- 商品货号:ZT1777
- 规格:10μl质粒
- 启动子:GAP
- 复制子:pUC ori
- 终止子:AOX1 TT
- 质粒大小:3251bp
- 质粒标签:C-Myc, C-6×His
- 原核抗性:Zeo
- 筛选标记:博来霉素Zeo
- 克隆菌株:DH5α
- 表达宿主:酵母细胞
- 5测序引物:pGAP-F:GTCCCTATTTCAATCAATTGAA
- 3测序引物:AOX3:GGCAAATGGCATTCTGACAT
- 质粒简介: 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pGAPZαB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。
- 质粒图谱1: png/4-16091014260V35.png
质粒序列: 点击查看序列LOCUS Exported 3152 bp ds-DNA circular SYN 10-SEP-2016DEFINITION synthetic circular DNAKEYWORDS Untitled 14SOURCE synthetic DNA constructORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 3152)AUTHORS .TITLE Direct SubmissionJOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4http://www.fenghbio.cnFEATURES Location/Qualifierssource 1..3152/organism="synthetic DNA construct"/mol_type="other DNA"promoter 7..483/note="GAP promoter"/note="promoter for Pichia pastorisglyceraldehyde-3-phosphate dehydrogenase"CDS 832..861/codon_start=1/product="Myc (human c-Myc oncogene) epitope tag"/note="Myc"/translation="EQKLISEEDL"CDS 877..894/codon_start=1/product="6xHis affinity tag"/note="6xHis"/translation="HHHHHH"terminator 973..1219/gene="Pichia pastoris AOX1"/note="AOX1 terminator"/note="transcription terminator for AOX1"promoter 1234..1645/gene="S. cerevisiae TEF1"/note="TEF1 promoter"/note="promoter for EF-1-alpha"promoter 1652..1699/note="EM7 promoter"/note="synthetic bacterial promoter "CDS 1718..2092/codon_start=1/gene="Sh ble from Streptoalloteichus hindustanus"/product="antibiotic-binding protein"/note="BleoR"/note="confers resistance to bleomycin, phleomycin, andZeocin(TM)"/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDDVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGREFALRDPAGNCVHFVAEEQD"terminator 2158..2405/gene="S. cerevisiae CYC1"/note="CYC1 terminator"/note="transcription terminator for CYC1"rep_origin complement(2480..3068)/direction=LEFT/note="ori"/note="high-copy-number ColE1/pMB1/pBR322/pUC origin ofreplication"ORIGIN1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac481 tatttcgaaa cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa721 gaagaagggg tatctctcga gaagagagag gctgaagcat cgatgaattc acgtggccca781 gccggccgtc tcggatcggt acctcgagcc gcggcggccg ccagctttct agaacaaaaa841 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt901 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt961 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt1021 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct1081 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg1141 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag1201 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa1261 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac1321 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt1381 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg1441 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt1501 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc1561 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa1621 agaaagcata gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat1681 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt1741 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg1801 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct1861 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt1921 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga1981 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct2041 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg2101 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca2161 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa2221 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag2281 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg2341 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa2401 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg2641 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag2881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact3121 cacgttaagg gattttggtc atgcatgaga tc//
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