| 出品公司: | Invitrogen |
|---|---|
| 載體名稱: | pGAPZ C, pGAPZ-C, pGAPZC |
| 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: | 2883 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 C載體 pGAPZC質粒基本信息
pGAPZ C質粒圖譜 pGAPZC載體圖譜和pGAPZ C載體序列 pGAPZC質粒序列多克隆位點信息
pGAPZ C質粒 pGAPZC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 C質粒序列 pGAPZC載體序列
LOCUS pGAPZ C 2883 bp ds-DNA circular SYN 05-二月-2013
DEFINITION Pichia pastoris vector for constitutive intracellular expression of
a protein.
ACCESSION .
VERSION .
KEYWORDS pGAPZ C
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..2883
/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..562
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 563..592
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 608..625
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 704..950
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 965..1376
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for?EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1383..1430
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1449..1823
/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 1889..2136
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2211..2799)
/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 ccagcttacg tagaacaaaa actcatctca gaagaggatc tgaatagcgc
601 cgtcgaccat catcatcatc atcattgagt tttagcctta gacatgactg ttcctcagtt
661 caagttgggc acttacgaga agaccggtct tgctagattc taatcaagag gatgtcagaa
721 tgccatttgc ctgagagatg caggcttcat ttttgatact tttttatttg taacctatat
781 agtataggat tttttttgtc attttgtttc ttctcgtacg agcttgctcc tgatcagcct
841 atctcgcagc tgatgaatat cttgtggtag gggtttggga aaatcattcg agtttgatgt
901 ttttcttggt atttcccact cctcttcaga gtacagaaga ttaagtgaga ccttcgtttg
961 tgcggatccc ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga
1021 ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
1081 ttttccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
1141 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
1201 gtttcttttt cttgaaattt ttttttttag tttttttctc tttcagtgac ctccattgat
1261 atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat
1321 tacaactttt tttacttctt gttcattaga aagaaagcat agcaatctaa tctaagggcg
1381 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
1441 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
1501 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
1561 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
1621 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
1681 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
1741 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
1801 tcgtggccga ggagcaggac tgacacgtcc gacggcggcc cacgggtccc aggcctcgga
1861 gatccgtccc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
1921 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
1981 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
2041 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
2101 gagaaggttt tgggacgctc gaaggcttta atttgcaagc tggagaccaa catgtgagca
2161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
2221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
2281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
2341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
2401 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
2461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
2521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
2581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
2641 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
2701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
2761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
2821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcatgag
2881 atc
//