pPIC9序列_分类序列

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载体基本信息

出品公司: 载体名称: 质粒类型: 表达水平: 启动子: 克隆方法: 载体大小: 5' 测序引物及序列: 3' 测序引物及序列: 载体标签: 载体抗性: 筛选标记: 备注: 产品目录号: 稳定性: 组成型: 病毒/非病毒: 载体质粒图谱和多克隆位点信息

Invitrogen pPIC9, pPIC 9

毕赤酵母蛋白表达载体 高拷贝 AOX1

多克隆位点，限制性内切酶 8023 bp

5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´；

alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´

/ Ampicillin HIS4 利用alpha分泌因子，分泌表达蛋白。K1710-01 稳定 Stable 组成型 Constitutive 非病毒

载体简介

pPIC9载体的详细信息如下：

• pPIC9载体大小8,023 bp，是融合表达载体。

• 载体构建过程中，可供使用的单一限制性内切酶位点为Xho I, SnaB I, EcoR I, Avr II, Not I.• 利用alpha因子分泌信号肽，分泌表达蛋白基因。

• 在载体构建过程中，你的基因必须保证与信号肽的起始密码子的读码框一致。

• 毕赤酵母中利用HIS4进行筛选。

• 为了将基因插入GS115或KM71的AOX区, 使用SacI限制性内切酶线性化质粒(GS115中产生His+Mut+基因型，KM71中产生His+ MutS基因型)

•为了将基因插入GS115或KM71的His4区, 使用Sal I或者Stu I限制性内切酶线性化质粒(GS115中产生His+ Mut+基因型，KM71中产生His+ MutS基因型)

