| 出品公司: | Invitrogen |
|---|---|
| 載體名稱: | pGAPZ A, pGAPZ-A, pGAPZA |
| 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: | 2884 bp |
| 5' 測序引物及序列: | 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
pGAPZ A載體 pGAPZA質粒基本信息
pGAPZ A質粒圖譜 pGAPZA載體圖譜和pGAPZ A載體序列 pGAPZA質粒序列多克隆位點信息
pGAPZ A質粒 pGAPZA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。
pGAPZ A質粒序列 pGAPZA載體序列
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
//gt12365.cn/
COMMENT VNTAUTHORNAME|gt12365.cn|
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
//