 4620
tgcgttgagc atgcgccagc acgaactgag caaaaccatg ccaagaggtt tcaccggacg 4680
cagccaagtg gtaaatccct gccaggtgac ggttgtcttg cccattgaag atttgccgca 4740
ggatgtgtgc agtaacgtcg gcgatcaggt cagcgcccgt gggtgcgcca aattggtctg 4800
ctaccacgct caacgtctca cgctccgccg ccaggcgcag catggtcttg gcaaaattgt 4860
gcccgcgcgc agcatacacc cagctggtgc gcagtaccac ggccttggcg ccgctggcga 4920
gaatggcatg ctcgccttcc agcttggtcc ggccgtagac cgaaaggggg ccggtaggcg 4980
cagtttcctc ccagcgctga ctgccgctgc cgtcaaatac ataatcggtg gaatagtgaa 5040
tcaaccaggc gcccaaagct gctgtttcac gtgctaatac agcaggagcc gcggcattga 5100
tcattgcagc cagtgcctga tcgctctcag ctttatccac tgcagtgtag gcagcagcgt 5160
tgacgatcac gtccggcgcc agctgacgaa tcgtagcggc caagccgtcc aggttggaca 5220
agtcgccaca taagccctcg gccccctgac gatccagcgc aatgacctca cccagcggcg 5280
ccaaggcgcg ctgtagctcc cagcctactt gcccgttttt ccccaacagc aggattttca 5340
cgctttattt gccccgtatt gttgtgccac ccagtcacgg tagctgccgt ccatgacacc 5400
ttttacccat ttctggttgg ccaagtacca agcgactgtc tttcgaatgc ccgtctcgaa 5460
ggtttcggca ggtttccagc cgagctcccg ctcgatcttg cgtgcatcga tggcataacg 5520
gcggtcatgg cctgggcggt cggttacgta ggcgatgagt tctgcatact gttcgacagg 5580
ctcgccggtc ttctgattga ttacctggcg cgatgccgca ggtgccatct cgtcgagaag 5640
gctgcagagt gtacgcacaa tgtcaatgtt ggctttttca ttccagccgc caatattgta 5700
cgtctcgccg aacgcaccgg cttccagtac gcgacggatg cccgagcagt gatcttcgac 5760
atacagccag tcgcggattt gctggccgtc gccatagaca ggcagcgcct taccggcgag 5820
tgcgttgacg atcatcagcg ggatcagttt ttccgggaag tggagcggcc cgtaattgtt 5880
ggagcagttg gtagtgagta ccggcatgcc gtaggtatgg aaatacgagc gtaccagatg 5940
gtcgctggct gccttgctgg cggagtatgg gctgttcggc gcgtacggcg tggtttcggt 6000
gaacgccggg tcgtttggcc ctagtgtgcc gtagacttcg tcggtagaga catggaggaa 6060
acggaaggcc tccttctctg caccttccaa actattccaa tgcgcccggg cggcttcaag 6120
caagcgaaac gtgcccatca cgttggtttc gacaaacgct tcggggccgg tgattgagcg 6180
gtctacatgg gattccgccg cgaagtgaac cacggcgcgc gggcggtgct ctgcgaacag 6240
cttggtcaga agcgcagcat cgcaaatatt gccttgcaca aagcgatgct gagggttgcc 6300
ttccagcggc tgcaggttgg ccaggttgcc tgcgtaggtc agggcgtcga ggttgaggac 6360
gggttcctca ttgtgcgcac accattgcag tacgaaattt gagccgatga agccggctcc 6420
gcctgttact agaatcataa tttggctcta attggacaaa aggtgttgtc gtagacagat 6480
gacgctgaga aacttcgccc cggcaacgca aggcgagctg ctagcgctgc cggaattaga 6540
ccaccctgat gtcgaaaaat acactgctgg gtgccggtac tgaccgtgcc gatctgctca 6600
cgctggaaga ggaggagccg ttgcagcggt ggaaggtgac ctcatggttg cctagacaca 6660
acagggagtc gtcaggtgtc catgcaagaa acagttgttt gagcgatgca actcacggag 6720
acaaatctgg gtgctgacaa gtctagaggg ccttaagcta ccgctatcct gggactctag 6780
ttcgctgggc atggcagcgc ggggtgcgca tctggaagtc gggttcgttt ccgggggttg 6840
gaaagtgtca tttttatttg tcagtattcc tgaaatagga attagagtga tggcagttct 6900
tgcattcatg ttgaacaacc caacaaaaat tataatagcc tcggggctcg gcaggttaga 6960
cgtcattctt catgtttcag cggtctggcg ctcttgaata ttcataggaa agtgaaaaat 7020
gtctgttaag gaaggcttgc cgagcgtaga tgtgcagccg agcatgcatg ttaagggaaa 7080
ttcagtgccc gtcccccaga aaaagactgg ctcaaggcga tttgtgctgg atctggctta 7140
ttatcggaat cgatacccag acctatcttc gctcacggac gatgcgctcg aacagcattg 7200
gcatgcgttc ggtcatcaag aaggccgtta tgcctcggct tcgcacgaaa gtggtgatgc 7260
ggagcttatc gctgaagcag ttctcaatgc ggccgacagc gccgttatgg tcaagccgga 7320
tgttgcgaaa gttgacctac cggtaaacca gcaatagaca taagcggcta tttaacgacc 7380
ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta 7440
ttatcactta ttcaggcgta gcaccaggcg tttaagggca ccaataactg ccttaaaaaa 7500
attacgcccc gccctgccac tcatcgcagt cggcctattg gttaaaaaat gagctgattt 7560
aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt 7620
caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 7680
ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc 7740
acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt 7800
ggagctccac cgcggtggcg gccgctctag agggaatgcg tttcgccgac taggccttgg 7860
ccttgccgga agctacggac gccacggccg ggccggcgag gcgcttcagc aggcgcgggc 7920
ggttggtctc cagcgcgccg ccgcgtcccc gcaggccgtc ccacaggccc cagcccaggc 7980
agcgcagctt gagcagcttg tcgcgttcga gcaggagcac cgcgaggccc tgggtcaggg 8040
tcggcaggtt cgccagcagg gccagcggcg aggaccgggc gtagcggcgc aggaccagca 8100
ggccgttgcg cgccaggtag tagcggcgca gcggggcgtg gttcatcgcg ctgaggctga 8160
gaccgccgag gcggcgggtc ttgcgcgtgc cgatgcggtg ctcgaggacc agccgcgggt 8220
cgacgtacag gggcacgtcc agcgcctggg cgcgcaggct gtattcggtg tccacgtggt 8280
cgatgaacag ttcctcgtcg aagtggccga ggcgctggta ggcctcgcgg gtcagcaggc 8340
agccggagga gatcaggaac gaggtgcgct gcggggtcgt caggccgtcc agagacaatt 8400
gcctgagcgt cagtccgtcg agatggatgg ccggcaggaa gcgccggtca ccccggtcga 8460
agatccgtgg gccgagcagg caggcctgac cgttgcgcgc ctgcaggttg cgccactggg 8520
cggcgaggaa ggcgccgccg ggacgggagt cctggtcgag cagcagcaca ccctgcacgc 8580
cacgccggaa tagcgcgtcg agtccctggt tgaaggcgcc ggcgatgccc tgccggttgc 8640
cgtggtgcag cacggcgatg ccttgcccgc gcagccgggc attgcgctgc ggatcgctgt 8700
gcggtgagtt gtcgacggca aggaagcgca gttgcggaaa cgccgccgcc agttcgccaa 8760
ggtgttccag gtcgtcgtcg ccaggattga acagtaccac cagcacgccc atgtctatcc 8820
ggtccatgat cgttcttctc ccgtaggtcg aagttgccag gccaggacca gcccggccag 8880
aacgagaagg gcgccggcga ggaatggcgc gccggccagg ggcagcggcg cgagcggacc 8940
gctgccccag tggaacaggc cgctcatcag cggcggaccg acgatcgcgg cgaggctcat 9000
caggctgctc agcacgccct gcaactcgcc ctggcggtcg accggcacgc gggccgagag 9060
cagcccctgc atggccgggg tggcgaggct gccgagcgcg aagggcagca gcgcgcagac 9120
cagccagaat gacgagtcga ccagggcgaa cagcagcagg ccgcagcctt gcagggcgag 9180
gcccaggcgc agcaggcggg cgtcgtccag gcgccgcttg cagaggttca cgccgagggt 9240
ctgggcgagc accgcgagca cgccgtagag ggccagcgag tagccgatcc aggcgctgct 9300
ccagtgaaac ttctcgatca cgaagaacgg ccagaccacc atcaccgcct gcaagccgag 9360
gaataccagg gcaagcaccg ccagcaggcg tccgaccccc ggttgccgag ccaggccgct 9420
gatcgagcgc aaggcattca tccgcctcgg gtccaggcgg cggcgtcgcg tcgggggcag 9480
ggtttcctcg aggaacaggc cggcgagcag ggcgttgagc aggcacaggc cggcggccag 9540
caacagcggc agcgtcgtgc cgtgcaccgc cagcagccca ccgagggcgg ggccgaggat 9600
catgcccagg gcgaggccgg cgtacagcca gccgaagtgc cgggtgcgct gcccgtgcgt 9660
gccgaggtca gccgcgcagg ccatcgcggt ggccacgctg gcgccggtga gcccggccag 9720
cgcgcgaccg aggaacagca tccagaggct gtcggccagc gccagcagca gatagctgag 9780
ggcgaagccg agcatcgcca ggaccaggac ggggcggcgt ccgaagcggt cgctgaggct 9840
gccgaggacc ggcgaaaaga acaattgcag cagcgcgaag gtcatcacca gggcggcgcc 9900
ccaggtggcc gcgtcgcgga ccgccagcgg cgccacgctg ccgatcagcg tcggcagcag 9960
gggcacgatc aggccgacgc cagcggcatc cagcaggcag gtgaggaaca gcagaggcag 10020
gacgcgtttc gcgccgggac cgtgttcccg cgtggcggag gggcagaggc tggtcgtgga 10080
cacgccagga tcctcccggc gtcaggacgc agccttcagc catcgcgcat ccccctccct 10140
atgacaacgt tcgaccacct gggccgcttt accgcaagcg atactgtgcg gttgtgacaa 10200
ttccatgaaa cgccgacagg ccgccgccat ggccgggtcc tcgagcaagc gccacagcgc 10260
cccgcgcaac tcctgctcgc gcaatggcac gcccaggcgc atcccgcagc cgagccggac 10320
cagccgttcg gcattgtcga actggtcgtg ggcgcagggc agcagcacct gcggcacccc 10380
cgccgccaag gccaggctca tggcgccgat accgcccgga tggaccagcc cggcgcacga 10440
tggcagcaag gctcccagtg gcgcgtaggc gcgctgcagc acgtggttcg gcaagccgcg 10500
cagcggttcc tggccggcgc cggtgaggaa gatcccacgc gcgccgaggc gttccagcgc 10560
gcgcagggcc atggcgtaga agtcgccctg caggtgttcg gtcgagccct gggtgaacac 10620
cagcggccgg ctgccctgat cgagaaagcg ttgcagttcg tcgtcgagcg gggtccccgg 10680
gatactgccg tcgaacagcg ggaagccggt catgtgcagg ggttgcggcc aatcctgctg 10740
gggcggcgcg aaccaggccg ggaacaggca gaccacgccc tgcggcgaat gcatccattg 10800
ggtgaagatg cgcttcaccg gcgtctccag gccgaccttg cgccgcaccg cgttgatatc 10860
cggcgcgcag gtgcgatcca gcttgaagcg ctcgatgcag cgccagagca gcttgcgcat 10920
cgccagcggc atctgctcgg gcacgttgaa cttggggtgt accggcggca ggtgcgccga 10980
caacaaggtc gatggcgaga cctgcgcgga caggtaggga atcccgtact tctcgtgagc 11040
gatgcgtgcg cccagcgccc agagcgagcc gaccaccacg atgtcgtcat ggcgctgcgc 11100
cgagacgtac tcgtagaccg gctcgatcat cccggcgatg gtttgccaga gcacgccgaa 11160
ggacgtcttg gggtcccaca ggcgcggatc gcccatggtc cggcggtagg tcagttcgtc 11220
gctcagcggg acgaacgcga tgccgtgctg ctccaccgcg tcgcgaaaca ccgggatggt 11280
gcagaggctc acgcggtgcc cgcgcaattt cagggtccgg gccaggccga tgaagggaaa 11340
tacgtcgccg gccgagccga tggcgatgag gatggcgtgc atggtgctac tccgtgcgtt 11400
atgcaaccgc aaagcccggc caggccgggt cttcgcaggt caagggttca ggcgtagccg 11460
atggccatct cgtggaatcc cgccgcgcgt tccgcccgct gcggctccgg ttgcttcagc 11520
aggtgctcga gcagggcgcg gtgcacgcgt accgcggcca gcttggactc caggtcgagg 11580
aaatgcccgg tgccctccac ccgcgagaaa ctgcagtgcg gcaggtagtc gcggaactgg 11640
cgggcgtcct cggcggtggt gtattcgtcc cagctgccgt tgatgaaatg cacgtggctc 11700
tggatccgct ccaggcaagc caggtagccc cgatcgttga gcgccagcac ctggtcgatg 11760
tgaaagcgcg cctgctcgta ttcgccggtg gccagcgaag ccatgtgctg atggttgctg 11820
gctttcaggc gcggcggcag gtatttgccg acggtctcgt tgagcagatg gccgatcgcc 11880
