- 商品货号:ZT1766
- 规格:10μl质粒
- 启动子:GAP
- 复制子:pUC ori
- 终止子:AOX1 TT
- 质粒分类:酵母系列,毕赤酵母表达载体
- 质粒大小:3151bp
- 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/4-160910142035395.png
质粒序列: 点击查看序列LOCUS Exported 3151 bp ds-DNA circular SYN 10-SEP-2016DEFINITION synthetic circular DNAACCESSION .VERSION .KEYWORDS Untitled 13SOURCE synthetic DNA constructORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 3151)AUTHORS .TITLE Direct SubmissionJOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4http://www.fenghbio.cnFEATURES Location/Qualifierssource 1..3151/organism="synthetic DNA construct"/mol_type="other DNA"promoter 7..483/note="GAP promoter"/note="promoter for Pichia pastorisglyceraldehyde-3-phosphate dehydrogenase"CDS 493..759/codon_start=1/gene="MF-alpha-1"/product="N-terminal secretion signal from S. cerevisiaealpha-factor"/note="alpha-factor secretion signal"/note="Cleavage by the Kex2 protease occurs after thedibasic KR sequence. The EA dipeptides are then removed bydipeptidyl aminopeptidase A."/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLEGDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"CDS 831..860/codon_start=1/product="Myc (human c-Myc oncogene) epitope tag"/note="Myc"/translation="EQKLISEEDL"CDS 876..893/codon_start=1/product="6xHis affinity tag"/note="6xHis"/translation="HHHHHH"terminator 972..1218/gene="Pichia pastoris AOX1"/note="AOX1 terminator"/note="transcription terminator for AOX1"promoter 1233..1644/gene="S. cerevisiae TEF1"/note="TEF1 promoter"/note="promoter for EF-1-alpha"promoter 1651..1698/note="EM7 promoter"/note="synthetic bacterial promoter "CDS 1717..2091/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 2157..2404/gene="S. cerevisiae CYC1"/note="CYC1 terminator"/note="transcription terminator for CYC1"rep_origin complement(2479..3067)/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 gaaaagagag gctgaagctg caggaattca cgtggcccag781 ccggccgtct cggatcggta cctcgagccg cggcggccgc cagctttcta gaacaaaaac841 tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt901 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg961 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt1021 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt1081 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg1141 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt1201 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat1261 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact1321 tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta1381 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt1441 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt1501 tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca1561 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa1621 gaaagcatag caatctaatc taagggcggt gttgacaatt aatcatcggc atagtatatc1681 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt1741 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg1801 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg1861 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg1921 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac1981 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg2041 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtccga2101 cggcggccca cgggtcccag gcctcggaga tccgtccccc ttttcctttg tcgatatcat2161 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa2221 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt2281 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc2341 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat2401 ttgcaagctg gagaccaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa2461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg2521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc2581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc2641 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc2701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg2761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc2821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga2881 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc2941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac3001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg3061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc3121 acgttaaggg attttggtca tgcatgagat c//
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