pPICZ B

pPICZ B

pPICZB载体基本信息

pPICZB载体质粒图谱和多克隆位点信息

pPICZB载体简介

pPICZ A，B和C是3.3 kb的毕赤酵母蛋白载体。表达的重组蛋白是融合蛋白，含有一个C-端多肽，多太重含c-myc和C-His标签。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系列载体包含以下元素：

•5'片段含有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）。

•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。

pPICZB载体序列

LOCUS       pPICZ B           3328 bp ds-DNA    circular 
DEFINITION  Pichia pastoris vector for methanol-inducible intracellular 
            expression of a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pPICZ B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     This file is created by Vector NTI
COMMENT     Linearize with SacI, PmeI, or BstXI for integration at the AOX1 
            locus.
FEATURES             Location/Qualifiers
     source          1..3328
                     /organism="synthetic DNA construct"
                     /lab_host="Pichia pastoris"
                     /mol_type="other DNA"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 promoter"
                     /note="inducible promoter, regulated by methanol"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    932..1011
                     /note="MCS"
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     CDS             1010..1039
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             1055..1072
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      1152..1398
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 terminator"
                     /note="transcription terminator for AOX1"
                     /note="color: #ffffff"
     promoter        1438..1824
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        1832..1879
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter?"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1898..2272
                     /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      2338..2585
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
     rep_origin      complement(2660..3248)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        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 aggaattcac gtggcccagc
      961 cggccgtctc ggatcggtac ctcgagccgc ggcggccgcc agctttctag aacaaaaact
     1021 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagtttg
     1081 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg
     1141 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt
     1201 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt
     1261 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg
     1321 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt
     1381 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat
     1441 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     1501 tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta
     1561 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     1621 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt
     1681 tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca
     1741 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa
     1801 gaaagcatag caatctaatc taaggggcgg tgttgacaat taatcatcgg catagtatat
     1861 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     1921 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     1981 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     2041 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     2101 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     2161 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     2221 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg
     2281 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca
     2341 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     2401 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     2461 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     2521 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     2581 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2821 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     2881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3061 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3301 cacgttaagg gattttggtc atgagatc
//

pPICZ B其他酵母表达载体：