gacttgtcgt ccagctcgat cagcgcctgc gcccgcccga cgtagtcgag catcgcctgg 11940
ttcagtccag gggcgaatgc catcaccacc gagctgcgga tgccgcgcgg attgcgcgac 12000
agcgccagca gcgtggagat accgccccag gacgcggaga ccaggtgatt gacctcgaag 12060
cgctcgatca gcgccaggag gatttccacc tcgtcgtcct tggtgatcaa cccccgctgc 12120
gggttgtgct gacgcgactg cccggcgaag ggcaggtcga acagcaccac gttgaaatgt 12180
tcggccaggc acttgcaggt ccgggcgaac gaggcggtgg tcgccatcgc gccgttgacc 12240
agcatcaccg tgctgcgccc gggatcctgc ccaacgcgct cgacatgtac ccgcaggccc 12300
ttgcaaaccg ataccaacag actttcgcgc cgcatttcac acctcccaaa aatgccagat 12360
cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc 12420
ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg 12480
tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 12540
ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 12600
ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 12660
ggccaacgcg cggggagagg cggtttgcgt attgggcgca tgcataaaaa ctgttgtaat 12720
tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc 12780
agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatgggggt gggcgaagaa 12840
ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc 12900
gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg 12960
cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg 13020
cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg 13080
tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga 13140
tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc 13200
accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc 13260
atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc 13320
agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt 13380
ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca 13440
tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc 13500
ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct 13560
gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca 13620
ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc 13680
cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc 13740
ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac 13800
gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc 13860
aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct 13920
ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc 13980
ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag 14040
tagctgacat tcatcccagg tggcactttt cggggaaatg tgcgcgcccg cgttcctgct 14100
ggcgctgggc ctgtttctgg cgctggactt cccgctgttc cgtcagcagc ttttcgccca 14160
cggccttgat gatcgcggcg gccttggcct gcatatcccg attcaacggc cccagggcgt 14220
ccagaacggg cttcaggcgc tcccgaaggt 14250
<210> 76
<211> 1011
<212> DNA
<213> Burkholderia thailandensis
<400> 76
tctagatacg ggagaagaac gatcatgacg atcctggggg cgctggtgat tctgtacgac 60
ccgacggacg agcagttgtc ggggctggag gcgctcgcgc gcgacagcga cgcgctcgtg 120
gtcgtggaca acacgccgca cgagcacgcg gcggcgcgcg agcgggtgcg tgcgctgtcg 180
gcgcggacga acacggtgtg gcgacaccac ggcaaccggg gcggggtcgc gggcgggtac 240
aacgcggggc tgtcggtgct gttcgcgcag ggcgtcgagg cggtcgcgct gttcgaccag 300
gactcgacgg tgccggccgg gtacttcgag cggatgcgcg aggcgtgcgc gcaactgggt 360
gagcaaccgg gcgcgcacgc gggcgcgttc atcgcgggcc cgcggatcta cgacgcgaac 420
gagcagcgct tcctgccgga gctgatgacg agcggggtga cggtgcgccg cgtgcgggtg 480
gagggcgaga cggcgccgca gcgctgcgcg ttcctgatct cgtcgggcag cgtgatttcg 540
cgggccgcgt acgcgcggct cggtcgattc gacgaggcgc tgttcatcga tcacgtcgac 600
accgagtatt gcctgcgcgc gctcgcgcac aacgtgccgc tgtacgtggt gccgccgctc 660
gtgctgacgc accggatcgg cgcgcggcgc cggcacaagg tggggccgtt cgagctgacg 720
gcgatgcatc acgggtggtt gcgccgatac tacggcgcgc gcaacgcgat gcaactgggg 780
ctgcagtacg gcttgcggtt tccggtggcg ctggtgccga atctgctgac gatatggcag 840
gtgatccagg tggtgctgtg cgagcgggag aagggcgcga agctgcgcgg gatcgcgctg 900
ggcgtgctcg acggcctgtt cgggcggctg ggatcgttcg acgatgcgcg cgcgggcgcg 960
gcggcgcgcg agccggtgcg gcaggaatga tcggcgaaac gcattgagct c 1011
<210> 77
<211> 888
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(888)
<400> 77
atg cgg cgc gaa agt ctg ttg gta tcg gtt tgc aag ggc ctg cgg gta 48
Met Arg Arg Glu Ser Leu Leu Val Ser Val Cys Lys Gly Leu Arg Val
1 5 10 15
cat gtc gag cgc gtt ggg cag gat ccc ggg cgc agc acg gtg atg ctg 96
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Ser Thr Val Met Leu
20 25 30
gtc aac ggc gcg atg gcg acc acc gcc tcg ttc gcc cgg acc tgc aag 144
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
tgc ctg gcc gaa cat ttc aac gtg gtg ctg ttc gac ctg ccc ttc gcc 192
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
ggg cag tcg cgt cag cac aac ccg cag cgc ggg ttg atc acc aag gac 240
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
gac gag gtg gaa atc ctc ctg gcg ctg atc gag cgc ttc gag gtc aat 288
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Glu Val Asn
85 90 95
cac ctg gtc tcc gcg tcg tgg ggc ggt atc tcc acg ctg ctg gcg ctg 336
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
tcg cgc aat ccg cgc ggc atc cgc agc tcg gtg gtg atg gca ttc gcc 384
Ser Arg Asn Pro Arg Gly Ile Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
cct gga ctg aac cag gcg atg ctc gac tac gtc ggg cgg gcg cag gcg 432
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Ala
130 135 140
ctg atc gag ctg gac gac aag tcg gcg atc ggc cat ctg ctc aac gag 480
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
acc gtc ggc aaa tac ctg ccg ccg cgc ctg aaa gcc agc aac cat cag 528
Thr Val Gly Lys Tyr Leu Pro Pro Arg Leu Lys Ala Ser Asn His Gln
165 170 175
cac atg gct tcg ctg gcc acc ggc gaa tac gag cag gcg cgc ttt cac 576
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
atc gac cag gtg ctg gcg ctc aac gat cgg ggc tac ctg gct tgc ctg 624
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ala Cys Leu
195 200 205
gag cgg atc cag agc cac gtg cat ttc atc aac ggc agc tgg gac gaa 672
Glu Arg Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
tac acc acc gcc gag gac gcc cgc cag ttc cgc gac tac ctg ccg cac 720
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
tgc agt ttc tcg cgg gtg gag ggc acc ggg cat ttc ctc gac ctg gag 768
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
tcc aag ctg gcc gcg gta cgc gtg cac cgc gcc ctg ctc gag cac ctg 816
Ser Lys Leu Ala Ala Val Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
ctg aag caa ccg gag ccg cag cgg gcg gaa cgc gcg gcg gga ttc cac 864
Leu Lys Gln Pro Glu Pro Gln Arg Ala Glu Arg Ala Ala Gly Phe His
275 280 285
gag atg gcc atc ggc tac gcc tga 888
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 78
<211> 295
<212> PRT
<213> Pseudomonas aeruginosa
<400> 78
Met Arg Arg Glu Ser Leu Leu Val Ser Val Cys Lys Gly Leu Arg Val
1 5 10 15
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Ser Thr Val Met Leu
20 25 30
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Glu Val Asn
85 90 95
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
Ser Arg Asn Pro Arg Gly Ile Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Ala
130 135 140
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
Thr Val Gly Lys Tyr Leu Pro Pro Arg Leu Lys Ala Ser Asn His Gln
165 170 175
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ala Cys Leu
195 200 205
Glu Arg Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
Ser Lys Leu Ala Ala Val Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
Leu Lys Gln Pro Glu Pro Gln Arg Ala Glu Arg Ala Ala Gly Phe His
275 280 285
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 79
<211> 888
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(888)
<400> 79
atg cgg cgc gaa agt ctg ttg gta tcg gtt tgc aag ggc ctg cgg gta 48
Met Arg Arg Glu Ser Leu Leu Val Ser Val Cys Lys Gly Leu Arg Val
1 5 10 15
cat gtc gag cgc gtt ggg cag gat ccc ggg cgc agc acg gtg atg ctg 96
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Ser Thr Val Met Leu
20 25 30
gtc aac ggc gcg atg gcg acc acc gcc tcg ttc gcc cgg acc tgc aag 144
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
tgc ctg gcc gaa cat ttc aac gtg gtg ctg ttc gac ctg ccc ttc gcc 192
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
ggg cag tcg cgt cag cac aac ccg cag cgc ggg ttg atc acc aag gac 240
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
gac gag gtg gaa atc ctc ctg gcg ctg atc gag cgc ttc gag gtc aat 288
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Glu Val Asn
85 90 95
cac ctg gtc tcc gcg tcc tgg ggc ggt atc tcc acg ctg ctg gcg ctg 336
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
tcg cgc aat ccg cgc ggc atc cgc agc tcg gtg gtg atg gca ttc gcc 384
Ser Arg Asn Pro Arg Gly Ile Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
cct gga ctg aac cag gcg atg ctc gac tac gtc ggg cgg gcg cag gcg 432
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Ala
130 135 140
