| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pGAPZ C, pGAPZ-C, pGAPZC |
| 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: | 2883 bp |
| 5' 测序引物及序列: | 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: | V200-20 |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
买家导航
pGAPZ C载体 pGAPZC质粒基本信息
pGAPZ C质粒图谱 pGAPZC载体图谱和pGAPZ C载体序列 pGAPZC质粒序列多克隆位点信息
pGAPZ C质粒 pGAPZC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。
pGAPZ C质粒序列 pGAPZC载体序列
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
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
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
//
