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pGAPZB载体基本信息
| Invitrogen |
| pGAPZ A, pGAPZ-A, pGAPZA |
| 毕赤酵母蛋白表达载体 |
| 高拷贝 |
| GAP |
| 多克隆位点，限制性内切酶 |
| 2882 bp |
| pGAP-F:5´-GTCCCTATTTCAATCAATTGAA-3´ |
| 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| C-Myc, C-His |
| Zeocin 博来霉素 |
| HIS4/ Zeocin |
插入基因是必需包含起始密码子ATG。 |
| V200-20 |
| 稳定 Stable |
| 组成型 Constitutive |
| 非病毒 |
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
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
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
//
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
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
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
//
pGAPZB载体质粒图谱pdf版和相关资料下载
pGAPZB载体应用举例
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