pPIC9k-His

价格:1250元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Invitrogen/ Biofeng
载体名称: pPIC9K-His, pPIC 9K His
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: 9.3 kb
5' 测序引物及序列: 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´; alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´
3' 测序引物及序列: 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´
载体标签: N-alpha factor, C-His
载体抗性: 氨苄和卡那
筛选标记: HIS4
备注:
利用alpha分泌因子,分泌表达带有His标签的蛋白。生物风销售SC-URA酵母表达筛选培养基,有需要和我们联系!
产品目录号: V175-20
稳定性: 稳定 Stable
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pPIC9K-His载体图谱






pPIC9k-His多克隆位点

载体简介

pPIC9K-His载体中存在卡那抗性基因,允许利用卡那抗性在酵母体内筛选多克隆拷贝,载体其余部分与pPIC9载体完全一样。pPIC9K-His载体能够使用的毕赤酵母宿主菌是KM71和GS115。
 
pPIC9K-His载体详细信息如下:
 
• pPIC9K-His载体大小9.3 bp,是融合表达载体, 含有C-His标签,表达出来的蛋白含有C端his标签,可以直接用于镍柱纯化。
 
• 载体构建过程中,可供使用的单一限制性内切酶位点为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Sac 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)
 
•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       pPIC9K-His              9294 bp    DNA     circular     25-DEC-2016
DEFINITION  pPIC9K-His.
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTDATE|-11561372|
COMMENT     VNTDBDATE|-11561372|
COMMENT     LSOWNER|
COMMENT     VNTNAME|pPIC9K-His|
COMMENT     VNTAUTHORNAME|Demo User|
COMMENT     VNTOAUTHORNAME|UNKNOWN|
FEATURES             Location/Qualifiers
     misc_feature    1152..1172
                     /vntifkey="21"
                     /label=alpha_factor_primer
     misc_feature    complement(1345..1365)
                     /vntifkey="21"
                     /label=3_AOX1_primer
     gene            complement(1668..1955)
                     /gene="tet(288-0)"
                     /vntifkey="60"
     CDS             complement(1998..4532)
                     /gene="HIS4"
                     /vntifkey="4"
                     /label=HIS4
                     /note="ORF frame 3"
     gene            complement(1998..4532)
                     /gene="HIS4"
                     /vntifkey="60"
     CDS             complement(4946..5761)
                     /gene="KanR2"
                     /vntifkey="4"
                     /label=Kana\Resistance
                     /note="ORF frame 2"
     gene            complement(4946..5761)
                     /gene="KanR2"
                     /vntifkey="60"
     misc_feature    complement(5926..5945)
                     /vntifkey="21"
                     /label=pBRrevBam_primer
     CDS             complement(5927..6649)
                     /vntifkey="4"
                     /label=ORF\frame\2
     misc_feature    complement(6931..6953)
                     /vntifkey="21"
                     /label=pGEX_3_primer
     rep_origin      complement(7306..7979)
                     /vntifkey="33"
                     /label=pBR322_origin
     CDS             complement(8124..8984)
                     /gene="Ampicillin"
                     /vntifkey="4"
                     /label=Ampicillin
                     /note="ORF frame 3"
     gene            complement(8124..8984)
                     /gene="Ampicillin"
                     /vntifkey="60"
     promoter        complement(9026..9054)
                     /vntifkey="30"
                     /label=AmpR_promoter
     promoter        1..948
                     /vntifkey="29"
                     /label=5'\AOX1\Promoter\Fragment
     primer          855..875
                     /vntifkey="27"
                     /label=5'\AOX1\Primer
     sig_peptide     949..1218
                     /vntifkey="94"
                     /label=alpha\Factor\Secretion\Signal
     misc_feature    1192..1241
                     /vntifkey="21"
                     /label=Multiple\Cloing\Site
     misc_feature    1271..1604
                     /vntifkey="21"
                     /label=3'\AOX1\transcription\termination\(TT)
     misc_feature    6140..6897
                     /vntifkey="21"
                     /label=3'\AOX1\Fragment
     misc_feature    1246..1263
                     /vntifkey="21"
                     /label=His\Tag
BASE COUNT     2499 a      2205 c      2078 g      2512 t 
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 cgaatcatca tcaccatcac 
     1261 cattaattcg ccttagacat gactgttcct cagttcaagt tgggcactta cgagaagacc 
     1321 ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag agatgcaggc 
     1381 ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt ttgtcatttt 
     1441 gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg aatatcttgt 
     1501 ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc ccactcctct 