ctg atc gag ctg gac gac aag tcg gcg atc ggc cat ctg ctc aac gag 480
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
acc gtc ggc aaa tac ctg ccg cag cgc ctg aaa gcc agc aac cat cag 528
Thr Val Gly Lys Tyr Leu Pro Gln Arg Leu Lys Ala Ser Asn His Gln
165 170 175
cac atg gct tcg ctg gcc acc ggc gaa tac gag cag gcg cgc ttt cac 576
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
atc gac cag gtg ctg gcg ctc aac gat cgg ggc tac ttg gct tgc ctg 624
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ala Cys Leu
195 200 205
gag cgg atc cag agc cac gtg cat ttc atc aac ggc agc tgg gac gaa 672
Glu Arg Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
tac acc acc gcc gag gac gcc cgc cag ttc cgc gac tac ctg ccg cac 720
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
tgc agt ttc tcg cgg gtg gag ggc acc ggg cat ttc ctc gac ctg gag 768
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
tcc aag ctg gca gcg gta cgc gtg cac cgc gcc ctg ctc gag cac ctg 816
Ser Lys Leu Ala Ala Val Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
ctg aag caa ccg gag ccg cag cgg gcg gaa cgc gcg gcg gga ttc cac 864
Leu Lys Gln Pro Glu Pro Gln Arg Ala Glu Arg Ala Ala Gly Phe His
275 280 285
gag atg gcc atc ggc tac gcc tga 888
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 80
<211> 295
<212> PRT
<213> Pseudomonas aeruginosa
<400> 80
Met Arg Arg Glu Ser Leu Leu Val Ser Val Cys Lys Gly Leu Arg Val
1 5 10 15
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Ser Thr Val Met Leu
20 25 30
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Glu Val Asn
85 90 95
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
Ser Arg Asn Pro Arg Gly Ile Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Ala
130 135 140
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
Thr Val Gly Lys Tyr Leu Pro Gln Arg Leu Lys Ala Ser Asn His Gln
165 170 175
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ala Cys Leu
195 200 205
Glu Arg Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
Ser Lys Leu Ala Ala Val Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
Leu Lys Gln Pro Glu Pro Gln Arg Ala Glu Arg Ala Ala Gly Phe His
275 280 285
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 81
<211> 888
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(888)
<400> 81
atg cgg cgc gaa agt ctg ttg gta acg gta tgc aag ggc ctg cgg gta 48
Met Arg Arg Glu Ser Leu Leu Val Thr Val Cys Lys Gly Leu Arg Val
1 5 10 15
cat gtc gag cgc gtg ggg cag gat ccc ggg cgc gac acg gtg atg ctg 96
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Asp Thr Val Met Leu
20 25 30
gtc aac ggc gcg atg gcg acc acc gcc tcg ttc gcc cgg acc tgc aag 144
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
tgc ctg gcc gaa cat ttc aac gtg gtg ctg ttc gac ctg ccc ttc gcc 192
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
ggg cag tcg cgg cag cac aat ccg cag cgc ggg ttg atc acc aag gac 240
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
gac gag gtg gag att ctc ctg gcg ctg atc gag cgc ttc gct gtc aac 288
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Ala Val Asn
85 90 95
cac ctg gtc tcg gcc tcc tgg ggc ggc atc tcc acg ctg ctg gcg ctg 336
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
tcg cgc aac ccg cgc ggg gtc cgc agc tcg gtg gtg atg gcg ttc gcg 384
Ser Arg Asn Pro Arg Gly Val Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
ccg ggg ctg aac cag gcg atg ctc gat tat gtc ggg cgg gcc cag gaa 432
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Glu
130 135 140
ctg atc gaa ctg gac gac aag tcg gcg atc ggc cac ctg ctc aac gag 480
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
acc gtc ggc aag tac ctg ccg ccg cgg ctg aag gcc agc aac cat cag 528
Thr Val Gly Lys Tyr Leu Pro Pro Arg Leu Lys Ala Ser Asn His Gln
165 170 175
cac atg gcc tcc ctg gcc act ggc gag tac gag cag gcg cgt ttc cac 576
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
atc gac cag gtg ctg gcg ctc aat gac cgt ggc tac ctg agc tgc ctg 624
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ser Cys Leu
195 200 205
ggg cag atc cag agt cac gtg cat ttc atc aac ggc agc tgg gac gag 672
Gly Gln Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
tac acc acc gcc gag gac gcc cgc cag ttc cgc gat tac ctg ccg cat 720
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
tgc agt ttt tcg cgg gtg gaa ggc acc ggg cac ttc ctc gac ctg gag 768
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
tcc aag ctg gcg gcg gcg cgt gtg cac cgg gcg ttg ctc gag cac ctg 816
Ser Lys Leu Ala Ala Ala Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
ctg gcg caa ccg gaa ccg tgg cgc tcc gag cag gcg gcg gga ttc cac 864
Leu Ala Gln Pro Glu Pro Trp Arg Ser Glu Gln Ala Ala Gly Phe His
275 280 285
gag atg gcc atc ggc tac gcc tga 888
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 82
<211> 295
<212> PRT
<213> Pseudomonas aeruginosa
<400> 82
Met Arg Arg Glu Ser Leu Leu Val Thr Val Cys Lys Gly Leu Arg Val
1 5 10 15
His Val Glu Arg Val Gly Gln Asp Pro Gly Arg Asp Thr Val Met Leu
20 25 30
Val Asn Gly Ala Met Ala Thr Thr Ala Ser Phe Ala Arg Thr Cys Lys
35 40 45
Cys Leu Ala Glu His Phe Asn Val Val Leu Phe Asp Leu Pro Phe Ala
50 55 60
Gly Gln Ser Arg Gln His Asn Pro Gln Arg Gly Leu Ile Thr Lys Asp
65 70 75 80
Asp Glu Val Glu Ile Leu Leu Ala Leu Ile Glu Arg Phe Ala Val Asn
85 90 95
His Leu Val Ser Ala Ser Trp Gly Gly Ile Ser Thr Leu Leu Ala Leu
100 105 110
Ser Arg Asn Pro Arg Gly Val Arg Ser Ser Val Val Met Ala Phe Ala
115 120 125
Pro Gly Leu Asn Gln Ala Met Leu Asp Tyr Val Gly Arg Ala Gln Glu
130 135 140
Leu Ile Glu Leu Asp Asp Lys Ser Ala Ile Gly His Leu Leu Asn Glu
145 150 155 160
Thr Val Gly Lys Tyr Leu Pro Pro Arg Leu Lys Ala Ser Asn His Gln
165 170 175
His Met Ala Ser Leu Ala Thr Gly Glu Tyr Glu Gln Ala Arg Phe His
180 185 190
Ile Asp Gln Val Leu Ala Leu Asn Asp Arg Gly Tyr Leu Ser Cys Leu
195 200 205
Gly Gln Ile Gln Ser His Val His Phe Ile Asn Gly Ser Trp Asp Glu
210 215 220
Tyr Thr Thr Ala Glu Asp Ala Arg Gln Phe Arg Asp Tyr Leu Pro His
225 230 235 240
Cys Ser Phe Ser Arg Val Glu Gly Thr Gly His Phe Leu Asp Leu Glu
245 250 255
Ser Lys Leu Ala Ala Ala Arg Val His Arg Ala Leu Leu Glu His Leu
260 265 270
Leu Ala Gln Pro Glu Pro Trp Arg Ser Glu Gln Ala Ala Gly Phe His
275 280 285
Glu Met Ala Ile Gly Tyr Ala
290 295
<210> 83
<211> 1281
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(1281)
<400> 83
atg cac gcc att ctc atc gcc atc ggt tcg gcc ggc gac gtg ttc ccc 48
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
ttc atc ggc ctg gcc cgc acc ctg aag ttg cgc ggc cac cgc gtc agc 96
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
ctg tgc acc att ccg gtg ttt cgc gcc gcg gtg gag cag cac ggc atc 144
Leu Cys Thr Ile Pro Val Phe Arg Ala Ala Val Glu Gln His Gly Ile
35 40 45
gag ttc gtc ccg ctc agc gac gaa ctg acc tac cgc cgg acc atg ggc 192
Glu Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
gac ccg cgc ctg tgg gat ccg aag acc tcg ttc gga gtg ctc tgg cag 240
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
gcc atc gcc ggg atg atc gag ccg gtc tac gag tac gtc tgc gca cag 288
Ala Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Cys Ala Gln
85 90 95
cgc cac gac gac atc gtg gtg gtc ggt tcg ctg tgg gcc ctg ggc gcg 336
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
cgg atc gcc cat gag aaa tac ggg att ccc tac ctg tcg gtg cag gtc 384
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Val Gln Val
115 120 125
tcg ccg tcg acc ctg ctg tcg gcg cac ctg ccg ccg gtc cac ccc agg 432
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Arg
130 135 140
ttc aac gtg ccc gag cag gtc ccg ctg gcg atg cgc aag ttg ctc tgg 480
Phe Asn Val Pro Glu Gln Val Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
cgc tgc atc gaa cgc ttc aag ctg gac cgc acc tgc gcc ccg gag atc 528
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Glu Ile
165 170 175
aac gcg gtg cgc cgc aag gtc ggc ctg gtc ggc ccg gcg aag cgc atc 576
Asn Ala Val Arg Arg Lys Val Gly Leu Val Gly Pro Ala Lys Arg Ile
180 185 190
ttc acc cag tgg atg cat tcg cca cag gga gtg ctc tgc ctg ttc ccg 624
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Leu Cys Leu Phe Pro
195 200 205
gcc tgg ttc gca ccg ccc cag cag gac tgg ccg caa ccg ctg cac atg 672
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
acc ggc ttc ccg ctg ttc gac ggc agc gtc ccg ggg acc cgc ctc gac 720
Thr Gly Phe Pro Leu Phe Asp Gly Ser Val Pro Gly Thr Arg Leu Asp
225 230 235 240
gac gag ttg cag cgc ttc ctc gag cag ggc agt cgg ccg ctg gtg ttc 768
Asp Glu Leu Gln Arg Phe Leu Glu Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
acc cag ggt tcg acc gag cac ctg cag gga gac ttc tat gcc atg gcc 816
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
