CN107630029B - 一种产朊假丝酵母游离型表达载体及其构建方法与应用 - Google Patents

Abstract

本发明涉及基因工程领域，公开了一种产朊假丝酵母(Candida utilis,C.utilis)游离型表达载体及其构建方法与应用，该表达载体以毕赤酵母表达型载体pGAPZαA为基本骨架，利用产朊假丝酵母的启动子、酿酒酵母的启动子或毕赤酵母的启动子替换pGAPZαA载体上的GAP启动子，利用G418抗性基因替换pGAPZαA载体上的博来霉素基因，将产朊假丝酵母的自主复制序列插入pGAPZαA载体的PscI位点中。本发明的所构建的表达载体分别含有不同启动子，旨在构建适合产朊假丝酵母的表达载体，应用于产朊假丝酵母工程菌高效表达外源蛋白。

Description

一种产朊假丝酵母游离型表达载体及其构建方法与应用

技术领域

本发明涉及基因工程领域，具体涉及一种产朊假丝酵母游离型表达载体及 其构建方法与应用。

背景技术

酵母具有完整的亚细胞结构和严密的真核基因调控机制，是理想的真核表 达系统。产朊假丝酵母是美国食品和药物管理局(FDA)认证的安全的食品酵 母，与酿酒酵母和毕赤酵母相比具有如下优势：

①属于克拉布特里阴性菌(Crabtree-negative)，在有氧条件下不产生乙醇， 能够快速生长；②在有效的连续培养条件下能够实现高密度培养；③能够利用廉 价的糖蜜作为养分生长；④能够同时利用戊糖和己糖作为碳源；⑤适应多种碳源 和氮源，且其分泌蛋白质组不含任何蛋白酶；⑥产物和菌体无需经过复杂的分 离纯化程序，可直接作为添加剂应用于食品等行业。

作为一种重要的工业微生物，产朊假丝酵母已经被用于生产多种有价值的 生物制剂，包括谷胱甘肽、类胡萝卜素、RNA、α-淀粉酶等。因此，开发更多 的表达元件应用于产朊假丝酵母，并进一步提高其作为宿主表达外源蛋白的价 值在该领域备受关注。然而，产朊假丝酵母是三倍体的单细胞微生物，利用整 合型载体进行同源重组时会出现回复突变，导致所有染色体不能同时整合成 功，不仅操作不便，而且不能实现稳定遗传。目前产朊假丝酵母表达载体尚未 商品化，构建可以高效表达外源蛋白的产朊假丝酵母表达载体，或探索一种可 行的产朊假丝酵母表达载体构建方法将具有重要意义。

发明内容

本发明旨在针对现有技术的不足，即缺乏产朊假丝酵母的成熟外源蛋白表 达体系的现状，提供了一种产朊假丝酵母游离型表达载体。

为实现以上目的，本发明采用的技术方案如下：

一种产朊假丝酵母游离型表达载体，以毕赤酵母表达型载体pGAPZαA为 基本骨架，利用G418抗性基因替换pGAPZαA载体上的博来霉素基因，将产 朊假丝酵母的自主复制序列插入pGAPZαA载体的CYCTT元件和pUCori元件 之间。

进一步地，该表达载体还利用产朊假丝酵母的启动子、酿酒酵母的启动子 或毕赤酵母的启动子替换pGAPZαA载体上的GAP启动子。

本发明分别含有来自酿酒酵母、产朊假丝酵母或毕赤酵母的不同启动子， 带有产朊假丝酵母的自主复制序列(CuARS)，从而实现在产朊假丝酵母中游 离表达，并采用单一抗性基因，实现在原核宿主中具有卡那霉素抗性、而酵母 宿主中具有G418抗性。本发明所述的一种产朊假丝酵母游离型表达载体以 G418抗性基因作为抗性筛选标记，该基因在原核系统中呈现卡那霉素抗性，而 在真核系统中呈现G481抗性，相比于博莱霉素、卡那霉素，G418的价格较 低，以其作为抗性基因替换博莱霉素抗性基因，有利于节约生产成本。

进一步地，所述产朊假丝酵母的启动子为glyceraldehyde-3-phosphatedehydrogenase promoter(CuGAPp)核苷酸序列如SEQ ID NO.1所示、 phosphoglyceratekinase promoter(CuPGKp)核苷酸序列如SEQ ID NO.2所示、 maltase promoter(CuMALp)核苷酸序列如SEQ ID NO.3所示。

进一步地，所述酿酒酵母的启动子为General amino-acid permease promoter(ScGAP1p)核苷酸序列如SEQ ID NO.4所示、transcriptional elongation factor EF-1αpromoter(ScTEF1p)核苷酸序列如SEQ ID NO.5所示、triose phosphate dehydrogenasepromoter(ScTDH3p)核苷酸序列如SEQ ID NO.6所示、 phosphoglycerate kinase1promoter(ScPGK1p)核苷酸序列如SEQ ID NO.7所 示、alcohol dehydrogenase promoter(ScADH1p)核苷酸序列如SEQ ID NO.8所 示、hexose transporter promoter(ScHXT7p)核苷酸序列如SEQ ID NO.9所示、 galactosidase promoter(ScGAL1p)核苷酸序列如SEQ IDNO.10所示。

进一步地，所述毕赤酵母的启动子为glyceraldehyde-3-phosphatedehydrogenase promoter(PsGAPp)核苷酸序列如SEQ ID NO.11所示、alcohol oxidase1promoter(PsAOX1p)核苷酸序列如SEQ ID NO.12所示、 formaldehyde dehydrogenase1promoter(PsFLD1p)核苷酸序列如SEQ ID NO.12所示。

启动子是影响宿主微生物中外源蛋白表达效率的关键因素之一，本发明中 含有来源不同强度不同的启动子，不仅提高表达外源蛋白的效率和灵活度，同 时还扩大了产朊假丝酵母表达系统的应用范围。

进一步地，所述产朊假丝酵母的自主复制序列(CuARS)的核苷酸序列如 SEQ IDNO.15所示，用于实现在产朊假丝酵母中的游离表达。

在载体上加入产朊假丝酵母的自主复制序列(CuARS)，使重组载体转化 前无需线性化，以及转化后不与产朊假丝酵母染色体整合，最终以游离的形式 存在于产朊假丝酵母细胞质中，方便操作，同时提高了重组载体的转化效率。

进一步地，所述表达载体的序列为SEQ ID NO.46-SEQ ID NO.58所示。

本发明的另一个目的是提供上述的一种产朊假丝酵母游离型表达载体的构 建方法，包括如下步骤：

S1.G418抗性基因片段的制备：为将毕赤酵母表达载体pGAPZαA的博来 霉素基因替换为G418抗性基因，需要将G418抗性基因的编码区与CYC终止 子的一部分利用重叠延伸PCR相连接，以便于后续的酶切与连接操作。根据载 体pScIKP的序列和pGAPZαA的序列，设计G418抗性基因编码区以及部分 CYC终止子片段的特异性扩增引物及重叠PCR引物，并加上相应的限制性内 切酶的酶切位点(引物序列如SEQ ID NO.16-SEQ ID NO.17所示)。所得G418- CYCTT片段长度为892bp，其序列如SEQ ID NO.14所示；

S2.产朊假丝酵母的自主复制序列的扩增：根据产朊假丝酵母基因组的 DNA序列设计产朊假丝酵母自主复制序列的特异性扩增引物，并在引物的5’端 加上相应的限制性内切酶位点(引物序列如SEQ ID NO.18所示)。以产朊假丝 酵母的基因组DNA为模板，扩增产朊假丝酵母的自主复制序列片段，长度约 为1900bp。采用重叠延伸PCR技术对自主复制序列中的PscI酶切位点进行定 点突变，PCR引物序列如SEQ ID NO.19所示，得到突变后的自主复制序列如 SEQ ID NO.15所示。

S3.多种启动子片段的制备：采用Gibson组装的方法进行克隆，需要在 PCR扩增引物两端添加15～30bp的同源短重叠序列。根据载体pGAPZαA的序 列，在启动子特异性扩增引物序列5’端加上同源短重叠序列(引物序列如SEQ ID NO.22-SEQ ID NO.45所示)。以酿酒酵母、产朊假丝酵母及毕赤酵母的基因 组DNA为模板，分别用对应的引物进行PCR扩增，得到两端分别带有Gibson 组装同源序列的启动子片段；

S4.将上述S1和S2获得的G418-CYCTT片段和产朊假丝酵母的自主复制 序列经纯化后分别与克隆载体pMD19-T连接，并转化大肠杆菌DH5α。经 DNA测序验证后获得阳性克隆；

S5.将上述S4经测序验证的G418-CYCTT片段阳性克隆和毕赤酵母表达载 体pGAPZαA分别用限制性内切酶进行双酶切，酶切片段经纯化回收后，通过 连接反应获得第一次连接产物pGAPGαA，即原毕赤酵母表达载体的博莱霉素 基因替换为G418抗性基因；

S6.将上述S4经测序验证的产朊假丝酵母的自主复制序列和上述S5所得第 一次连接产物pGAPGαA分别用相同的限制性内切酶进行单酶切，酶切片段经 纯化回收后，通过连接反应获得第二次连接产物pCuGAPGαA，即原毕赤酵母 表达载体加入了产朊假丝酵母的自主复制序列。

S7.将上述S6所得的第二次连接产物pCuGAPGαA用限制性内切酶进行双 酶切，酶切片段经纯化回收后，与上述S3所得的启动子片段纯化后分别进行连 接反应并转化大肠杆菌DH5α。经DNA测序验证后获得阳性克隆，即将原毕赤 酵母表达载体的GAP启动子替换为其他启动子，获得最终的一种产朊假丝酵母 游离型表达载体。

进一步地，上述S1和S5所述的限制性内切酶为EcoRV、BbsI和NcoI，或 与其具有相同识别序列或产生相同粘性末端的限制性内切酶。

上述S2和S6所述的限制性内切酶为PscI，或与其具有相同识别序列或产 生相同粘性末端的限制性内切酶。

上述S7所述的限制性内切酶为Bsp119I和BglII，或与其具有相同识别序 列或产生相同粘性末端的限制性内切酶。

进一步地，上述步骤S4～S6所述的连接反应采用T4连接酶连接；上述S7 所述的连接反应为Gibson组装。

本发明的另一个目的是提供上述一种产朊假丝酵母游离型表达载体在表达 外源基因方面的应用，以包括但不限于产朊假丝酵母在内的酵母细胞为宿主， 表达包括但不限于绿色荧光蛋白、木聚糖酶在内的外源蛋白。

本发明可用于外源基因的表达，通过构建含有不同启动子的适合产朊假丝 酵母的表达载体，将外源基因克隆至表达载体的多克隆位点，并转化产朊假丝 酵母；通过含G418的平板筛选转化子，从而实现外源蛋白在产朊假丝酵母表 达体系的表达，应用于产朊假丝酵母工程菌高效表达外源蛋白。

本发明通过实施具体的技术指标，实现本发明内容，达到以下有益效果：

本发明以毕赤酵母表达型载体pGAPZαA为基本骨架，分别含有酿酒酵母 启动子、产朊假丝酵母启动子或毕赤酵母启动子，采用单一抗性基因，实现在 原核宿主中具有卡那霉素抗性、而酵母宿主中具有G418抗性，并带有产朊假 丝酵母的自主复制序列(CuARS)，从而实现在产朊假丝酵母中游离表达，构 建该游离型表达载体，不仅能高效表达，且操作方便、在选择压力下能够稳定 遗传。此外，本发明所采用的Gibson组装法能够同时将多种启动子分别构建至 表达载体进行筛选，这种新兴的克隆方案能够同时处理多个DNA片段，使实 验过程操作简单，快速高效。本发明对将来实现高效、通用的产朊假丝酵母表 达载体的商业化具有重要意义。

附图说明

图1为产朊假丝酵母游离型表达载体pCuXGαA构建流程(其中：100启动 子片段)；

图2为产朊假丝酵母游离型表达载体pCuXGαA表达绿色荧光蛋白及木聚 糖酶表达载体构建示意图；

图3为产朊假丝酵母转化子pCuXGA-GFP表达绿色荧光蛋白的平均荧光强 度(A含来源于酿酒酵母启动子的重组酵母、B含来源于毕赤酵母启动子的重组 酵母、C含来源于产朊假丝酵母启动子的重组酵母)；

图4为重组木聚糖酶酶解半纤维素所得产物的薄层层析分析(其中：1低聚 木糖、2木糖、3重组木聚糖酶为0.5U/g底物时的酶解产物、4重组木聚糖酶为 1.0U/g底物时的酶解产物、5重组木聚糖酶为2.5U/g底物时的酶解产物、6重 组木聚糖酶为4.0U/g底物时的酶解产物)。

具体实施例

以下内容为本发明的进一步说明，不应理解为对本发明的限制。在没有违背 本发明所涉及的技术方法实质下，对本发明的中的方法、条件和步骤进行简单的 改动均属于本发明要求保护的范围。

实施例1：载体构建所需各元件的获得

步骤1、G418抗性基因片段的获得

通过研究毕赤酵母载体pGAPZαA的序列图谱，为便于将G418抗性基因替 换原载体上的博莱霉素抗性基因，本发明先将G418抗性基因的编码区与CYC 终止子的一部分分别扩增，然后利用重叠延伸PCR将二者连接成为一个完整的 片段，再通过双酶切和连接反应进行基因替换。

(1)G418抗性基因编码区及部分CYC终止子片段的扩增

根据载体pScIKP上的G418抗性基因编码区的序列和pGAPZαA的序列分 别设计G418抗性基因编码区以及部分CYC终止子片段的特异性扩增引物，并 加上相应的限制性内切酶的酶切位点。其中，为将G418抗性基因编码区与CYC 终止子部分片段通过重叠延伸PCR连接成为一个片段，G418抗性基因编码区的 下游引物与CYC终止子部分片段的上游引物被设计为反向互补序列。

分别以质粒pScIKP和pGAPZαA为模板，使用高保真DNA聚合酶，按照 如下反应体系和反应条件，分别扩增G418抗性基因编码区和部分CYC终止子 片段：

反应体系：

反应条件：

(2)G418抗性基因编码区和部分CYC终止子片段进行重叠延伸PCR

将上述步骤(1)所得的G418抗性基因编码区和部分CYC终止子片段经 PCR产物纯化回收试剂盒纯化后，按照如下反应体系和反应条件进行第一轮PCR 反应，将二者连接成为一个大片段：

反应体系：

反应条件：

取出2μl上述反应后的PCR产物作为模板，以G418抗性基因编码区的上游 引物和CYC终止子的下游引物为扩增引物，加入下述反应体系中，继续进行第 二轮PCR反应：

以第一轮PCR反应相同的反应条件进行30个循环，获得连接后的G418- CYCTT基因片段。

(3)G418-CYCTT基因片段克隆进大肠杆菌DH5α

将上述步骤(2)所得的G418-CYCTT基因片段经PCR产物纯化试剂盒纯 化后，与pMD19-T载体连接。连接反应按照pMD19-T载体的说明书进行。连接 产物转化进大肠杆菌DH5α，挑选阳性克隆进行DNA测序，所获得的G418- CYCTT片段的核酸序列如SEQ ID NO.14所示。

步骤2、产朊假丝酵母的自主复制序列的获得

根据产朊假丝酵母基因组的DNA序列设计产朊假丝酵母自主复制序列的特 异性扩增引物，并在引物的5’端加上PscI限制性内切酶位点(引物序列如SEQ ID NO.18所示)。以产朊假丝酵母的基因组DNA为模板，扩增产朊假丝酵母的 自主复制序列片段。为了后续的PscI酶切和连接反应，采用重叠延伸PCR技术 对自主复制序列中的PscI酶切位点进行定点突变，PCR引物序列如SEQ ID NO.19所示。

(1)产朊假丝酵母自主复制序列的扩增

将产朊假丝酵母GIM2.176的基因组作为模板，以产朊假丝酵母自主复制序 列两端特异性引物为扩增引物，使用高保真DNA聚合酶进行PCR扩增。

反应条件：

(2)自主复制序列中PscI酶切位点的定点突变

根据上述步骤(1)中所得的产朊假丝酵母自主复制序列设计重叠互补引物， 并以自主复制序列为模板，分别将自主复制序列两端特异性引物与中间引物配对 作为扩增引物，使用高保真DNA聚合酶进行PCR扩增。

反应条件：

将上述步骤胶回收后的PCR产物作为重叠延伸PCR第一步反应的引物和模 板。

反应体系：

反应条件：

将重叠延伸PCR第一步反应的PCR产物作为模板，以产朊假丝酵母自主复 制序列两端特异性引物为扩增引物进行重叠延伸PCR第二步反应。

反应体系：

将该PCR反应体系加至第一步PCR扩增反应结束后的产物中，混合成50 μl的反应体系，以第一步PCR反应相同的反应条件进行30个循环，获得PscI 酶切位点突变后的产朊假丝酵母自主复制序列。

(3)将突变后的产朊假丝酵母自主复制序列经PCR产物纯化试剂盒纯化 后，与pMD19-T载体连接。连接反应按照pMD19-T载体的说明书进行。连接产 物转化进大肠杆菌DH5α，挑选阳性克隆进行DNA测序，所获得的突变后的产 朊假丝酵母自主复制序列如SEQID NO.15所示。

步骤3、启动子片段的获得

根据载体pGAPZαA的序列，在启动子特异性扩增引物正反向序列5’端分别 加上同源的短重叠序列GCTATACGAAGTTATAGATCT和 CCGTCGACTGCAGAATTCGAA(引物序列如SEQ IDNO.22-SEQ ID NO.45所 示)。以酿酒酵母、产朊假丝酵母及毕赤酵母的基因组为模板，分别将对应的特 异性引物作为扩增引物，使用高保真DNA聚合酶进行PCR扩增，得到两端分别带有Gibson组装同源序列的启动子片段。

