pGAPZ A

pGAPZ A

pGAPZA载体基本信息

pGAPZA载体质粒图谱和多克隆位点信息

pGAPZA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。

pGAPZA载体序列

LOCUS       pGAPZ A           2884 bp ds-DNA    circular SYN 05-二月-2013
DEFINITION  Pichia pastoris vector for constitutive intracellular expression of 
            a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGAPZ A
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..2884
                     /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             564..593
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             609..626
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      705..951
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 terminator"
                     /note="transcription terminator for AOX1"
                     /note="color: #ffffff"
     promoter        966..1377
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for?EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        1384..1431
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter?"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1450..1824
                     /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      1890..2137
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
     rep_origin      complement(2212..2800)
                     /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 ccagcttggg cccgaacaaa aactcatctc agaagaggat ctgaatagcg
      601 ccgtcgacca tcatcatcat catcattgag ttttagcctt agacatgact gttcctcagt
      661 tcaagttggg cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga
      721 atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata
      781 tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc
      841 tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg
      901 tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt
      961 gtgcggatcc cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag
     1021 attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa
     1081 attttccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag
     1141 aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca
     1201 cgtttctttt tcttgaaatt ttttttttta gtttttttct ctttcagtga cctccattga
     1261 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
     1321 ttacaacttt ttttacttct tgttcattag aaagaaagca tagcaatcta atctaagggc
     1381 ggtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     1441 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     1501 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     1561 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     1621 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     1681 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     1741 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     1801 ttcgtggccg aggagcagga ctgacacgtc cgacggcggc ccacgggtcc caggcctcgg
     1861 agatccgtcc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     1921 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1981 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     2041 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     2101 tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctggagacca acatgtgagc
     2161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2401 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2641 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcatga
     2881 gatc
//

pGAPZ A其他酵母表达载体：