ttg cgc gcg ctg gag cgt ctc ggc gcc cgc ggc atc ttc ctc acc ggc 864
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
gcc ggc cag gag ccg ctg cgt ggc ttg ccg agc cac gtg ctg caa cgc 912
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Ser His Val Leu Gln Arg
290 295 300
tcg tac gtg ccg ttg ggg gcc ttg ctg ccg gcg tgc gcc ggg ctg gtc 960
Ser Tyr Val Pro Leu Gly Ala Leu Leu Pro Ala Cys Ala Gly Leu Val
305 310 315 320
cac ccg gcc ggc atc ggc gcc atg agc ctg gcg ctg gcg gcg ggg gtg 1008
His Pro Ala Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
ccg cag gtg ctg ctg cct tgc gcc cac gac cag ttc gac aac gcc gaa 1056
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
cgc ctg gtc cgc ctc ggc tgc ggt atc cgc ctg ggc ctg ccg cta cgc 1104
Arg Leu Val Arg Leu Gly Cys Gly Ile Arg Leu Gly Leu Pro Leu Arg
355 360 365
gag cag gcg ctg cgc gag tcg ctc tgg cgg ctg ctc gag gac ccg gcg 1152
Glu Gln Ala Leu Arg Glu Ser Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
ctg gcg gcg gcc tgt cgg cgt ttc atg gaa ttg tca caa ccg cac agt 1200
Leu Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
atc gct tgc ggt aaa gcg gcc caa gtg gtc gaa cgt tgt cat agg gag 1248
Ile Ala Cys Gly Lys Ala Ala Gln Val Val Glu Arg Cys His Arg Glu
405 410 415
ggg gat gtg cga tgg ctg aaa gcc gcg tcc tga 1281
Gly Asp Val Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 84
<211> 426
<212> PRT
<213> Pseudomonas aeruginosa
<400> 84
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
Leu Cys Thr Ile Pro Val Phe Arg Ala Ala Val Glu Gln His Gly Ile
35 40 45
Glu Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
Ala Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Cys Ala Gln
85 90 95
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Val Gln Val
115 120 125
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Arg
130 135 140
Phe Asn Val Pro Glu Gln Val Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Glu Ile
165 170 175
Asn Ala Val Arg Arg Lys Val Gly Leu Val Gly Pro Ala Lys Arg Ile
180 185 190
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Leu Cys Leu Phe Pro
195 200 205
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
Thr Gly Phe Pro Leu Phe Asp Gly Ser Val Pro Gly Thr Arg Leu Asp
225 230 235 240
Asp Glu Leu Gln Arg Phe Leu Glu Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Ser His Val Leu Gln Arg
290 295 300
Ser Tyr Val Pro Leu Gly Ala Leu Leu Pro Ala Cys Ala Gly Leu Val
305 310 315 320
His Pro Ala Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
Arg Leu Val Arg Leu Gly Cys Gly Ile Arg Leu Gly Leu Pro Leu Arg
355 360 365
Glu Gln Ala Leu Arg Glu Ser Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
Leu Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
Ile Ala Cys Gly Lys Ala Ala Gln Val Val Glu Arg Cys His Arg Glu
405 410 415
Gly Asp Val Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 85
<211> 1281
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(1281)
<400> 85
atg cac gcc atc ctc atc gcc atc ggc tcg gcc ggc gac gta ttt ccc 48
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
ttc atc ggc ctg gcc cgg acc ctg aaa ctg cgc ggg cac cgc gtg agc 96
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
ctc tgc acc atc ccg gtg ttt cgc gac gcg gtg gag cag cac ggc atc 144
Leu Cys Thr Ile Pro Val Phe Arg Asp Ala Val Glu Gln His Gly Ile
35 40 45
gcg ttc gtc ccg ctg agc gac gaa ctg acc tac cgc cgg acc atg ggc 192
Ala Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
gat ccg cgc ctg tgg gac ccc aag acg tcc ttc ggc gtg ctc tgg caa 240
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
gcc atc gcc ggg atg atc gag ccg gtc tac gag tac gtc tcg gcg cag 288
Ala Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Ser Ala Gln
85 90 95
cgc cat gac gac atc gtg gtg gtc ggc tcg cta tgg gcg ctg ggc gca 336
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
cgc atc gct cac gag aag tac ggg att ccc tac ctg tcc gcg cag gtc 384
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Ala Gln Val
115 120 125
tcg cca tcg acc ctg ttg tcg gcg cac ctg ccg ccg gta cac ccc aag 432
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Lys
130 135 140
ttc aac gtg ccc gag cag atg ccg ctg gcg atg cgc aag ctg ctc tgg 480
Phe Asn Val Pro Glu Gln Met Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
cgc tgc atc gag cgc ttc aag ctg gat cgc acc tgc gcg ccg gag atc 528
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Glu Ile
165 170 175
aac gcg gtg cgc cgc aag gtc ggc ctg gaa acg ccg gtg aag cgc atc 576
Asn Ala Val Arg Arg Lys Val Gly Leu Glu Thr Pro Val Lys Arg Ile
180 185 190
ttc acc caa tgg atg cat tcg ccg cag ggc gtg gtc tgc ctg ttc ccg 624
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Val Cys Leu Phe Pro
195 200 205
gcc tgg ttc gcg ccg ccc cag cag gat tgg ccg caa ccc ctg cac atg 672
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
acc ggc ttc ccg ctg ttc gac ggc agt atc ccg ggg acc ccg ctc gac 720
Thr Gly Phe Pro Leu Phe Asp Gly Ser Ile Pro Gly Thr Pro Leu Asp
225 230 235 240
gac gaa ctg caa cgc ttt ctc gat cag ggc agc cgg ccg ctg gtg ttc 768
Asp Glu Leu Gln Arg Phe Leu Asp Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
acc cag ggc tcg acc gaa cac ctg cag ggc gac ttc tac gcc atg gcc 816
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
ctg cgc gcg ctg gaa cgc ctc ggc gcg cgt ggg atc ttc ctc acc ggc 864
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
gcc ggc cag gaa ccg ctg cgc ggc ttg ccg aac cac gtg ctg cag cgc 912
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Asn His Val Leu Gln Arg
290 295 300
gcc tac gcg cca ctg gga gcc ttg ctg cca tcg tgc gcc ggg ctg gtc 960
Ala Tyr Ala Pro Leu Gly Ala Leu Leu Pro Ser Cys Ala Gly Leu Val
305 310 315 320
cat ccg ggc ggt atc ggc gcc atg agc cta gcc ttg gcg gcg ggg gtg 1008
His Pro Gly Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
ccg cag gtg ctg ctg ccc tgt gcc cac gac cag ttc gac aat gcc gaa 1056
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
cgg ctg gtc cgg ctc ggc tgc ggg atg cgc ctg ggc gtg ccg ttg cgc 1104
Arg Leu Val Arg Leu Gly Cys Gly Met Arg Leu Gly Val Pro Leu Arg
355 360 365
gag cag gag ttg cgc ggg gcg ctg tgg cgc ttg ctc gag gac ccg gcc 1152
Glu Gln Glu Leu Arg Gly Ala Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
atg gcg gcg gcc tgt cgg cgt ttc atg gaa ttg tca caa ccg cac agt 1200
Met Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
atc gct tgc ggt aaa gcg gcc cag gtg gtc gaa cgt tgt cat agg gag 1248
Ile Ala Cys Gly Lys Ala Ala Gln Val Val Glu Arg Cys His Arg Glu
405 410 415
ggg gat gct cga tgg ctg aag gct gcg tcc tga 1281
Gly Asp Ala Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 86
<211> 426
<212> PRT
<213> Pseudomonas aeruginosa
<400> 86
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
Leu Cys Thr Ile Pro Val Phe Arg Asp Ala Val Glu Gln His Gly Ile
35 40 45
Ala Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
Ala Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Ser Ala Gln
85 90 95
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Ala Gln Val
115 120 125
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Lys
130 135 140
Phe Asn Val Pro Glu Gln Met Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Glu Ile
165 170 175
Asn Ala Val Arg Arg Lys Val Gly Leu Glu Thr Pro Val Lys Arg Ile
180 185 190
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Val Cys Leu Phe Pro
195 200 205
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
Thr Gly Phe Pro Leu Phe Asp Gly Ser Ile Pro Gly Thr Pro Leu Asp
225 230 235 240
Asp Glu Leu Gln Arg Phe Leu Asp Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Asn His Val Leu Gln Arg
290 295 300
Ala Tyr Ala Pro Leu Gly Ala Leu Leu Pro Ser Cys Ala Gly Leu Val
305 310 315 320
His Pro Gly Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
Arg Leu Val Arg Leu Gly Cys Gly Met Arg Leu Gly Val Pro Leu Arg
355 360 365
Glu Gln Glu Leu Arg Gly Ala Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
Met Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
Ile Ala Cys Gly Lys Ala Ala Gln Val Val Glu Arg Cys His Arg Glu
405 410 415
Gly Asp Ala Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 87
<211> 1281
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(1281)
<400> 87
atg cac gcc atc ctc atc gcc atc ggc tcg gcc ggc gac gta ttt ccc 48
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
ttc atc ggc ttg gcc cgg acc ctg aaa ttg cgc ggg cac cgc gtg agc 96
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
ctc tgc acc atc ccg gtg ttt cgc gac gcg gtg gag cag cac ggc atc 144
Leu Cys Thr Ile Pro Val Phe Arg Asp Ala Val Glu Gln His Gly Ile
35 40 45
