pYES3/CT酿酒酵母质粒

基本信息

启动子:	T7，GAL1，TRP1
复制子:	2μ ori,ori,FI ori
终止子:	CYC1
质粒分类:	酵母系列，酿酒酵母表达载体
质粒大小:	5870bp
原核抗性:	Amp
筛选标记:	TRP1
克隆菌株:	DH5α
培养条件:	37℃，有氧 LB
表达宿主:	酵母细胞
诱导方式:	半乳糖诱导
5'测序引物:	GAL1-F: ATTTTCGGTTTGTATTACTTC
3'测序引物:	GALI-R: GTTCTTAATACTAACATAACT

质粒简介

    pYES3/CT and pYES6/CT are S. cerevisiae expression vectors derived from the parental pYES2 vector. Vectors feature a C-terminal V5 epitope for detection with an Anti-V5 Antibody. pYES3/CT carries the TRP1 selection marker for selection in yeast. pYES6/CT carries the bsd resistance gene for selection with the antibiotic Blasticidin, a potent selection agent that can be used at very low concentrations and in any strain regardless of auxotrophic marker.

质粒图谱

质粒序列

LOCUS       Exported                5870 bp ds-DNA     circular SYN 29-NOV-2013
DEFINITION  High-copy episomal vector with a TRP1 marker for galactose-inducible
            expression of C-terminally tagged proteins in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    pYES3 CT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, September 11, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..5870
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /note="GAL1 promoter"
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /bound_moiety="Gal4"
                     /note="UAS"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        475..493
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    501..600
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             607..648
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /note="V5 tag"
                     /translation="GKPIPNPLLGLDST"
     CDS             658..675
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      709..956
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(1204..1792)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1963..2823)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2824..2928)
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        3031..3132
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     CDS             3133..3807
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      4449..5329
                     /note="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5398..5853)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagcttggta ccgagctcgg atccactagt aacggccgcc
      541 agtgtgctgg aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
      601 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat
      661 catcaccatc accattgagt ttaaacccgc tgatcctaga gggccgcatc atgtaattag
      721 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
      781 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
      841 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
      901 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
      961 tgcggccctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1141 gaacatgtga gcaaaaggcc agcaaaagcc caggaaccgt aaaaaggccg cgttgctggc
     1201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1621 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     1681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2041 cgtgtagata actacgatac gggagcgctt accatctggc cccagtgctg caatgatacc
     2101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     2581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2941 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3001 gcgtatcacg aggccctttc gtcttcaaga aattcggtcg aaaaaagaaa aggagagggc
     3061 caagagggag ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag
     3121 gcagcttgga gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt
     3181 tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg
     3241 ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt
     3301 tcaagtcttg taaaagcata taaaaatagt tcaggcactc cgaaatactt ggttggcgtg
     3361 tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc
     3421 gtccaactgc acggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt
     3481 attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa
     3541 cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat
     3601 tggaactcga tttctgactg ggttggaagg caagagagcc ccgagagctt acattttatg
     3661 ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt
     3721 ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca
     3781 aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt
     3841 ttgtgcactt gccctagctt atcgatgata agctgtcaaa gatgagaatt aattccacgg
     3901 actatagact atactagata ctccgtctac tgtacgatac acttccgctc aggtccttgt
     3961 cctttaacga ggccttacca ctcttttgtt actctattga tccagctcag caaaggcagt
     4021 gtgatctaag attctatctt cgcgatgtag taaaactagc tagaccgaga aagagactag
     4081 aaatgcaaaa ggcacttcta caatggctgc catcattatt atccgatgtg acgctgcagc
     4141 ttctcaatga tattcgaata cgctttgagg agatacagcc taatatccga caaactgttt
     4201 tacagattta cgatcgtact tgttacccat cattgaattt tgaacatccg aacctgggag
     4261 ttttccctga aacagatagt atatttgaac ctgtataata atatatagtc tagcgcttta
     4321 cggaagacaa tgtatgtatt tcggttcctg gagaaactat tgcatctatt gcataggtaa
     4381 tcttgcacgt cgcatccccg gttcattttc tgcgtttcca tcttgcactt caatagcata
     4441 tctttgttaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc
     4501 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag
     4561 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgac
     4621 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     4681 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     4741 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     4801 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     4861 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     4921 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     4981 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     5041 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     5101 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     5161 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     5221 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     5281 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaatg ggaagctcca
     5341 ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa
     5401 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5461 ttaacgaata gcccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5521 ggttgagtgt tgttccagtt tccaacaaga gtccactatt aaagaacgtg gactccaacg
     5581 tcaaagggcg aaaaagggtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5641 caagtttttt ggggtcgagg tgccgtaaag cagtaaatcg gaagggtaaa cggatgcccc
     5701 catttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5761 aaggagcggg ggctagggcg gtgggaagtg taggggtcac gctgggcgta accaccacac
     5821 ccgccgcgct taatggggcg ctacagggcg cgtggggatg atccactagt