     1561 tcagagtaca gaagattaag tgagaagttc gtttgtgcaa gcttatcgat aagctttaat 
     1621 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca 
     1681 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta 
     1741 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact 
     1801 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag 
     1861 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta 
     1921 tcgactacgc gatcatggcg accacacccg tcctgtggat ctatcgaatc taaatgtaag 
     1981 ttaaaatctc taaataatta aataagtccc agtttctcca tacgaacctt aacagcattg 
     2041 cggtgagcat ctagaccttc aacagcagcc agatccatca ctgcttggcc aatatgtttc 
     2101 agtccctcag gagttacgtc ttgtgaagtg atgaacttct ggaaggttgc agtgttaact 
     2161 ccgctgtatt gacgggcata tccgtacgtt ggcaaagtgt ggttggtacc ggaggagtaa 
     2221 tctccacaac tctctggaga gtaggcacca acaaacacag atccagcgtg ttgtacttga 
     2281 tcaacataag aagaagcatt ctcgatttgc aggatcaagt gttcaggagc gtactgattg 
     2341 gacatttcca aagcctgctc gtaggttgca accgataggg ttgtagagtg tgcaatacac 
     2401 ttgcgtacaa tttcaaccct tggcaactgc acagcttggt tgtgaacagc atcttcaatt 
     2461 ctggcaagct ccttgtctgt catatcgaca gccaacagaa tcacctggga atcaatacca 
     2521 tgttcagctt gagacagaag gtctgaggca acgaaatctg gatcagcgta tttatcagca 
     2581 ataactagaa cttcagaagg cccagcaggc atgtcaatac tacacagggc tgatgtgtca 
     2641 ttttgaacca tcatcttggc agcagtaacg aactggtttc ctggaccaaa tattttgtca 
     2701 cacttaggaa cagtttctgt tccgtaagcc atagcagcta ctgcctgggc gcctcctgct 
     2761 agcacgatac acttagcacc aaccttgtgg gcaacgtaga tgacttctgg ggtaagggta 
     2821 ccatccttct taggtggaga tgcaaaaaca atttctttgc aaccagcaac tttggcagga 
     2881 acacccagca tcagggaagt ggaaggcaga attgcggttc caccaggaat atagaggcca 
     2941 actttctcaa taggtcttgc aaaacgagag cagactacac cagggcaagt ctcaacttgc 
     3001 aacgtctccg ttagttgagc ttcatggaat ttcctgacgt tatctataga gagatcaatg 
     3061 gctctcttaa cgttatctgg caattgcata agttcctctg ggaaaggagc ttctaacaca 
     3121 ggtgtcttca aagcgactcc atcaaacttg gcagttagtt ctaaaagggc tttgtcacca 
     3181 ttttgacgaa cattgtcgac aattggtttg actaattcca taatctgttc cgttttctgg 
     3241 ataggacgac gaagggcatc ttcaatttct tgtgaggagg ccttagaaac gtcaattttg 
     3301 cacaattcaa tacgaccttc agaagggact tctttaggtt tggattcttc tttaggttgt 
     3361 tccttggtgt atcctggctt ggcatctcct ttccttctag tgacctttag ggacttcata 
     3421 tccaggtttc tctccacctc gtccaacgtc acaccgtact tggcacatct aactaatgca 
     3481 aaataaaata agtcagcaca ttcccaggct atatcttcct tggatttagc ttctgcaagt 
     3541 tcatcagctt cctccctaat tttagcgttc aacaaaactt cgtcgtcaaa taaccgtttg 
     3601 gtataagaac cttctggagc attgctctta cgatcccaca aggtggcttc catggctcta 
     3661 agaccctttg attggccaaa acaggaagtg cgttccaagt gacagaaacc aacacctgtt 
     3721 tgttcaacca caaatttcaa gcagtctcca tcacaatcca attcgatacc cagcaacttt 
     3781 tgagttgctc cagatgtagc acctttatac cacaaaccgt gacgacgaga ttggtagact 
     3841 ccagtttgtg tccttatagc ctccggaata gactttttgg acgagtacac caggcccaac 
     3901 gagtaattag aagagtcagc caccaaagta gtgaatagac catcggggcg gtcagtagtc 
     3961 aaagacgcca acaaaatttc actgacaggg aactttttga catcttcaga aagttcgtat 
     4021 tcagtagtca attgccgagc atcaataatg gggattatac cagaagcaac agtggaagtc 
     4081 acatctacca actttgcggt ctcagaaaaa gcataaacag ttctactacc gccattagtg 
     4141 aaacttttca aatcgcccag tggagaagaa aaaggcacag cgatactagc attagcgggc 
     4201 aaggatgcaa ctttatcaac cagggtccta tagataaccc tagcgcctgg gatcatcctt 
     4261 tggacaactc tttctgccaa atctaggtcc aaaatcactt cattgatacc attattgtac 
     4321 aacttgagca agttgtcgat cagctcctca aattggtcct ctgtaacgga tgactcaact 
     4381 tgcacattaa cttgaagctc agtcgattga gtgaacttga tcaggttgtg cagctggtca 
     4441 gcagcatagg gaaacacggc ttttcctacc aaactcaagg aattatcaaa ctctgcaaca 
     4501 cttgcgtatg caggtagcaa gggaaatgtc atacttgaag tcggacagtg agtgtagtct 
     4561 tgagaaattc tgaagccgta tttttattat cagtgagtca gtcatcagga gatcctctac 
     4621 gccggacgca tcgtggccga cctgcagggg gggggggggc gctgaggtct gcctcgtgaa 
     4681 gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg 
     4741 gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc 
     4801 tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca 
     4861 aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt 
     4921 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca 
     4981 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag 
     5041 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc 
     5101 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa 
     5161 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt 
     5221 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa 