gcg ttc gtc ccg ctg agc gac gaa ctg acc tac cgc cgg acc atg ggc 192
Ala Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
gat ccg cgc ctg tgg gac ccc aag acg tcc ttc ggc gtg ctc tgg caa 240
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
gcc atc gcc ggg atg atc gag ccg gtc tac gag tac gtc tcg gcg cag 288
Ala Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Ser Ala Gln
85 90 95
cgc cat gac gac atc gtg gtg gtc ggc tcg ctc tgg gcg ctg ggc gca 336
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
cgc atc gct cac gag aag tac ggg att ccc tac ctg tcc gcg cag gtc 384
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Ala Gln Val
115 120 125
tcg cca tcg acc ttg ttg tcg gcg cac ctg ccg ccg gta cac ccc aag 432
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Lys
130 135 140
ttc aac gtg ccc gag cag atg ccg ctg gcg atg cgc aag ctg ctc tgg 480
Phe Asn Val Pro Glu Gln Met Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
cgc tgc atc gag cgc ttc aag ctg gat cgc acc tgc gcg ccg gag atc 528
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Glu Ile
165 170 175
aac gcg gtg cgc cgc aag gtc ggc ctg gag acg ccg gtg aag cgc atc 576
Asn Ala Val Arg Arg Lys Val Gly Leu Glu Thr Pro Val Lys Arg Ile
180 185 190
ttc acc caa tgg atg cat tcg ccg cag ggc gtg gtc tgc ctg ttc ccg 624
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Val Cys Leu Phe Pro
195 200 205
gcc tgg ttc gcg ccg ccc cag cag gat tgg ccg caa ccc ctg cac atg 672
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
acc ggc ttc ccg ctg ttc gac ggc agt atc ccg ggg acc ccg ctc gac 720
Thr Gly Phe Pro Leu Phe Asp Gly Ser Ile Pro Gly Thr Pro Leu Asp
225 230 235 240
gac gaa ctg caa cgc ttt ctc gat cag ggc agc cgg ccg ctg gtg ttc 768
Asp Glu Leu Gln Arg Phe Leu Asp Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
acc cag ggc tcg acc gaa cac ctg cag ggc gac ttc tac gcc atg gcc 816
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
ctg cgc gcg ctg gaa cgc ctc ggc gcg cgt ggg atc ttc ctc acc ggc 864
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
gcc ggc cag gaa ccg ctg cgc ggc ttg ccg aat cac gtg ctg cag cgc 912
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Asn His Val Leu Gln Arg
290 295 300
gcc tac gcg cca ctg gga gcc ttg ctg cca tcg tgc gcc ggg ctg gtc 960
Ala Tyr Ala Pro Leu Gly Ala Leu Leu Pro Ser Cys Ala Gly Leu Val
305 310 315 320
cat ccg ggc ggt atc ggc gcc atg agc ctg gcc ttg gcg gcg ggg gtg 1008
His Pro Gly Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
ccg cag gtg ctg ctg ccc tgc gcc cac gac cag ttc gac aat gcc gaa 1056
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
cgg ctg gtc cgg ctc ggc tgc ggg atg cgc ctg ggc gtg ccg ttg cgc 1104
Arg Leu Val Arg Leu Gly Cys Gly Met Arg Leu Gly Val Pro Leu Arg
355 360 365
gag cag gag ttg cgc ggg gcg ctg tgg cgc ttg ctc gag gac ccg gcc 1152
Glu Gln Glu Leu Arg Gly Ala Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
atg gcg gcg gcc tgt cgg cgt ttc atg gaa ttg tca caa ccg cac agt 1200
Met Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
atc gct tgc ggt aaa gcg gcc cac gtg gtc gaa cgt tgt cat agg gag 1248
Ile Ala Cys Gly Lys Ala Ala His Val Val Glu Arg Cys His Arg Glu
405 410 415
ggg gat gcg cga tgg ctg aag gct gcg tcc tga 1281
Gly Asp Ala Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 88
<211> 426
<212> PRT
<213> Pseudomonas aeruginosa
<400> 88
Met His Ala Ile Leu Ile Ala Ile Gly Ser Ala Gly Asp Val Phe Pro
1 5 10 15
Phe Ile Gly Leu Ala Arg Thr Leu Lys Leu Arg Gly His Arg Val Ser
20 25 30
Leu Cys Thr Ile Pro Val Phe Arg Asp Ala Val Glu Gln His Gly Ile
35 40 45
Ala Phe Val Pro Leu Ser Asp Glu Leu Thr Tyr Arg Arg Thr Met Gly
50 55 60
Asp Pro Arg Leu Trp Asp Pro Lys Thr Ser Phe Gly Val Leu Trp Gln
65 70 75 80
Ala Ile Ala Gly Met Ile Glu Pro Val Tyr Glu Tyr Val Ser Ala Gln
85 90 95
Arg His Asp Asp Ile Val Val Val Gly Ser Leu Trp Ala Leu Gly Ala
100 105 110
Arg Ile Ala His Glu Lys Tyr Gly Ile Pro Tyr Leu Ser Ala Gln Val
115 120 125
Ser Pro Ser Thr Leu Leu Ser Ala His Leu Pro Pro Val His Pro Lys
130 135 140
Phe Asn Val Pro Glu Gln Met Pro Leu Ala Met Arg Lys Leu Leu Trp
145 150 155 160
Arg Cys Ile Glu Arg Phe Lys Leu Asp Arg Thr Cys Ala Pro Glu Ile
165 170 175
Asn Ala Val Arg Arg Lys Val Gly Leu Glu Thr Pro Val Lys Arg Ile
180 185 190
Phe Thr Gln Trp Met His Ser Pro Gln Gly Val Val Cys Leu Phe Pro
195 200 205
Ala Trp Phe Ala Pro Pro Gln Gln Asp Trp Pro Gln Pro Leu His Met
210 215 220
Thr Gly Phe Pro Leu Phe Asp Gly Ser Ile Pro Gly Thr Pro Leu Asp
225 230 235 240
Asp Glu Leu Gln Arg Phe Leu Asp Gln Gly Ser Arg Pro Leu Val Phe
245 250 255
Thr Gln Gly Ser Thr Glu His Leu Gln Gly Asp Phe Tyr Ala Met Ala
260 265 270
Leu Arg Ala Leu Glu Arg Leu Gly Ala Arg Gly Ile Phe Leu Thr Gly
275 280 285
Ala Gly Gln Glu Pro Leu Arg Gly Leu Pro Asn His Val Leu Gln Arg
290 295 300
Ala Tyr Ala Pro Leu Gly Ala Leu Leu Pro Ser Cys Ala Gly Leu Val
305 310 315 320
His Pro Gly Gly Ile Gly Ala Met Ser Leu Ala Leu Ala Ala Gly Val
325 330 335
Pro Gln Val Leu Leu Pro Cys Ala His Asp Gln Phe Asp Asn Ala Glu
340 345 350
Arg Leu Val Arg Leu Gly Cys Gly Met Arg Leu Gly Val Pro Leu Arg
355 360 365
Glu Gln Glu Leu Arg Gly Ala Leu Trp Arg Leu Leu Glu Asp Pro Ala
370 375 380
Met Ala Ala Ala Cys Arg Arg Phe Met Glu Leu Ser Gln Pro His Ser
385 390 395 400
Ile Ala Cys Gly Lys Ala Ala His Val Val Glu Arg Cys His Arg Glu
405 410 415
Gly Asp Ala Arg Trp Leu Lys Ala Ala Ser
420 425
<210> 89
<211> 978
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(978)
<400> 89
atg gac cgg ata gac atg ggc gtg ctg gtg gtg ctg ttc aat cct ggc 48
Met Asp Arg Ile Asp Met Gly Val Leu Val Val Leu Phe Asn Pro Gly
1 5 10 15
gac gac gac ctg gaa cac ctt ggc gaa ctg gcg gcg gcc ttt ccg caa 96
Asp Asp Asp Leu Glu His Leu Gly Glu Leu Ala Ala Ala Phe Pro Gln
20 25 30
ctg cgc ttc ctc gcc gtc gac aac tcg ccg cac agc gat ccg cag cgc 144
Leu Arg Phe Leu Ala Val Asp Asn Ser Pro His Ser Asp Pro Gln Arg
35 40 45
aac gcc cgg ctg cgc ggg caa ggc atc gcc gtg ctc tac cac ggc aac 192
Asn Ala Arg Leu Arg Gly Gln Gly Ile Ala Val Leu Tyr His Gly Asn
50 55 60
cgg cag ggc atc gcc ggc gcc ttc aac cag ggg ctc gac acg ctg ttc 240
Arg Gln Gly Ile Ala Gly Ala Phe Asn Gln Gly Leu Asp Thr Leu Phe
65 70 75 80
cgg cgc ggc ctg cag ggt gtg ctg ctg ctc gac cag gac tcc cgt ccc 288
Arg Arg Gly Leu Gln Gly Val Leu Leu Leu Asp Gln Asp Ser Arg Pro
85 90 95
ggc ggc gcc ttc ctc gcc gcc cag tgg cgc aac ctg cag gca tgc aac 336
Gly Gly Ala Phe Leu Ala Ala Gln Trp Arg Asn Leu Gln Ala Cys Asn
100 105 110
ggc cag gcc tgc ctg ctc ggc cca cgg atc ttc gac cgg ggc gac cgg 384
Gly Gln Ala Cys Leu Leu Gly Pro Arg Ile Phe Asp Arg Gly Asp Arg
115 120 125
cgc ttc ctg ccg gcc atc cac ctc gac ggg ctg gcg ctc agg caa ctg 432
Arg Phe Leu Pro Ala Ile His Leu Asp Gly Leu Ala Leu Arg Gln Leu
130 135 140
tcc ctg gac ggc ctg acg acc cca cag cgc acc tcg ttc ctg atc tcc 480
Ser Leu Asp Gly Leu Thr Thr Pro Gln Arg Thr Ser Phe Leu Ile Ser
145 150 155 160
tcc ggc tgc ctg ctg acc cgc gag gcc tac cag cgc ctc ggc cac ttc 528
Ser Gly Cys Leu Leu Thr Arg Glu Ala Tyr Gln Arg Leu Gly His Phe
165 170 175
gac gag gaa ctg ttc atc gac cac gtg gac acc gag tac agc ctg cgc 576
Asp Glu Glu Leu Phe Ile Asp His Val Asp Thr Glu Tyr Ser Leu Arg
180 185 190
gcc cag gcg ctg gac gtg ccc ctg tac gtc gac ccg cgg ctg gtc ctc 624
Ala Gln Ala Leu Asp Val Pro Leu Tyr Val Asp Pro Arg Leu Val Leu
195 200 205
gag cac cgc atc ggc acg cgc aag acc cgc cgc ctc ggc ggt ctc agc 672
Glu His Arg Ile Gly Thr Arg Lys Thr Arg Arg Leu Gly Gly Leu Ser
210 215 220
ctc agc gcg atg aac cac gcc cca ctg cgc cgc tac tac ctg gcg cgc 720
Leu Ser Ala Met Asn His Ala Pro Leu Arg Arg Tyr Tyr Leu Ala Arg
225 230 235 240
aac ggc ctg ctg gtc ctg cgc cgc tac gcc cgg tcc tcg ccg ctg gcc 768
Asn Gly Leu Leu Val Leu Arg Arg Tyr Ala Arg Ser Ser Pro Leu Ala
245 250 255
ctg ctg gcg aac ctg ccg acc ctg acc cag ggc ctc gcg gtg ctc ctg 816
Leu Leu Ala Asn Leu Pro Thr Leu Thr Gln Gly Leu Ala Val Leu Leu
260 265 270
ctc gaa cgc gac aag ctg ctc aag ctg cgc tgc ctg ggc tgg ggc ctg 864
Leu Glu Arg Asp Lys Leu Leu Lys Leu Arg Cys Leu Gly Trp Gly Leu
275 280 285
tgg gac ggc ctg cgg ggg cgc ggc ggc gcg ctg gag cgc aac cgc ccg 912
Trp Asp Gly Leu Arg Gly Arg Gly Gly Ala Leu Glu Arg Asn Arg Pro
290 295 300
cgc ctg ctg aag cgc ctc gcc ggt ccg gcg gtg gcg ccc aca gtt ccc 960
Arg Leu Leu Lys Arg Leu Ala Gly Pro Ala Val Ala Pro Thr Val Pro
305 310 315 320
ggc aag gcc aag gcc tag 978
Gly Lys Ala Lys Ala
325
<210> 90
<211> 325
<212> PRT
<213> Pseudomonas aeruginosa
<400> 90
Met Asp Arg Ile Asp Met Gly Val Leu Val Val Leu Phe Asn Pro Gly
1 5 10 15
Asp Asp Asp Leu Glu His Leu Gly Glu Leu Ala Ala Ala Phe Pro Gln
20 25 30