反应条件：

实施例2：产朊假丝酵母游离型表达载体的构建

本发明所述的一种产朊假丝酵母游离型表达载体的详细构建流程图如图1 所示，具体步骤如下：

(1)将毕赤酵母表达载体pGAPZαA用NcoI和EcoRV双酶切，得到去掉 博来霉素抗性基因的骨架；同时将由实施例1中步骤1所得的含重叠的G418抗 性基因片段的pMD19-T质粒用BbsI和EcoRV双酶切，回收重叠的G418抗性基 因片段。由于在本发明的设计中，BbsI与NcoI可以产生相同的粘性末端，因此 可以将所回收的重叠的G418抗性基因片段和去掉博来霉素抗性基因的毕赤酵母 表达载体pGAPZαA骨架用T4连接酶连接起来，得到表达载体1，命名为 pGAPGαA。

(2)将上一步骤所得的表达载体1和由实施例1中步骤2所得的含产朊假 丝酵母的自主复制序列的pMD19-T质粒分别用PscI单酶切，然后将回收的产朊 假丝酵母的自主复制序列片段与去磷酸化的表达载体1用T4连接酶连接，得到 表达载体2，命名为pCuGAPGαA。

(3)将上一步骤所得的表达载体2用Bsp119I和BglII双酶切，得到去掉启 动子的表达载体2骨架。将所得的去掉启动子的表达载体2骨架和由实施例1中 步骤3所得的启动子片段pEASY-Uni Seamless Cloning and Assembly Kit试剂盒 分别进行Gibson组装，按照如下反应体系和反应条件，连接后获得一种产朊假 丝酵母游离型表达载体3，命名为pCuXGαA(X代表各启动子片段)。

反应体系：

其中，载体与各个插入片段的最佳摩尔比为1：2。将反应体系于50℃反应 15分钟，反应结束后，将连接产物置于冰上冷却数秒，转化进大肠杆菌DH5α， 挑选阳性克隆进行DNA测序检测，所获得的一种产朊假丝酵母游离型表达载体 如SEQ ID NO.1-SEQ ID NO.13所示。

实施例3：绿色荧光蛋白基因表达性能的检测

(1)如图2所示，将载体pEGFP–N1和实施例2中步骤(3)所得的表达载 体3同时用Bsp119I和NotI双酶切，分别获得绿色荧光蛋白基因和去掉α信号 肽和多克隆位点的表达载体3骨架。然后将两者用T4连接酶连接，得到表达载 体4，命名为pCuXGA-GFP(X代表各启动子片段)。

(2)将上一步骤所得的表达载体4分别用电穿孔法转入产朊假丝酵母 GIM2.176中，涂布于含G418(300μg/ml)的YPD琼脂平板上培养3-4天后， 挑取正常生长的菌落，经菌落PCR鉴定确定为含有上述重组质粒的转化子。

(3)将上一步骤所得的产朊假丝酵母阳性转化子接种于含G418的YPD液 体培养基中，30℃，200rpm条件下培养60h。每隔12h取适量菌液于4℃， 12000rpm离心1min后，用1×PBS重悬菌体，使菌液OD600在0.1-0.2。以产 朊假丝酵母原始菌为对照，用流式细胞仪检测酵母细胞的平均荧光强度，如图3 所示。由此可证明上述步骤(1)中得到的表达载体pCuXGA-GFP在产朊假丝酵 母中表达，且含不同启动子的表达载体pCuXGA-GFP表达性能不同。

实施例4：木聚糖酶基因的表达和应用

(1)如图2所示，将从里氏木霉cDNA文库中扩增所得木聚糖酶基因xyn2 的编码序列(引物序列如SEQ ID NO.20所示，xyn2序列如SEQ ID NO.21所示) 和实施例2中步骤(3)所得的表达载体3同时用XhoI和XbaI双酶切，将回收 的xyn2基因片段和表达载体3骨架用T4连接酶连接，获得表达载体5，命名为pCuXGαA-xyn2(X代表各启动子片段)。

(2)将上一步骤所得的表达载体5分别用电穿孔法转入产朊假丝酵母 GIM2.176中，涂布于含G418(300μg/ml)的YPD琼脂平板上培养3-4天后， 挑取正常生长的菌落，经菌落PCR鉴定确定为含有上述重组质粒的转化子。

(3)将上一步骤中得到的产朊假丝酵母阳性转化子分别用含有G418的 YPD液体培养基在30℃，250rpm培养60h后，4℃，12000rpm离心5min，收 取上清。将上清液按照酶活浓度梯度与甘蔗渣提取的半纤维素在50℃，200rpm 下反应6h，取适量样品于沸水中煮10min终止酶解，采用薄层层析鉴定鉴定酶 解产物，如图4所示。由此可证明上述步骤(1)中得到的表达载体pCuXGαA- xyn2在产朊假丝酵母中分泌表达了木聚糖酶基因xyn2，进一步证明本研究所构 建的产朊假丝酵母高效游离型表达载体在实际应用中具有可行性。

