| 出品公司: | Invitrogen |
|---|---|
| 載體名稱: | pGAPZalpha A, pGAPZα A, pGAPZalphaA, pGAPZαA |
| 質粒類型: | 畢赤酵母蛋白表達載體 |
| 表達水平: | 高拷貝 |
| 啟動子: | GAP |
| 克隆方法: | 多克隆位點，限制性內切酶 |
| 載體大小: | 3147 bp |
| 5' 測序引物及序列: | alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´; pGAP-F:5´-GTCCCTATTTCAATCAATTGAA-3´ |
| 3' 測序引物及序列: | 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| 載體標簽: | C-Myc, C-His |
| 載體抗性: | Zeocin 博來霉素 |
| 篩選標記: | HIS4/ Zeocin |
| 備注: |
插入基因是必需包含起始密碼子ATG。
|
| 產品目錄號: | V200-20 |
| 穩定性: | 穩定 Stable |
| 組成型: | 組成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
買家導航
pGAPZα A載體 pGAPZalpha A質粒基本信息
pGAPZα A質粒圖譜 pGAPZalpha A載體圖譜和pGAPZα A載體序列 pGAPZalpha A質粒序列多克隆位點信息
pGAPZα A質粒 pGAPZalpha A載體簡介
甘油醛-3-磷酸脫氫酶（GAPDH）的酶在許多生物細胞中包括畢赤酵母，穩定高水平表達。 最近，GAPDH蛋白編碼基因啟動子（GAP）被深入研究，并在畢赤酵母中能夠高水表達重組蛋白，這取決于使用的碳源（Waterham等 人，，1997）。GAP啟動子（PGAP）的重組蛋白表達水平略高于AOX1啟動子。
pGAPZ A，B，和C載體（2.9 KB）和pGAPZα，B，和C（3.1 KB）載體使用GAP啟動子在畢赤酵母中穩定表達重組蛋白。重組蛋白表達為為帶有C-末端myc抗原表位和C端His標簽的融合蛋白。此外，pGAPZα 表達的融合蛋白在N-端帶有釀酒酵母（Saccharomyces cerevisiae）的α-因子分泌信號肽。這兩個系列的載體都提供三個不同讀碼框的載體，以方便克隆的帶有C-端標簽和/或N-末端分泌信號肽的載體 中。
這些載體是基于顯性選擇標記，Zeocin（博萊霉素）的篩選標記可以同時使用于畢赤酵母和大腸桿菌。
pGAPZα A質粒序列 pGAPZalpha A載體序列
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
//gt12365.cn/
COMMENT VNTAUTHORNAME|gt12365.cn|
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
//