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pGAPZalpha C载体基本信息
| Invitrogen |
| pGAPZalpha C, pGAPZα C, pGAPZalphaC, pGAPZαC |
| 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 |
| 3152 bp |
| alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´; 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 |
pGAPZalpha C载体质粒图谱和多克隆位点信息
pGAPZalpha C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。
pGAPZalpha C载体序列
LOCUS pGAPZα C 3152 bp ds-DNA circular SYN 05-二月-2013
DEFINITION Pichia pastoris vector for constitutive expression of a secreted
protein.
ACCESSION .
VERSION .
KEYWORDS pGAPZ(alpha) C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3152)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
JOURNAL Exported 2013年7月30日 from SnapGene Viewer 1.5.3
http://www.snapgene.com
COMMENT Linearize within the GAP promoter for integration.
FEATURES Location/Qualifiers
source 1..3152
/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"
CDS 493..759
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/note="This feature has 3 segments:
???1:?493?..?549?/?#993366?/?presequence
???2:?550?..?747?/?#993366?/?pro?region
???3:?748?..?759?/?#993366
Cleavage sites after bases 549, 747"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 759..833
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 832..861
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 877..894
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 973..1219
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1234..1645
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for?EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1652..1699
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1718..2092
/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 2158..2405
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2480..3068)
/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 cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc
541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa
601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc
661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa
721 gaagaagggg tatctctcga gaagagagag gctgaagcat cgatgaattc acgtggccca
781 gccggccgtc tcggatcggt acctcgagcc gcggcggccg ccagctttct agaacaaaaa
841 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt
901 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt
961 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt
1021 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct
1081 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg
1141 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag
1201 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa
1261 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac
1321 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt
1381 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg
1441 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt
1501 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc
1561 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa
1621 agaaagcata gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat
1681 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
1741 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
1801 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
1861 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
1921 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
1981 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
2041 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg
2101 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca
2161 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
2221 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
2281 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
2341 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
2401 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
2641 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
2881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
3121 cacgttaagg gattttggtc atgcatgaga tc
//
DEFINITION Pichia pastoris vector for constitutive expression of a secreted
protein.
ACCESSION .
VERSION .
KEYWORDS pGAPZ(alpha) C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3152)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
JOURNAL Exported 2013年7月30日 from SnapGene Viewer 1.5.3
http://www.snapgene.com
COMMENT Linearize within the GAP promoter for integration.
FEATURES Location/Qualifiers
source 1..3152
/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"
CDS 493..759
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/note="This feature has 3 segments:
???1:?493?..?549?/?#993366?/?presequence
???2:?550?..?747?/?#993366?/?pro?region
???3:?748?..?759?/?#993366
Cleavage sites after bases 549, 747"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 759..833
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 832..861
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 877..894
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 973..1219
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1234..1645
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for?EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1652..1699
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1718..2092
/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 2158..2405
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2480..3068)
/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 cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc
541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa
601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc
661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa
721 gaagaagggg tatctctcga gaagagagag gctgaagcat cgatgaattc acgtggccca
781 gccggccgtc tcggatcggt acctcgagcc gcggcggccg ccagctttct agaacaaaaa
841 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt
901 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt
961 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt
1021 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct
1081 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg
1141 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag
1201 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa
1261 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac
1321 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt
1381 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg
1441 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt
1501 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc
1561 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa
1621 agaaagcata gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat
1681 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
1741 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
1801 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
1861 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
1921 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
1981 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
2041 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg
2101 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca
2161 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
2221 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
2281 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
2341 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
2401 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
2641 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
2881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
3121 cacgttaagg gattttggtc atgcatgaga tc
//
pGAPZalpha C载体质粒图谱pdf版和相关资料下载
pGAPZalpha C载体应用举例
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