Leu Arg Phe Leu Ala Val Asp Asn Ser Pro His Ser Asp Pro Gln Arg
35 40 45
Asn Ala Arg Leu Arg Gly Gln Gly Ile Ala Val Leu Tyr His Gly Asn
50 55 60
Arg Gln Gly Ile Ala Gly Ala Phe Asn Gln Gly Leu Asp Thr Leu Phe
65 70 75 80
Arg Arg Gly Leu Gln Gly Val Leu Leu Leu Asp Gln Asp Ser Arg Pro
85 90 95
Gly Gly Ala Phe Leu Ala Ala Gln Trp Arg Asn Leu Gln Ala Cys Asn
100 105 110
Gly Gln Ala Cys Leu Leu Gly Pro Arg Ile Phe Asp Arg Gly Asp Arg
115 120 125
Arg Phe Leu Pro Ala Ile His Leu Asp Gly Leu Ala Leu Arg Gln Leu
130 135 140
Ser Leu Asp Gly Leu Thr Thr Pro Gln Arg Thr Ser Phe Leu Ile Ser
145 150 155 160
Ser Gly Cys Leu Leu Thr Arg Glu Ala Tyr Gln Arg Leu Gly His Phe
165 170 175
Asp Glu Glu Leu Phe Ile Asp His Val Asp Thr Glu Tyr Ser Leu Arg
180 185 190
Ala Gln Ala Leu Asp Val Pro Leu Tyr Val Asp Pro Arg Leu Val Leu
195 200 205
Glu His Arg Ile Gly Thr Arg Lys Thr Arg Arg Leu Gly Gly Leu Ser
210 215 220
Leu Ser Ala Met Asn His Ala Pro Leu Arg Arg Tyr Tyr Leu Ala Arg
225 230 235 240
Asn Gly Leu Leu Val Leu Arg Arg Tyr Ala Arg Ser Ser Pro Leu Ala
245 250 255
Leu Leu Ala Asn Leu Pro Thr Leu Thr Gln Gly Leu Ala Val Leu Leu
260 265 270
Leu Glu Arg Asp Lys Leu Leu Lys Leu Arg Cys Leu Gly Trp Gly Leu
275 280 285
Trp Asp Gly Leu Arg Gly Arg Gly Gly Ala Leu Glu Arg Asn Arg Pro
290 295 300
Arg Leu Leu Lys Arg Leu Ala Gly Pro Ala Val Ala Pro Thr Val Pro
305 310 315 320
Gly Lys Ala Lys Ala
325
<210> 91
<211> 978
<212> DNA
<213> Pseudomonas aeruginosa
<220>
<221> CDS
<222> (1)..(978)
<400> 91
atg gac cgg ata gac atg ggc gtg ctg gtg gta ctg ttc aat cct ggc 48
Met Asp Arg Ile Asp Met Gly Val Leu Val Val Leu Phe Asn Pro Gly
1 5 10 15
gac gac gac ctg gaa cac ctt ggc gaa ctg gcg gcg gcg ttt ccg caa 96
Asp Asp Asp Leu Glu His Leu Gly Glu Leu Ala Ala Ala Phe Pro Gln
20 25 30
ctg cgc ttc ctc gcc gtc gac aac tca ccg cac agc gat ccg cag cgc 144
Leu Arg Phe Leu Ala Val Asp Asn Ser Pro His Ser Asp Pro Gln Arg
35 40 45
aat gcc cgg ctg cgc ggg caa ggc atc gcc gtg ctg cac cac ggc aac 192
Asn Ala Arg Leu Arg Gly Gln Gly Ile Ala Val Leu His His Gly Asn
50 55 60
cgg cag ggc atc gcc ggc gcc ttc aac cag ggg ctc gac gcg ctg ttc 240
Arg Gln Gly Ile Ala Gly Ala Phe Asn Gln Gly Leu Asp Ala Leu Phe
65 70 75 80
cgg cgt ggc gtg cag ggt gtg ctg ctg ctc gac cag gac tcc cgt ccc 288
Arg Arg Gly Val Gln Gly Val Leu Leu Leu Asp Gln Asp Ser Arg Pro
85 90 95
ggc ggc gcc ttc ctc gcc gcc cag tgg cgc aac ctg cag gcg cgc aac 336
Gly Gly Ala Phe Leu Ala Ala Gln Trp Arg Asn Leu Gln Ala Arg Asn
100 105 110
ggt cag gcc tgc ctg ctc ggc cca cgg atc ttc gac cgg ggt gac cgg 384
Gly Gln Ala Cys Leu Leu Gly Pro Arg Ile Phe Asp Arg Gly Asp Arg
115 120 125
cgc ttc ctg ccg gcc atc cat ctc gac gga ctg acg ctc agg caa ttg 432
Arg Phe Leu Pro Ala Ile His Leu Asp Gly Leu Thr Leu Arg Gln Leu
130 135 140
tct ctg gac ggc ctg acg acc ccg cag cgc acc tcg ttc ctg atc tcc 480
Ser Leu Asp Gly Leu Thr Thr Pro Gln Arg Thr Ser Phe Leu Ile Ser
145 150 155 160
tcc ggc tgc ctg ctg acc cgc gag gcc tac cag cgc ctc ggc cac ttc 528
Ser Gly Cys Leu Leu Thr Arg Glu Ala Tyr Gln Arg Leu Gly His Phe
165 170 175
gac gag gaa ctg ttc atc gac cac gtg gac acc gaa tac agc ctg cgc 576
Asp Glu Glu Leu Phe Ile Asp His Val Asp Thr Glu Tyr Ser Leu Arg
180 185 190
gcc cag gcg ctg gac gtg ccc ctg tac gtc gac ccg cgg ctg gtc ctc 624
Ala Gln Ala Leu Asp Val Pro Leu Tyr Val Asp Pro Arg Leu Val Leu
195 200 205
gag cac cgc atc ggc acg cgc aag acc cgc cgc ctc ggc ggt ctc agc 672
Glu His Arg Ile Gly Thr Arg Lys Thr Arg Arg Leu Gly Gly Leu Ser
210 215 220
ctc agc gcg atg aac cac gcc ccg ctg cgc cgc tac tac ctg gcg cgc 720
Leu Ser Ala Met Asn His Ala Pro Leu Arg Arg Tyr Tyr Leu Ala Arg
225 230 235 240
aac ggc ctg ctg gtc ctg cgc cgc tac gcc cgg tcc tcg ccg ctg gcc 768
Asn Gly Leu Leu Val Leu Arg Arg Tyr Ala Arg Ser Ser Pro Leu Ala
245 250 255
ctg ctg gcg aac ctg ccg acc ctg acc cag ggc ctc gcg gtg ctc ctg 816
Leu Leu Ala Asn Leu Pro Thr Leu Thr Gln Gly Leu Ala Val Leu Leu
260 265 270
ctc gaa cgc gac aag ctg ctc aag ctg cgc tgc ctg ggc tgg ggc ctg 864
Leu Glu Arg Asp Lys Leu Leu Lys Leu Arg Cys Leu Gly Trp Gly Leu
275 280 285
tgg gac ggc ctg cgg gga cgc ggc ggc gcg ctg gag cgc aac cgc ccg 912
Trp Asp Gly Leu Arg Gly Arg Gly Gly Ala Leu Glu Arg Asn Arg Pro
290 295 300
cgc ctg ctg aag cgc ctc gcc ggc ccg gcc gtg gcg tcc gta gct tcc 960
Arg Leu Leu Lys Arg Leu Ala Gly Pro Ala Val Ala Ser Val Ala Ser
305 310 315 320
ggc aag gcc aag gcc tag 978
Gly Lys Ala Lys Ala
325
<210> 92
<211> 325
<212> PRT
<213> Pseudomonas aeruginosa
<400> 92
Met Asp Arg Ile Asp Met Gly Val Leu Val Val Leu Phe Asn Pro Gly
1 5 10 15
Asp Asp Asp Leu Glu His Leu Gly Glu Leu Ala Ala Ala Phe Pro Gln
20 25 30
Leu Arg Phe Leu Ala Val Asp Asn Ser Pro His Ser Asp Pro Gln Arg
35 40 45
Asn Ala Arg Leu Arg Gly Gln Gly Ile Ala Val Leu His His Gly Asn
50 55 60
Arg Gln Gly Ile Ala Gly Ala Phe Asn Gln Gly Leu Asp Ala Leu Phe
65 70 75 80
Arg Arg Gly Val Gln Gly Val Leu Leu Leu Asp Gln Asp Ser Arg Pro
85 90 95
Gly Gly Ala Phe Leu Ala Ala Gln Trp Arg Asn Leu Gln Ala Arg Asn
100 105 110
Gly Gln Ala Cys Leu Leu Gly Pro Arg Ile Phe Asp Arg Gly Asp Arg
115 120 125
Arg Phe Leu Pro Ala Ile His Leu Asp Gly Leu Thr Leu Arg Gln Leu
130 135 140
Ser Leu Asp Gly Leu Thr Thr Pro Gln Arg Thr Ser Phe Leu Ile Ser
145 150 155 160
Ser Gly Cys Leu Leu Thr Arg Glu Ala Tyr Gln Arg Leu Gly His Phe
165 170 175
Asp Glu Glu Leu Phe Ile Asp His Val Asp Thr Glu Tyr Ser Leu Arg
180 185 190
Ala Gln Ala Leu Asp Val Pro Leu Tyr Val Asp Pro Arg Leu Val Leu
195 200 205
Glu His Arg Ile Gly Thr Arg Lys Thr Arg Arg Leu Gly Gly Leu Ser
210 215 220
Leu Ser Ala Met Asn His Ala Pro Leu Arg Arg Tyr Tyr Leu Ala Arg
225 230 235 240
Asn Gly Leu Leu Val Leu Arg Arg Tyr Ala Arg Ser Ser Pro Leu Ala
245 250 255
Leu Leu Ala Asn Leu Pro Thr Leu Thr Gln Gly Leu Ala Val Leu Leu
260 265 270
Leu Glu Arg Asp Lys Leu Leu Lys Leu Arg Cys Leu Gly Trp Gly Leu
275 280 285
Trp Asp Gly Leu Arg Gly Arg Gly Gly Ala Leu Glu Arg Asn Arg Pro
290 295 300
Arg Leu Leu Lys Arg Leu Ala Gly Pro Ala Val Ala Ser Val Ala Ser
305 310 315 320
Gly Lys Ala Lys Ala
325

Claims (14)

1.细胞,其能够形成至少一种通式(I)的鼠李糖脂,
其中
m=2、1或0,特别为1或0,
n=1或0,特别为1,
R1和R2=彼此独立地为具有2-24个的相同或不同的有机基团,特别是任选地分支的,任选地取代的、特别是羟基取代的,任选地不饱和的,特别是任选地单-、双-或三-不饱和的烃基,
其特征在于所述细胞已被遗传学修饰,从而使得与其野生型相比,所述细胞中酶E1、E2和E3中的至少一种的活性是增加的,其中酶E1能够催化3-羟基链烷酰-ACP经由3-羟基链烷酰-3-羟基链烷酸-ACP转化为羟基链烷酰-3-羟基链烷酸;酶E2是鼠李糖基转移酶I,并能够催化dTDP-鼠李糖和3-羟基链烷酰-3-羟基链烷酸酯转化为α-L-吡喃鼠李糖基-3-羟基链烷酰-3-羟基链烷酸酯;而酶E3是鼠李糖基转移酶II,并能够催化dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基链烷酰-3-羟基链烷酸酯转化为α-L-吡喃鼠李糖基-(1-2)-α-L-吡喃鼠李糖基-3-羟基链烷酰-3-羟基链烷酸酯,
与其野生型相比,所述细胞具有增加的至少酶E8的活性,所述酶E8催化通式(I)的鼠李糖脂从细胞输送至周围介质中,其优选具有多肽序列Seq ID No.8、Seq ID No.24、Seq IDNo.26或Seq ID No.28,或者其具有的多肽序列与相应的参照序列Seq ID No.8、Seq IDNo.24、Seq ID No.26或Seq ID No.28相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有相应的参照序列Seq ID No.8、Seq IDNo.24、Seq ID No.26或Seq ID No.28的酶的酶活性。
2.权利要求1的细胞,其特征在于酶E1、E2和E3选自以下一组:
至少一种酶E1,其选自
酶E1a,其具有多肽序列Seq ID No.2,或者其具有的多肽序列与参照序列Seq ID No.2相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.2的酶的酶活性,
酶E1b,其具有多肽序列Seq ID No.18,或者其具有的多肽序列与参照序列Seq IDNo.18相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.18的酶的酶活性,
酶E1c,其具有多肽序列Seq ID No.78,或者其具有的多肽序列与参照序列Seq IDNo.78相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.78的酶的酶活性,
酶E1d,其具有多肽序列Seq ID No.80,或者其具有的多肽序列与参照序列Seq IDNo.80相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.80的酶的酶活性,以及
酶E1e,其具有多肽序列Seq ID No.82,或者其具有的多肽序列与参照序列Seq IDNo.82相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.82的酶的酶活性,
至少一种酶E2,其选自
酶E2a,其具有多肽序列Seq ID No.4,或者其具有的多肽序列与参照序列Seq ID No.4相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.4的酶的酶活性,
酶E2b,其具有多肽序列Seq ID No.20,或者其具有的多肽序列与参照序列Seq IDNo.20相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.20的酶的酶活性,
酶E2c,其具有多肽序列Seq ID No.84,或者其具有的多肽序列与参照序列Seq IDNo.84相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.84的酶的酶活性,
酶E2d,其具有多肽序列Seq ID No.86,或者其具有的多肽序列与参照序列Seq IDNo.86相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.86的酶的酶活性,以及
酶E2e,其具有多肽序列Seq ID No.88,或者其具有的多肽序列与参照序列Seq IDNo.88相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.88的酶的酶活性,以及
至少一种酶E3,其选自
酶E3a,其具有多肽序列Seq ID No.6,或者其具有的多肽序列与参照序列Seq ID No.6相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.6的酶的酶活性,
酶E3b,其具有多肽序列Seq ID No.22,或者其具有的多肽序列与参照序列Seq IDNo.22相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.22的酶的酶活性,
酶E3c,其具有多肽序列Seq ID No.90,或者其具有的多肽序列与参照序列Seq IDNo.90相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.90的酶的酶活性,以及
酶E3d,其具有多肽序列Seq ID No.92,或者其具有的多肽序列与参照序列Seq IDNo.92相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.92的酶的酶活性。
3.权利要求1或2的细胞,其特征在于它具有增加的酶组合的活性,所述酶组合选自E2、E2E3和E1E2E3
4.前述权利要求中任一项的细胞,其特征在于它具有增加的酶组合E1E2E3的活性,并且n为1。
