| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pGAPZalpha A, pGAPZα A, pGAPZalphaA, pGAPZαA |
| 质粒类型: | 毕赤酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 启动子: | GAP |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 3147 bp |
| 5' 测序引物及序列: | alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´; pGAP-F:5´-GTCCCTATTTCAATCAATTGAA-3´ |
| 3' 测序引物及序列: | 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| 载体标签: | C-Myc, C-His |
| 载体抗性: | Zeocin 博来霉素 |
| 筛选标记: | HIS4/ Zeocin |
| 备注: |
插入基因是必需包含起始密码子ATG。
|
| 产品目录号: | ZY200-20 |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pGAPZα A
- 库存：
20
- 供应商：
泽叶生物
pGAPZalpha A载体基本信息
pGAPZalpha A载体质粒图谱和多克隆位点信息
pGAPZalpha A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 A载体序列
LOCUS pGAPZα A 3147 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) A
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..3147
/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 760..828
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 827..856
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 872..889
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 968..1214
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1229..1640
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for?EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1647..1694
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1713..2087
/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 2153..2400
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2475..3063)
/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 gaaaagagag gctgaagctg aattcacgtg gcccagccgg
781 ccgtctcgga tcggtacctc gagccgcggc ggccgccagc tttctagaac aaaaactcat
841 ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt gagttttagc
901 cttagacatg actgttcctc agttcaagtt gggcacttac gagaagaccg gtcttgctag
961 attctaatca agaggatgtc agaatgccat ttgcctgaga gatgcaggct tcatttttga
1021 tactttttta tttgtaacct atatagtata ggattttttt tgtcattttg tttcttctcg
1081 tacgagcttg ctcctgatca gcctatctcg cagctgatga atatcttgtg gtaggggttt
1141 gggaaaatca ttcgagtttg atgtttttct tggtatttcc cactcctctt cagagtacag
1201 aagattaagt gagaccttcg tttgtgcgga tcccccacac accatagctt caaaatgttt
1261 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
1321 aacacccaag cacagcatac taaattttcc ctctttcttc ctctagggtg tcgttaatta
1381 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
1441 aaaggcaata aaaattttta tcacgtttct ttttcttgaa attttttttt ttagtttttt
1501 tctctttcag tgacctccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
1561 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgttcat tagaaagaaa
1621 gcatagcaat ctaatctaag ggcggtgttg acaattaatc atcggcatag tatatcggca
1681 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
1741 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
1801 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
1861 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
1921 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
1981 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
2041 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtccgacggc
2101 ggcccacggg tcccaggcct cggagatccg tccccctttt cctttgtcga tatcatgtaa
2161 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
2221 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
2281 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
2341 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
2401 aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
2461 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
2521 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
2581 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
2641 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
2701 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
2761 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
2821 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
2881 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
2941 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
3001 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
3061 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
3121 taagggattt tggtcatgca tgagatc
//
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pGAPZα A载体
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