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基本信息

注意：

客戶在我公司選擇構建載體服務項目時，請自行提供母源載體。

如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。 

載體描述

pPICZB載體簡介

pPICZ A，B和C是3.3 kb的畢赤酵母蛋白載體。表達的重組蛋白是融合蛋白，含有一個C-端多肽，多太重含c-myc和C-His標簽。載體能夠在畢赤酵母中利用甲醇誘導的高水平的表達目的蛋白，并且可以用在任何畢赤酵母中，包括X33，GS115菌株，SMD1168H，KM71H。

pPICZ系列載體包含以下元素：

?5'片段含有AOX1啟動子的嚴格調控，利用甲醇誘導表達任何感興趣的基因（Ellis等，1985; Koutz等人，1989;tschopp等人，1987A）。

?Zeocin抗性基因在大腸桿菌和畢赤酵母都能用于篩選（Baron等人,1992; Drocourt等人，1990）。

?C-端含Myc和His標簽，可以用于檢測和純化重組蛋白。

?pPICZ A, B, C三種讀碼框使得可以將基因克隆入載體而不發生任何移碼突變。

載體序列

LOCUS       Exported                3328 bp ds-DNA     circular SYN 04-APR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pPICZ B

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 3328)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Thursday, Apr 4, 2019 from SnapGene Viewer 4.3.6

FEATURES             Location/Qualifiers

     source          1..3328

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        2..940

                     /gene="Pichia pastoris AOX1"

                     /label=AOX1 promoter

                     /note="inducible promoter, regulated by methanol"

     misc_feature    943..1009

                     /label=MCS

     CDS             1010..1039

                     /codon_start=1

                     /product="Myc (human c-Myc proto-oncogene) epitope tag"

                     /label=Myc

                     /translation="EQKLISEEDL"

     CDS             1055..1072

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     terminator      1152..1398

                     /gene="Pichia pastoris AOX1"

                     /label=AOX1 terminator

                     /note="transcription terminator for AOX1"

     promoter        1413..1824

                     /gene="S. cerevisiae TEF1"

                     /label=TEF1 promoter

                     /note="promoter for EF-1-alpha"

     promoter        1832..1879

                     /label=EM7 promoter

                     /note="synthetic bacterial promoter "

     CDS             1898..2272

                     /codon_start=1

                     /gene="Sh ble from Streptoalloteichus hindustanus"

                     /product="antibiotic-binding protein"

                     /label=BleoR

                     /note="confers resistance to bleomycin, phleomycin, and

                     Zeocin(TM)"

                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD

                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR

                     EFALRDPAGNCVHFVAEEQD"

     terminator      2338..2585

                     /gene="S. cerevisiae CYC1"

                     /label=CYC1 terminator

                     /note="transcription terminator for CYC1"

     rep_origin      complement(2660..3248)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

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 aggaattcac gtggcccagc

      961 cggccgtctc ggatcggtac ctcgagccgc ggcggccgcc agctttctag aacaaaaact

     1021 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagtttg

     1081 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg

     1141 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt

     1201 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt

     1261 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg

     1321 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt

     1381 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat

     1441 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact

     1501 tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta

     1561 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt

     1621 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt

     1681 tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca

     1741 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa

     1801 gaaagcatag caatctaatc taaggggcgg tgttgacaat taatcatcgg catagtatat

     1861 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt

     1921 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg

     1981 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct

     2041 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt

     2101 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga

     2161 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct

     2221 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg

     2281 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca

     2341 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa

     2401 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag

     2461 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg

     2521 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa

     2581 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa

     2641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc

     2701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc

     2761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg

     2821 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt

     2881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc

     2941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc

     3001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag

     3061 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg

     3121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa

     3181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag

     3241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact

     3301 cacgttaagg gattttggtc atgagatc

//