5.前述权利要求中任一项的细胞,其特征在于它选自以下菌属:曲霉属(Aspergillus)、棒状杆菌属(Corynebacterium)、短杆菌属(Brevibacterium)、芽孢杆菌属(Bacillus)、不动杆菌属(Acinetobacter)、产碱杆菌属(Alcaligenes)、乳杆菌属(Lactobacillus)、副球菌属(Paracoccus)、乳球菌属(Lactococcus)、念珠菌属(Candida)、毕赤酵母属(Pichia)、汉逊酵母属(Hansenula)、克鲁维酵母菌属(Kluyveromyces)、酵母菌属(Saccharomyces)、埃希氏杆菌属(Escherichia)、单胞发酵菌属(Zymomonas)、耶罗威亚酵母属(Yarrowia)、甲基杆菌属(Methylobacterium)、青枯菌属(Ralstonia)、假单胞菌属(Pseudomonas)、红螺菌属(Rhodospirillum)、红杆菌属(Rhodobacter)、伯克氏菌属(Burkholderia)、梭菌属(Clostridium)和贪铜菌属(Cupriavidus)。
6.前述权利要求中任一项的细胞,其特征在于它能够如同野生型那样形成具有C6至C16链长度的聚羟基链烷酸酯,特别在于它已经过遗传学修饰,从而使得与其野生型相比,它能够形成较少的聚羟基链烷酸酯。
7.权利要求6的细胞,其特征在于所述细胞与其野生型相比,具有降低的至少一种酶E9或E10的活性,
其中E9是聚羟基链烷酸酯合成酶,EC:2.3.1.-,其具有将3-羟基链烷酰-辅酶A转化为聚-3-羟基链烷酸的能力,特别是具有多肽序列Seq ID No.30或Seq ID No.32,或者其具有的多肽序列与相应的参照序列Seq ID No.30或Seq ID No.32相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且其仍具有至少10%的具有相应的参照序列Seq ID No.30或Seq ID No.32的酶的酶活性,以及
E10是3-羟基链烷酰-ACP:辅酶A转移酶,其具有将3-羟基链烷酰-ACP转化为3-羟基链烷酰-辅酶A的能力,特别具有多肽序列Seq ID No.34或Seq ID No.36,或者其具有的多肽序列与相应的参照序列Seq ID No.34或Seq ID No.36相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且其仍具有至少10%的具有相应的参照序列Seq IDNo.34或Seq ID No.36的酶的酶活性。
8.权利要求5的细胞,其特征在于所述细胞选自恶臭假单胞菌(P.putida)GPp121、恶臭假单胞菌GPp122、恶臭假单胞菌GPp123、恶臭假单胞菌GPp124和恶臭假单胞菌GPp104、恶臭假单胞菌KT42C1、恶臭假单胞菌KTOY01或恶臭假单胞菌KTOY02。
9.前述权利要求中任一项的细胞,其特征在于:与其野生型相比,所述细胞中选自以下一组中的至少一种酶具有增加的活性:
至少一种酶E4,即dTTP:α-D-葡萄糖-1-磷酸胸苷转移酶,EC 2.7.7.24,特别是所述酶具有多肽序列Seq ID No.10,或者其具有的多肽序列与参照序列Seq ID No.10相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.10的酶的酶活性,
至少一种酶E5,即dTTP-葡萄糖-4,6-水解酶,EC 4.2.1.46,特别是所述酶具有多肽序列Seq ID No.12,或者其具有的多肽序列与参照序列Seq ID No.12相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列SeqID No.12的酶的酶活性,
至少一种酶E6,即dTDP-4-脱氢鼠李糖-3,5-差向异构酶,EC 5.1.3.13,特别是所述酶具有多肽序列Seq ID No.14,或者其具有的多肽序列与参照序列Seq ID No.14相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.14的酶的酶活性,
至少一种酶E7,即dTDP-4-脱氢鼠李糖-还原酶,EC 1.1.1.133,特别是所述酶具有多肽序列Seq ID No.16,或者其具有的多肽序列与参照序列Seq ID No.16相比其中有高达25%的氨基酸基通过缺失、插入、取代或其组合而被修饰,并且仍具有至少10%的具有参照序列Seq ID No.16的酶的酶活性。
10.分离的核酸,其在每种情况下都含有选自[A1-G1]、[A2-G2]和[A3-G3]三组中的至少一个序列,其中
组[A1-G1]由以下序列组成:
A1a)Seq ID No.1的序列,其中该序列编码蛋白,所述蛋白能够将3-羟基癸酰基-ACP经由3-羟基癸酰基-3-羟基癸酰基-ACP转化为3-羟基癸酰基-3-羟基癸酸,
B1a)无内含子的序列,其来源于A1a)的序列,并与Seq ID No.1的序列编码相同的蛋白或肽,
C1a)编码包含Seq ID No.2的氨基酸序列的蛋白或肽的序列,
D1a)与组A1a)-C1a)之一的序列有至少70%相同性的序列,
E1a)与组A1a)-D1a)之一的序列的互补链杂交的序列,或者考虑到遗传密码子的简并性,会与所述互补链杂交的序列,
F1a)通过组A1a)-E1a)之一的序列的至少一个碱基的取代、添加、倒位和/或缺失获得的衍生物,
G1a)组A1a)-F1a)之一的序列的互补序列,
A1b)Seq ID No.17的序列,其中该序列编码蛋白,所述蛋白能够将3-羟基十四烷酰基-ACP经由3-羟基十四烷酰基-3-羟基十四烷酰基-ACP转化为3-羟基十四烷酰基-3-羟基十四烷酸,
B1b)无内含子的序列,其来源于A1b)的序列,并与Seq ID No.17的序列编码相同的蛋白或肽,
C1b)编码包含Seq ID No.18的氨基酸序列的蛋白或肽的序列,
D1b)与组A1b)-C1b)之一的序列有至少70%相同性的序列,
E1b)与组A1b)-D1b)之一的序列的互补链杂交的序列,或者考虑到遗传密码子的简并性,会与所述互补链杂交的序列,
F1b)通过组A1b)-E1b)之一的序列的至少一个碱基的取代、添加、倒位和/或缺失获得的衍生物,和
G1b)组A1b)-F1b)之一的序列的互补序列,以及
组[A2-G2]由以下序列组成:
A2a)Seq ID No.3的序列,其中该序列编码蛋白,所述蛋白能够将dTDP-鼠李糖和3-羟基癸酰基-3-羟基癸酸转化为α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸,
B2a)无内含子的序列,其来源于A2a)的序列,并与Seq ID No.3的序列编码相同的蛋白或肽,
C2a)编码包含Seq ID No.4的氨基酸序列的蛋白或肽的序列,
D2a)与组A2a)-C2a)之一的序列有至少80%相同性的序列,
E2a)与组A2a)-D2a)之一的序列的互补链杂交的序列,或者考虑到遗传密码子的简并性,会与所述互补链杂交的序列,
F2a)通过组A2a)-E2a)之一的序列的至少一个碱基的取代、添加、倒位和/或缺失获得的衍生物,
G2a)组A2a)-F2a)之一的序列的互补序列,
A2b)Seq ID No.19的序列,其中该序列编码蛋白,所述蛋白能够将dTDP-鼠李糖和3-羟基十四烷酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸,
B2b)无内含子的序列,其来源于A2b)的序列,并与Seq ID No.19的序列编码相同的蛋白或肽,
C2b)编码包含Seq ID No.20的氨基酸序列的蛋白或肽的序列,
D2b)与组A2b)-C2b)之一的序列有至少70%相同性的序列,
E2b)与组A2b)-D2b)之一的序列的互补链杂交的序列,或者考虑到遗传密码子的简并性,会与所述互补链杂交的序列,
F2b)通过组A2b)-E2b)之一的序列的至少一个碱基的取代、添加、倒位和/或缺失获得的衍生物,和
G2b)组A2b)-F2b)之一的序列的互补序列,以及
组[A3-G3]由以下序列组成:
A3a)Seq ID No.5的序列,其中该序列编码蛋白,所述蛋白能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸转化为α-L-吡喃鼠李糖基-(1–2)-α-L-吡喃鼠李糖基-3-羟基癸酰基-3-羟基癸酸,
B3a)无内含子的序列,其来源于A3a)的序列,并与Seq ID No.5的序列编码相同的蛋白或肽,
C3a)编码包含Seq ID No.6的氨基酸序列的蛋白或肽的序列,
D3a)与组A3a)-C3a)之一的序列有至少80%相同性的序列,
E3a)与组A3a)-D3a)之一的序列的互补链杂交的序列,或者考虑到遗传密码子的简并性,会与所述互补链杂交的序列,
F3a)通过组A3a)-E3a)之一的序列的至少一个碱基的取代、添加、倒位和/或缺失获得的衍生物,
G3a)组A3a)-F3a)之一的序列的互补序列,
A3b)Seq ID No.21的序列,其中该序列编码蛋白,所述蛋白能够将dTDP-鼠李糖和α-L-吡喃鼠李糖基-3-羟基十四烷酰基-3-羟基十四烷酸转化为α-L-吡喃鼠李糖基-(1-2)-α-L-吡喃鼠李糖基-3-羟基十四烷酰-3-羟基十四烷酸,
B3b)无内含子的序列,其来源于A3b)的序列,并与Seq ID No.21的序列编码相同的蛋白或肽,
C3b)编码包含Seq ID No.22的氨基酸序列的蛋白或肽的序列,
D3b)与组A3b)-C3b)之一的序列有至少60%相同性的序列,
E3b)与组A3b)-D3b)之一的序列的互补链杂交的序列,或者考虑到遗传密码子的简并性,会与所述互补链杂交的序列,
F3b)通过组A3b)-E3b)之一的序列的至少一个碱基的取代、添加、倒位和/或缺失获得的衍生物,和
G3b)组A3b)-F3b)之一的序列的互补序列。
11.载体,特别是表达载体或基因过表达盒,其包含选自Seq ID No.38、Seq ID No.40、Seq ID No.42、Seq ID No.45、Seq ID No.47和权利要求10所述的核酸中的至少一种核酸序列。
12.权利要求1-9中任一项的细胞,其特征在于它含有权利要求10所述的至少一种核酸或权利要求11所述的至少一种载体。
13.本发明的另外主题是用于生产通式(I)的鼠李糖脂的方法,所述方法包括以下处理步骤:
I)使权利要求1-9和12中任一项所述的细胞与含碳源的介质接触,
II)在可以使细胞从所述碳源形成鼠李糖脂的条件下培养细胞,以及
III)任选地分离形成的鼠李糖脂。
14.利用权利要求13所述的方法获得的鼠李糖脂用于生产化妆品、皮肤制剂或药物制剂、用于生产植物保护制剂以及用于生产护理和清洁剂以及表面活性剂浓缩物的用途。
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Families Citing this family (55)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102006025821A1 (de) * 2006-06-02 2007-12-06 Degussa Gmbh Ein Enzym zur Herstellung von Mehylmalonatsemialdehyd oder Malonatsemialdehyd
DE102007052463A1 (de) * 2007-11-02 2009-05-07 Evonik Degussa Gmbh Fermentative Gewinnung von Aceton aus erneuerbaren Rohstoffen mittels neuen Stoffwechselweges
DE102007060705A1 (de) * 2007-12-17 2009-06-18 Evonik Degussa Gmbh ω-Aminocarbonsäuren oder ihre Lactame, herstellende, rekombinante Zellen
DE102008002715A1 (de) * 2008-06-27 2009-12-31 Evonik Röhm Gmbh 2-Hydroxyisobuttersäure produzierende rekombinante Zelle
DE102009009580A1 (de) 2009-02-19 2010-08-26 Evonik Degussa Gmbh Verfahren zur Herstellung freier Säuren aus ihren Salzen
DE102009046626A1 (de) 2009-11-11 2011-05-12 Evonik Degussa Gmbh Candida tropicalis Zellen und deren Verwendung
DE102009046623A1 (de) 2009-11-11 2011-05-12 Evonik Röhm Gmbh Verwendung eines zu einem MeaB-Protein homologen Proteins zur Erhöhung der enzymatischen Aktivität einer 3-Hydroxycarbonsäure-CoA-Mutase
DE102010014680A1 (de) 2009-11-18 2011-08-18 Evonik Degussa GmbH, 45128 Zellen, Nukleinsäuren, Enzyme und deren Verwendung sowie Verfahren zur Herstellung von Sophorolipiden
DE102010015807A1 (de) 2010-04-20 2011-10-20 Evonik Degussa Gmbh Biokatalytisches Oxidationsverfahren mit alkL-Genprodukt
DE102011004465A1 (de) 2010-09-10 2012-03-15 Evonik Degussa Gmbh Verfahren zur direkten Aminierung sekundärer Alkohole mit Ammoniak zu primären Aminen
DE102010043470A1 (de) 2010-11-05 2012-05-10 Evonik Degussa Gmbh Zusammensetzung aus Polyamiden mit niedriger Konzentration an Carbonsäureamidgruppen und elektrisch leitfähigem Kohlenstoff
DE102011075162A1 (de) 2010-12-08 2012-06-14 Evonik Degussa Gmbh Verfahren zur homogen-katalysierte, hochselektiven direkten Aminierung von primären Alkoholen mit Ammoniak zu primären Aminen bei hohem Volumenverhältnis von Flüssig- zu Gasphase und/oder hohen Drücken
CN103370302B (zh) 2011-02-21 2015-06-17 赢创德固赛有限公司 借助Xantphos催化剂体系用氨将醇直接胺化成伯胺的方法
RU2014106109A (ru) 2011-07-20 2015-08-27 Эвоник Дегусса Гмбх Окисление и аминирование первичных спиртов
EP2573172A1 (en) 2011-09-21 2013-03-27 Heinrich-Heine-Universität Düsseldorf Means and methods for rhamnolipid production
DE102011084518A1 (de) 2011-10-14 2013-04-18 Evonik Industries Ag Verwendung einer Mehrschichtfolie mit Polyamid- und Polyesterschichten fürdie Herstellung photovoltaischer Module
EP2602328A1 (de) 2011-12-05 2013-06-12 Evonik Industries AG Verfahren zur Oxidation von Alkanen unter Verwendung einer AlkB Alkan 1-Monooxygenase
EP2607479A1 (en) 2011-12-22 2013-06-26 Evonik Industries AG Biotechnological production of alcohols and derivatives thereof
EP2631298A1 (en) 2012-02-22 2013-08-28 Evonik Industries AG Biotechnological method for producing butanol and butyric acid
EP2639308A1 (de) 2012-03-12 2013-09-18 Evonik Industries AG Enzymatische omega-Oxidation und -Aminierung von Fettsäuren
EP2647696A1 (de) 2012-04-02 2013-10-09 Evonik Degussa GmbH Verfahren zur aeroben Herstellung von Alanin oder einer unter Verbrauch von Alanin entstehenden Verbindung
WO2014039940A1 (en) * 2012-09-10 2014-03-13 Logos Technologies Llc Cells and methods for the production of rhamnolipids
DE102012221519A1 (de) 2012-11-26 2014-05-28 Evonik Industries Ag Verfahren zur Isolierung von Rhamnolipiden
EP2746400A1 (de) 2012-12-21 2014-06-25 Evonik Industries AG Herstellung von Aminen und Diaminen aus einer Carbonsäure oder Dicarbonsäure oder eines Monoesters davon
DE102013205755A1 (de) 2013-04-02 2014-10-02 Evonik Industries Ag Waschmittelformulierung für Textilien enthaltend Rhamnolipide mit einem überwiegenden Gehalt an di-Rhamnolipiden
DE102013205756A1 (de) 2013-04-02 2014-10-02 Evonik Industries Ag Mischungszusammensetzung enthaltend Rhamnolipide