SEQUENCE LISTING

<110> 广东启智生物科技有限公司

<120> 一种产朊假丝酵母游离型表达载体及其构建方法与应用

<130>

<160> 63

<170> PatentIn version 3.3

<210> 1

<211> 985

<212> DNA

<213> CuGAPp

<400> 1

agatctctta cagcgagcac tcaaatctgc cctccgagcc ctccggccct ctcttcaaca 60

aactcgcgct gcacttcgtc gtcagtggtg ccaatcaccc aacgtggagg tatcaagagg 120

tgctccagcc cacaaagcga catcaaagac aacaaccctg ccggcctacg tcctacacac 180

cctggtgatc gcagacattg tacaaggtgc cacgcaataa cctacaggca ccgcacatga 240

cgatggcctt ggttgtgcaa ccagtgactt ccacggtcca cgcagcaaca tgaaccacac 300

cacccagaat cgatgcgcgc aacaacagtt gttccggttc actcagcccc acagcgagtc 360

gctggcagaa cacgagcctg agggcggaaa gagggtagag gaaagcgcaa ggacagggga 420

caacctggcc caattgatgt catataaacc ctctcgatca attgagcaca ctcatccgcc 480

aattgacccc tgttcgcagc tccacgcccc atgttcctcg tccctggtgt agcttctccc 540

ctaaattcca gcgcttggtt ccgccctccc tgtctcccgg gtttaacgaa cgtgtgtacc 600

atctgatggt aatccgctcc cgtccgcgca acacaactca caagcagatc acacctctac 660

acgccgctgc tgatgcgccc aatttaattt ttttttctct caatgtaggg gagaagcctt 720

gggagctccc gactcccagt tgggcacagc tgccacctca tgacttttcc tgtgtgtgcc 780

tgtctgacgt tacgtgtgat gtagtggccc ccgtccggtg tgttttcgcc tgttgcgctg 840

tgcccccctt aaaagtataa aaggaagtgc aattgctgtt tgtgttgatt gttgatcctt 900

gtttcctctg ttttctcctc atcacacaag aaaggtttct tctttccaac agatacaaaa 960

cacacttaca aacaacatat tcgaa 985

<210> 2

<211> 1358

<212> DNA

<213> CuPGKp

<400> 2

agatctaagc ttttgtcttt taggagcctt cttttcaccc tggctttctt cagactccac 60

gcctctcgcc cgtttgttgt tgatcttctt ctgttgcttc ttcgtgagct taccagtatc 120

cagatgcgtt gtcagggcaa gagggtcatg ttcaagctcc tctttcactt tcagtccaat 180

acgtttccag gcagggatgt gttcgctcat cgttccagac tcgagtggtg aaaactatgg 240

caacctctac ttcctttcca aacacacagc gtgctttgta gtgtgtgcct aagagctgaa 300

ttttttttcc ttccatgctg cgctgcgatg agctctgccc gcccgcagcc tcggaggcta 360

gcgacgtata aaaaaggcct gtgaaaattt tatcctcctc cttaacgacc cttctttctc 420

ttcttcacat tcaaaaactt caagcagctg tctctgttcc tttgctgtgt tctaccattg 480

gatattccca ttccccgtgg agaaccgaac tggagtctag cagcatgcga gatcaatatt 540

acacggtttg agtctgatac gcttgagcag ccattttttg gcttctcctg gtgtgtatcc 600

agatatagaa gttcgtatac atttcccata gcgattgtaa aatgattctg caatggaacc 660

atccgtaatt gtaggcctgc tgagatggca ctcgcaatgc ctctgtgtct ggttttttgc 720

cttctccgtc catcagcacc agtggcttct tagggcataa cgagacggct ccttggtgaa 780

agatgccctg ctccgtctgt ctgcctgttg ctacaaccac tgcgtagtca gatgacccgg 840

tctgtgtgct gtggaatcac cgggagcgaa attccggttt cgctggcaga tgagctcatc 900

aaccacatca actggagcaa cctcaccaga ggacacgtaa cctgcccggt tgaattctgt 960

caaaccgtac atcacacaac aacagcagca gcaacaacaa caacgtcagt tgtcgttcgc 1020

atggcgacgt tacctaacgg caccaacatc gtctcgtcct cgccagtgcc tgtttcccct 1080

acccggagtg gcccggccca cctgtcgttc ttttttcgtc aattgtgtcc agctggtgcc 1140

atcaccatat gttcaagtgc gtggcctgta ctagcgcagt ctgctgcagt ataaaaggga 1200

ttgctgaggc cccctttagc gtttccaatt aacaattgat tcccttttcc ccatagtccg 1260

tttgtactac atcctacata acaaaagtga gtgttacaag acaagtgtgg cggtcaattg 1320

gatcatttgg actaacatac tggcggataa agttcgaa 1358

<210> 3

<211> 1475

<212> DNA

<213> CuMALp

<400> 3

agatctgtat gaaggtaggt tttaaggagg tgtctaatag tccttgtatt gactggtttt 60

gttttttgaa tctctaaagg gaggctcgag tccttccaat tggggtggtt tttaatgagc 120

attagagatg tacaattgcg gggtgtatgt agaggagatc acctttggat gatgttttct 180

caaaaactgt tcaacatttc ttcccaagag caattgactt ttaattgttt ctccgacacc 240

ccaaattaat tctccgtctt ctgcagccat ttgcgtcggt ctagatgaag tgatccttgt 300

acatgggaag tttgtgtgca ctggaatttg gaatctacac taacccctcg taagagatcg 360

atcagcataa gataatgcaa attggggggg gggggaaatt atcaattgta aactcccgtt 420

tgaattgatc caattggcaa ttggatatgt tcttatttag ggatgtggga taagtttcac 480

tgtagaactg gcttctcgct cttagaacgg gtcatcagtt tgaagccttt tcgatcactt 540

ggcatttaac acttgaactg ttgaacatag tctgacaact aacacgttgt ttcggaaaac 600

tcatgactgt ttttcatgtt acatatttgc tgcatgtatt tatgcagatg ttcatattat 660

tccaccccga gcctttattt gtagaacttg aacggaatag attgtaagga aatgacactt 720

atgtgcatga atcaaggatt aaattctatg atcaaggcgt agacactcca gtagaaaaca 780

ttgctatggt tgtgtgctgc acttccataa aatcacccga ctaagcatcg tactctaaat 840

ttacgtccac taatgaggag aagtagggag agattttgga tacgaaaaga tattttcaac 900

cccatgcaca tggggcggtt aaattttcca acaattaatc caaagcgcta aaatattaac 960

ccacccgaaa ctgttgatac aacagggata ataattgttg aaaagaaccg caacggttat 1020

ccgttgcgac gaatacgtcg agtaaggagc ctagcacaac cccttttgga ttgacaaatt 1080

tggaattatc ttctgtctcc tagttttaga cagaggacca tctcggggct tggaaacaag 1140

gtgctatgta ccaagctgca cctccattat tccgtttgca taaaatttca aacaaggttc 1200

aacttggtgt ccagccgttc atgacatggt actgatttcc ccaagcctct atcctgaacg 1260

ggtgtgtgac aacctcatag agattctcat gtaaatcttc gatcattcat gaatgttgct 1320

gccccaagaa ttgatgccat gtgatcgttg aattccagcg ataattcttc cgattgcgtt 1380

gtcaactcag cgtatatatt gcaattagat cacaatagta ttttctaacc ctccaaagtg 1440

aatcaaaaac aagaacaagt tctcgcttat tcgaa 1475

<210> 4

<211> 412

<212> DNA

<213> ScGAP1p

<400> 4

agatctaaag tcacatcgcc catatctctc gagtgcgata acggggaact tgatttggta 60

actgataaga ttgttaaatg tcagtttgga tgctttttct tacgtccgat tagcttatct 120

tctggagcaa ccggctattt acctcctcat agtaaaataa acatgataag cgcatagttg 180

gggcaacaca cctttcttcc ggaattcgct ctggatgaga catataaaga tgaaggtgaa 240

gtccacttaa atgaatgtca atgagacgat gttttttctc ctagattgat ttttgaattc 300

cttgtataca aagtcttgtt ttcttattgt cctcaacaaa acaaaagtag aaaagaacag 360

atcaaggaca gcaacattta taagaaacaa aaaaaagaaa taaaaattcg aa 412

<210> 5

<211> 431

<212> DNA

<213> ScTEF1p

<400> 5

agatctgcac acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt 60

ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaattcc 120

ccctctttct tcctctaggg tggcgttaat tacccgtact aaaggtttgg aaaagaaaaa 180

agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt 240

ctttttcttg aaaatttttt ttttggtttt tttctctttc gatgacctcc cattgatatt 300

taagttaata aatggtcttc aatttctcaa gtttcagttt catttttctt gttctattac 360

aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct aagttttaat 420

tacaattcga a 431

<210> 6

<211> 552

<212> DNA

<213> ScTDH3p

<400> 6

agatctaaat agggggcggg ttacacagaa tatataacat cataggtgtc tgggtgaaca 60

gtttattcct ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa 120

aaaaaagaat cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca 180

ttctcttagc gcaactacag agaacaggga cacaaacagg caaaaaacgg gcacaacctc 240

aatggagtga tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg 300

tatctatctc attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc 360

tgaaaaaaaa ggttgaaacc agttccctga aattattccc ctacttgact aataagtata 420

taaagacggt aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct 480

acttttagag ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgacggatt 540

ctagaattcg aa 552

<210> 7

<211> 996

<212> DNA

<213> ScPGK1p

<400> 7

agatctggaa gtaccttcaa agaatggggt ctcatcttgt tttgcaagta ccactgagca 60

ggataataat agaaatgata atatactata gtagagataa cgtcgatgac ttcccatact 120

gtaattgctt ttagttgtgt atttttagtg tgcaagtttc tgtaaatcga ttaatttttt 180

tttctttcct ctttttatta accttaattt ttattttaga ttcctgactt caactcaaga 240

cgcacagata ttataacatc tgcacaatag gcatttgcaa gaattactcg tgagtaagga 300

aagagtgagg aactatcgca tacctgcatt taaagatgcc gatttgggcg cgaatccttt 360

attttggctt caccctcata ctattatcag ggccagaaaa aggaagtgtt tccctccttc 420

ttgaattgat gttaccctca taaaacacgt ggcctcttat caagaaagaa attaccgtcg 480

ctcgtgattt gtttgcaaag agaacaaaac tgaaaaaacc cagacacgct cgacttcctg 540

tcttcctatt gattgcagct tccaatttcg tcacacaaca aggtcctagc gacggctcac 600

aggttttgta acaagcaatc gaaggttctg gaatggcggg aaagggttta gtaccacatg 660

ctatgatgcc cactgtgatc tccagagcaa agttcgttcg atcgtactgt tactctctct 720

ctttcaaaca gaaatgtccg aatcgtgtga caacaacagc ctgttctcac acactctttt 780

cttctaacca agggggtggt ttagtttagt agaacctcgt gaaacttaca tttacatata 840

tataaacttg cataaattgg tcaatgcaag aaatacatat ttggtctttt ctaattcgta 900

gtttttcaag ttcttagatg ctttcttttt ctctttttta cagatcatca aggaagtaat 960

tatctacttt ttacaacaaa tataaaacaa ttcgaa 996

<210> 8

<211> 1478

<212> DNA

<213> ScADH1p

<400> 8

agatctaggg ggatcgaaga aatgatggta aatgaaatag aaaatcaagg agcatgaagg 60

caaaagacaa atataagggt caaacgaaaa ataaagtgaa aagtgttgat atgatgtatt 120

tggctttgcg gcgccgaaaa aacgagttta cgcaattgca caatcatgct gactctgtgg 180

cggacccgcg ctcttgccgg cccggcgata acgctgggcg tgaggctgtg cccggcggag 240

ttttttgcgc ctgcattttc caaggtttac cctgcgctaa ggggcgagat tggagaagca 300

ataagaatgc cggttggggt tgcgatgatg acgaccacga caactggtgt cattatttaa 360

tttgccgaaa gaacctgagt gcatttgcaa catgagtata ctagaagaat gagccaagac 420

ttgcgagacg cgagtttgcc ggtggtgcga acaatagagc gaccatgacc ctgaaggtga 480

gacgcgcata accgctagag tactttgaag aggaaacagc aatagggttg ctaccagtat 540

aaatagacag gtacatacga cactggaaat ggttgtctgt ttgagtacgc tttcaattca 600

tttgggtgtg cactttatta tgttacaata tggaagggaa ctttacactt ctcctatgca 660

catatattaa ttaaagtcca atgctagtag agaagggggg taacacccct ccgcgctctt 720

ttccgatttt tttctaaacc gtggaatatt tcggatatcc tcttgttgtt tccgggtgta 780

caatatggac ttcctctttt ctggcaacca aacccataca tcgggattcc tataatacct 840

tcgttggtct ccctaacatg taggtggcgg aggggagata tacaatagaa cagataccag 900

acaagacata atgggctaaa caagactaca ccaattacac tgcctcattg atggtggtac 960

ataacgaact aatactgtag ccctagactt gatagccatc atcatatcga agtttcacta 1020

ccctttttcc atttgccatc tattgaagta ataataggcg catgcaactc cttttctttt 1080

tttttttttt ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat 1140

ggaagacact aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca 1200

gagctgatga ggggtatctc gaagcacacg aaactttttc cctccttcat tcacgcacac 1260

tactctctaa tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa 1320

aaagtttgct gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc 1380

tcgtcattgt tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta 1440

taccaagcat acaatcaact atctcatata cattcgaa 1478

<210> 9

<211> 605

<212> DNA

<213> ScHXT7p

<400> 9

agatcttttt accccgtgga aatgaggggt atgcaggaat ttgtgcgggg taggaaatct 60

tttttttttt aggaggaaca actggtggaa gaatgcccac acttctcaga aatgcatgca 120

gtggcagcac gctaattcga aaaaattctc cagaaaggca acgcaaaatt ttttttccag 180

ggaataaact ttttatgacc cactacttct cgtaggaaca atttcggtca cctgcgtgtt 240

cttctgaggt tcatctttta catttgcttc tgctggataa ttttcagagg caacaaggaa 300

aaattagatg gcaaaaagtc gtctttcaag gaaaaatccc caccatcttt cgagatcccc 360

tgtaacttat tggcaactga aagaatgaaa aggaggaaaa tacaaaatat actagaactg 420

aaaaaaaaaa agtataaata gagacgatat atgccaatac ttcacaatgt tcgaatctat 480

tcttcatttg cagctattgt aaaataataa aacatcaaga acaaacaagc tcaacttgtc 540

ttttctaaga acaaagaata aacacaaaaa caaaaagttt ttttaatttt aatcaaaaat 600

tcgaa 605

<210> 10

<211> 612

<212> DNA

<213> SEQ ID NO.10

<400> 10

agatctacat ggcattacca ccatatacat atccatatct aatcttactt atatgttgtg 60

gaaatgtaaa gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta 120

atacgcttaa ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggcgac 180

agccctccga cggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa 240

acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt 300

tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc aaattaacga 360

atcaaattaa caaccatagg acgataatgc gattagtttt ttagccttat ttctggggta 420

attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc aaaagctgca 480

taaccacttt aactaatact ttcaacattt tcggtttgtg ttacttctta ttcaaatgtc 540

ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa 600

actatattcg aa 612

<210> 11

<211> 489

<212> DNA

<213> PsGAPp

<400> 11

agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat 60

atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa 120

cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc 180

gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc 240

aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc 300

ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga 360

gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt 420

ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac 480

tatttcgaa 489

<210> 12

<211> 937

<212> DNA

<213> PsAOX1p

<400> 12

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaa 937

<210> 13

<211> 609

<212> DNA

<213> PsFLD1p

<400> 13

agatctgcat gcaggaatct ctggcacggt gctaatggta gttatccaac ggagctgagg 60

tagtcgatat atctggatat gccgcctata ggataaaaac aggagagggt gaaccttgct 120

tatggctact agattgttct tgtactctga attctcatta tgggaaacta aactaatctc 180

atctgtgtgt tgcagtacta ttgaatcgtt gtagtatcta cctggagggc attccatgaa 240

ttagtgagat aacagagttg ggtaactaga gagaataata gacgtatgca tgattactac 300

acaacggatg tcgcactctt tccttagtta aaactatcat ccaatcacaa gatgcgggct 360

ggaaagactt gctcccgaag gataatcttc tgcttctatc tcccttcctc atatggtttc 420

gcagggctca tgccccttct tccttcgaac tgcccgatga ggaagtcctt agcctatcaa 480

agaattcggg accatcatcg atttttagag ccttacctga tcgcaatcag gatttcacta 540

ctcatataaa tacatcgctc aaagctccaa ctttgcttgt tcatacaatt cttgatattc 600

acattcgaa 609

<210> 14

<211> 886

<212> DNA

<213> G418-CYCTT

<400> 14

atgagccata ttcaacggga aacgtcttgt tcgaggccgc gattaaattc caacatggat 60

gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc 120

tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc 180

gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct 240

cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg 300

atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt 360

gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct 420

tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg 480

gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa 540

gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca 600

cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc 660

ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct 720

ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa 780

ttgcagtttc atttgatgct cgatgagttt ttctaacacg tccgacggcg gcccacgggt 840

cccaggcctc ggagatccgt cccccttttc ctttgtcgat atcatg 886

<210> 15

<211> 1874

<212> DNA

<213> CuARS

<400> 15

cgtctcacat gtgatcatgt ttcgtacaaa acaccgccac tccgttatga gaactcatag 60

ttatatttcg gggaaactta acttgcttgc aagcaataat agacagacaa atgttttacg 120

aaactggagg attaatacaa ttgatgcaaa aacaaaaaaa aaacaaaaaa aaaaaacaca 180

cgagaacata caaaaaaaga ctacaaaata tatatttgct aataccatca ccctcataca 240

aaaaaaagaa caaaacggaa ggagcacgta tatcttttct atgatctttg gatatggaat 300

agataagcaa cccctctagt gaagtacaca tcaagatact tggggaacaa ctgagaagac 360

atgctataca cgaaaaccat catctaatca tatttacata tatgaaacat gaaaagttat 420

tcatctattg gattcacttt tatatgtttt catgcattgt gcactctatg cttctgcttt 480

ttctctgttc ttttatactc ctctactatc aaattgagta tttcaattat caaagaaatt 540

tgagctggat cagaacttta gatattcatt tctgttttcg atatatttaa acgatgttct 600

aatgtcaaat ctcaatatcg ttgctaaaaa atctaacttt atagctttgt gcattaatgg 660

aaaaaaatac gtggatcttt tgaatgttta aaaacgagat actcttttgg aaaccagaaa 720

ataattttga aatacaaatt ggtgaatgta agcaagcaaa aattggatgc tgaagaaaat 780

aaatttatga actaaattat ctctgaacaa acatttagtc aaccttttct ccttgctcct 840

caatcaaagg tttttcgtat agatatatac gcaggttgct ttttgtttcc gctcataaaa 900

tcaagtttcg ataccaatgg agatttattc tttcgttttg attccataat aatcgaaatt 960

cttttaaaac actttgaagc gaactgaaat cccaatatct ttctgaccac taaaacaatt 1020

ataaagtact agatgttcaa aaagctttga gaattgccta gcttcaaaat atatttgttt 1080

gcaaatacta ttttattttt tcctttcttt gtcttctttg gttatttaaa atacgaagtt 1140

cgctacgttg gttttgagga accattactc tatttactca aatttattat caaagtgatt 1200

ttttttgctt gatctatcta aaggctgtgg atatgcccca gaaacaccat aaaagtcaaa 1260

tttctaaagc cagctcttac tataaagatt agagttcatt cgacttaatt gcaagtatgt 1320

agctcaaacc aaggttactc tataattata aaaagactag gaaccaatgc aagatcccct 1380

tatttcatac aaatttagta taactcaaat tcacattttc atatgcaaaa acttaaagaa 1440

aagcatcatt gtaaagagcc agttgttgta tattcaagtg ggcactgaac ttgtggttag 1500

attttatgga catcttgtca tcgaagtttt ttgatagaag ttgtgccatt gagaattgaa 1560

atataaaatg atattggatt gagaatatac gaatgtctta ttactcgtaa taataggaga 1620

actcatcgaa tatgtaaaat ataggctgat taaggtacat acagcatcct gttcatcatt 1680

tcaactattt aagtatcaaa ttagtgaaga aacgtattat gaaccattca ttgttcaata 1740

agtttactct tttttgtact tttttgtttt tgcttttcgg acattgtaat ttttactcat 1800

ttattcgatg tctctttagg tttttttttg tttttaaatc ttcgcggaat tgagataaga 1860

tcacatgtga gacg 1874

<210> 16

<211> 33

<212> DNA

<213> G418F-BbsI

<400> 16

gaagacgtca tgagccatat tcaacgggaa acg 33

<210> 17

<211> 47

<212> DNA

<213> G418R-CYCTTF

<400> 17

gtgggccgcc gtcggacgtg ttagaaaaac tcatcgagca tcaaatg 47

<210> 18

<211> 47

<212> DNA

<213> CYCTTF-G418R

<400> 18

catttgatgc tcgatgagtt tttctaacac gtccgacggc ggcccac 47

<210> 19

<211> 24

<212> DNA

<213> CYCTTR EcoRV

<400> 19

catgatatcg acaaaggaaa aggg 24

<210> 20

<211> 40

<212> DNA

<213> CuARSF

<400> 20

cgtctcacat gtgatcatgt ttcgtacaaa acaccgccac 40

<210> 21

<211> 38

<212> DNA

<213> CuARSR

<400> 21

cgtctcacat gtgatcttat ctcaattccg cgaagatt 38

<210> 22

<211> 33

<212> DNA

<213> CuARS-PscImutF

<400> 22

ggagaactca tcgaatatgt aaaatatagg ctg 33

<210> 23

<211> 33

<212> DNA

<213> CuARS-PscImutR

<400> 23

cagcctatat tttacatatt cgatgagttc tcc 33

<210> 24

<211> 43

<212> DNA

<213> Mxyn2-F

<400> 24

ctcgagaaaa gagaggctga agctatgccc acaggcctag agc 43

<210> 25

<211> 32

<212> DNA

<213> Mxyn2-R

<400> 25

catatgttag ttgctgacac tctgtgaggc ag 32

<210> 26

<211> 633

<212> DNA

<213> xyn2

<400> 26

atgcccacag gcctagagcc tgagagcagt gtcaacgtca cagagcgtgg catgtacgac 60

tttgttcttg gtgctcacaa tgatcatcgc cgtcgtgcaa gcatcaacta cgaccaaaac 120

taccaaactg gcggacaagt cagctattcg ccttccaaca ctggcttctc agtgaactgg 180

aacactcaag atgactttgt tgtgggcgtt ggttggacga ctggatcttc tgctcccatc 240

aactttggcg gctcttttag tgtcaacagc ggaactggcc tgctttccgt ctatggctgg 300

agcaccaacc cactggttga gtactacatc atggaggaca accacaacta cccagcacag 360

ggtaccgtca agggaaccgt caccagcgac ggagccactt acaccatctg ggagaatacc 420

cgtgtcaacg agccttccat ccagggcaca gcgaccttca accagtacat ttccgtgcgg 480

