pYES2/CT

价格:2000元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Invitrogen
载体名称: pYES2-CT, pYES2/CT
质粒类型: 酿酒酵母表达载体
表达水平: 高拷贝
诱导方法: 半乳糖
启动子: GAL1
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5963 bp
5' 测序引物及序列: GAL1-F: AATATACCTCTATACTTTAACGTC
3' 测序引物及序列: CYC1-R: GCGTGAATGTAAGCGTGAC
载体标签: C-His, C-V5
载体抗性: 氨苄青霉素
筛选标记: URA3报告基因
克隆菌株: TOP10, E.coli
表达菌株: INVSc1, Saccharomyces cerevisiae
备注: 酿酒酵母表达载体pYES2/CT含有URA3报告基因;
GAL1启动子驱动目的基因高水平表达;
半乳糖诱导目基因表达,葡萄糖阻遏目的基因表达,二者起到分子开关作用。
产品目录号: V8251-20
稳定性: 稳表达
组成型/诱导型: 诱导型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pYES2-CT载体图谱

pYES2-CT多克隆位点
pYES2-CT 载体特征
pYES2-CT 载体特征

载体简介


pYES2/CT酿酒酵母表达载体
pYES2/CT载体大小为6.0 kb左右,用于重组蛋白在酿酒酵母细胞中的诱导型表达。诱导剂为半乳糖。重组蛋白的C-端融合了6XHis标签,用于重组蛋白的纯化和检测。
 
pYES2/CT载体含有以下元件:

  Yeast GAL1 promoter for high level inducible protein expression in yeast by galactose and repression by glucose (Giniger et al., 1985; West et al., 1984)
  Multiple cloning site (MCS) with 8 or 9 unique sites (plus two BstX I sites) to facilitate in-frame cloning with the C-terminal peptide
  C-terminal peptide encoding the V5 epitope and a polyhistidine (6xHis) tag for detection and purification of your recombinant fusion protein
  2μ origin for episomal maintenance and high copy replication or CEN6/ARSH4 sequence for non-integrative centromeric maintenance and low copy replication (pYC2/CT)
  URA3 auxotrophic marker for selection of yeast transformants 
  Ampicillin resistance gene for selection in E. coli


使用pYES2/CT载体表达目的基因的实验流程如下:

1 Consult the multiple cloning site described on page 7 to determine a strategy to clone your gene in frame with the C-terminal peptide.
2 Ligate your insert into the appropriate vector and transform into E. coli. Select transformants on LB plates containing 50 to 100 μg/mL ampicillin.
3 Analyze your transformants for the presence of insert by restriction digestion.
4 Select a transformant with the correct restriction pattern and sequence to confirm that your gene is cloned in frame with the C-terminal peptide.
5 Transform your construct into competent INVSc1 cells and select for the appropriate amino acid prototrophy.
6 Test for expression of your recombinant protein by western blot analysis or functional assay.
7 Use metal-chelating resin such as ProBond to purify your recombinant protein.

