北京华越洋生物
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首页    载体及质粒    pPICZα A
bjcn3scmz_6830_635930338740636762

pPICZα A

pPICZα A

pPICZα A

编号

载体名称

北京华越洋生物VECT2580

pPICZα A

 

pPICZα A载体基本信息

载体名称:pPICZα A
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:3593 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:

该载体本身含有signal alpha 分泌信号肽,该信号肽本身含有ATG启动子,所以插入基因可以不包含起始密码子ATG。

稳定性:稳定 Stable
组成型:组成型 Constitutive
病毒/非病毒:非病毒

pPICZα A载体质粒图谱和多克隆位点信息

pPICZalpha A载体图谱

pPICZalpha A多克隆位点

pPICZaA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。

pPICZaA载体序列

LOCUS       pPICZα\A               3593 bp    DNA     circular SYN  2-SEP-2003
SOURCE      
  ORGANISM  
COMMENT     This file is created by Vector NTI
FEATURES             Location/Qualifiers
     source          1..3593
                     /invitrogen="25"
                     /vntifkey="98"
     primer          complement(1423..1443)
                     /invitrogen="1880000"
                     /vntifkey="27"
                     /label=3'\AOX1\primer
     primer          855..875
                     /invitrogen="1890000"
                     /vntifkey="27"
                     /label=5'\AOX1\primer
     CDS             1320..1337
                     /invitrogen="1010000"
                     /vntifkey="4"
                     /label=6xHis
     primer          1144..1164
                     /invitrogen="1900000"
                     /vntifkey="27"
                     /label=alpha-factor\primer
     terminator      1341..1682
                     /invitrogen="2550000"
                     /vntifkey="43"
                     /label=AOX1\transcription\terminator
     CDS             1275..1304
                     /invitrogen="1090000"
                     /vntifkey="4"
                     /label=c-myc\epitope
     terminator      2538..2855
                     /invitrogen="2560000"
                     /vntifkey="43"
                     /label=CYC1\transcription\terminator
     promoter        2095..2162
                     /invitrogen="2340000"
                     /vntifkey="30"
                     /label=EM7\promoter
     misc_feature    1208..1274
                     /invitrogen="1460000"
                     /vntifkey="21"
                     /label=MCS
     rep_origin      complement(2866..3539)
                     /invitrogen="2540000"
                     /vntifkey="33"
                     /label=pUC\origin
     CDS             941..1207
                     /invitrogen="1280000"
                     /vntifkey="4"
                     /label=alpha-factor\signal\peptide
     promoter        1683..2094
                     /invitrogen="2300000"
                     /vntifkey="29"
                     /label=TEF1\promoter
     CDS             2163..2537
                     /invitrogen="1370000"
                     /vntifkey="4"
                     /label=Zeo(R)
     promoter        1..940
                     /invitrogen="2160000"
                     /vntifkey="29"
                     /label=AOX1\promoter
BASE COUNT      892 a       886 c       801 g      1014 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 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 aaagagaggc
     1201 tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg
     1261 ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
     1321 atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg
     1381 ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt
     1441 tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag
     1501 gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc
     1561 agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt
     1621 ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat
     1681 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
     1741 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaattttccc
     1801 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
     1861 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
     1921 tttcttgaaa tttttttttt tagttttttt ctctttcagt gacctccatt gatatttaag
     1981 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
     2041 ttttttactt cttgttcatt agaaagaaag catagcaatc taatctaagg ggcggtgttg
     2101 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     2161 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     2221 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     2281 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     2341 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     2401 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     2461 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     2521 ccgaggagca ggactgacac gtccgacggc ggcccacggg tcccaggcct cggagatccg
     2581 tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     2641 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     2701 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     2761 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     2821 gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg agcaaaaggc
     2881 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2941 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3001 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3061 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     3121 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3181 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3241 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3301 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3361 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3421 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3481 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3541 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag atc
//

pPICZaA其他酵母表达载体:

pPIC6αA

pYD1

pGAG424

pSos-MAFB

pRS406

p53his

pGADT7-T

pBridge

pRS416

pCL1

pHis2

pACT2-AD

pSEP1

pSos

pGBKT7

pFLD/CAT

pHisSi-1

pGBKT7-53

pFLD

pYES2-EGFP

pMyr

pFLDα

Ycp22lac-EGFP

pPIC9k-His

pGBKT7-Lam

pADH2

SUMO protease

pPICZ A

pDisplay

pYIP5

pGADT7

LA503

pPinkα-HC

pDR195

BFD15

pPIC ZαFC

pPIC3.5K

pYES2

pPIC ZαGB

pYEPlac112

pAUR123

pPIC ZαD

pYEPlac195

pYRP7

pPICZ C

Ycplac33

pYX212

pFA6a-FGP(S65T)-kanMX6

pHIC-PI

pYIP211

pRS41H

pESC-URA

pRS426gal

pYES2-kan

pRS316

pYES6/CT

p416GFD

pPICZα B

pESC-His

pYES2-NTB

pYCP211

pESC-Leu

pYES2-NTA

pMET C

pRS426

pYES2-NTC

pMETαA

pPink-HC

pESC-TRP

pMETαB

pPink-LC

pGAPZ B

pYES3/CT

pMET A

pGAPZα C

pPICZα A

pMETαC

pHIL-S1

pPICZ B

pYC2/CT

pMET B

pPICZα C

pPIC9

pRS405

pHIL-D2

pPIC3.5

pGAPZ A

pRS414

pGAPZα A

pUG66

pAD-GAL4-2.1

pSEP2

pAO815

pDEST32

pYC2/NT B

p53blue

pGAPZ C

pYC2/NT A

pPIC6 C

pRS415

pYC2/NT C

pYES-DEST52

pPIC6αB

pSEP3

pYES2/CT

pPIC6αC

pBD-GAL4-Cam

pPIC6 A

pRS403

pRS413