•将基因插入GS115的AOX1区域，需要使用Bgl II线性化质粒(产生His+MutS基因型)。载体序列

LOCUS pPIC9 8023 bp ds-DNA circular SYN DEFINITION Pichia pastoris HIS4 vector for methanol-inducible expreion of a secreted protein.ACCESSION.VERSION.KEYWORDS pPIC9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct COMMENT This file is created by Vector NTI http://www.daodoc.com/ COMMENT VNTAUTHORNAME|biofeng.com| FEATURES Location/Qualifiers source 1..8023 /organism=“synthetic DNA construct” /lab_host=“Pichia pastoris” /mol_type=“other DNA” promoter 2..937 /gene=“Pichia pastoris AOX1” /note=“AOX1 promoter” /note=“inducible promoter, regulated by methanol” /note=“color: #ffffff;direction: RIGHT” CDS 949..1215 /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:??949?..?1005?/?#993366?/?presequence ???2:?1006?..?1203?/?#993366?/?pro?region ???3:?1204?..?1215?/?#993366 Cleavage sites after bases 1005, 1203” /translation=“MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA” misc_feature 1192..1241 /note=“MCS” /note=“multiple cloning site” /note=“color: #99ccff” terminator 1321..1567 /gene=“Pichia pastoris AOX1” /note=“AOX1 terminator” /note=“transcription terminator for AOX1” /note=“color: #ffffff” CDS complement(1980..4514)/codon_start=1 /gene=“Pichia pastoris HIS4” /locus_tag=“ ” /product=“multifunctional enzyme, required for histidine biosynthesis” /note=“PpHIS4” /note=“auxotrophic marker for Pichia pastoris” /note=“color: #ff7f50” /protein_id=“ ” /translation=“MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI” misc_feature 4869..5625 /note=“AOX1 3' fragment” /note=“region downstream of Pichia pastoris AOX1 gene” /note=“color: #ffffff” rep_origin complement(6094..6682)/direction=LEFT /note=“ori” /note=“high-copy-number ColE1/pMB1/pBR322/pUC origin of replication” /note=“color: #ffff00” CDS complement(6853..7713)/codon_start=1 /gene=“bla” /product=“beta-lactamase” /note=“AmpR” /note=“confers resistance to ampicillin, carbenicillin, and related antibiotics” /note=“This feature has 2 segments: ???1:?6853?..?7644?/?#ccffcc ???2:?7645?..?7713?/?#ccffcc?/?signal?sequence Cleavage site after base 7644” /translation=“MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW” promoter complement(7714..7818)/gene=“bla” /note=“AmpR promoter” /note=“color: #ffffff;direction: LEFT” 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 aaagtcgcca 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 attcgaagga tccaaacgat gagatttcct 961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact 1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta 1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg 1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa 1201 agagaggctg aagcttacgt agaattccct agggcggccg cgaattaatt cgccttagac 1261 atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa 1321 tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt 1381 ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc 1441 ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa 1501 tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta 1561 agtgagaagt tcgtttgtgc aagcttatcg ataagcttta atgcggtagt ttatcacagt 1621 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc 1681 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc 1741 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc 1801 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg 1861 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg 1921 cgaccacacc cgtcctgtgg atctatcgaa tctaaatgta agttaaaatc tctaaataat 1981 taaataagtc ccagtttctc catacgaacc ttaacagcat tgcggtgagc atctagacct 2041 tcaacagcag ccagatccat cactgcttgg ccaatatgtt tcagtccctc aggagttacg 2101 tcttgtgaag tgatgaactt ctggaaggtt gcagtgttaa ctccgctgta ttgacgggca 2161 tatccgtacg ttggcaaagt gtggttggta ccggaggagt aatctccaca actctctgga 2221 gagtaggcac caacaaacac agatccagcg tgttgtactt gatcaacata agaagaagca 2281 ttctcgattt gcaggatcaa gtgttcagga gcgtactgat tggacatttc caaagcctgc 2341 tcgtaggttg caaccgatag ggttgtagag tgtgcaatac acttgcgtac aatttcaacc 2401 cttggcaact gcacagcttg gttgtgaaca gcatcttcaa ttctggcaag ctccttgtct 2461 gtcatatcga cagccaacag aatcacctgg gaatcaatac catgttcagc ttgagacaga 2521 aggtctgagg caacgaaatc tggatcagcg tatttatcag caataactag aacttcagaa 2581 ggcccagcag gcatgtcaat actacacagg gctgatgtgt cattttgaac catcatcttg 2641 gcagcagtaa cgaactggtt tcctggacca aatattttgt cacacttagg aacagtttct 