载体序列

LOCUS       pYES2/CT	5963 bp 	DNA     circular	SYN
DEFINITION  pYES2/CT
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     source          1..5963
                     /organism="pYES2/CT"
                     /mol_type="other DNA"
     promoter        1..451
                     /label="GAL1_promoter"
     misc_feature    414..437
                     /label="GAL1_primer"
     promoter        475..493
                     /label="T7_promoter"
     terminator      716..961
                     /label="CYC1_terminator"
     misc_feature    complement(725..743)
                     /label="pYESTrp_rev_primer"
     misc_feature    complement(725..743)
                     /label="CYC1_primer"
     rep_origin      complement(1189..1808)
                     /label="pBR322_origin"
     CDS             complement(1963..2823)
                     /label="ORF frame 3"
     CDS             complement(2919..3722)
                     /label="ORF frame 1"
     gene            complement(2922..3722)
                     /label="URA3"
                     /gene="URA3"
     CDS             3211..3687
                     /label="ORF frame 1"
     promoter        complement(3724..3949)
                     /label="URA3_promoter"
     rep_origin      3952..5423
                     /label="2micron_origin"
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 GCATTGCTAC
 2281 AGGCATCGTG GTGTCACTCT 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 CGGCGTCAAT
 2581 ACGGGATAAT AGTGTATCAC 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 GGGTAATAAC TGATATAATT AAATTGAAGC TCTAATTTGT
 2881 GAGTTTAGTA TACATGCATT TACTTATAAT ACAGTTTTTT AGTTTTGCTG GCCGCATCTT
 2941 CTCAAATATG CTTCCCAGCC TGCTTTTCTG TAACGTTCAC CCTCTACCTT AGCATCCCTT
 3001 CCCTTTGCAA ATAGTCCTCT TCCAACAATA ATAATGTCAG ATCCTGTAGA GACCACATCA
 3061 TCCACGGTTC TATACTGTTG ACCCAATGCG TCTCCCTTGT CATCTAAACC CACACCGGGT
 3121 GTCATAATCA ACCAATCGTA ACCTTCATCT CTTCCACCCA TGTCTCTTTG AGCAATAAAG
 3181 CCGATAACAA AATCTTTGTC GCTCTTCGCA ATGTCAACAG TACCCTTAGT ATATTCTCCA
 3241 GTAGATAGGG AGCCCTTGCA TGACAATTCT GCTAACATCA AAAGGCCTCT AGGTTCCTTT
 3301 GTTACTTCTT CTGCCGCCTG CTTCAAACCG CTAACAATAC CTGGGCCCAC CACACCGTGT
 3361 GCATTCGTAA TGTCTGCCCA TTCTGCTATT CTGTATACAC CCGCAGAGTA CTGCAATTTG
 3421 ACTGTATTAC CAATGTCAGC AAATTTTCTG TCTTCGAAGA GTAAAAAATT GTACTTGGCG
 3481 GATAATGCCT TTAGCGGCTT AACTGTGCCC TCCATGGAAA AATCAGTCAA GATATCCACA
 3541 TGTGTTTTTA GTAAACAAAT TTTGGGACCT AATGCTTCAA CTAACTCCAG TAATTCCTTG
 3601 GTGGTACGAA CATCCAATGA AGCACACAAG TTTGTTTGCT TTTCGTGCAT GATATTAAAT
 3661 AGCTTGGCAG CAACAGGACT AGGATGAGTA GCAGCACGTT CCTTATATGT AGCTTTCGAC
 3721 ATGATTTATC TTCGTTTCCT GCAGGTTTTT GTTCTGTGCA GTTGGGTTAA GAATACTGGG
