pPICZaA图谱及序列

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出品公司:Invitrogen载体名称:pPICZalpha A,pPICZαA,pPICZalphaA,pPICZαA, pPICZaA,pPICZa-A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:3593bp5'测序引物及序列:5´AOX1:5´-GACTGGTTCCAATTGACAAGC-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备注:插入基因是必需包含起始密码子ATG。

产品目录号:V195-20稳定性:稳定Stable组成型:组成型Constitutive病毒/非病毒:非病毒pPICZaa载体简介pPICZαA,B和C是3.6kb的毕赤酵母蛋白分泌表达载体。

表达的重组蛋白是融合蛋白,含有一个N-端多肽,编码酿酒酵母(Saccharomyces cerevisiae)α-因子分泌信号。

载体能够在毕赤酵母中利用甲醇诱导的高水平的表达目的蛋白,并且可以用在任何毕赤酵母中,包括X33,GS115菌株,SMD1168H,KM71H。

pPICZαA,B和C系列载体包含以下元素:•5'端含有AOX1启动子的严格调控,利用甲醇诱导表达任何感兴趣的基因(Ellis等,1985;Koutz等人,1989;tschopp等人,1987A)。

•α-因子分泌信号能够分泌性表达目的蛋白。

•Zeocin抗性基因在大肠杆菌和毕赤酵母都能用于筛选(Baron等人,1992;Drocourt等人,1990)。

•C-端含Myc和His标签,可以用于检测和纯化重组蛋白。

•pPICZ A,B,C三种读码框使得可以将基因克隆入载体而不发生任何移码突变。

pPICZaa载体序列LOCUS pPICZα\A3593bp DNA circular SYN2-SEP-2003SOURCEORGANISMCOMMENT This file is created by Vector NTI/COMMENT VNTAUTHORNAME||FEATURES Location/Qualifierssource 1..3593/invitrogen="25"/vntifkey="98"primer complement(1423..1443)/invitrogen="1880000"/vntifkey="27"/label=3'\AOX1\primerprimer855..875/invitrogen="1890000"/vntifkey="27"/label=5'\AOX1\primerCDS1320..1337/invitrogen="1010000"/vntifkey="4"/label=6xHisprimer1144..1164/invitrogen="1900000"/vntifkey="27"/label=alpha-factor\primer terminator1341..1682/invitrogen="2550000"/vntifkey="43"/label=AOX1\transcription\terminator CDS1275..1304/invitrogen="1090000"/vntifkey="4"/label=c-myc\epitopeterminator2538..2855/invitrogen="2560000"/vntifkey="43"/label=CYC1\transcription\terminator promoter2095..2162/invitrogen="2340000"/vntifkey="30"/label=EM7\promotermisc_feature1208..1274/invitrogen="1460000"/vntifkey="21"/label=MCSrep_origin complement(2866..3539)/invitrogen="2540000"/vntifkey="33"/label=pUC\originCDS941..1207/invitrogen="1280000"/vntifkey="4"/label=alpha-factor\signal\peptidepromoter1683..2094/invitrogen="2300000"/vntifkey="29"/label=TEF1\promoterCDS2163..2537/invitrogen="1370000"/vntifkey="4"/label=Zeo(R)promoter 1..940/invitrogen="2160000"/vntifkey="29"/label=AOX1\promoterBASE COUNT892a886c801g1014tORIGIN1agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 61gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 121tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 181agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 241acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 301tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 361agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 421gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 481ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 541cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct601ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 661ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 721gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 781atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 841actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 901caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 961tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1021agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1081tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1141tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1201tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg 1261ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc 1321atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg 1381ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt 1441tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag 1501gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc 1561agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt 1621ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat 1681cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc 1741ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaattttccc 1801tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga 1861gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt 1921tttcttgaaa tttttttttt tagttttttt ctctttcagt gacctccatt gatatttaag 1981ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact 2041ttttttactt cttgttcatt agaaagaaag catagcaatc taatctaagg ggcggtgttg 2101acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa 2161ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg 2221tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg 2281gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg 2341acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg 2401aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc 2461agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg 2521ccgaggagca ggactgacac gtccgacggc ggcccacggg tcccaggcct cggagatccg 2581tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct 2641ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct 2701atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt 2761ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag 2821gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg agcaaaaggc 2881cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 2941ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 3001ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 3061ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 3121tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 3181cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 3241aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 3301gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 3361agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 3421ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 3481cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 3541tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag atc//。