pGAPZα C

保存条件 ：-20℃

保质期 ：6个月

英文名 ：pGAPZα C

库存 ：大量

供应商 ：钰博生物

规格 ：1ug/5ug

pGAPZα C

pGAPZalpha C载体基本信息

出品公司:	Ybscience
载体名称:	pGAPZalpha C, pGAPZα C, pGAPZalphaC, pGAPZαC
质粒类型:	毕赤酵母蛋白表达载体
表达水平:	高拷贝
启动子:	GAP
克隆方法:	多克隆位点，限制性内切酶
载体大小:	3152 bp
5' 测序引物及序列:	alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´;  pGAP-F:5´-GTCCCTATTTCAATCAATTGAA-3´
3' 测序引物及序列:	3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´
载体标签:	C-Myc, C-His
载体抗性:	Zeocin 博来霉素
筛选标记:	HIS4/ Zeocin
备注:	插入基因是必需包含起始密码子ATG。
稳定性:	稳定 Stable
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pGAPZalpha C载体质粒图谱和多克隆位点信息

pGAPZalpha C载体简介

甘油醛-3-磷酸脱氢酶（GAPDH）的酶在许多生物细胞中包括毕赤酵母，稳定高水平表达。 最近，GAPDH蛋白编码基因启动子（GAP）被深入研究，并在毕赤酵母中能够高水表达重组蛋白，这取决于使用的碳源（Waterham等 人，，1997）。GAP启动子（PGAP）的重组蛋白表达水平略高于AOX1启动子。

pGAPZ A，B，和C载体（2.9 KB）和pGAPZα，B，和C（3.1 KB）载体使用GAP启动子在毕赤酵母中稳定表达重组蛋白。重组蛋白表达为为带有C-末端myc抗原表位和C端His标签的融合蛋白。此外，pGAPZα 表达的融合蛋白在N-端带有酿酒酵母（Saccharomyces cerevisiae）的α-因子分泌信号肽。这两个系列的载体都提供三个不同读码框的载体，以方便克隆的带有C-端标签和/或N-末端分泌信号肽的载体 中。

这些载体是基于显性选择标记，Zeocin（博莱霉素）的筛选标记可以同时使用于毕赤酵母和大肠杆菌。

pGAPZalpha C载体序列

LOCUS       pGAPZα C           3152 bp ds-DNA    circular SYN 05-二月-2013DEFINITION  Pichia pastoris vector for constitutive expression of a secreted             protein.ACCESSION   .VERSION     .KEYWORDS    pGAPZ(alpha) CSOURCE      synthetic DNA construct  ORGANISM  synthetic DNA constructREFERENCE   1  (bases 1 to 3152)  AUTHORS   Invitrogen (Life Technologies)  TITLE     Direct Submission  JOURNAL   Exported 2013年7月30日 from SnapGene Viewer 1.5.3            http://www.snapgene.comCOMMENT     Linearize within the GAP promoter for integration.FEATURES             Location/Qualifiers     source          1..3152                     /organism="synthetic DNA construct"                     /lab_host="Pichia pastoris"                     /mol_type="other DNA"     promoter        7..483                     /note="GAP promoter"                     /note="promoter for Pichia pastoris                      glyceraldehyde-3-phosphate dehydrogenase"                     /note="color: #ffffff; direction: RIGHT"     CDS             493..759                     /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 removed by                      dipeptidyl aminopeptidase A."                     /note="This feature has 3 segments:                     ???1:?493?..?549?/?#993366?/?presequence                     ???2:?550?..?747?/?#993366?/?pro?region                     ???3:?748?..?759?/?#993366                     Cleavage sites after bases 549, 747"                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"     misc_feature    759..833                     /note="MCS"                     /note="multiple cloning site"                     /note="color: #99ccff"     CDS             832..861                     /codon_start=1                     /product="Myc (human c-Myc oncogene) epitope tag"                     /note="Myc"                     /note="color: #cc99b2"                     /translation="EQKLISEEDL"     CDS             877..894                     /codon_start=1                     /product="6xHis affinity tag"                     /note="6xHis"                     /note="color: #cc99b2"                     /translation="HHHHHH"     terminator      973..1219                     /gene="Pichia pastoris AOX1"                     /note="AOX1 terminator"                     /note="transcription terminator for AOX1"                     /note="color: #ffffff"     promoter        1234..1645                     /gene="S. cerevisiae TEF1"                     /note="TEF1 promoter"                     /note="promoter for?EF-1-alpha"                     /note="color: #ffffff; direction: RIGHT"     promoter        1652..1699                     /note="EM7 promoter"                     /note="synthetic bacterial promoter?"                     /note="color: #ffffff; direction: RIGHT"     CDS             1718..2092                     /codon_start=1                     /gene="Sh?ble from?Streptoalloteichus hindustanus"                     /product="antibiotic-binding protein"                     /note="BleoR"                     /note="confers resistance to bleomycin, phleomycin, and                      Zeocin(TM)"                     /note="color: #ccffcc"                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR                     EFALRDPAGNCVHFVAEEQD"     terminator      2158..2405                     /gene="S. cerevisiae CYC1"                     /note="CYC1 terminator"                     /note="transcription terminator for CYC1"                     /note="color: #ffffff"     rep_origin      complement(2480..3068)                     /direction=LEFT                     /note="ori"                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of                      replication"                     /note="color: #ffff00"ORIGIN        1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat       61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa      121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc      181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc      241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc      301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga      361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt      421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac      481 tatttcgaaa cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc      541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa      601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc      661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa      721 gaagaagggg tatctctcga gaagagagag gctgaagcat cgatgaattc acgtggccca      781 gccggccgtc tcggatcggt acctcgagcc gcggcggccg ccagctttct agaacaaaaa      841 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt      901 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt      961 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt     1021 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct     1081 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg     1141 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag     1201 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa     1261 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac     1321 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt     1381 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg     1441 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt     1501 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc     1561 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa     1621 agaaagcata gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat     1681 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt     1741 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg     1801 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct     1861 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt     1921 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga     1981 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct     2041 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg     2101 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca     2161 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa     2221 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag     2281 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg     2341 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa     2401 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa     2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc     2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc     2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg     2641 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt     2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc     2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc     2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag     2881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg     2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa     3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag     3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact     3121 cacgttaagg gattttggtc atgcatgaga tc//