2701 gttccgtaag ccatagcagc tactgcctgg gcgcctcctg ctagcacgat acacttagca 2761 ccaaccttgt gggcaacgta gatgacttct ggggtaaggg taccatcctt cttaggtgga 2821 gatgcaaaaa caatttcttt gcaaccagca actttggcag gaacacccag catcagggaa 2881 gtggaaggca gaattgcggt tccaccagga atatagaggc caactttctc aataggtctt 2941 gcaaaacgag agcagactac accagggcaa gtctcaactt gcaacgtctc cgttagttga 3001 gcttcatgga atttcctgac gttatctata gagagatcaa tggctctctt aacgttatct 3061 ggcaattgca taagttcctc tgggaaagga gcttctaaca caggtgtctt caaagcgact 3121 ccatcaaact tggcagttag ttctaaaagg gctttgtcac cattttgacg aacattgtcg 3181 acaattggtt tgactaattc cataatctgt tccgttttct ggataggacg acgaagggca 3241 tcttcaattt cttgtgagga ggccttagaa acgtcaattt tgcacaattc aatacgacct 3301 tcagaaggga cttctttagg tttggattct tctttaggtt gttccttggt gtatcctggc 3361 ttggcatctc ctttccttct agtgaccttt agggacttca tatccaggtt tctctccacc 3421 tcgtccaacg tcacaccgta cttggcacat ctaactaatg caaaataaaa taagtcagca 3481 cattcccagg ctatatcttc cttggattta gcttctgcaa gttcatcagc ttcctcccta 3541 attttagcgt tcaacaaaac ttcgtcgtca aataaccgtt tggtataaga accttctgga 3601 gcattgctct tacgatccca caaggtggct tccatggctc taagaccctt tgattggcca 3661 aaacaggaag tgcgttccaa gtgacagaaa ccaacacctg tttgttcaac cacaaatttc 3721 aagcagtctc catcacaatc caattcgata cccagcaact tttgagttgc tccagatgta 3781 gcacctttat accacaaacc gtgacgacga gattggtaga ctccagtttg tgtccttata 3841 gcctccggaa tagacttttt ggacgagtac accaggccca acgagtaatt agaagagtca 3901 gccaccaaag tagtgaatag accatcgggg cggtcagtag tcaaagacgc caacaaaatt 3961 tcactgacag ggaacttttt gacatcttca gaaagttcgt attcagtagt caattgccga 4021 gcatcaataa tggggattat accagaagca acagtggaag tcacatctac caactttgcg 4081 gtctcagaaa aagcataaac agttctacta ccgccattag tgaaactttt caaatcgccc 4141 agtggagaag aaaaaggcac agcgatacta gcattagcgg gcaaggatgc aactttatca 4201 accagggtcc tatagataac cctagcgcct gggatcatcc tttggacaac tctttctgcc 4261 aaatctaggt ccaaaatcac ttcattgata ccattattgt acaacttgag caagttgtcg 4321 atcagctcct caaattggtc ctctgtaacg gatgactcaa cttgcacatt aacttgaagc 4381 tcagtcgatt gagtgaactt gatcaggttg tgcagctggt cagcagcata gggaaacacg 4441 gcttttccta ccaaactcaa ggaattatca aactctgcaa cacttgcgta tgcaggtagc 4501 aagggaaatg tcatacttga agtcggacag tgagtgtagt cttgagaaat tctgaagccg 4561 tatttttatt atcagtgagt cagtcatcag gagatcctct acgccggacg catcgtggcc 4621 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg 4681 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca 4741 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg 4801 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag 4861 ggagagcgtc gagtatctat gattggaagt atgggaatgg tgatacccgc attcttcagt 4921 gtcttgaggt ctcctatcag attatgccca actaaagcaa ccggaggagg agatttcatg 4981 gtaaatttct ctgacttttg gtcatcagta gactcgaact gtgagactat ctcggttatg 5041 acagcagaaa tgtccttctt ggagacagta aatgaagtcc caccaataaa gaaatccttg 5101 ttatcaggaa caaacttctt gtttcgaact ttttcggtgc cttgaactat aaaatgtaga 5161 gtggatatgt cgggtaggaa tggagcgggc aaatgcttac cttctggacc ttcaagaggt 5221 atgtagggtt tgtagatact gatgccaact tcagtgacaa cgttgctatt tcgttcaaac 5281 cattccgaat ccagagaaat caaagttgtt tgtctactat tgatccaagc cagtgcggtc 5341 ttgaaactga caatagtgtg ctcgtgtttt gaggtcatct ttgtatgaat aaatctagtc 5401 tttgatctaa ataatcttga cgagccaagg cgataaatac ccaaatctaa aactctttta 5461 aaacgttaaa aggacaagta tgtctgcctg tattaaaccc caaatcagct cgtagtctga 5521 tcctcatcaa cttgaggggc actatcttgt tttagagaaa tttgcggaga tgcgatatcg 5581 agaaaaaggt acgctgattt taaacgtgaa atttatctca agatctctgc ctcgcgcgtt 5641 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc 5701 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt 5761 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta 5821 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 5881 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct 5941 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 6001 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 6601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 7141 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg 7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 7441 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac 7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 7861 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga 7921 attaattctc atgtttgaca gcttatcatc gataagctga ctcatgttgg tattgtgaaa 7981 tagacgcaga tcgggaacac tgaaaaataa cagttattat tcg