aactcgccca ggaccagcgg aactgttact gtgcagaacc acttcaatgc ttgggcctcg 540

cttggcctgc accttgggca gatgaactac caggttgtcg ctgtcgaagg ctggggtggt 600

agtggttctg cctcacagag tgtcagcaac taa 633

<210> 27

<211> 41

<212> DNA

<213> ScTEF1F

<400> 27

gctatacgaa gttatagatc tgcacacacc atagcttcaa a 41

<210> 28

<211> 43

<212> DNA

<213> ScTEF1R

<400> 28

ccgtcgactg cagaattcga attgtaatta aaacttagat tag 43

<210> 29

<211> 41

<212> DNA

<213> ScPGK1F

<400> 29

gctatacgaa gttatagatc tggaagtacc ttcaaagaat g 41

<210> 30

<211> 45

<212> DNA

<213> ScPGK1R

<400> 30

ccgtcgactg cagaattcga attgttttat atttgttgta aaaag 45

<210> 31

<211> 42

<212> DNA

<213> ScADH1F

<400> 31

gctatacgaa gttatagatc tagggggatc gaagaaatga tg 42

<210> 32

<211> 42

<212> DNA

<213> ScADH1R

<400> 32

ccgtcgactg cagaattcga atgtatatga gatagttgat tg 42

<210> 33

<211> 45

<212> DNA

<213> ScGAP1F

<400> 33

gctatacgaa gttatagatc taaagtcaca tcgcccatat ctctc 45

<210> 34

<211> 44

<212> DNA

<213> ScGAP1R

<400> 34

ccgtcgactg cagaattcga atttttattt cttttttttg tttc 44

<210> 35

<211> 41

<212> DNA

<213> ScHXT7F

<400> 35

gctatacgaa gttatagatc tttttacccc gtggaaatga g 41

<210> 36

<211> 44

<212> DNA

<213> ScHXT7R

<400> 36

ccgtcgactg cagaattcga atttttgatt aaaattaaaa aaac 44

<210> 37

<211> 43

<212> DNA

<213> ScGAL1F

<400> 37

gctatacgaa gttatagatc tacatggcat taccaccata tac 43

<210> 38

<211> 42

<212> DNA

<213> ScGAL1R

<400> 38

ccgtcgactg cagaattcga atatagtttt ttctccttga cg 42

<210> 39

<211> 43

<212> DNA

<213> ScTDH3F

<400> 39

gctatacgaa gttatagatc taaatagggg gcgggttaca cag 43

<210> 40

<211> 44

<212> DNA

<213> ScTDH3R

<400> 40

ccgtcgactg cagaattcga attctagaat ccgtcgaaac taag 44

<210> 41

<211> 39

<212> DNA

<213> PsFLD1F

<400> 41

gctatacgaa gttatagatc tgcatgcagg aatctctgg 39

<210> 42

<211> 50

<212> DNA

<213> PsFLD1R

<400> 42

ccgtcgactg cagaattcga atgtgaatat caagaattgt atgaacaagc 50

<210> 43

<211> 32

<212> DNA

<213> PsAOX1F

<400> 43

aaggtaccag atctaacatc caaagacgaa ag 32

<210> 44

<211> 33

<212> DNA

<213> PsAOX1R

<400> 44

gttgaattct ttcgaataat tagttgtttt ttg 33

<210> 45

<211> 47

<212> DNA

<213> CuMALF

<400> 45

gctatacgaa gttatagatc tgtatgaagg taggttttaa ggaggtg 47

<210> 46

<211> 46

<212> DNA

<213> CuMALR

<400> 46

ccgtcgactg cagaattcga ataagcgaga acttgttctt gttttt 46

<210> 47

<211> 46

<212> DNA

<213> CuPGKF

<400> 47

gctatacgaa gttatagatc taagcttttg tcttttagga gccttc 46

<210> 48

<211> 46

<212> DNA

<213> CuPGKR

<400> 48

ccgtcgactg cagaattcga actttatccg ccagtatgtt agtcca 46

<210> 49

<211> 42

<212> DNA

<213> CuGAPF

<400> 49

gctatacgaa gttatagatc tcttacagcg agcactcaaa tc 42

<210> 50

<211> 40

<212> DNA

<213> CuGAPR

<400> 50

ccgtcgactg cagaattcga atatgttgtt tgtaagtgtg 40

<210> 51

<211> 5940

<212> DNA

<213> pCuARS-CuGAP

<400> 51

atgcatgaga tcagatctct tacagcgagc actcaaatct gccctccgag ccctccggcc 60

ctctcttcaa caaactcgcg ctgcacttcg tcgtcagtgg tgccaatcac ccaacgtgga 120

ggtatcaaga ggtgctccag cccacaaagc gacatcaaag acaacaaccc tgccggccta 180

cgtcctacac accctggtga tcgcagacat tgtacaaggt gccacgcaat aacctacagg 240

caccgcacat gacgatggcc ttggttgtgc aaccagtgac ttccacggtc cacgcagcaa 300

catgaaccac accacccaga atcgatgcgc gcaacaacag ttgttccggt tcactcagcc 360

ccacagcgag tcgctggcag aacacgagcc tgagggcgga aagagggtag aggaaagcgc 420

aaggacaggg gacaacctgg cccaattgat gtcatataaa ccctctcgat caattgagca 480

cactcatccg ccaattgacc cctgttcgca gctccacgcc ccatgttcct cgtccctggt 540

gtagcttctc ccctaaattc cagcgcttgg ttccgccctc cctgtctccc gggtttaacg 600

aacgtgtgta ccatctgatg gtaatccgct cccgtccgcg caacacaact cacaagcaga 660

tcacacctct acacgccgct gctgatgcgc ccaatttaat ttttttttct ctcaatgtag 720

gggagaagcc ttgggagctc ccgactccca gttgggcaca gctgccacct catgactttt 780

cctgtgtgtg cctgtctgac gttacgtgtg atgtagtggc ccccgtccgg tgtgttttcg 840

cctgttgcgc tgtgcccccc ttaaaagtat aaaaggaagt gcaattgctg tttgtgttga 900

ttgttgatcc ttgtttcctc tgttttctcc tcatcacaca agaaaggttt cttctttcca 960

acagatacaa aacacactta caaacaacat attcgaaacg atgagatttc cttcaatttt 1020

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1080

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1140

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1200

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1260

tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg 1320

ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc 1380

atcatcatca tcatcattga gttttagcct tagacatgac tgttcctcag ttcaagttgg 1440

gcacttacga gaagaccggt cttgctagat tctaatcaag aggatgtcag aatgccattt 1500

gcctgagaga tgcaggcttc atttttgata cttttttatt tgtaacctat atagtatagg 1560

attttttttg tcattttgtt tcttctcgta cgagcttgct cctgatcagc ctatctcgca 1620

gctgatgaat atcttgtggt aggggtttgg gaaaatcatt cgagtttgat gtttttcttg 1680

gtatttccca ctcctcttca gagtacagaa gattaagtga gaccttcgtt tgtgcggatc 1740

ccccacacac catagcttca aaatgtttct actccttttt tactcttcca gattttctcg 1800

gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta aattttccct 1860

ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag 1920

accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt 1980

ttcttgaaat tttttttttt agtttttttc tctttcagtg acctccattg atatttaagt 2040

taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt 2100

tttttacttc ttgttcatta gaaagaaagc atagcaatct aatctaaggg cggtgttgac 2160

aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc 2220

atgagccata ttcaacggga aacgtcttgt tcgaggccgc gattaaattc caacatggat 2280

gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc 2340

tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc 2400

gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct 2460

cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg 2520

atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt 2580

gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct 2640

tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg 2700

gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa 2760

gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca 2820

cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc 2880

ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct 2940

ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa 3000

ttgcagtttc atttgatgct cgatgagttt ttctaacacg tccgacggcg gcccacgggt 3060

cccaggcctc ggagatccgt cccccttttc ctttgtcgat atcatgtaat tagttatgtc 3120

acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca 3180

acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt 3240

tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact 3300

gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca agctggagac 3360

caacatgtga tcatgtttcg tacaaaacac cgccactccg ttatgagaac tcatagttat 3420

atttcgggga aacttaactt gcttgcaagc aataatagac agacaaatgt tttacgaaac 3480

tggaggatta atacaattga tgcaaaaaca aaaaaaaaac aaaaaaaaaa aacacacgag 3540

aacatacaaa aaaagactac aaaatatata tttgctaata ccatcaccct catacaaaaa 3600

aaagaacaaa acggaaggag cacgtatatc ttttctatga tctttggata tggaatagat 3660

aagcaacccc tctagtgaag tacacatcaa gatacttggg gaacaactga gaagacatgc 3720

tatacacgaa aaccatcatc taatcatatt tacatatatg aaacatgaaa agttattcat 3780

ctattggatt cacttttata tgttttcatg cattgtgcac tctatgcttc tgctttttct 3840

ctgttctttt atactcctct actatcaaat tgagtatttc aattatcaaa gaaatttgag 3900

ctggatcaga actttagata ttcatttctg ttttcgatat atttaaacga tgttctaatg 3960

tcaaatctca atatcgttgc taaaaaatct aactttatag ctttgtgcat taatggaaaa 4020

aaatacgtgg atcttttgaa tgtttaaaaa cgagatactc ttttggaaac cagaaaataa 4080

ttttgaaata caaattggtg aatgtaagca agcaaaaatt ggatgctgaa gaaaataaat 4140

ttatgaacta aattatctct gaacaaacat ttagtcaacc ttttctcctt gctcctcaat 4200

caaaggtttt tcgtatagat atatacgcag gttgcttttt gtttccgctc ataaaatcaa 4260

gtttcgatac caatggagat ttattctttc gttttgattc cataataatc gaaattcttt 4320

taaaacactt tgaagcgaac tgaaatccca atatctttct gaccactaaa acaattataa 4380

agtactagat gttcaaaaag ctttgagaat tgcctagctt caaaatatat ttgtttgcaa 4440

atactatttt attttttcct ttctttgtct tctttggtta tttaaaatac gaagttcgct 4500

acgttggttt tgaggaacca ttactctatt tactcaaatt tattatcaaa gtgatttttt 4560

ttgcttgatc tatctaaagg ctgtggatat gccccagaaa caccataaaa gtcaaatttc 4620

taaagccagc tcttactata aagattagag ttcattcgac ttaattgcaa gtatgtagct 4680

caaaccaagg ttactctata attataaaaa gactaggaac caatgcaaga tccccttatt 4740

tcatacaaat ttagtataac tcaaattcac attttcatat gcaaaaactt aaagaaaagc 4800

atcattgtaa agagccagtt gttgtatatt caagtgggca ctgaacttgt ggttagattt 4860

tatggacatc ttgtcatcga agttttttga tagaagttgt gccattgaga attgaaatat 4920

aaaatgatat tggattgaga atatacgaat gtcttattac tcgtaataat aggagaactc 4980

atcgaatatg taaaatatag gctgattaag gtacatacag catcctgttc atcatttcaa 5040

ctatttaagt atcaaattag tgaagaaacg tattatgaac cattcattgt tcaataagtt 5100

tactcttttt tgtacttttt tgtttttgct tttcggacat tgtaattttt actcatttat 5160

tcgatgtctc tttaggtttt tttttgtttt taaatcttcg cggaattgag ataagatcac 5220

atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 5280

ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 5340

cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 5400

tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 5460

gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 5520

aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 5580

tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 5640

aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 5700

aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 5760

ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 5820

ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 5880

atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 5940

<210> 52

<211> 6313

<212> DNA

<213> pCuARS-CuPGK

<400> 52

aagggatttt ggtcatgcat gagatcagat ctaagctttt gtcttttagg agccttcttt 60

tcaccctggc tttcttcaga ctccacgcct ctcgcccgtt tgttgttgat cttcttctgt 120

tgcttcttcg tgagcttacc agtatccaga tgcgttgtca gggcaagagg gtcatgttca 180

agctcctctt tcactttcag tccaatacgt ttccaggcag ggatgtgttc gctcatcgtt 240

ccagactcga gtggtgaaaa ctatggcaac ctctacttcc tttccaaaca cacagcgtgc 300

tttgtagtgt gtgcctaaga gctgaatttt ttttccttcc atgctgcgct gcgatgagct 360

ctgcccgccc gcagcctcgg aggctagcga cgtataaaaa aggcctgtga aaattttatc 420

ctcctcctta acgacccttc tttctcttct tcacattcaa aaacttcaag cagctgtctc 480

tgttcctttg ctgtgttcta ccattggata ttcccattcc ccgtggagaa ccgaactgga 540

gtctagcagc atgcgagatc aatattacac ggtttgagtc tgatacgctt gagcagccat 600

tttttggctt ctcctggtgt gtatccagat atagaagttc gtatacattt cccatagcga 660

ttgtaaaatg attctgcaat ggaaccatcc gtaattgtag gcctgctgag atggcactcg 720

caatgcctct gtgtctggtt ttttgccttc tccgtccatc agcaccagtg gcttcttagg 780

gcataacgag acggctcctt ggtgaaagat gccctgctcc gtctgtctgc ctgttgctac 840

aaccactgcg tagtcagatg acccggtctg tgtgctgtgg aatcaccggg agcgaaattc 900

cggtttcgct ggcagatgag ctcatcaacc acatcaactg gagcaacctc accagaggac 960

acgtaacctg cccggttgaa ttctgtcaaa ccgtacatca cacaacaaca gcagcagcaa 1020

caacaacaac gtcagttgtc gttcgcatgg cgacgttacc taacggcacc aacatcgtct 1080

cgtcctcgcc agtgcctgtt tcccctaccc ggagtggccc ggcccacctg tcgttctttt 1140

ttcgtcaatt gtgtccagct ggtgccatca ccatatgttc aagtgcgtgg cctgtactag 1200

cgcagtctgc tgcagtataa aagggattgc tgaggccccc tttagcgttt ccaattaaca 1260

attgattccc ttttccccat agtccgtttg tactacatcc tacataacaa aagtgagtgt 1320

tacaagacaa gtgtggcggt caattggatc atttggacta acatactggc ggataaagtt 1380

cgaaacgatg agatttcctt caatttttac tgctgtttta ttcgcagcat cctccgcatt 1440

agctgctcca gtcaacacta caacagaaga tgaaacggca caaattccgg ctgaagctgt 1500

catcggttac tcagatttag aaggggattt cgatgttgct gttttgccat tttccaacag 1560

cacaaataac gggttattgt ttataaatac tactattgcc agcattgctg ctaaagaaga 1620

aggggtatct ctcgagaaaa gagaggctga agctgaattc acgtggccca gccggccgtc 1680

tcggatcggt acctcgagcc gcggcggccg ccagctttct agaacaaaaa ctcatctcag 1740

aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt ttagccttag 1800

acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt gctagattct 1860

aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt tttgatactt 1920

ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct tctcgtacga 1980

gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa 2040

aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag tacagaagat 2100

taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa tgtttctact 2160

ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac 2220

ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt aattacccgt 2280

actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg 2340

caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt ttttttctct 2400

ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt 2460

ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa agaaagcata 2520

gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat cggcatagta 2580

taatacgaca aggtgaggaa ctaaaccatg agccatattc aacgggaaac gtcttgttcg 2640

aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat 2700

aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag 2760

ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga 2820

ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct 2880

gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa 2940

gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg 3000

cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag 3060

gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat 3120

ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat 3180

tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta 3240

ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc 3300

ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat 3360

ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc 3420

taacacgtcc gacggcggcc cacgggtccc aggcctcgga gatccgtccc ccttttcctt 3480

tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc 3540

tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat 3600

agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag 3660

acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc 3720

gaaggcttta atttgcaagc tggagaccaa catgtgatca tgtttcgtac aaaacaccgc 3780

cactccgtta tgagaactca tagttatatt tcggggaaac ttaacttgct tgcaagcaat 3840

aatagacaga caaatgtttt acgaaactgg aggattaata caattgatgc aaaaacaaaa 3900

aaaaaacaaa aaaaaaaaac acacgagaac atacaaaaaa agactacaaa atatatattt 3960

gctaatacca tcaccctcat acaaaaaaaa gaacaaaacg gaaggagcac gtatatcttt 4020

tctatgatct ttggatatgg aatagataag caacccctct agtgaagtac acatcaagat 4080

acttggggaa caactgagaa gacatgctat acacgaaaac catcatctaa tcatatttac 4140

atatatgaaa catgaaaagt tattcatcta ttggattcac ttttatatgt tttcatgcat 4200

tgtgcactct atgcttctgc tttttctctg ttcttttata ctcctctact atcaaattga 4260

gtatttcaat tatcaaagaa atttgagctg gatcagaact ttagatattc atttctgttt 4320

tcgatatatt taaacgatgt tctaatgtca aatctcaata tcgttgctaa aaaatctaac 4380

tttatagctt tgtgcattaa tggaaaaaaa tacgtggatc ttttgaatgt ttaaaaacga 4440

gatactcttt tggaaaccag aaaataattt tgaaatacaa attggtgaat gtaagcaagc 4500

aaaaattgga tgctgaagaa aataaattta tgaactaaat tatctctgaa caaacattta 4560

gtcaaccttt tctccttgct cctcaatcaa aggtttttcg tatagatata tacgcaggtt 4620

gctttttgtt tccgctcata aaatcaagtt tcgataccaa tggagattta ttctttcgtt 4680

ttgattccat aataatcgaa attcttttaa aacactttga agcgaactga aatcccaata 4740

tctttctgac cactaaaaca attataaagt actagatgtt caaaaagctt tgagaattgc 4800

ctagcttcaa aatatatttg tttgcaaata ctattttatt ttttcctttc tttgtcttct 4860

ttggttattt aaaatacgaa gttcgctacg ttggttttga ggaaccatta ctctatttac 4920

tcaaatttat tatcaaagtg attttttttg cttgatctat ctaaaggctg tggatatgcc 4980

ccagaaacac cataaaagtc aaatttctaa agccagctct tactataaag attagagttc 5040

attcgactta attgcaagta tgtagctcaa accaaggtta ctctataatt ataaaaagac 5100

taggaaccaa tgcaagatcc ccttatttca tacaaattta gtataactca aattcacatt 5160

ttcatatgca aaaacttaaa gaaaagcatc attgtaaaga gccagttgtt gtatattcaa 5220

gtgggcactg aacttgtggt tagattttat ggacatcttg tcatcgaagt tttttgatag 5280

aagttgtgcc attgagaatt gaaatataaa atgatattgg attgagaata tacgaatgtc 5340

ttattactcg taataatagg agaactcatc gaatatgtaa aatataggct gattaaggta 5400

catacagcat cctgttcatc atttcaacta tttaagtatc aaattagtga agaaacgtat 5460

tatgaaccat tcattgttca ataagtttac tcttttttgt acttttttgt ttttgctttt 5520

cggacattgt aatttttact catttattcg atgtctcttt aggttttttt ttgtttttaa 5580

atcttcgcgg aattgagata agatcacatg tgagcaaaag gccagcaaaa ggccaggaac 5640

cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 5700

aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 5760

tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 5820

ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 5880

ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 5940

cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 6000

ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 6060

gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 6120

atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 6180

aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 6240

aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 6300

gaaaactcac gtt 6313

<210> 53

<211> 6433

<212> DNA

<213> pCuARS-CuMAL

<400> 53

taagggattt tggtcatgca tgagatcaga tctgtatgaa ggtaggtttt aaggaggtgt 60

ctaatagtcc ttgtattgac tggttttgtt ttttgaatct ctaaagggag gctcgagtcc 120

ttccaattgg ggtggttttt aatgagcatt agagatgtac aattgcgggg tgtatgtaga 180

ggagatcacc tttggatgat gttttctcaa aaactgttca acatttcttc ccaagagcaa 240

ttgactttta attgtttctc cgacacccca aattaattct ccgtcttctg cagccatttg 300

cgtcggtcta gatgaagtga tccttgtaca tgggaagttt gtgtgcactg gaatttggaa 360

tctacactaa cccctcgtaa gagatcgatc agcataagat aatgcaaatt gggggggggg 420

ggaaattatc aattgtaaac tcccgtttga attgatccaa ttggcaattg gatatgttct 480

tatttaggga tgtgggataa gtttcactgt agaactggct tctcgctctt agaacgggtc 540

atcagtttga agccttttcg atcacttggc atttaacact tgaactgttg aacatagtct 600

