pPICZαA质粒图谱.docx

pPICZaa载体基本信息出品公司:Invitrogen载体名称:pPICZalpha A, pPICZ A, pPICZalphaA, pPICZA, 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:3593 bp5 测序引物及序列:5 AOX1:5-GACTGGTTCCAATTGACAAGC-33 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载体质粒图谱和多克隆位点信息pPICZaa载体简介pPICZ A，B和C是3.6 kb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）-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。pPICZ A，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）。-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(Escherichia coli)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 pPICZA 3593 bp DNA circular SYN 2-SEP-2003SOURCE ORGANISM COMMENT This file is created by Vector NTI /COMMENT VNTAUTHORNAME||FEATURES Location/Qualifiers source 1.3593 /invitrogen=25 /vntifkey=98 primer complement(1423.1443) /invitrogen=1880000 /vntifkey=27 /label=3AOX1primer primer 855.875 /invitrogen=1890000 /vntifkey=27 /label=5AOX1primer CDS 1320.1337 /invitrogen=1010000 /vntifkey=4 /label=6xHis primer 1144.1164 /invitrogen=1900000 /vntifkey=27 /label=alpha-factorprimer terminator 1341.1682 /invitrogen=2550000 /vntifkey=43 /label=AOX1transcriptionterminator CDS 1275.1304 /invitrogen=1090000 /vntifkey=4 /label=c-mycepitope terminator 2538.2855 /invitrogen=2560000 /vntifkey=43 /label=CYC1transcriptionterminator promoter 2095.2162 /invitrogen=2340000 /vntifkey=30 /label=EM7promoter misc_feature 1208.1274 /invitrogen=1460000 /vntifkey=21 /label=MCS rep_origin complement(2866.3539) /invitrogen=2540000 /vntifkey=33 /label=pUCorigin CDS 941.1207 /invitrogen=1280000 /vntifkey=4 /label=alpha-factorsignalpeptide promoter 1683.2094 /invitrogen=2300000 /vntifkey=29 /label=TEF1promoter CDS 2163.2537 /invitrogen=1370000 /vntifkey=4 /label=Zeo(R) promoter 1.940 /invitrogen=2160000 /vntifkey=29 /label=AOX1promoterBASE COUNT 892 a 886 c 801 g 1014 tORIGIN 1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1201 tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg 1261 ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc 1321 atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg 1381 ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt 1441 tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag 1501 gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc 1561 agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt 1621 ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat 1681 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc 1741 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaattttccc 1801 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga 1861 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt 1921 tttcttgaaa tttttttttt tagttttttt ctctttcagt gacctccatt gatatttaag 1981 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact 2041 ttttttactt cttgttcatt agaaagaaag catagcaatc taatctaagg ggcggtgttg 2101 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa 2161 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg 2221 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg 2281 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg 2341 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg 2401 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc 2461 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg 2521 ccgaggagca ggactgacac gtccgacggc ggcccacggg tcccaggcct cggagatccg 2581 tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct 2641 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct 2701 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt 2761 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag 2821 gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg agcaaaaggc 2881 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 2941 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 3001 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttcc