pPIC9序列.docxVIP

载体基本信息出品公司:Invitrogen载体名称:pPIC9, pPIC 9质粒类型:毕赤酵母蛋白表达载体表达水平:高拷贝启动子:AOX1克隆方法:多克隆位点，限制性内切酶载体大小:8023 bp5 测序引物及序列:5′ AOX1:5′-GACTGGTTCCAATTGACAAGC-3′； alpha-Factor-F: 5′-TACTATTGCCAGCATTGCTGC-3′3 测序引物及序列:3′ AOX1:5′-GCAAATGGCATTCTGACATCC-3′载体标签:/载体抗性:Ampicillin筛选标记:HIS4备注:利用alpha分泌因子，分泌表达蛋白。产品目录号:K1710-01稳定性:稳定 Stable组成型:组成型 Constitutive病毒/非病毒:非病毒载体质粒图谱和多克隆位点信息载体简介pPIC9载体的详细信息如下：? pPIC9载体大小8,023 bp，是融合表达载体。? 载体构建过程中，可供使用的单一限制性内切酶位点为Xho I, SnaB I, EcoR I, Avr II, Not I.? 利用alpha因子分泌信号肽，分泌表达蛋白基因。? 在载体构建过程中，你的基因必须保证与信号肽的起始密码子的读码框一致。? 毕赤酵母中利用HIS4进行筛选。? 为了将基因插入GS115或KM71的AOX区, 使用SacI限制性内切酶线性化质粒(GS115中产生His+Mut+基因型，KM71中产生His+ MutS基因型)?为了将基因插入GS115或KM71的His4区, 使用Sal I或者Stu I限制性内切酶线性化质粒(GS115中产生His+ Mut+基因型，KM71中产生His+ MutS基因型)?将基因插入GS115的AOX1区域，需要使用Bgl II线性化质粒 (产生His+MutS基因型)。载体序列LOCUS?????? pPIC9 ????????? 8023 bp ds-DNA??? circular SYNDEFINITION? Pichia pastoris HIS4 vector for methanol-inducible expression of a??????????? secreted protein.ACCESSION?? .VERSION???? .KEYWORDS??? pPIC9SOURCE????? synthetic DNA construct? ORGANISM? synthetic DNA constructCOMMENT???? This file is created by Vector NTI??????????? /COMMENT???? VNTAUTHORNAME||FEATURES???????????? Location/Qualifiers???? source????????? 1..8023???????????????????? /organism=synthetic DNA construct???????????????????? /lab_host=Pichia pastoris???????????????????? /mol_type=other DNA???? promoter??????? 2..937???????????????????? /gene=Pichia pastoris AOX1???????????????????? /note=AOX1 promoter???????????????????? /note=inducible promoter, regulated by methanol???????????????????? /note=color: #ffffff; direction: RIGHT???? CDS???????????? 949..1215???????????????????? /codon_start=1???????????????????? /gene=MF-alpha-1???????????????????? /product=N-terminal secretion signal from S. cerevisiae???????????????????? alpha-factor???????????????????? /note=alpha-factor secretion signal???????????????????? /note=Cleavage by the Kex2 protease occurs after the???????????????????? dibasic KR sequence. The EA dipeptides are then