pPICZα C

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pPICZalpha C载体基本信息

出品公司: Invitrogen
载体名称: pPICZalpha C, pPICZα C, pPICZalphaC, pPICZαC
质粒类型: 毕赤酵母蛋白表达载体
表达水平: 高拷贝
启动子: AOX1
克隆方法: 多克隆位点,限制性内切酶
载体大小: 3598 bp
5' 测序引物及序列: 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-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
备注:
插入基因是必需包含起始密码子ATG。
产品目录号: V195-20
稳定性: 稳定 Stable
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pPICZalpha C载体质粒图谱和多克隆位点信息

pPICZalph C载体图谱

pPICZalph C多克隆位点

pPICZalpha C载体简介

pPICZα  A,B和C是3.6 kb的毕赤酵母蛋白分泌表达载体。表达的重组蛋白是融合蛋白,含有一个N-端多肽,编码酿酒酵母(Saccharomyces cerevisiae)α-因子分泌信号。载体能够在毕赤酵母中利用甲醇诱导的高水 平的表达目的蛋白,并且可以用在任何毕赤酵母中,包括X33,GS115菌株,SMD1168H,KM71H。

pPICZα  A,B和C系列载体包含以下元素:

•5'端含有AOX1启动子的严格调控,利用甲醇诱导表达任何感兴趣的基因(Ellis等,1985; Koutz等人,1989;tschopp等人,1987A)。

α-因子分泌信号能够分泌性表达目的蛋白。

•Zeocin抗性基因在大肠杆菌和毕赤酵母都能用于筛选(Baron等人,1992; Drocourt等人,1990)。

•C-端含Myc和His标签,可以用于检测和纯化重组蛋白。

•pPICZ A, B, C三种读码框使得可以将基因克隆入载体而不发生任何移码突变。

pPICZalpha C载体序列

LOCUS       pPICZα C           3598 bp ds-DNA    circular SYN 05-二月-2013
DEFINITION  Pichia pastoris vector for methanol-inducible expression of a 
            secreted protein.
ACCESSION   .
VERSION     .
KEYWORDS    pPICZ(alpha) 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 with SacI, PmeI, or BstXI for integration at the AOX1 
            locus.
FEATURES             Location/Qualifiers
     source          1..3598
                     /organism="synthetic DNA construct"
                     /lab_host="Pichia pastoris"
                     /mol_type="other DNA"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 promoter"
                     /note="inducible promoter, regulated by methanol"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             941..1207
                     /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."
                     /note="This feature has 3 segments:
                     ???1:??941?..??997?/?#993366?/?presequence
                     ???2:??998?..?1195?/?#993366?/?pro?region
                     ???3:?1196?..?1207?/?#993366
                     Cleavage sites after bases 997, 1195"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     misc_feature    1207..1281
                     /note="MCS"
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     CDS             1280..1309
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             1325..1342
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      1422..1668
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 terminator"
                     /note="transcription terminator for AOX1"
                     /note="color: #ffffff"
     promoter        1708..2094
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        2102..2149
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter?"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2168..2542
                     /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      2608..2855
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
     rep_origin      complement(2930..3518)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt
      961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
     1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
     1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
     1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga agagagaggc
     1201 tgaagcatcg atgaattcac gtggcccagc cggccgtctc ggatcggtac ctcgagccgc
     1261 ggcggccgcc agctttctag aacaaaaact catctcagaa gaggatctga atagcgccgt
     1321 cgaccatcat catcatcatc attgagtttg tagccttaga catgactgtt cctcagttca
     1381 agttgggcac ttacgagaag accggtcttg ctagattcta atcaagagga tgtcagaatg
     1441 ccatttgcct gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag
     1501 tataggattt tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat
     1561 ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt
     1621 ttcttggtat ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg
     1681 cggatccccc acacaccata gcttcaaaat gtttctactc cttttttact cttccagatt
     1741 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     1801 ttccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     1861 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     1921 ttctttttct tgaaattttt ttttttagtt tttttctctt tcagtgacct ccattgatat
     1981 ttaagttaat aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta
     2041 caactttttt tacttcttgt tcattagaaa gaaagcatag caatctaatc taaggggcgg
     2101 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     2161 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     2221 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     2281 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     2341 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     2401 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     2461 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     2521 cgtggccgag gagcaggact gacacgtccg acggcggccc acgggtccca ggcctcggag
     2581 atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     2641 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     2701 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     2761 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     2821 agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgagcaa
     2881 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2941 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3001 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3061 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3121 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3181 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3241 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3301 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3361 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3421 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3481 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3541 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagatc
//

pPICZalpha C载体质粒图谱pdf版和相关资料下载

pPICZalpha C载体应用举例

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