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基本信息

質粒圖譜和多克隆位點信息
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

pPICZA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                3329 bp ds-DNA     circular SYN 04-APR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pPICZ A

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 3329)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

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

FEATURES             Location/Qualifiers

     source          1..3329

                     /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..1011

                     /label=MCS

     CDS             1012..1041

                     /codon_start=1

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

                     /label=Myc

                     /translation="EQKLISEEDL"

     CDS             1057..1074

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     terminator      1153..1399

                     /gene="Pichia pastoris AOX1"

                     /label=AOX1 terminator

                     /note="transcription terminator for AOX1"

     promoter        1414..1825

                     /gene="S. cerevisiae TEF1"

                     /label=TEF1 promoter

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

     promoter        1833..1880

                     /label=EM7 promoter

                     /note="synthetic bacterial promoter "

     CDS             1899..2273

                     /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      2339..2586

                     /gene="S. cerevisiae CYC1"

                     /label=CYC1 terminator

                     /note="transcription terminator for CYC1"

     rep_origin      complement(2661..3249)

                     /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 agcttgggcc cgaacaaaaa

     1021 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt

     1081 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt

     1141 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt

     1201 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct

     1261 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg

     1321 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag

     1381 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa

     1441 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac

     1501 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt

     1561 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg

     1621 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt

     1681 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc

     1741 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa

     1801 agaaagcata gcaatctaat ctaaggggcg gtgttgacaa ttaatcatcg gcatagtata

     1861 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg

     1921 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg

     1981 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc

     2041 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg

     2101 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg

     2161 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc

     2221 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtcc

     2281 gacggcggcc cacgggtccc aggcctcgga gatccgtccc ccttttcctt tgtcgatatc

     2341 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa

     2401 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta

     2461 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac

     2521 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta

     2581 atttgcaagc tggagaccaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa

     2641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat

     2701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc

     2761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc

     2821 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt

     2881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac

     2941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg

     3001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca

     3061 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc

     3121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa

     3181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa

     3241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac

     3301 tcacgttaag ggattttggt catgagatc

//