     5281 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa 
     5341 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa 
     5401 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga 
     5461 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa 
     5521 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa 
     5581 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat 
     5641 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag 
     5701 catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca 
     5761 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat 
     5821 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tccccccccc 
     5881 ccctgcaggt cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca 
     5941 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg 
     6001 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat 
     6061 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg 
     6121 agtcgcataa gggagagcgt cgagtatcta tgattggaag tatgggaatg gtgatacccg 
     6181 cattcttcag tgtcttgagg tctcctatca gattatgccc aactaaagca accggaggag 
     6241 gagatttcat ggtaaatttc tctgactttt ggtcatcagt agactcgaac tgtgagacta 
     6301 tctcggttat gacagcagaa atgtccttct tggagacagt aaatgaagtc ccaccaataa 
     6361 agaaatcctt gttatcagga acaaacttct tgtttcgaac tttttcggtg ccttgaacta 
     6421 taaaatgtag agtggatatg tcgggtagga atggagcggg caaatgctta ccttctggac 
     6481 cttcaagagg tatgtagggt ttgtagatac tgatgccaac ttcagtgaca acgttgctat 
     6541 ttcgttcaaa ccattccgaa tccagagaaa tcaaagttgt ttgtctacta ttgatccaag 
     6601 ccagtgcggt cttgaaactg acaatagtgt gctcgtgttt tgaggtcatc tttgtatgaa 
     6661 taaatctagt ctttgatcta aataatcttg acgagccaag gcgataaata cccaaatcta 
     6721 aaactctttt aaaacgttaa aaggacaagt atgtctgcct gtattaaacc ccaaatcagc 
     6781 tcgtagtctg atcctcatca acttgagggg cactatcttg ttttagagaa atttgcggag 
     6841 atgcgatatc gagaaaaagg tacgctgatt ttaaacgtga aatttatctc aagatctctg 
     6901 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 
     6961 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 
     7021 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac 
     7081 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 
     7141 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc 
     7201 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 
     7261 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 
     7321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 
     7381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 
     7441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 
     7501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 
     7561 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 
     7621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 
     7681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 
     7741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 
     7801 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 
     7861 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 
     7921 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 
     7981 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 
     8041 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 
     8101 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 
     8161 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 
     8221 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 
     8281 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 
     8341 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 
     8401 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc 
     8461 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 
     8521 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 
     8581 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 
     8641 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 
     8701 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca 
     8761 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 
     8821 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 
     8881 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 
     8941 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 
     9001 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 
     9061 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 
     9121 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt 
     9181 cgtcttcaag aattaattct catgtttgac agcttatcat cgataagctg actcatgttg 
     9241 gtattgtgaa atagacgcag atcgggaaca ctgaaaaata acagttatta ttcg 
//

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