 3781 CAATTTCATG TTTCTTCAAC ACTACATATG CGTATATATA CCAATCTAAG TCTGTGCTCC
 3841 TTCCTTCGTT CTTCCTTCTG TTCGGAGATT ACCGAATCAA AAAAATTTCA AAGAAACCGA
 3901 AATCAAAAAA AAGAATAAAA AAAAAATGAT GAATTGAATT GAAAAGCTAG CTTATCGATG
 3961 ATAAGCTGTC AAAGATGAGA ATTAATTCCA CGGACTATAG ACTATACTAG ATACTCCGTC
 4021 TACTGTACGA TACACTTCCG CTCAGGTCCT TGTCCTTTAA CGAGGCCTTA CCACTCTTTT
 4081 GTTACTCTAT TGATCCAGCT CAGCAAAGGC AGTGTGATCT AAGATTCTAT CTTCGCGATG
 4141 TAGTAAAACT AGCTAGACCG AGAAAGAGAC TAGAAATGCA AAAGGCACTT CTACAATGGC
 4201 TGCCATCATT ATTATCCGAT GTGACGCTGC AGCTTCTCAA TGATATTCGA ATACGCTTTG
 4261 AGGAGATACA GCCTAATATC CGACAAACTG TTTTACAGAT TTACGATCGT ACTTGTTACC
 4321 CATCATTGAA TTTTGAACAT CCGAACCTGG GAGTTTTCCC TGAAACAGAT AGTATATTTG
 4381 AACCTGTATA ATAATATATA GTCTAGCGCT TTACGGAAGA CAATGTATGT ATTTCGGTTC
 4441 CTGGAGAAAC TATTGCATCT ATTGCATAGG TAATCTTGCA CGTCGCATCC CCGGTTCATT
 4501 TTCTGCGTTT CCATCTTGCA CTTCAATAGC ATATCTTTGT TAACGAAGCA TCTGTGCTTC
 4561 ATTTTGTAGA ACAAAAATGC AACGCGAGAG CGCTAATTTT TCAAACAAAG AATCTGAGCT
 4621 GCATTTTTAC AGAACAGAAA TGCAACGCGA AAGCGCTATT TTACCAACGA AGAATCTGTG
 4681 CTTCATTTTT GTAAAACAAA AATGCAACGC GACGAGAGCG CTAATTTTTC AAACAAAGAA
 4741 TCTGAGCTGC ATTTTTACAG AACAGAAATG CAACGCGAGA GCGCTATTTT ACCAACAAAG
 4801 AATCTATACT TCTTTTTTGT TCTACAAAAA TGCATCCCGA GAGCGCTATT TTTCTAACAA
 4861 AGCATCTTAG ATTACTTTTT TTCTCCTTTG TGCGCTCTAT AATGCAGTCT CTTGATAACT
 4921 TTTTGCACTG TAGGTCCGTT AAGGTTAGAA GAAGGCTACT TTGGTGTCTA TTTTCTCTTC
 4981 CATAAAAAAA GCCTGACTCC ACTTCCCGCG TTTACTGATT ACTAGCGAAG CTGCGGGTGC
 5041 ATTTTTTCAA GATAAAGGCA TCCCCGATTA TATTCTATAC CGATGTGGAT TGCGCATACT
 5101 TTGTGAACAG AAAGTGATAG CGTTGATGAT TCTTCATTGG TCAGAAAATT ATGAACGGTT
 5161 TCTTCTATTT TGTCTCTATA TACTACGTAT AGGAAATGTT TACATTTTCG TATTGTTTTC
 5221 GATTCACTCT ATGAATAGTT CTTACTACAA TTTTTTTGTC TAAAGAGTAA TACTAGAGAT
 5281 AAACATAAAA AATGTAGAGG TCGAGTTTAG ATGCAAGTTC AAGGAGCGAA AGGTGGATGG
 5341 GTAGGTTATA TAGGGATATA GCACAGAGAT ATATAGCAAA GAGATACTTT TGAGCAATGT
 5401 TTGTGGAAGC GGTATTCGCA ATGGGAAGCT CCACCCCGGT TGATAATCAG AAAAGCCCCA
 5461 AAAACAGGAA GATTGTATAA GCAAATATTT AAATTGTAAA CGTTAATATT TTGTTAAAAT
 5521 TCGCGTTAAA TTTTTGTTAA ATCAGCTCAT TTTTTAACGA ATAGCCCGAA ATCGGCAAAA
 5581 TCCCTTATAA ATCAAAAGAA TAGACCGAGA TAGGGTTGAG TGTTGTTCCA GTTTCCAACA
 5641 AGAGTCCACT ATTAAAGAAC GTGGACTCCA ACGTCAAAGG GCGAAAAAGG GTCTATCAGG
 5701 GCGATGGCCC ACTACGTGAA CCATCACCCT AATCAAGTTT TTTGGGGTCG AGGTGCCGTA
 5761 AAGCAGTAAA TCGGAAGGGT AAACGGATGC CCCCATTTAG AGCTTGACGG GGAAAGCCGG
 5821 CGAACGTGGC GAGAAAGGAA GGGAAGAAAG CGAAAGGAGC GGGGGCTAGG GCGGTGGGAA
 5881 GTGTAGGGGT CACGCTGGGC GTAACCACCA CACCCGCCGC GCTTAATGGG GCGCTACAGG
 5941 GCGCGTGGGG ATGATCCACT AGT
//

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