pGAPZ C

pGAPZ C

pGAPZC载体基本信息

pGAPZC载体质粒图谱和多克隆位点信息

pGAPZC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。

pGAPZC载体序列

LOCUS       pGAPZ C           2883 bp ds-DNA    circular SYN 05-二月-2013
DEFINITION  Pichia pastoris vector for constitutive intracellular expression of 
            a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGAPZ C
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..2883
                     /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..562
                     /note="MCS"
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     CDS             563..592
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             608..625
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      704..950
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 terminator"
                     /note="transcription terminator for AOX1"
                     /note="color: #ffffff"
     promoter        965..1376
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for?EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        1383..1430
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter?"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1449..1823
                     /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      1889..2136
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
     rep_origin      complement(2211..2799)
                     /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 ccagcttacg tagaacaaaa actcatctca gaagaggatc tgaatagcgc
      601 cgtcgaccat catcatcatc atcattgagt tttagcctta gacatgactg ttcctcagtt
      661 caagttgggc acttacgaga agaccggtct tgctagattc taatcaagag gatgtcagaa
      721 tgccatttgc ctgagagatg caggcttcat ttttgatact tttttatttg taacctatat
      781 agtataggat tttttttgtc attttgtttc ttctcgtacg agcttgctcc tgatcagcct
      841 atctcgcagc tgatgaatat cttgtggtag gggtttggga aaatcattcg agtttgatgt
      901 ttttcttggt atttcccact cctcttcaga gtacagaaga ttaagtgaga ccttcgtttg
      961 tgcggatccc ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga
     1021 ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
     1081 ttttccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
     1141 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
     1201 gtttcttttt cttgaaattt ttttttttag tttttttctc tttcagtgac ctccattgat
     1261 atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat
     1321 tacaactttt tttacttctt gttcattaga aagaaagcat agcaatctaa tctaagggcg
     1381 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     1441 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     1501 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     1561 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     1621 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     1681 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     1741 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     1801 tcgtggccga ggagcaggac tgacacgtcc gacggcggcc cacgggtccc aggcctcgga
     1861 gatccgtccc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     1921 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     1981 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     2041 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     2101 gagaaggttt tgggacgctc gaaggcttta atttgcaagc tggagaccaa catgtgagca
     2161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2401 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2641 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcatgag
     2881 atc
//

 pGAPZ C其他酵母表达载体：