pGAPZ B

pGAPZ B

pGAPZB载体基本信息

pGAPZB载体质粒图谱和多克隆位点信息

pGAPZB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。

pGAPZB载体序列

LOCUS       pGAPZ B           2882 bp ds-DNA    circular SYN 05-二月-2013
DEFINITION  Pichia pastoris vector for constitutive intracellular expression of 
            a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGAPZ B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     This file is created by Vector NTI
COMMENT     Linearize within the GAP promoter for integration.
FEATURES             Location/Qualifiers
     source          1..2882
                     /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"
     misc_feature    484..563
                     /note="MCS"
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     CDS             562..591
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             607..624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      703..949
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 terminator"
                     /note="transcription terminator for AOX1"
                     /note="color: #ffffff"
     promoter        964..1375
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for?EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        1382..1429
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter?"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1448..1822
                     /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      1888..2135
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
     rep_origin      complement(2210..2798)
                     /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 cgaggaattc acgtggccca gccggccgtc tcggatcggt acctcgagcc
      541 gcggcggccg ccagctttct agaacaaaaa ctcatctcag aagaggatct gaatagcgcc
      601 gtcgaccatc atcatcatca tcattgagtt ttagccttag acatgactgt tcctcagttc
      661 aagttgggca cttacgagaa gaccggtctt gctagattct aatcaagagg atgtcagaat
      721 gccatttgcc tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata
      781 gtataggatt ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta
      841 tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt
      901 tttcttggta tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt
      961 gcggatcccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
     1021 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
     1081 tttccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
     1141 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
     1201 tttctttttc ttgaaatttt tttttttagt ttttttctct ttcagtgacc tccattgata
     1261 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     1321 acaacttttt ttacttcttg ttcattagaa agaaagcata gcaatctaat ctaagggcgg
     1381 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     1441 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     1501 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     1561 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     1621 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     1681 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     1741 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     1801 cgtggccgag gagcaggact gacacgtccg acggcggccc acgggtccca ggcctcggag
     1861 atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     1921 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1981 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     2041 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     2101 agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgagcaa
     2161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2401 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2641 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcatgaga
     2881 tc
//

pGAPZ B其他酵母表达载体：