gacaactaac acgttgtttc ggaaaactca tgactgtttt tcatgttaca tatttgctgc 660

atgtatttat gcagatgttc atattattcc accccgagcc tttatttgta gaacttgaac 720

ggaatagatt gtaaggaaat gacacttatg tgcatgaatc aaggattaaa ttctatgatc 780

aaggcgtaga cactccagta gaaaacattg ctatggttgt gtgctgcact tccataaaat 840

cacccgacta agcatcgtac tctaaattta cgtccactaa tgaggagaag tagggagaga 900

ttttggatac gaaaagatat tttcaacccc atgcacatgg ggcggttaaa ttttccaaca 960

attaatccaa agcgctaaaa tattaaccca cccgaaactg ttgatacaac agggataata 1020

attgttgaaa agaaccgcaa cggttatccg ttgcgacgaa tacgtcgagt aaggagccta 1080

gcacaacccc ttttggattg acaaatttgg aattatcttc tgtctcctag ttttagacag 1140

aggaccatct cggggcttgg aaacaaggtg ctatgtacca agctgcacct ccattattcc 1200

gtttgcataa aatttcaaac aaggttcaac ttggtgtcca gccgttcatg acatggtact 1260

gatttcccca agcctctatc ctgaacgggt gtgtgacaac ctcatagaga ttctcatgta 1320

aatcttcgat cattcatgaa tgttgctgcc ccaagaattg atgccatgtg atcgttgaat 1380

tccagcgata attcttccga ttgcgttgtc aactcagcgt atatattgca attagatcac 1440

aatagtattt tctaaccctc caaagtgaat caaaaacaag aacaagttct cgcttattat 1500

tcgaaacgat gagatttcct tcaattttta ctgctgtttt attcgcagca tcctccgcat 1560

tagctgctcc agtcaacact acaacagaag atgaaacggc acaaattccg gctgaagctg 1620

tcatcggtta ctcagattta gaaggggatt tcgatgttgc tgttttgcca ttttccaaca 1680

gcacaaataa cgggttattg tttataaata ctactattgc cagcattgct gctaaagaag 1740

aaggggtatc tctcgagaaa agagaggctg aagctgaatt cacgtggccc agccggccgt 1800

ctcggatcgg tacctcgagc cgcggcggcc gccagctttc tagaacaaaa actcatctca 1860

gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt tttagcctta 1920

gacatgactg ttcctcagtt caagttgggc acttacgaga agaccggtct tgctagattc 1980

taatcaagag gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact 2040

tttttatttg taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg 2100

agcttgctcc tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga 2160

aaatcattcg agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga 2220

ttaagtgaga ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac 2280

tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca 2340

cccaagcaca gcatactaaa ttttccctct ttcttcctct agggtgtcgt taattacccg 2400

tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag 2460

gcaataaaaa tttttatcac gtttcttttt cttgaaattt ttttttttag tttttttctc 2520

tttcagtgac ctccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag 2580

tttcattttt cttgttctat tacaactttt tttacttctt gttcattaga aagaaagcat 2640

agcaatctaa tctaagggcg gtgttgacaa ttaatcatcg gcatagtata tcggcatagt 2700

ataatacgac aaggtgagga actaaaccat gagccatatt caacgggaaa cgtcttgttc 2760

gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga 2820

taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga 2880

gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag 2940

actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc 3000

tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga 3060

agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt 3120

gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca 3180

ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa 3240

tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga 3300

ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt 3360

aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat 3420

cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata 3480

tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt 3540

ctaacacgtc cgacggcggc ccacgggtcc caggcctcgg agatccgtcc cccttttcct 3600

ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg 3660

ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta 3720

tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca 3780

gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct 3840

cgaaggcttt aatttgcaag ctggagacca acatgtgatc atgtttcgta caaaacaccg 3900

ccactccgtt atgagaactc atagttatat ttcggggaaa cttaacttgc ttgcaagcaa 3960

taatagacag acaaatgttt tacgaaactg gaggattaat acaattgatg caaaaacaaa 4020

aaaaaaacaa aaaaaaaaaa cacacgagaa catacaaaaa aagactacaa aatatatatt 4080

tgctaatacc atcaccctca tacaaaaaaa agaacaaaac ggaaggagca cgtatatctt 4140

ttctatgatc tttggatatg gaatagataa gcaacccctc tagtgaagta cacatcaaga 4200

tacttgggga acaactgaga agacatgcta tacacgaaaa ccatcatcta atcatattta 4260

catatatgaa acatgaaaag ttattcatct attggattca cttttatatg ttttcatgca 4320

ttgtgcactc tatgcttctg ctttttctct gttcttttat actcctctac tatcaaattg 4380

agtatttcaa ttatcaaaga aatttgagct ggatcagaac tttagatatt catttctgtt 4440

ttcgatatat ttaaacgatg ttctaatgtc aaatctcaat atcgttgcta aaaaatctaa 4500

ctttatagct ttgtgcatta atggaaaaaa atacgtggat cttttgaatg tttaaaaacg 4560

agatactctt ttggaaacca gaaaataatt ttgaaataca aattggtgaa tgtaagcaag 4620

caaaaattgg atgctgaaga aaataaattt atgaactaaa ttatctctga acaaacattt 4680

agtcaacctt ttctccttgc tcctcaatca aaggtttttc gtatagatat atacgcaggt 4740

tgctttttgt ttccgctcat aaaatcaagt ttcgatacca atggagattt attctttcgt 4800

tttgattcca taataatcga aattctttta aaacactttg aagcgaactg aaatcccaat 4860

atctttctga ccactaaaac aattataaag tactagatgt tcaaaaagct ttgagaattg 4920

cctagcttca aaatatattt gtttgcaaat actattttat tttttccttt ctttgtcttc 4980

tttggttatt taaaatacga agttcgctac gttggttttg aggaaccatt actctattta 5040

ctcaaattta ttatcaaagt gatttttttt gcttgatcta tctaaaggct gtggatatgc 5100

cccagaaaca ccataaaagt caaatttcta aagccagctc ttactataaa gattagagtt 5160

cattcgactt aattgcaagt atgtagctca aaccaaggtt actctataat tataaaaaga 5220

ctaggaacca atgcaagatc cccttatttc atacaaattt agtataactc aaattcacat 5280

tttcatatgc aaaaacttaa agaaaagcat cattgtaaag agccagttgt tgtatattca 5340

agtgggcact gaacttgtgg ttagatttta tggacatctt gtcatcgaag ttttttgata 5400

gaagttgtgc cattgagaat tgaaatataa aatgatattg gattgagaat atacgaatgt 5460

cttattactc gtaataatag gagaactcat cgaatatgta aaatataggc tgattaaggt 5520

acatacagca tcctgttcat catttcaact atttaagtat caaattagtg aagaaacgta 5580

ttatgaacca ttcattgttc aataagttta ctcttttttg tacttttttg tttttgcttt 5640

tcggacattg taatttttac tcatttattc gatgtctctt taggtttttt tttgttttta 5700

aatcttcgcg gaattgagat aagatcacat gtgagcaaaa ggccagcaaa aggccaggaa 5760

ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5820

caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 5880

gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5940

cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 6000

tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6060

gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6120

cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6180

tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6240

tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6300

caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6360

aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6420

cgaaaactca cgt 6433

<210> 54

<211> 5367

<212> DNA

<213> pCuARS-ScGAP1

<400> 54

ggtcatgcat gagatcagat ctaaagtcac atcgcccata tctctcgagt gcgataacgg 60

ggaacttgat ttggtaactg ataagattgt taaatgtcag tttggatgct ttttcttacg 120

tccgattagc ttatcttctg gagcaaccgg ctatttacct cctcatagta aaataaacat 180

gataagcgca tagttggggc aacacacctt tcttccggaa ttcgctctgg atgagacata 240

taaagatgaa ggtgaagtcc acttaaatga atgtcaatga gacgatgttt tttctcctag 300

attgattttt gaattccttg tatacaaagt cttgttttct tattgtcctc aacaaaacaa 360

aagtagaaaa gaacagatca aggacagcaa catttataag aaacaaaaaa aagaaataaa 420

aattcgaaac gatgagattt ccttcaattt ttactgctgt tttattcgca gcatcctccg 480

cattagctgc tccagtcaac actacaacag aagatgaaac ggcacaaatt ccggctgaag 540

ctgtcatcgg ttactcagat ttagaagggg atttcgatgt tgctgttttg ccattttcca 600

acagcacaaa taacgggtta ttgtttataa atactactat tgccagcatt gctgctaaag 660

aagaaggggt atctctcgag aaaagagagg ctgaagctga attcacgtgg cccagccggc 720

cgtctcggat cggtacctcg agccgcggcg gccgccagct ttctagaaca aaaactcatc 780

tcagaagagg atctgaatag cgccgtcgac catcatcatc atcatcattg agttttagcc 840

ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg tcttgctaga 900

ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt catttttgat 960

acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt ttcttctcgt 1020

acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg taggggtttg 1080

ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc agagtacaga 1140

agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc aaaatgtttc 1200

tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa 1260

acacccaagc acagcatact aaattttccc tctttcttcc tctagggtgt cgttaattac 1320

ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa 1380

aaggcaataa aaatttttat cacgtttctt tttcttgaaa tttttttttt tagttttttt 1440

ctctttcagt gacctccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt 1500

cagtttcatt tttcttgttc tattacaact ttttttactt cttgttcatt agaaagaaag 1560

catagcaatc taatctaagg gcggtgttga caattaatca tcggcatagt atatcggcat 1620

agtataatac gacaaggtga ggaactaaac catgagccat attcaacggg aaacgtcttg 1680

ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg 1740

cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc 1800

agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt 1860

cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac 1920

tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt 1980

agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg 2040

gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc 2100

tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg 2160

taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc 2220

ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa 2280

attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc 2340

catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa 2400

atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt 2460

tttctaacac gtccgacggc ggcccacggg tcccaggcct cggagatccg tccccctttt 2520

cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat 2580

ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt 2640

ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt 2700

acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac 2760

gctcgaaggc tttaatttgc aagctggaga ccaacatgtg atcatgtttc gtacaaaaca 2820

ccgccactcc gttatgagaa ctcatagtta tatttcgggg aaacttaact tgcttgcaag 2880

caataataga cagacaaatg ttttacgaaa ctggaggatt aatacaattg atgcaaaaac 2940

aaaaaaaaaa caaaaaaaaa aaacacacga gaacatacaa aaaaagacta caaaatatat 3000

atttgctaat accatcaccc tcatacaaaa aaaagaacaa aacggaagga gcacgtatat 3060

cttttctatg atctttggat atggaataga taagcaaccc ctctagtgaa gtacacatca 3120

agatacttgg ggaacaactg agaagacatg ctatacacga aaaccatcat ctaatcatat 3180

ttacatatat gaaacatgaa aagttattca tctattggat tcacttttat atgttttcat 3240

gcattgtgca ctctatgctt ctgctttttc tctgttcttt tatactcctc tactatcaaa 3300

ttgagtattt caattatcaa agaaatttga gctggatcag aactttagat attcatttct 3360

gttttcgata tatttaaacg atgttctaat gtcaaatctc aatatcgttg ctaaaaaatc 3420

taactttata gctttgtgca ttaatggaaa aaaatacgtg gatcttttga atgtttaaaa 3480

acgagatact cttttggaaa ccagaaaata attttgaaat acaaattggt gaatgtaagc 3540

aagcaaaaat tggatgctga agaaaataaa tttatgaact aaattatctc tgaacaaaca 3600

tttagtcaac cttttctcct tgctcctcaa tcaaaggttt ttcgtataga tatatacgca 3660

ggttgctttt tgtttccgct cataaaatca agtttcgata ccaatggaga tttattcttt 3720

cgttttgatt ccataataat cgaaattctt ttaaaacact ttgaagcgaa ctgaaatccc 3780

aatatctttc tgaccactaa aacaattata aagtactaga tgttcaaaaa gctttgagaa 3840

ttgcctagct tcaaaatata tttgtttgca aatactattt tattttttcc tttctttgtc 3900

ttctttggtt atttaaaata cgaagttcgc tacgttggtt ttgaggaacc attactctat 3960

ttactcaaat ttattatcaa agtgattttt tttgcttgat ctatctaaag gctgtggata 4020

tgccccagaa acaccataaa agtcaaattt ctaaagccag ctcttactat aaagattaga 4080

gttcattcga cttaattgca agtatgtagc tcaaaccaag gttactctat aattataaaa 4140

agactaggaa ccaatgcaag atccccttat ttcatacaaa tttagtataa ctcaaattca 4200

cattttcata tgcaaaaact taaagaaaag catcattgta aagagccagt tgttgtatat 4260

tcaagtgggc actgaacttg tggttagatt ttatggacat cttgtcatcg aagttttttg 4320

atagaagttg tgccattgag aattgaaata taaaatgata ttggattgag aatatacgaa 4380

tgtcttatta ctcgtaataa taggagaact catcgaatat gtaaaatata ggctgattaa 4440

ggtacataca gcatcctgtt catcatttca actatttaag tatcaaatta gtgaagaaac 4500

gtattatgaa ccattcattg ttcaataagt ttactctttt ttgtactttt ttgtttttgc 4560

ttttcggaca ttgtaatttt tactcattta ttcgatgtct ctttaggttt ttttttgttt 4620

ttaaatcttc gcggaattga gataagatca catgtgagca aaaggccagc aaaaggccag 4680

gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4740

tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 4800

ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 4860

atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag 4920

gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 4980

tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5040

cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5100

cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 5160

tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 5220

cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 5280

cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5340

gaacgaaaac tcacgttaag ggatttt 5367

<210> 55

<211> 5386

<212> DNA

<213> pCuARS-ScTEF1

<400> 55

tcacgttaag ggattttggt catgcatgag atcagatctg cacacaccat agcttcaaaa 60

tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac 120

ttcaaaacac ccaagcacag catactaaat tccccctctt tcttcctcta gggtggcgtt 180

aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg 240

tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt tttttttggt 300

ttttttctct ttcgatgacc tcccattgat atttaagtta ataaatggtc ttcaatttct 360

caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga 420

aagaaagcat agcaatctaa tctaagtttt aattacaatt cgaaacgatg agatttcctt 480

caatttttac tgctgtttta ttcgcagcat cctccgcatt agctgctcca gtcaacacta 540

caacagaaga tgaaacggca caaattccgg ctgaagctgt catcggttac tcagatttag 600

aaggggattt cgatgttgct gttttgccat tttccaacag cacaaataac gggttattgt 660

ttataaatac tactattgcc agcattgctg ctaaagaaga aggggtatct ctcgagaaaa 720

gagaggctga agctgaattc acgtggccca gccggccgtc tcggatcggt acctcgagcc 780

gcggcggccg ccagctttct agaacaaaaa ctcatctcag aagaggatct gaatagcgcc 840

gtcgaccatc atcatcatca tcattgagtt ttagccttag acatgactgt tcctcagttc 900

aagttgggca cttacgagaa gaccggtctt gctagattct aatcaagagg atgtcagaat 960

gccatttgcc tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata 1020

gtataggatt ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta 1080

tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt 1140

tttcttggta tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt 1200

gcggatcccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat 1260

tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat 1320

tttccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa 1380

aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg 1440

tttctttttc ttgaaatttt tttttttagt ttttttctct ttcagtgacc tccattgata 1500

tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt 1560

acaacttttt ttacttcttg ttcattagaa agaaagcata gcaatctaat ctaagggcgg 1620

tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa 1680

ctaaaccatg agccatattc aacgggaaac gtcttgttcg aggccgcgat taaattccaa 1740

catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc 1800

gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa 1860

aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt 1920

tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac 1980

cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga 2040

aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa 2100

ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa 2160

cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt 2220

ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga 2280

tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg 2340

acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga 2400

gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat 2460

gaataaattg cagtttcatt tgatgctcga tgagtttttc taacacgtcc gacggcggcc 2520

cacgggtccc aggcctcgga gatccgtccc ccttttcctt tgtcgatatc atgtaattag 2580

ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag 2640

ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa 2700

cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca 2760

ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc 2820

tggagaccaa catgtgatca tgtttcgtac aaaacaccgc cactccgtta tgagaactca 2880

tagttatatt tcggggaaac ttaacttgct tgcaagcaat aatagacaga caaatgtttt 2940

acgaaactgg aggattaata caattgatgc aaaaacaaaa aaaaaacaaa aaaaaaaaac 3000

acacgagaac atacaaaaaa agactacaaa atatatattt gctaatacca tcaccctcat 3060

acaaaaaaaa gaacaaaacg gaaggagcac gtatatcttt tctatgatct ttggatatgg 3120

aatagataag caacccctct agtgaagtac acatcaagat acttggggaa caactgagaa 3180

gacatgctat acacgaaaac catcatctaa tcatatttac atatatgaaa catgaaaagt 3240

tattcatcta ttggattcac ttttatatgt tttcatgcat tgtgcactct atgcttctgc 3300

tttttctctg ttcttttata ctcctctact atcaaattga gtatttcaat tatcaaagaa 3360

atttgagctg gatcagaact ttagatattc atttctgttt tcgatatatt taaacgatgt 3420

tctaatgtca aatctcaata tcgttgctaa aaaatctaac tttatagctt tgtgcattaa 3480

tggaaaaaaa tacgtggatc ttttgaatgt ttaaaaacga gatactcttt tggaaaccag 3540

aaaataattt tgaaatacaa attggtgaat gtaagcaagc aaaaattgga tgctgaagaa 3600

aataaattta tgaactaaat tatctctgaa caaacattta gtcaaccttt tctccttgct 3660

cctcaatcaa aggtttttcg tatagatata tacgcaggtt gctttttgtt tccgctcata 3720

aaatcaagtt tcgataccaa tggagattta ttctttcgtt ttgattccat aataatcgaa 3780

attcttttaa aacactttga agcgaactga aatcccaata tctttctgac cactaaaaca 3840

attataaagt actagatgtt caaaaagctt tgagaattgc ctagcttcaa aatatatttg 3900

tttgcaaata ctattttatt ttttcctttc tttgtcttct ttggttattt aaaatacgaa 3960

gttcgctacg ttggttttga ggaaccatta ctctatttac tcaaatttat tatcaaagtg 4020

attttttttg cttgatctat ctaaaggctg tggatatgcc ccagaaacac cataaaagtc 4080

aaatttctaa agccagctct tactataaag attagagttc attcgactta attgcaagta 4140

