• 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-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 13

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 3151)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4

            http://www.fenghbio.cn

FEATURES             Location/Qualifiers

     source          1..3151

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        7..483

                     /note="GAP promoter"

                     /note="promoter for Pichia pastoris 

                     glyceraldehyde-3-phosphate dehydrogenase"

     CDS             493..759

                     /codon_start=1

                     /gene="MF-alpha-1"

                     /product="N-terminal secretion signal from S. cerevisiae 

                     alpha-factor"

                     /note="alpha-factor secretion signal"

                     /note="Cleavage by the Kex2 protease occurs after the 

                     dibasic KR sequence. The EA dipeptides are then removed by 

                     dipeptidyl aminopeptidase A."

                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE

                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"

     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, and 

                     Zeocin(TM)"

                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD

                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR

                     EFALRDPAGNCVHFVAEEQD"

     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 of 

                     replication"

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 cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc

      541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa

      601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc

      661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa

      721 gaagaagggg tatctctcga gaaaagagag gctgaagctg caggaattca cgtggcccag

      781 ccggccgtct cggatcggta cctcgagccg cggcggccgc cagctttcta gaacaaaaac

      841 tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt

      901 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg

      961 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt

     1021 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt

     1081 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg

     1141 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt

     1201 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat

     1261 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact

     1321 tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta

     1381 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt

     1441 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt

     1501 tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca

     1561 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa

     1621 gaaagcatag caatctaatc taagggcggt gttgacaatt aatcatcggc atagtatatc

     1681 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt

     1741 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg

     1801 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg

     1861 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg

     1921 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac

     1981 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg

     2041 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtccga

     2101 cggcggccca cgggtcccag gcctcggaga tccgtccccc ttttcctttg tcgatatcat

     2161 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa

     2221 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt

     2281 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc

     2341 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat

     2401 ttgcaagctg gagaccaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa

     2461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg

     2521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc

     2581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc

     2641 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc

     2701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg

     2761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc

     2821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga

     2881 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc

     2941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac

     3001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg

     3061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc

     3121 acgttaaggg attttggtca tgcatgagat c

//