WO2014197457A1 (en) * 2013-06-06 2014-12-11 The United States Of America, As Represented By The Secretary Of Agriculture Production of dirhamnose-lipid in recombinant nonpathogenic bacterium pseudomonas chlororaphis
EP2949214A1 (en) 2014-05-26 2015-12-02 Evonik Degussa GmbH Methods of producing rhamnolipids
EP3023431B1 (de) 2014-11-19 2017-01-04 Evonik Degussa GmbH Konzentrierte, niedrigviskose Rhamnolipid-Zusammensetzungen
US9884883B2 (en) * 2015-01-12 2018-02-06 Logos Technologies, Llc Production of rhamnolipid compositions
EP3259363A1 (en) 2015-02-19 2017-12-27 Evonik Degussa GmbH Rhamnolipid synthesis
CA2937594A1 (en) 2015-02-26 2016-08-26 Evonik Degussa Gmbh Alkene production
EP3061442A1 (de) 2015-02-27 2016-08-31 Evonik Degussa GmbH Zusammensetzung enthaltend Rhamnolipid und Siloxan
DK3292210T3 (da) * 2015-05-05 2020-05-25 Stepan Co Semikontinuerlig fremgangsmåde til fremstilling af rhamnolipider med højt udbytte og høj titer
BR112018000257A2 (pt) 2015-07-08 2018-09-04 Rheinisch Westfaelische Technische Hochschule Aachen Rwth célula hospedeira, método para produzir um haa, uso de uma célula hospedeira, método para produzir uma célula hospedeira, célula, preparação de haa, método para produzir uma composição de ácido graxo e composição de ácido graxo
EP3419985A1 (de) 2016-02-22 2019-01-02 Evonik Degussa GmbH Rhamnolipidester als nichtionische tenside zur kosmetischen anwendung
US10941173B2 (en) 2016-02-22 2021-03-09 Evonik Operations Gmbh Rhamnolipid amides for hair scent retention
RU2018132032A (ru) 2016-03-18 2020-03-10 Эвоник Дегусса Гмбх Гранулят, содержащий неорганический твердый носитель с по меньшей мере одним содержащимся на нем поверхностно-активным веществом биологического происхождения
WO2018077700A1 (en) * 2016-10-24 2018-05-03 Evonik Degussa Gmbh Rhamnolipid-producing cell having reduced glucose dehydrogenase activity
CN109843909B (zh) * 2016-10-24 2022-07-12 赢创运营有限公司 利用替代的葡萄糖转运蛋白产生鼠李糖脂的细胞和方法
WO2018144053A1 (en) 2017-02-06 2018-08-09 Logos Technologies, Llc Decolorization of concentrated rhamnolipid composition
AU2018309664B2 (en) * 2017-07-31 2023-09-28 Stepan Company Enhanced production of rhamnolipids using at least two carbon sources
CN110997069B (zh) 2017-08-24 2022-12-30 赢创运营有限公司 作为乳化剂和分散助剂的鼠李糖脂衍生物
CN108060111B (zh) * 2017-10-27 2021-06-04 中国科学院微生物研究所 一种提高鼠李糖脂产量的铜绿假单胞菌及其构建方法
EP3749679A1 (en) * 2018-02-09 2020-12-16 Evonik Operations GmbH Mixture composition comprising glucolipids
US11541105B2 (en) 2018-06-01 2023-01-03 The Research Foundation For The State University Of New York Compositions and methods for disrupting biofilm formation and maintenance
CN112481335A (zh) * 2019-09-11 2021-03-12 万华化学集团股份有限公司 一种鼠李糖脂发酵方法
CN115190792A (zh) 2020-03-11 2022-10-14 赢创运营有限公司 包含糖脂和柠檬酸三乙酯的混合物组合物
CN115175660A (zh) 2020-03-24 2022-10-11 赢创运营有限公司 包含鼠李糖脂、烷基多糖苷和酰基乳酸盐的组合物
CN116249706A (zh) 2020-07-22 2023-06-09 赢创运营有限公司 新型鼠李糖脂低聚酯
CN114438000B (zh) * 2020-11-05 2024-02-27 万华化学(四川)有限公司 一株铜绿假单胞菌及其构建方法与应用
WO2023198511A1 (en) * 2022-04-13 2023-10-19 Evonik Operations Gmbh Process for the fermentative production of a biosurfactant
CN115290765B (zh) * 2022-04-28 2024-02-13 华东理工大学 一种羧酸标记hplc-uv定量检测鼠李糖脂的方法
EP4155371A1 (en) 2022-08-29 2023-03-29 Evonik Operations GmbH Composition rich in mono-rhamnolipids
CH720165A2 (de) 2022-10-26 2024-04-30 Chemtek Ug Zusammensetzungen mit N-Acylglycaminen

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1295127A (zh) * 1999-11-05 2001-05-16 德古萨-于尔斯股份公司 来自谷氨酸棒杆菌的质粒及其用途
US6551795B1 (en) * 1998-02-18 2003-04-22 Genome Therapeutics Corporation Nucleic acid and amino acid sequences relating to pseudomonas aeruginosa for diagnostics and therapeutics
US20040033549A1 (en) * 1999-09-03 2004-02-19 Greenberg E. Peter Quorum sensing signaling in bacteria
EP1567649A1 (en) * 2002-12-05 2005-08-31 Plant Bioscience Limited Bioremediation with transgenic plants
WO2009018780A1 (en) * 2007-08-08 2009-02-12 Panorama Research Inc. Andrographolide derivatives and use thereof in manufacture of medicaments
WO2014039940A1 (en) * 2012-09-10 2014-03-13 Logos Technologies Llc Cells and methods for the production of rhamnolipids
CN104830889A (zh) * 2015-03-06 2015-08-12 西安海格生物技术研究所有限公司 一种铜绿假单胞菌高产鼠李糖脂的基因重组方法
CN106414758A (zh) * 2014-05-26 2017-02-15 赢创德固赛有限公司 生产鼠李糖脂的方法
CN106755031A (zh) * 2016-11-14 2017-05-31 国家海洋局天津海水淡化与综合利用研究所 鼠李糖脂生产质粒及其构建方法与大肠杆菌工程菌和应用

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US601494A (en) 1898-03-29 Spar for vessels
NL301993A (zh) 1962-12-18
US4601893A (en) 1984-02-08 1986-07-22 Pfizer Inc. Laminate device for controlled and prolonged release of substances to an ambient environment and method of use
EP0473869A1 (de) 1990-08-28 1992-03-11 Landis & Gyr Business Support AG Verfahren zur automatischen Sendewiederholung eines Telegramms bei dessen fehlerhaftem Empfang
DE4027453A1 (de) 1990-08-30 1992-03-05 Degussa Neue plasmide aus corynebacterium glutamicum und davon abgeleitete plasmidvektoren
DE4440118C1 (de) 1994-11-11 1995-11-09 Forschungszentrum Juelich Gmbh Die Genexpression in coryneformen Bakterien regulierende DNA
CN1236063C (zh) 1996-11-13 2006-01-11 纳幕尔杜邦公司 用重组生物体生产1,3-丙二醇的方法
JPH10229891A (ja) 1997-02-20 1998-09-02 Mitsubishi Rayon Co Ltd マロン酸誘導体の製造法
DE10031999A1 (de) 1999-09-09 2001-04-19 Degussa Verfahren zur fermentativen Herstellung von D-Pantothensäure unter Verwendung coryneformer Bakterien
EP1083225A1 (de) 1999-09-09 2001-03-14 Degussa-Hüls Aktiengesellschaft Verfahren zur fermentativen Herstellung von D-Pantothensäure unter Verwendung coryneformer Bakterien
US6713289B2 (en) 1999-10-05 2004-03-30 Degussa Ag Nucleotide sequences which code for the eno gene

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6551795B1 (en) * 1998-02-18 2003-04-22 Genome Therapeutics Corporation Nucleic acid and amino acid sequences relating to pseudomonas aeruginosa for diagnostics and therapeutics
US20070020624A1 (en) * 1998-02-18 2007-01-25 Genome Therapeutics Corporation Nucleic acid and amino acid sequences relating to Pseudomonas aeruginosa for diagnostics and therapeutics
US20040033549A1 (en) * 1999-09-03 2004-02-19 Greenberg E. Peter Quorum sensing signaling in bacteria
CN1295127A (zh) * 1999-11-05 2001-05-16 德古萨-于尔斯股份公司 来自谷氨酸棒杆菌的质粒及其用途
EP1567649A1 (en) * 2002-12-05 2005-08-31 Plant Bioscience Limited Bioremediation with transgenic plants
US20060150279A1 (en) * 2002-12-05 2006-07-06 Sorokin Alexander P Bioremediation with transgenic plants
WO2009018780A1 (en) * 2007-08-08 2009-02-12 Panorama Research Inc. Andrographolide derivatives and use thereof in manufacture of medicaments
WO2014039940A1 (en) * 2012-09-10 2014-03-13 Logos Technologies Llc Cells and methods for the production of rhamnolipids
CN106414758A (zh) * 2014-05-26 2017-02-15 赢创德固赛有限公司 生产鼠李糖脂的方法
CN104830889A (zh) * 2015-03-06 2015-08-12 西安海格生物技术研究所有限公司 一种铜绿假单胞菌高产鼠李糖脂的基因重组方法
CN106755031A (zh) * 2016-11-14 2017-05-31 国家海洋局天津海水淡化与综合利用研究所 鼠李糖脂生产质粒及其构建方法与大肠杆菌工程菌和应用

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
MISUN CHA等: ""Heterologous production of Pseudomonas aeruginosa EMS1 Biosurfactant in Pseudomonas putida"", 《BIORESOUR TECHNOL》 *
RUSSELL J.COLEMAN等: ""Identification and organization of genes for diutan polysaccharide synthesis from Sphingomonas sp.ATCC 53159"", 《JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY》 *
U A OCHSNER等: ""Autoinducer-mediated regulation of rhamnolipid biosurfactant synthesis in Pseudomonas aeruginosa"", 《PNAS》 *
U A OCHSNER等: "production of pseudomonas aeruginosa rhamnolipid biosurfactants in heterologous hosts", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 *
XIANGDONG FANG等: "Final repirt:bio-enginering high performance microbial strains for meor by directed protein evolution technology", 《OIL & NATURAL GAS TECHNOLOGY PROJECTS》 *
郝东辉: ""采油微生物筛选、鼠李糖脂产脂性能及关键酶基因克隆与表达研究"", 《中国博士学位论文全文数据库(电子期刊)》 *
陈桂秋等: ""鼠李糖脂的表面化学和生物合成及其在垃圾堆肥中的应用展望"", 《中国生物工程杂志》 *

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US20130130319A1 (en) 2013-05-23
ES2837485T3 (es) 2021-06-30
US20150247151A1 (en) 2015-09-03
HUE053232T2 (hu) 2021-06-28
EP2598646A1 (de) 2013-06-05
JP6461078B2 (ja) 2019-01-30
RU2619877C2 (ru) 2017-05-18
CN103038357A (zh) 2013-04-10
CA2806430A1 (en) 2012-02-02
US9580720B2 (en) 2017-02-28
JP2017060523A (ja) 2017-03-30
DE102010032484A1 (de) 2012-02-02
EP3418388A1 (de) 2018-12-26
BR112013002124A2 (pt) 2016-09-20
JP2013537411A (ja) 2013-10-03
BR112013002124B1 (pt) 2021-08-31
HUE042573T2 (hu) 2019-07-29
CN108587995B (zh) 2022-04-01
WO2012013554A1 (de) 2012-02-02
EP2598646B1 (de) 2018-11-07
CA2806430C (en) 2019-06-25
BR122020023808B1 (pt) 2022-01-11
CN103038357B (zh) 2018-05-25
RU2013108700A (ru) 2014-09-10
JP6066908B2 (ja) 2017-01-25

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