tgtagctcaa accaaggtta ctctataatt ataaaaagac taggaaccaa tgcaagatcc 4200

ccttatttca tacaaattta gtataactca aattcacatt ttcatatgca aaaacttaaa 4260

gaaaagcatc attgtaaaga gccagttgtt gtatattcaa gtgggcactg aacttgtggt 4320

tagattttat ggacatcttg tcatcgaagt tttttgatag aagttgtgcc attgagaatt 4380

gaaatataaa atgatattgg attgagaata tacgaatgtc ttattactcg taataatagg 4440

agaactcatc gaatatgtaa aatataggct gattaaggta catacagcat cctgttcatc 4500

atttcaacta tttaagtatc aaattagtga agaaacgtat tatgaaccat tcattgttca 4560

ataagtttac tcttttttgt acttttttgt ttttgctttt cggacattgt aatttttact 4620

catttattcg atgtctcttt aggttttttt ttgtttttaa atcttcgcgg aattgagata 4680

agatcacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 4740

ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 4800

gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 4860

cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 4920

gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 4980

tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 5040

cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 5100

cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 5160

gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 5220

agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 5280

cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 5340

tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaac 5386

<210> 56

<211> 5507

<212> DNA

<213> pCUARS-ScTDH3

<400> 56

agatcagatc taaatagggg gcgggttaca cagaatatat aacatcatag gtgtctgggt 60

gaacagttta ttcctggcat ccactaaata taatggagcc cgctttttaa gctggcatcc 120

agaaaaaaaa agaatcccag caccaaaata ttgttttctt caccaaccat cagttcatag 180

gtccattctc ttagcgcaac tacagagaac agggacacaa acaggcaaaa aacgggcaca 240

acctcaatgg agtgatgcaa cctgcctgga gtaaatgatg acacaaggca attgacccac 300

gcatgtatct atctcatttt cttacacctt ctattacctt ctgctctctc tgatttggaa 360

aaagctgaaa aaaaaggttg aaaccagttc cctgaaatta ttcccctact tgactaataa 420

gtatataaag acggtaggta ttgattgtaa ttctgtaaat ctatttctta aacttcttaa 480

attctacttt tagagttagt ctttttttta gttttaaaac accaagaact tagtttcgac 540

ggattctaga attcgaaacg atgagatttc cttcaatttt tactgctgtt ttattcgcag 600

catcctccgc attagctgct ccagtcaaca ctacaacaga agatgaaacg gcacaaattc 660

cggctgaagc tgtcatcggt tactcagatt tagaagggga tttcgatgtt gctgttttgc 720

cattttccaa cagcacaaat aacgggttat tgtttataaa tactactatt gccagcattg 780

ctgctaaaga agaaggggta tctctcgaga aaagagaggc tgaagctgaa ttcacgtggc 840

ccagccggcc gtctcggatc ggtacctcga gccgcggcgg ccgccagctt tctagaacaa 900

aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga 960

gttttagcct tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt 1020

cttgctagat tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc 1080

atttttgata cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt 1140

tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt 1200

aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca 1260

gagtacagaa gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca 1320

aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac 1380

cacttcaaaa cacccaagca cagcatacta aattttccct ctttcttcct ctagggtgtc 1440

gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct 1500

tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaat tttttttttt 1560

agtttttttc tctttcagtg acctccattg atatttaagt taataaacgg tcttcaattt 1620

ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc ttgttcatta 1680

gaaagaaagc atagcaatct aatctaaggg cggtgttgac aattaatcat cggcatagta 1740

tatcggcata gtataatacg acaaggtgag gaactaaacc atgagccata ttcaacggga 1800

aacgtcttgt tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa 1860

atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc 1920

cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga 1980

tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt 2040

tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt 2100

ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt 2160

cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt 2220

tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga 2280

tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc 2340

attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga 2400

cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca 2460

ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct 2520

ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct 2580

cgatgagttt ttctaacacg tccgacggcg gcccacgggt cccaggcctc ggagatccgt 2640

cccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc 2700

cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta 2760

tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt 2820

tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg 2880

ttttgggacg ctcgaaggct ttaatttgca agctggagac caacatgtga tcatgtttcg 2940

tacaaaacac cgccactccg ttatgagaac tcatagttat atttcgggga aacttaactt 3000

gcttgcaagc aataatagac agacaaatgt tttacgaaac tggaggatta atacaattga 3060

tgcaaaaaca aaaaaaaaac aaaaaaaaaa aacacacgag aacatacaaa aaaagactac 3120

aaaatatata tttgctaata ccatcaccct catacaaaaa aaagaacaaa acggaaggag 3180

cacgtatatc ttttctatga tctttggata tggaatagat aagcaacccc tctagtgaag 3240

tacacatcaa gatacttggg gaacaactga gaagacatgc tatacacgaa aaccatcatc 3300

taatcatatt tacatatatg aaacatgaaa agttattcat ctattggatt cacttttata 3360

tgttttcatg cattgtgcac tctatgcttc tgctttttct ctgttctttt atactcctct 3420

actatcaaat tgagtatttc aattatcaaa gaaatttgag ctggatcaga actttagata 3480

ttcatttctg ttttcgatat atttaaacga tgttctaatg tcaaatctca atatcgttgc 3540

taaaaaatct aactttatag ctttgtgcat taatggaaaa aaatacgtgg atcttttgaa 3600

tgtttaaaaa cgagatactc ttttggaaac cagaaaataa ttttgaaata caaattggtg 3660

aatgtaagca agcaaaaatt ggatgctgaa gaaaataaat ttatgaacta aattatctct 3720

gaacaaacat ttagtcaacc ttttctcctt gctcctcaat caaaggtttt tcgtatagat 3780

atatacgcag gttgcttttt gtttccgctc ataaaatcaa gtttcgatac caatggagat 3840

ttattctttc gttttgattc cataataatc gaaattcttt taaaacactt tgaagcgaac 3900

tgaaatccca atatctttct gaccactaaa acaattataa agtactagat gttcaaaaag 3960

ctttgagaat tgcctagctt caaaatatat ttgtttgcaa atactatttt attttttcct 4020

ttctttgtct tctttggtta tttaaaatac gaagttcgct acgttggttt tgaggaacca 4080

ttactctatt tactcaaatt tattatcaaa gtgatttttt ttgcttgatc tatctaaagg 4140

ctgtggatat gccccagaaa caccataaaa gtcaaatttc taaagccagc tcttactata 4200

aagattagag ttcattcgac ttaattgcaa gtatgtagct caaaccaagg ttactctata 4260

attataaaaa gactaggaac caatgcaaga tccccttatt tcatacaaat ttagtataac 4320

tcaaattcac attttcatat gcaaaaactt aaagaaaagc atcattgtaa agagccagtt 4380

gttgtatatt caagtgggca ctgaacttgt ggttagattt tatggacatc ttgtcatcga 4440

agttttttga tagaagttgt gccattgaga attgaaatat aaaatgatat tggattgaga 4500

atatacgaat gtcttattac tcgtaataat aggagaactc atcgaatatg taaaatatag 4560

gctgattaag gtacatacag catcctgttc atcatttcaa ctatttaagt atcaaattag 4620

tgaagaaacg tattatgaac cattcattgt tcaataagtt tactcttttt tgtacttttt 4680

tgtttttgct tttcggacat tgtaattttt actcatttat tcgatgtctc tttaggtttt 4740

tttttgtttt taaatcttcg cggaattgag ataagatcac atgtgagcaa aaggccagca 4800

aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4860

tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4920

aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4980

gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 5040

acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5100

accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5160

ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5220

gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5280

gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5340

ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5400

gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5460

cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcatg 5507

<210> 57

<211> 5951

<212> DNA

<213> pCuARS-ScPGK1

<400> 57

aactcacgtt aagggatttt ggtcatgcat gagatcagat ctggaagtac cttcaaagaa 60

tggggtctca tcttgttttg caagtaccac tgagcaggat aataatagaa atgataatat 120

actatagtag agataacgtc gatgacttcc catactgtaa ttgcttttag ttgtgtattt 180

ttagtgtgca agtttctgta aatcgattaa tttttttttc tttcctcttt ttattaacct 240

taatttttat tttagattcc tgacttcaac tcaagacgca cagatattat aacatctgca 300

caataggcat ttgcaagaat tactcgtgag taaggaaaga gtgaggaact atcgcatacc 360

tgcatttaaa gatgccgatt tgggcgcgaa tcctttattt tggcttcacc ctcatactat 420

tatcagggcc agaaaaagga agtgtttccc tccttcttga attgatgtta ccctcataaa 480

acacgtggcc tcttatcaag aaagaaatta ccgtcgctcg tgatttgttt gcaaagagaa 540

caaaactgaa aaaacccaga cacgctcgac ttcctgtctt cctattgatt gcagcttcca 600

atttcgtcac acaacaaggt cctagcgacg gctcacaggt tttgtaacaa gcaatcgaag 660

gttctggaat ggcgggaaag ggtttagtac cacatgctat gatgcccact gtgatctcca 720

gagcaaagtt cgttcgatcg tactgttact ctctctcttt caaacagaaa tgtccgaatc 780

gtgtgacaac aacagcctgt tctcacacac tcttttcttc taaccaaggg ggtggtttag 840

tttagtagaa cctcgtgaaa cttacattta catatatata aacttgcata aattggtcaa 900

tgcaagaaat acatatttgg tcttttctaa ttcgtagttt ttcaagttct tagatgcttt 960

ctttttctct tttttacaga tcatcaagga agtaattatc tactttttac aacaaatata 1020

aaacaattcg aaacgatgag atttccttca atttttactg ctgttttatt cgcagcatcc 1080

tccgcattag ctgctccagt caacactaca acagaagatg aaacggcaca aattccggct 1140

gaagctgtca tcggttactc agatttagaa ggggatttcg atgttgctgt tttgccattt 1200

tccaacagca caaataacgg gttattgttt ataaatacta ctattgccag cattgctgct 1260

aaagaagaag gggtatctct cgagaaaaga gaggctgaag ctgaattcac gtggcccagc 1320

cggccgtctc ggatcggtac ctcgagccgc ggcggccgcc agctttctag aacaaaaact 1380

catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagtttt 1440

agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc 1500

tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt 1560

tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc 1620

tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg 1680

tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta 1740

cagaagatta agtgagacct tcgtttgtgc ggatccccca cacaccatag cttcaaaatg 1800

tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt 1860

caaaacaccc aagcacagca tactaaattt tccctctttc ttcctctagg gtgtcgttaa 1920

ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc 1980

gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaatttttt tttttagttt 2040

ttttctcttt cagtgacctc cattgatatt taagttaata aacggtcttc aatttctcaa 2100

gtttcagttt catttttctt gttctattac aacttttttt acttcttgtt cattagaaag 2160

aaagcatagc aatctaatct aagggcggtg ttgacaatta atcatcggca tagtatatcg 2220

gcatagtata atacgacaag gtgaggaact aaaccatgag ccatattcaa cgggaaacgt 2280

cttgttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 2340

ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg 2400

cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 2460

tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 2520

gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg 2580

tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc 2640

gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 2700

tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 2760

agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct 2820

caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 2880

ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc 2940

ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc 3000

aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg 3060

agtttttcta acacgtccga cggcggccca cgggtcccag gcctcggaga tccgtccccc 3120

ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcccccc 3180

acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat 3240

ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt 3300

ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg 3360

ggacgctcga aggctttaat ttgcaagctg gagaccaaca tgtgatcatg tttcgtacaa 3420

aacaccgcca ctccgttatg agaactcata gttatatttc ggggaaactt aacttgcttg 3480

caagcaataa tagacagaca aatgttttac gaaactggag gattaataca attgatgcaa 3540

aaacaaaaaa aaaacaaaaa aaaaaaacac acgagaacat acaaaaaaag actacaaaat 3600

atatatttgc taataccatc accctcatac aaaaaaaaga acaaaacgga aggagcacgt 3660

atatcttttc tatgatcttt ggatatggaa tagataagca acccctctag tgaagtacac 3720

atcaagatac ttggggaaca actgagaaga catgctatac acgaaaacca tcatctaatc 3780

atatttacat atatgaaaca tgaaaagtta ttcatctatt ggattcactt ttatatgttt 3840

tcatgcattg tgcactctat gcttctgctt tttctctgtt cttttatact cctctactat 3900

caaattgagt atttcaatta tcaaagaaat ttgagctgga tcagaacttt agatattcat 3960

ttctgttttc gatatattta aacgatgttc taatgtcaaa tctcaatatc gttgctaaaa 4020

aatctaactt tatagctttg tgcattaatg gaaaaaaata cgtggatctt ttgaatgttt 4080

aaaaacgaga tactcttttg gaaaccagaa aataattttg aaatacaaat tggtgaatgt 4140

aagcaagcaa aaattggatg ctgaagaaaa taaatttatg aactaaatta tctctgaaca 4200

aacatttagt caaccttttc tccttgctcc tcaatcaaag gtttttcgta tagatatata 4260

cgcaggttgc tttttgtttc cgctcataaa atcaagtttc gataccaatg gagatttatt 4320

ctttcgtttt gattccataa taatcgaaat tcttttaaaa cactttgaag cgaactgaaa 4380

tcccaatatc tttctgacca ctaaaacaat tataaagtac tagatgttca aaaagctttg 4440

agaattgcct agcttcaaaa tatatttgtt tgcaaatact attttatttt ttcctttctt 4500

tgtcttcttt ggttatttaa aatacgaagt tcgctacgtt ggttttgagg aaccattact 4560

ctatttactc aaatttatta tcaaagtgat tttttttgct tgatctatct aaaggctgtg 4620

gatatgcccc agaaacacca taaaagtcaa atttctaaag ccagctctta ctataaagat 4680

tagagttcat tcgacttaat tgcaagtatg tagctcaaac caaggttact ctataattat 4740

aaaaagacta ggaaccaatg caagatcccc ttatttcata caaatttagt ataactcaaa 4800

ttcacatttt catatgcaaa aacttaaaga aaagcatcat tgtaaagagc cagttgttgt 4860

atattcaagt gggcactgaa cttgtggtta gattttatgg acatcttgtc atcgaagttt 4920

tttgatagaa gttgtgccat tgagaattga aatataaaat gatattggat tgagaatata 4980

cgaatgtctt attactcgta ataataggag aactcatcga atatgtaaaa tataggctga 5040

ttaaggtaca tacagcatcc tgttcatcat ttcaactatt taagtatcaa attagtgaag 5100

aaacgtatta tgaaccattc attgttcaat aagtttactc ttttttgtac ttttttgttt 5160

ttgcttttcg gacattgtaa tttttactca tttattcgat gtctctttag gttttttttt 5220

gtttttaaat cttcgcggaa ttgagataag atcacatgtg agcaaaaggc cagcaaaagg 5280

ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 5340

agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5400

accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5460

ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct 5520

gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5580

ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5640

gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5700

taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 5760

tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5820

gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5880

cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5940

agtggaacga a 5951

<210> 58

<211> 6433

<212> DNA

<213> pCuARS-ScADH1

<400> 58

tcacgttaag ggattttggt catgcatgag atcagatcta gggggatcga agaaatgatg 60

gtaaatgaaa tagaaaatca aggagcatga aggcaaaaga caaatataag ggtcaaacga 120

aaaataaagt gaaaagtgtt gatatgatgt atttggcttt gcggcgccga aaaaacgagt 180

ttacgcaatt gcacaatcat gctgactctg tggcggaccc gcgctcttgc cggcccggcg 240

ataacgctgg gcgtgaggct gtgcccggcg gagttttttg cgcctgcatt ttccaaggtt 300

taccctgcgc taaggggcga gattggagaa gcaataagaa tgccggttgg ggttgcgatg 360

atgacgacca cgacaactgg tgtcattatt taatttgccg aaagaacctg agtgcatttg 420

caacatgagt atactagaag aatgagccaa gacttgcgag acgcgagttt gccggtggtg 480

cgaacaatag agcgaccatg accctgaagg tgagacgcgc ataaccgcta gagtactttg 540

aagaggaaac agcaataggg ttgctaccag tataaataga caggtacata cgacactgga 600

aatggttgtc tgtttgagta cgctttcaat tcatttgggt gtgcacttta ttatgttaca 660

atatggaagg gaactttaca cttctcctat gcacatatat taattaaagt ccaatgctag 720

tagagaaggg gggtaacacc cctccgcgct cttttccgat ttttttctaa accgtggaat 780

atttcggata tcctcttgtt gtttccgggt gtacaatatg gacttcctct tttctggcaa 840

ccaaacccat acatcgggat tcctataata ccttcgttgg tctccctaac atgtaggtgg 900

cggaggggag atatacaata gaacagatac cagacaagac ataatgggct aaacaagact 960

acaccaatta cactgcctca ttgatggtgg tacataacga actaatactg tagccctaga 1020

cttgatagcc atcatcatat cgaagtttca ctaccctttt tccatttgcc atctattgaa 1080

gtaataatag gcgcatgcaa ctccttttct tttttttttt tttctctctc ccccgttgtt 1140

gtctcaccat atccgcaatg acaaaaaaat gatggaagac actaaaggaa aaaattaacg 1200

acaaagacag caccaacaga tgtcgttgtt ccagagctga tgaggggtat ctcgaagcac 1260

acgaaacttt ttccctcctt cattcacgca cactactctc taatgagcaa cggtatacgg 1320

ccttccttcc agttacttga atttgaaata aaaaaagttt gctgtcttgc tatcaagtat 1380

aaatagacct gcaattatta atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc 1440

cttgtttctt tttctgcaca atatttcaag ctataccaag catacaatca actatctcat 1500

atacattcga aacgatgaga tttccttcaa tttttactgc tgttttattc gcagcatcct 1560

ccgcattagc tgctccagtc aacactacaa cagaagatga aacggcacaa attccggctg 1620

aagctgtcat cggttactca gatttagaag gggatttcga tgttgctgtt ttgccatttt 1680

ccaacagcac aaataacggg ttattgttta taaatactac tattgccagc attgctgcta 1740

aagaagaagg ggtatctctc gagaaaagag aggctgaagc tgaattcacg tggcccagcc 1800

ggccgtctcg gatcggtacc tcgagccgcg gcggccgcca gctttctaga acaaaaactc 1860

atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca ttgagtttta 1920

gccttagaca tgactgttcc tcagttcaag ttgggcactt acgagaagac cggtcttgct 1980

agattctaat caagaggatg tcagaatgcc atttgcctga gagatgcagg cttcattttt 2040

gatacttttt tatttgtaac ctatatagta taggattttt tttgtcattt tgtttcttct 2100

cgtacgagct tgctcctgat cagcctatct cgcagctgat gaatatcttg tggtaggggt 2160

ttgggaaaat cattcgagtt tgatgttttt cttggtattt cccactcctc ttcagagtac 2220

agaagattaa gtgagacctt cgtttgtgcg gatcccccac acaccatagc ttcaaaatgt 2280

ttctactcct tttttactct tccagatttt ctcggactcc gcgcatcgcc gtaccacttc 2340

aaaacaccca agcacagcat actaaatttt ccctctttct tcctctaggg tgtcgttaat 2400

tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg 2460

aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg aaattttttt ttttagtttt 2520

tttctctttc agtgacctcc attgatattt aagttaataa acggtcttca atttctcaag 2580

tttcagtttc atttttcttg ttctattaca acttttttta cttcttgttc attagaaaga 2640

aagcatagca atctaatcta agggcggtgt tgacaattaa tcatcggcat agtatatcgg 2700

catagtataa tacgacaagg tgaggaacta aaccatgagc catattcaac gggaaacgtc 2760

ttgttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc 2820

tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc 2880

gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat 2940

ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg 3000

tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt 3060

attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg 3120

ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct 3180

cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga 3240

gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc 3300

accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg 3360

gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct 3420

tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca 3480

aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga 3540

gtttttctaa cacgtccgac ggcggcccac gggtcccagg cctcggagat ccgtccccct 3600

tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcccccca 3660

catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt 3720

tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc 3780

tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg 3840

gacgctcgaa ggctttaatt tgcaagctgg agaccaacat gtgatcatgt ttcgtacaaa 3900

acaccgccac tccgttatga gaactcatag ttatatttcg gggaaactta acttgcttgc 3960

aagcaataat agacagacaa atgttttacg aaactggagg attaatacaa ttgatgcaaa 4020

aacaaaaaaa aaacaaaaaa aaaaaacaca cgagaacata caaaaaaaga ctacaaaata 4080

tatatttgct aataccatca ccctcataca aaaaaaagaa caaaacggaa ggagcacgta 4140

tatcttttct atgatctttg gatatggaat agataagcaa cccctctagt gaagtacaca 4200

tcaagatact tggggaacaa ctgagaagac atgctataca cgaaaaccat catctaatca 4260

tatttacata tatgaaacat gaaaagttat tcatctattg gattcacttt tatatgtttt 4320

catgcattgt gcactctatg cttctgcttt ttctctgttc ttttatactc ctctactatc 4380

aaattgagta tttcaattat caaagaaatt tgagctggat cagaacttta gatattcatt 4440

tctgttttcg atatatttaa acgatgttct aatgtcaaat ctcaatatcg ttgctaaaaa 4500

atctaacttt atagctttgt gcattaatgg aaaaaaatac gtggatcttt tgaatgttta 4560

aaaacgagat actcttttgg aaaccagaaa ataattttga aatacaaatt ggtgaatgta 4620

agcaagcaaa aattggatgc tgaagaaaat aaatttatga actaaattat ctctgaacaa 4680

acatttagtc aaccttttct ccttgctcct caatcaaagg tttttcgtat agatatatac 4740

gcaggttgct ttttgtttcc gctcataaaa tcaagtttcg ataccaatgg agatttattc 4800

tttcgttttg attccataat aatcgaaatt cttttaaaac actttgaagc gaactgaaat 4860

cccaatatct ttctgaccac taaaacaatt ataaagtact agatgttcaa aaagctttga 4920

gaattgccta gcttcaaaat atatttgttt gcaaatacta ttttattttt tcctttcttt 4980

gtcttctttg gttatttaaa atacgaagtt cgctacgttg gttttgagga accattactc 5040

tatttactca aatttattat caaagtgatt ttttttgctt gatctatcta aaggctgtgg 5100

atatgcccca gaaacaccat aaaagtcaaa tttctaaagc cagctcttac tataaagatt 5160

agagttcatt cgacttaatt gcaagtatgt agctcaaacc aaggttactc tataattata 5220

aaaagactag gaaccaatgc aagatcccct tatttcatac aaatttagta taactcaaat 5280

tcacattttc atatgcaaaa acttaaagaa aagcatcatt gtaaagagcc agttgttgta 5340

tattcaagtg ggcactgaac ttgtggttag attttatgga catcttgtca tcgaagtttt 5400

ttgatagaag ttgtgccatt gagaattgaa atataaaatg atattggatt gagaatatac 5460

gaatgtctta ttactcgtaa taataggaga actcatcgaa tatgtaaaat ataggctgat 5520

taaggtacat acagcatcct gttcatcatt tcaactattt aagtatcaaa ttagtgaaga 5580

aacgtattat gaaccattca ttgttcaata agtttactct tttttgtact tttttgtttt 5640

tgcttttcgg acattgtaat ttttactcat ttattcgatg tctctttagg tttttttttg 5700

tttttaaatc ttcgcggaat tgagataaga tcacatgtga gcaaaaggcc agcaaaaggc 5760

caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 5820

gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 5880

ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5940

cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 6000

taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 6060

cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 6120

acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 6180

aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 6240

atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 6300

atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 6360

gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6420

gtggaacgaa aac 6433

<210> 59

<211> 5560

<212> DNA

<213> pCuARS-ScHXT7

<400> 59

tgcatgagat cagatctttt taccccgtgg aaatgagggg tatgcaggaa tttgtgcggg 60

gtaggaaatc tttttttttt taggaggaac aactggtgga agaatgccca cacttctcag 120

aaatgcatgc agtggcagca cgctaattcg aaaaaattct ccagaaaggc aacgcaaaat 180

tttttttcca gggaataaac tttttatgac ccactacttc tcgtaggaac aatttcggtc 240

acctgcgtgt tcttctgagg ttcatctttt acatttgctt ctgctggata attttcagag 300

gcaacaagga aaaattagat ggcaaaaagt cgtctttcaa ggaaaaatcc ccaccatctt 360

tcgagatccc ctgtaactta ttggcaactg aaagaatgaa aaggaggaaa atacaaaata 420

tactagaact gaaaaaaaaa aagtataaat agagacgata tatgccaata cttcacaatg 480

ttcgaatcta ttcttcattt gcagctattg taaaataata aaacatcaag aacaaacaag 540

ctcaacttgt cttttctaag aacaaagaat aaacacaaaa acaaaaagtt tttttaattt 600

taatcaaaaa ttcgaaacga tgagatttcc ttcaattttt actgctgttt tattcgcagc 660

atcctccgca ttagctgctc cagtcaacac tacaacagaa gatgaaacgg cacaaattcc 720

ggctgaagct gtcatcggtt actcagattt agaaggggat ttcgatgttg ctgttttgcc 780

attttccaac agcacaaata acgggttatt gtttataaat actactattg ccagcattgc 840

tgctaaagaa gaaggggtat ctctcgagaa aagagaggct gaagctgaat tcacgtggcc 900

cagccggccg tctcggatcg gtacctcgag ccgcggcggc cgccagcttt ctagaacaaa 960

aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag 1020

ttttagcctt agacatgact gttcctcagt tcaagttggg cacttacgag aagaccggtc 1080

ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca 1140

tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt cattttgttt 1200

cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata tcttgtggta 1260

ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac tcctcttcag 1320

agtacagaag attaagtgag accttcgttt gtgcggatcc cccacacacc atagcttcaa 1380

aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc 1440

acttcaaaac acccaagcac agcatactaa attttccctc tttcttcctc tagggtgtcg 1500

ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt 1560

cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaatt ttttttttta 1620

gtttttttct ctttcagtga cctccattga tatttaagtt aataaacggt cttcaatttc 1680

tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct tgttcattag 1740

aaagaaagca tagcaatcta atctaagggc ggtgttgaca attaatcatc ggcatagtat 1800

atcggcatag tataatacga caaggtgagg aactaaacca tgagccatat tcaacgggaa 1860

acgtcttgtt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa 1920

tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc 1980

gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat 2040

gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt 2100

atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc 2160

caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc 2220

ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt 2280

cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat 2340

gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca 2400

ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac 2460

gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag 2520

gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt 2580

tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc 2640

gatgagtttt tctaacacgt ccgacggcgg cccacgggtc ccaggcctcg gagatccgtc 2700

ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc 2760

ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat 2820

ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt 2880

ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt 2940

tttgggacgc tcgaaggctt taatttgcaa gctggagacc aacatgtgat catgtttcgt 3000

acaaaacacc gccactccgt tatgagaact catagttata tttcggggaa acttaacttg 3060

cttgcaagca ataatagaca gacaaatgtt ttacgaaact ggaggattaa tacaattgat 3120

gcaaaaacaa aaaaaaaaca aaaaaaaaaa acacacgaga acatacaaaa aaagactaca 3180

aaatatatat ttgctaatac catcaccctc atacaaaaaa aagaacaaaa cggaaggagc 3240

acgtatatct tttctatgat ctttggatat ggaatagata agcaacccct ctagtgaagt 3300

acacatcaag atacttgggg aacaactgag aagacatgct atacacgaaa accatcatct 3360

aatcatattt acatatatga aacatgaaaa gttattcatc tattggattc acttttatat 3420

gttttcatgc attgtgcact ctatgcttct gctttttctc tgttctttta tactcctcta 3480

ctatcaaatt gagtatttca attatcaaag aaatttgagc tggatcagaa ctttagatat 3540

tcatttctgt tttcgatata tttaaacgat gttctaatgt caaatctcaa tatcgttgct 3600

aaaaaatcta actttatagc tttgtgcatt aatggaaaaa aatacgtgga tcttttgaat 3660

gtttaaaaac gagatactct tttggaaacc agaaaataat tttgaaatac aaattggtga 3720

atgtaagcaa gcaaaaattg gatgctgaag aaaataaatt tatgaactaa attatctctg 3780

aacaaacatt tagtcaacct tttctccttg ctcctcaatc aaaggttttt cgtatagata 3840

tatacgcagg ttgctttttg tttccgctca taaaatcaag tttcgatacc aatggagatt 3900

tattctttcg ttttgattcc ataataatcg aaattctttt aaaacacttt gaagcgaact 3960

gaaatcccaa tatctttctg accactaaaa caattataaa gtactagatg ttcaaaaagc 4020

tttgagaatt gcctagcttc aaaatatatt tgtttgcaaa tactatttta ttttttcctt 4080

tctttgtctt ctttggttat ttaaaatacg aagttcgcta cgttggtttt gaggaaccat 4140

tactctattt actcaaattt attatcaaag tgattttttt tgcttgatct atctaaaggc 4200

tgtggatatg ccccagaaac accataaaag tcaaatttct aaagccagct cttactataa 4260

agattagagt tcattcgact taattgcaag tatgtagctc aaaccaaggt tactctataa 4320

ttataaaaag actaggaacc aatgcaagat ccccttattt catacaaatt tagtataact 4380

caaattcaca ttttcatatg caaaaactta aagaaaagca tcattgtaaa gagccagttg 4440

ttgtatattc aagtgggcac tgaacttgtg gttagatttt atggacatct tgtcatcgaa 4500

gttttttgat agaagttgtg ccattgagaa ttgaaatata aaatgatatt ggattgagaa 4560

tatacgaatg tcttattact cgtaataata ggagaactca tcgaatatgt aaaatatagg 4620

ctgattaagg tacatacagc atcctgttca tcatttcaac tatttaagta tcaaattagt 4680

gaagaaacgt attatgaacc attcattgtt caataagttt actctttttt gtactttttt 4740

gtttttgctt ttcggacatt gtaattttta ctcatttatt cgatgtctct ttaggttttt 4800

ttttgttttt aaatcttcgc ggaattgaga taagatcaca tgtgagcaaa aggccagcaa 4860

aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4920

gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4980

agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5040

cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5100

cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5160

ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5220

gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5280

tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5340

acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5400

tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5460

attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5520

gctcagtgga acgaaaactc acgttaaggg attttggtca 5560

<210> 60

<211> 5567

<212> DNA

<213> pCuARS-ScGAL1

<400> 60

tgcatgagat cagatctaca tggcattacc accatataca tatccatatc taatcttact 60

tatatgttgt ggaaatgtaa agagccccat tatcttagcc taaaaaaacc ttctctttgg 120

aactttcagt aatacgctta actgctcatt gctatattga agtacggatt agaagccgcc 180

gagcgggcga cagccctccg acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc 240

gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca 300

atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt 360

caaattaacg aatcaaatta acaaccatag gacgataatg cgattagttt tttagcctta 420

tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg 480

caaaagctgc ataaccactt taactaatac tttcaacatt ttcggtttgt gttacttctt 540

attcaaatgt cataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc 600

aaggagaaaa aactatattc gaaacgatga gatttccttc aatttttact gctgttttat 660

tcgcagcatc ctccgcatta gctgctccag tcaacactac aacagaagat gaaacggcac 720

aaattccggc tgaagctgtc atcggttact cagatttaga aggggatttc gatgttgctg 780

ttttgccatt ttccaacagc acaaataacg ggttattgtt tataaatact actattgcca 840

gcattgctgc taaagaagaa ggggtatctc tcgagaaaag agaggctgaa gctgaattca 900

cgtggcccag ccggccgtct cggatcggta cctcgagccg cggcggccgc cagctttcta 960

gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat 1020

cattgagttt tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag 1080

accggtcttg ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca 1140

ggcttcattt ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat 1200

tttgtttctt ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct 1260

tgtggtaggg gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc 1320

tcttcagagt acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata 1380

gcttcaaaat gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg 1440

ccgtaccact tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag 1500

ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct 1560

ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt 1620

ttttttagtt tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt 1680

caatttctca agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt 1740

tcattagaaa gaaagcatag caatctaatc taagggcggt gttgacaatt aatcatcggc 1800

atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga gccatattca 1860

acgggaaacg tcttgttcga ggccgcgatt aaattccaac atggatgctg atttatatgg 1920

gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg 1980

gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt 2040

tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa 2100

gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac 2160

agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc 2220

agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg 2280

cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga 2340

ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct 2400

tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat 2460

ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg 2520

ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa 2580

acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt 2640

gatgctcgat gagtttttct aacacgtccg acggcggccc acgggtccca ggcctcggag 2700

atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac 2760

gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg 2820

tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt 2880

ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg 2940

agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgatcat 3000

gtttcgtaca aaacaccgcc actccgttat gagaactcat agttatattt cggggaaact 3060

taacttgctt gcaagcaata atagacagac aaatgtttta cgaaactgga ggattaatac 3120

aattgatgca aaaacaaaaa aaaaacaaaa aaaaaaaaca cacgagaaca tacaaaaaaa 3180

gactacaaaa tatatatttg ctaataccat caccctcata caaaaaaaag aacaaaacgg 3240

aaggagcacg tatatctttt ctatgatctt tggatatgga atagataagc aacccctcta 3300

gtgaagtaca catcaagata cttggggaac aactgagaag acatgctata cacgaaaacc 3360

atcatctaat catatttaca tatatgaaac atgaaaagtt attcatctat tggattcact 3420

tttatatgtt ttcatgcatt gtgcactcta tgcttctgct ttttctctgt tcttttatac 3480

tcctctacta tcaaattgag tatttcaatt atcaaagaaa tttgagctgg atcagaactt 3540

tagatattca tttctgtttt cgatatattt aaacgatgtt ctaatgtcaa atctcaatat 3600

cgttgctaaa aaatctaact ttatagcttt gtgcattaat ggaaaaaaat acgtggatct 3660

tttgaatgtt taaaaacgag atactctttt ggaaaccaga aaataatttt gaaatacaaa 3720

ttggtgaatg taagcaagca aaaattggat gctgaagaaa ataaatttat gaactaaatt 3780

atctctgaac aaacatttag tcaacctttt ctccttgctc ctcaatcaaa ggtttttcgt 3840

atagatatat acgcaggttg ctttttgttt ccgctcataa aatcaagttt cgataccaat 3900

ggagatttat tctttcgttt tgattccata ataatcgaaa ttcttttaaa acactttgaa 3960

gcgaactgaa atcccaatat ctttctgacc actaaaacaa ttataaagta ctagatgttc 4020

aaaaagcttt gagaattgcc tagcttcaaa atatatttgt ttgcaaatac tattttattt 4080

tttcctttct ttgtcttctt tggttattta aaatacgaag ttcgctacgt tggttttgag 4140

gaaccattac tctatttact caaatttatt atcaaagtga ttttttttgc ttgatctatc 4200

taaaggctgt ggatatgccc cagaaacacc ataaaagtca aatttctaaa gccagctctt 4260

actataaaga ttagagttca ttcgacttaa ttgcaagtat gtagctcaaa ccaaggttac 4320

tctataatta taaaaagact aggaaccaat gcaagatccc cttatttcat acaaatttag 4380

tataactcaa attcacattt tcatatgcaa aaacttaaag aaaagcatca ttgtaaagag 4440

ccagttgttg tatattcaag tgggcactga acttgtggtt agattttatg gacatcttgt 4500

catcgaagtt ttttgataga agttgtgcca ttgagaattg aaatataaaa tgatattgga 4560

ttgagaatat acgaatgtct tattactcgt aataatagga gaactcatcg aatatgtaaa 4620

atataggctg attaaggtac atacagcatc ctgttcatca tttcaactat ttaagtatca 4680

aattagtgaa gaaacgtatt atgaaccatt cattgttcaa taagtttact cttttttgta 4740

cttttttgtt tttgcttttc ggacattgta atttttactc atttattcga tgtctcttta 4800

ggtttttttt tgtttttaaa tcttcgcgga attgagataa gatcacatgt gagcaaaagg 4860

ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 4920

cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 4980

actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5040

cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5100

atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5160

gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5220

caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5280

agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5340

tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 5400

tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5460

gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5520

gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtca 5567

<210> 61

<211> 5444

<212> DNA

<213> pCuARS-PsGAPp

<400> 61

agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat 60

atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa 120

cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc 180

gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc 240

aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc 300

ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga 360

gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt 420

ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac 480

tatttcgaaa cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc 540

gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa 600

gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc 660

aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa 720

gaagaagggg tatctctcga gaaaagagag gctgaagctg aattcacgtg gcccagccgg 780

ccgtctcgga tcggtacctc gagccgcggc ggccgccagc tttctagaac aaaaactcat 840

ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt gagttttagc 900

cttagacatg actgttcctc agttcaagtt gggcacttac gagaagaccg gtcttgctag 960

attctaatca agaggatgtc agaatgccat ttgcctgaga gatgcaggct tcatttttga 1020

tactttttta tttgtaacct atatagtata ggattttttt tgtcattttg tttcttctcg 1080

tacgagcttg ctcctgatca gcctatctcg cagctgatga atatcttgtg gtaggggttt 1140

gggaaaatca ttcgagtttg atgtttttct tggtatttcc cactcctctt cagagtacag 1200

aagattaagt gagaccttcg tttgtgcgga tcccccacac accatagctt caaaatgttt 1260

ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa 1320

aacacccaag cacagcatac taaattttcc ctctttcttc ctctagggtg tcgttaatta 1380

cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa 1440

aaaggcaata aaaattttta tcacgtttct ttttcttgaa attttttttt ttagtttttt 1500

tctctttcag tgacctccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt 1560

tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgttcat tagaaagaaa 1620

gcatagcaat ctaatctaag ggcggtgttg acaattaatc atcggcatag tatatcggca 1680

tagtataata cgacaaggtg aggaactaaa ccatgagcca tattcaacgg gaaacgtctt 1740

gttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc 1800

gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc 1860

cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg 1920

tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta 1980

ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat 2040

tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc 2100

ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg 2160

ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc 2220

gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac 2280

cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga 2340

aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg 2400

ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa 2460

aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt 2520

ttttctaaca cgtccgacgg cggcccacgg gtcccaggcc tcggagatcc gtcccccttt 2580

tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca 2640

tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt 2700

tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg 2760

tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga 2820

cgctcgaagg ctttaatttg caagctggag accaacatgt gatcatgttt cgtacaaaac 2880

accgccactc cgttatgaga actcatagtt atatttcggg gaaacttaac ttgcttgcaa 2940

gcaataatag acagacaaat gttttacgaa actggaggat taatacaatt gatgcaaaaa 3000

caaaaaaaaa acaaaaaaaa aaaacacacg agaacataca aaaaaagact acaaaatata 3060

tatttgctaa taccatcacc ctcatacaaa aaaaagaaca aaacggaagg agcacgtata 3120

tcttttctat gatctttgga tatggaatag ataagcaacc cctctagtga agtacacatc 3180

aagatacttg gggaacaact gagaagacat gctatacacg aaaaccatca tctaatcata 3240

tttacatata tgaaacatga aaagttattc atctattgga ttcactttta tatgttttca 3300

tgcattgtgc actctatgct tctgcttttt ctctgttctt ttatactcct ctactatcaa 3360

attgagtatt tcaattatca aagaaatttg agctggatca gaactttaga tattcatttc 3420

tgttttcgat atatttaaac gatgttctaa tgtcaaatct caatatcgtt gctaaaaaat 3480

ctaactttat agctttgtgc attaatggaa aaaaatacgt ggatcttttg aatgtttaaa 3540

aacgagatac tcttttggaa accagaaaat aattttgaaa tacaaattgg tgaatgtaag 3600

caagcaaaaa ttggatgctg aagaaaataa atttatgaac taaattatct ctgaacaaac 3660

atttagtcaa ccttttctcc ttgctcctca atcaaaggtt tttcgtatag atatatacgc 3720

aggttgcttt ttgtttccgc tcataaaatc aagtttcgat accaatggag atttattctt 3780

tcgttttgat tccataataa tcgaaattct tttaaaacac tttgaagcga actgaaatcc 3840

caatatcttt ctgaccacta aaacaattat aaagtactag atgttcaaaa agctttgaga 3900

attgcctagc ttcaaaatat atttgtttgc aaatactatt ttattttttc ctttctttgt 3960

cttctttggt tatttaaaat acgaagttcg ctacgttggt tttgaggaac cattactcta 4020

tttactcaaa tttattatca aagtgatttt ttttgcttga tctatctaaa ggctgtggat 4080

atgccccaga aacaccataa aagtcaaatt tctaaagcca gctcttacta taaagattag 4140

agttcattcg acttaattgc aagtatgtag ctcaaaccaa ggttactcta taattataaa 4200

aagactagga accaatgcaa gatcccctta tttcatacaa atttagtata actcaaattc 4260

acattttcat atgcaaaaac ttaaagaaaa gcatcattgt aaagagccag ttgttgtata 4320

ttcaagtggg cactgaactt gtggttagat tttatggaca tcttgtcatc gaagtttttt 4380

gatagaagtt gtgccattga gaattgaaat ataaaatgat attggattga gaatatacga 4440

atgtcttatt actcgtaata ataggagaac tcatcgaata tgtaaaatat aggctgatta 4500

aggtacatac agcatcctgt tcatcatttc aactatttaa gtatcaaatt agtgaagaaa 4560

cgtattatga accattcatt gttcaataag tttactcttt tttgtacttt tttgtttttg 4620

cttttcggac attgtaattt ttactcattt attcgatgtc tctttaggtt tttttttgtt 4680

tttaaatctt cgcggaattg agataagatc acatgtgagc aaaaggccag caaaaggcca 4740

ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4800

atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4860

aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4920

gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta 4980

ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5040

ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5100

acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5160

gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 5220

ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5280

ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5340

gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5400

ggaacgaaaa ctcacgttaa gggattttgg tcatgcatga gatc 5444

<210> 62

<211> 5892

<212> DNA

<213> pCuARS-PsAOX1

<400> 62

ttaagggatt ttggtcatgc atgagatcag atctaacatc caaagacgaa aggttgaatg 60

aaaccttttt gccatccgac atccacaggt ccattctcac acataagtgc caaacgcaac 120

aggaggggat acactagcag cagaccgttg caaacgcagg acctccactc ctcttctcct 180

caacacccac ttttgccatc gaaaaaccag cccagttatt gggcttgatt ggagctcgct 240

cattccaatt ccttctatta ggctactaac accatgactt tattagcctg tctatcctgg 300

cccccctggc gaggttcatg tttgtttatt tccgaatgca acaagctccg cattacaccc 360

gaacatcact ccagatgagg gctttctgag tgtggggtca aatagtttca tgttccccaa 420

atggcccaaa actgacagtt taaacgctgt cttggaacct aatatgacaa aagcgtgatc 480

tcatccaaga tgaactaagt ttggttcgtt gaaatgctaa cggccagttg gtcaaaaaga 540

aacttccaaa agtcggcata ccgtttgtct tgtttggtat tgattgacga atgctcaaaa 600

ataatctcat taatgcttag cgcagtctct ctatcgcttc tgaaccccgg tgcacctgtg 660

ccgaaacgca aatggggaaa cacccgcttt ttggatgatt atgcattgtc tccacattgt 720

atgcttccaa gattctggtg ggaatactgc tgatagccta acgttcatga tcaaaattta 780

actgttctaa cccctacttg acagcaatat ataaacagaa ggaagctgcc ctgtcttaaa 840

cctttttttt tatcatcatt attagcttac tttcataatt gcgactggtt ccaattgaca 900

agcttttgat tttaacgact tttaacgaca acttgagaag atcaaaaaac aactaattat 960

tcgaaacgat gagatttcct tcaattttta ctgctgtttt attcgcagca tcctccgcat 1020

tagctgctcc agtcaacact acaacagaag atgaaacggc acaaattccg gctgaagctg 1080

tcatcggtta ctcagattta gaaggggatt tcgatgttgc tgttttgcca ttttccaaca 1140

gcacaaataa cgggttattg tttataaata ctactattgc cagcattgct gctaaagaag 1200

aaggggtatc tctcgagaaa agagaggctg aagctgaatt cacgtggccc agccggccgt 1260

ctcggatcgg tacctcgagc cgcggcggcc gccagctttc tagaacaaaa actcatctca 1320

gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt tttagcctta 1380

gacatgactg ttcctcagtt caagttgggc acttacgaga agaccggtct tgctagattc 1440

taatcaagag gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact 1500

tttttatttg taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg 1560

agcttgctcc tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga 1620

aaatcattcg agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga 1680

ttaagtgaga ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac 1740

tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca 1800

cccaagcaca gcatactaaa ttttccctct ttcttcctct agggtgtcgt taattacccg 1860

tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag 1920

gcaataaaaa tttttatcac gtttcttttt cttgaaattt ttttttttag tttttttctc 1980

tttcagtgac ctccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag 2040

tttcattttt cttgttctat tacaactttt tttacttctt gttcattaga aagaaagcat 2100

agcaatctaa tctaagggcg gtgttgacaa ttaatcatcg gcatagtata tcggcatagt 2160

ataatacgac aaggtgagga actaaaccat gagccatatt caacgggaaa cgtcttgttc 2220

gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga 2280

taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga 2340

gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag 2400

actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc 2460

tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga 2520

agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt 2580

gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca 2640

ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa 2700

tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga 2760

ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt 2820

aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat 2880

cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata 2940

tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt 3000

ctaacacgtc cgacggcggc ccacgggtcc caggcctcgg agatccgtcc cccttttcct 3060

ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg 3120

ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta 3180

tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca 3240

gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct 3300

cgaaggcttt aatttgcaag ctggagacca acatgtgatc atgtttcgta caaaacaccg 3360

ccactccgtt atgagaactc atagttatat ttcggggaaa cttaacttgc ttgcaagcaa 3420

taatagacag acaaatgttt tacgaaactg gaggattaat acaattgatg caaaaacaaa 3480

aaaaaaacaa aaaaaaaaaa cacacgagaa catacaaaaa aagactacaa aatatatatt 3540

tgctaatacc atcaccctca tacaaaaaaa agaacaaaac ggaaggagca cgtatatctt 3600

ttctatgatc tttggatatg gaatagataa gcaacccctc tagtgaagta cacatcaaga 3660

tacttgggga acaactgaga agacatgcta tacacgaaaa ccatcatcta atcatattta 3720

catatatgaa acatgaaaag ttattcatct attggattca cttttatatg ttttcatgca 3780

ttgtgcactc tatgcttctg ctttttctct gttcttttat actcctctac tatcaaattg 3840

agtatttcaa ttatcaaaga aatttgagct ggatcagaac tttagatatt catttctgtt 3900

ttcgatatat ttaaacgatg ttctaatgtc aaatctcaat atcgttgcta aaaaatctaa 3960

ctttatagct ttgtgcatta atggaaaaaa atacgtggat cttttgaatg tttaaaaacg 4020

agatactctt ttggaaacca gaaaataatt ttgaaataca aattggtgaa tgtaagcaag 4080

caaaaattgg atgctgaaga aaataaattt atgaactaaa ttatctctga acaaacattt 4140

agtcaacctt ttctccttgc tcctcaatca aaggtttttc gtatagatat atacgcaggt 4200

tgctttttgt ttccgctcat aaaatcaagt ttcgatacca atggagattt attctttcgt 4260

tttgattcca taataatcga aattctttta aaacactttg aagcgaactg aaatcccaat 4320

atctttctga ccactaaaac aattataaag tactagatgt tcaaaaagct ttgagaattg 4380

cctagcttca aaatatattt gtttgcaaat actattttat tttttccttt ctttgtcttc 4440

tttggttatt taaaatacga agttcgctac gttggttttg aggaaccatt actctattta 4500

ctcaaattta ttatcaaagt gatttttttt gcttgatcta tctaaaggct gtggatatgc 4560

cccagaaaca ccataaaagt caaatttcta aagccagctc ttactataaa gattagagtt 4620

cattcgactt aattgcaagt atgtagctca aaccaaggtt actctataat tataaaaaga 4680

ctaggaacca atgcaagatc cccttatttc atacaaattt agtataactc aaattcacat 4740

tttcatatgc aaaaacttaa agaaaagcat cattgtaaag agccagttgt tgtatattca 4800

agtgggcact gaacttgtgg ttagatttta tggacatctt gtcatcgaag ttttttgata 4860

gaagttgtgc cattgagaat tgaaatataa aatgatattg gattgagaat atacgaatgt 4920

cttattactc gtaataatag gagaactcat cgaatatgta aaatataggc tgattaaggt 4980

acatacagca tcctgttcat catttcaact atttaagtat caaattagtg aagaaacgta 5040

ttatgaacca ttcattgttc aataagttta ctcttttttg tacttttttg tttttgcttt 5100

tcggacattg taatttttac tcatttattc gatgtctctt taggtttttt tttgttttta 5160

aatcttcgcg gaattgagat aagatcacat gtgagcaaaa ggccagcaaa aggccaggaa 5220

ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5280

caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 5340

gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5400

cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 5460

tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5520

gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 5580

cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 5640

tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 5700

tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 5760

caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 5820

aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5880

cgaaaactca cg 5892

<210> 63

<211> 5564

<212> DNA

<213> pCuARS-PsFLD1

<400> 63

gattttggtc atgcatgaga tcagatctgc atgcaggaat ctctggcacg gtgctaatgg 60

tagttatcca acggagctga ggtagtcgat atatctggat atgccgccta taggataaaa 120

acaggagagg gtgaaccttg cttatggcta ctagattgtt cttgtactct gaattctcat 180

tatgggaaac taaactaatc tcatctgtgt gttgcagtac tattgaatcg ttgtagtatc 240

tacctggagg gcattccatg aattagtgag ataacagagt tgggtaacta gagagaataa 300

tagacgtatg catgattact acacaacgga tgtcgcactc tttccttagt taaaactatc 360

atccaatcac aagatgcggg ctggaaagac ttgctcccga aggataatct tctgcttcta 420

tctcccttcc tcatatggtt tcgcagggct catgcccctt cttccttcga actgcccgat 480

gaggaagtcc ttagcctatc aaagaattcg ggaccatcat cgatttttag agccttacct 540

gatcgcaatc aggatttcac tactcatata aatacatcgc tcaaagctcc aactttgctt 600

gttcatacaa ttcttgatat tcacattcga aacgatgaga tttccttcaa tttttactgc 660

tgttttattc gcagcatcct ccgcattagc tgctccagtc aacactacaa cagaagatga 720

aacggcacaa attccggctg aagctgtcat cggttactca gatttagaag gggatttcga 780

tgttgctgtt ttgccatttt ccaacagcac aaataacggg ttattgttta taaatactac 840

tattgccagc attgctgcta aagaagaagg ggtatctctc gagaaaagag aggctgaagc 900

tgaattcacg tggcccagcc ggccgtctcg gatcggtacc tcgagccgcg gcggccgcca 960

gctttctaga acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc 1020

atcatcatca ttgagtttta gccttagaca tgactgttcc tcagttcaag ttgggcactt 1080

acgagaagac cggtcttgct agattctaat caagaggatg tcagaatgcc atttgcctga 1140

gagatgcagg cttcattttt gatacttttt tatttgtaac ctatatagta taggattttt 1200

tttgtcattt tgtttcttct cgtacgagct tgctcctgat cagcctatct cgcagctgat 1260

gaatatcttg tggtaggggt ttgggaaaat cattcgagtt tgatgttttt cttggtattt 1320

cccactcctc ttcagagtac agaagattaa gtgagacctt cgtttgtgcg gatcccccac 1380

acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc 1440

gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttt ccctctttct 1500

tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc 1560

tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg 1620

aaattttttt ttttagtttt tttctctttc agtgacctcc attgatattt aagttaataa 1680

acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta 1740

cttcttgttc attagaaaga aagcatagca atctaatcta agggcggtgt tgacaattaa 1800

tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatgagc 1860

catattcaac gggaaacgtc ttgttcgagg ccgcgattaa attccaacat ggatgctgat 1920

ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga 1980

ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc 2040

aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg 2100

accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc 2160

gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat 2220

gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac 2280

agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat 2340

gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg 2400

cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat 2460

aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc 2520

gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca 2580

ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag 2640

tttcatttga tgctcgatga gtttttctaa cacgtccgac ggcggcccac gggtcccagg 2700

cctcggagat ccgtccccct tttcctttgt cgatatcatg taattagtta tgtcacgctt 2760

acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga 2820

agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt 2880

tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac 2940

cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcaagctgg agaccaacat 3000

gtgatcatgt ttcgtacaaa acaccgccac tccgttatga gaactcatag ttatatttcg 3060

gggaaactta acttgcttgc aagcaataat agacagacaa atgttttacg aaactggagg 3120

attaatacaa ttgatgcaaa aacaaaaaaa aaacaaaaaa aaaaaacaca cgagaacata 3180

caaaaaaaga ctacaaaata tatatttgct aataccatca ccctcataca aaaaaaagaa 3240

caaaacggaa ggagcacgta tatcttttct atgatctttg gatatggaat agataagcaa 3300

cccctctagt gaagtacaca tcaagatact tggggaacaa ctgagaagac atgctataca 3360

cgaaaaccat catctaatca tatttacata tatgaaacat gaaaagttat tcatctattg 3420

gattcacttt tatatgtttt catgcattgt gcactctatg cttctgcttt ttctctgttc 3480

ttttatactc ctctactatc aaattgagta tttcaattat caaagaaatt tgagctggat 3540

cagaacttta gatattcatt tctgttttcg atatatttaa acgatgttct aatgtcaaat 3600

ctcaatatcg ttgctaaaaa atctaacttt atagctttgt gcattaatgg aaaaaaatac 3660

gtggatcttt tgaatgttta aaaacgagat actcttttgg aaaccagaaa ataattttga 3720

aatacaaatt ggtgaatgta agcaagcaaa aattggatgc tgaagaaaat aaatttatga 3780

actaaattat ctctgaacaa acatttagtc aaccttttct ccttgctcct caatcaaagg 3840

tttttcgtat agatatatac gcaggttgct ttttgtttcc gctcataaaa tcaagtttcg 3900

ataccaatgg agatttattc tttcgttttg attccataat aatcgaaatt cttttaaaac 3960

actttgaagc gaactgaaat cccaatatct ttctgaccac taaaacaatt ataaagtact 4020

agatgttcaa aaagctttga gaattgccta gcttcaaaat atatttgttt gcaaatacta 4080

ttttattttt tcctttcttt gtcttctttg gttatttaaa atacgaagtt cgctacgttg 4140

gttttgagga accattactc tatttactca aatttattat caaagtgatt ttttttgctt 4200

gatctatcta aaggctgtgg atatgcccca gaaacaccat aaaagtcaaa tttctaaagc 4260

cagctcttac tataaagatt agagttcatt cgacttaatt gcaagtatgt agctcaaacc 4320

aaggttactc tataattata aaaagactag gaaccaatgc aagatcccct tatttcatac 4380

aaatttagta taactcaaat tcacattttc atatgcaaaa acttaaagaa aagcatcatt 4440

gtaaagagcc agttgttgta tattcaagtg ggcactgaac ttgtggttag attttatgga 4500

catcttgtca tcgaagtttt ttgatagaag ttgtgccatt gagaattgaa atataaaatg 4560

atattggatt gagaatatac gaatgtctta ttactcgtaa taataggaga actcatcgaa 4620

tatgtaaaat ataggctgat taaggtacat acagcatcct gttcatcatt tcaactattt 4680

aagtatcaaa ttagtgaaga aacgtattat gaaccattca ttgttcaata agtttactct 4740

tttttgtact tttttgtttt tgcttttcgg acattgtaat ttttactcat ttattcgatg 4800

tctctttagg tttttttttg tttttaaatc ttcgcggaat tgagataaga tcacatgtga 4860

gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 4920

aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 4980

ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 5040

gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5100

ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5160

ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5220

cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 5280

attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5340

ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5400

aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5460

gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 5520

tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagg 5564

Claims (8)

1.一种产朊假丝酵母游离型表达载体，其特征在于，以毕赤酵母表达型载体pGAPZαA为基本骨架，利用G418抗性基因替换pGAPZαA载体上的博来霉素基因，将产朊假丝酵母的自主复制序列插入pGAPZαA载体的PscI位点中；还利用产朊假丝酵母的启动子、酿酒酵母的启动子或毕赤酵母的启动子替换pGAPZαA载体上的GAP启动子；所述产朊假丝酵母的自主复制序列的核苷酸序列如SEQ ID NO.15所示。

2.根据权利要求1所述的一种产朊假丝酵母游离型表达载体，其特征在于，所述产朊假丝酵母的启动子为甘油醛-3-磷酸脱氢酶启动子、磷酸甘油酸激酶启动子或麦芽糖启动子。

3.根据权利要求1所述的一种产朊假丝酵母游离型表达载体，其特征在于，所述酿酒酵母的启动子为氨基酸通透酶启动子、转录伸长因子EF-1α启动子、磷酸丙糖脱氢酶启动子、磷酸甘油酸激酶启动子、乙醇脱氢酶启动子、己糖转运蛋白启动子或β-半乳糖激酶启动子。

4.根据权利要求1所述的一种产朊假丝酵母游离型表达载体，其特征在于，所述毕赤酵母的启动子为甘油醛-3-磷酸脱氢酶启动子、醇氧化酶1启动子或甲醛脱氢酶启动子。

5.根据权利要求1所述的一种产朊假丝酵母游离型表达载体，其特征在于，所述表达载体的序列为SEQ ID NO.46-SEQ ID NO.58所示。

6.根据权利要求1所述的一种产朊假丝酵母游离型表达载体的构建方法，其特征在于，该方法包括如下步骤：

S1.以质粒pScIKP和酿酒酵母基因组为模板，运用PCR技术分别扩增G418抗性基因编码区以及CYC终止子的部分片段，然后利用重叠PCR将G418基因的编码区与CYCTT部分片段连接，得到G418-CYCTT片段；

S2.以产朊假丝酵母GIM2.176基因组DNA为模板，运用PCR扩增获得产朊假丝酵母的自主复制序列，然后利用定点突变技术，消除序列中的PscI位点，得到所述产朊假丝酵母的自主复制序列；

S3.根据产朊假丝酵母的启动子、酿酒酵母的启动子和毕赤酵母的启动子的已知序列设计扩增引物，并分别在上下游引物5’端附加20bp的加上同源短重叠序列；

S4.分别以产朊假丝酵母GIM2.176、酿酒酵母As2.489、毕赤酵母x33的基因组DNA为模板，利用上述步骤S3所得的引物，通过PCR扩增获得所述产朊假丝酵母的启动子、酿酒酵母的启动子和毕赤酵母的启动子的片段；

S5.使用EcoRV和NcoI、或与其具有相同识别序列或产生相同粘性末端的限制性内切酶，将上述步骤S1所得G418-CYCTT片段及现有载体pGAPZαA进行双酶切后，进行连接并转化大肠杆菌DH5α，得到载体pGAPGαA；

S6.使用PscI或与其具有相同识别序列或产生相同粘性末端的限制性内切酶，将上述步骤S2所得的产朊假丝酵母的自主复制序列片段及上述步骤S5所得pGAPGαA进行单酶切后，进行连接并转化大肠杆菌DH5α，得到载体pCuGAPGαA；

S7.使用Bsp119I和BglII、或与其具有相同识别序列或产生相同粘性末端的限制性内切酶，将上述步骤S6所得载体pCuGAPGαA进行双酶切，然后运用Gibson组装技术，将上述步骤S4所得各启动子片段与酶切后的载体相连接并转化大肠杆菌DH5α，即得到一系列产朊假丝酵母游离表达载体。

7.根据权利要求5所述的一种产朊假丝酵母游离型表达载体的构建方法，其特征在于，步骤S5所述限制性内切酶为EcoRV和NcoI，或与其具有相同识别序列或产生相同粘性末端的限制性内切酶。

8.根据权利要求5所述的一种产朊假丝酵母游离型表达载体的构建方法，其特征在于，步骤S7所述限制性内切酶为Bsp119I和BglII，或与其具有相同识别序列或产生相同粘性末端的限制性内切酶。

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