pBD-GAL4-Cam载体

pBD-GAL4-Cam载体

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  • ¥1500
  • ZYbscience
  • 中国/美国
  • ZY838401-13
  • 2025年07月15日
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    • 详细信息
    • 询价记录
    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      pBD-GAL4-Cam

    • 库存

      20

    • 供应商

      泽叶生物

    载体基本信息

    出品公司: ZYbscience
    载体名称: pBD-GAL4-Cam
    质粒类型: 酵母双杂交载体;酵母噬粒载体
    克隆方法: 多克隆位点,限制性内切酶
    启动子: ADH1
    载体大小: 6491 bp
    5' 测序引物及序列: --
    3' 测序引物及序列: --
    载体标签:
    载体抗性: 氯霉素Chloramphenicol
    真核筛选标记: TRP1
    克隆菌株: XL1-Blue MRF'
    表达菌株: YRG-2
    备注: 含a beta-gal报告基因。配套载体为pAD-GAL4-2.1
    产品目录号: ZY838401-13
    组成型/诱导型: 组成型

    载体质粒图谱和多克隆位点信息

    pBD-GAL4-Cam载体图谱



    pBD-GAL4-Cam 多克隆位点

    pBD-GAL4-Cam 载体特征

    载体简介

    酵母双杂交系统
    
    Protein–protein interactions occur in many biological processes including replication, transcription, secretion, signal transduction, and metabolism. A fundamental question in the study of any protein is to identify proteins that interact with a given protein in vivo. Intense research efforts are focused on the identification of these proteins.
    The GAL4 two-hybrid phagemid vector system, a eukaryotic
    system to detect protein–protein interactions in vivo, provides a method for the rapid identification of genes encoding proteins that interact with a given protein (i.e., a bait protein).1,2 The system is based on the ability to separate eukaryotic transcriptional activators into two separate domains, the DNAbinding domain (BD) and the transcriptional activation domain (AD).
     In theGAL4 two-hybrid phagemid vector system, proteins that interact with the bait protein are identified by generating hybrids of the yeast GAL4 BD and the bait protein (X) and the GAL4 AD and a library of proteins (Y). Neither hybrid protein is capable of initiating specific transcription of reporter genes in yeast in the absence of a specific interaction with the other hybrid protein. 
    When the hybrid protein X is expressed in yeast, the GAL4 BD binds X to specific DNA sequences in the yeast chromosome defined by the GAL1 or GAL4 upstream activating sequences (UASGAL1 or UASGAL4, respectively), which regulate the expression of a reporter gene. Binding of X to the UAS is not sufficient to initiate transcription of the reporter gene. When Y is expressed in yeast, the AD interacts with other components of the transcription machinery required to initiate transcription of the reporter gene. However, Y alone is not localized to the reporter gene UAS and therefore does not activate transcription of the reporter gene. When a specific interaction between X and Y localizes both the GAL4 BD and GAL4 AD to the reporter gene UAS, transcriptional activation of the reporter gene occurs (Figure 2B). The reporter genes in the GAL4 twohybrid phagemid vector system are β-galactosidase (lacZ) and histidine (HIS3). 
    
    双杂交载体pAD-GAL4-2.1和pBD-GAL4-Cam
    
    The pAD-GAL4-2.1 phagemid vector contains a multiple cloning site (MCS) with BamH I, Nhe I, EcoR I, Xho I, Sal I, Xba I, Pst I, and Bgl II restriction sites. The pBD-GAL4-Cam phagemid vector contains an MCS with EcoR I, Srf I, Sma I, Xho I, Sal I, Xba I, and Pst I restriction sites. The unique EcoR I and Xho I cloning sites in the pAD-GAL4-2.1 vector make this vector compatible with the Agilent cDNA Synthesis Kit for the preparation of unidirectional cDNA libraries.
    The unique EcoR I and Sal I cloning sites are used for the preparation of cDNA libraries in the pBD-GAL4 Cam phagemid vector because the Xho I site in the MCS is not unique. The unique BamH I, Nhe I, and EcoR I sites at the 5′ end and the Xho I, Sal I, Xba I, and Bgl II sites at the 3′ end of the DNA insert facilitate the transfer of DNA encoding the target protein into commonly used protein expression/purification vectors. Genomic DNA digested with Mbo I, BamH I, or Sau3A I and partially filled-in can be inserted into a partially filled-in Xho I site in the pAD-GAL4-2.1 phagemid vector. The Xba I site in the pAD-GAL4-2.1 and pBD-GAL4-Cam phagemid vectors is not unique and contains the UAG amber suppressor in the same translational reading frame as the GAL4 domain. DNA should therefore be inserted such that the Xba I site is not between the GAL4 domain and the DNA insert.
    The pAD-GAL4-2.1 and pBD-GAL4 Cam phagemid vectors contain the pUC origin for replication and an f1 origin for production of single-stranded DNA (ssDNA) in E. coli. Single-stranded DNA can be used for DNA sequencing or site-directed mutagenesis. The pAD-GAL4-2.1 and pBDGAL4-Cam phagemid vectors contain ampicillin-resistance gene [β-lactamase (bla)] and chloramphenicol acetyltransferase genes, respectively, for selection with ampicillin and chloramphenicol in E. coli. The pADGAL4-2.1 and pBD-GAL4 Cam phagemid vectors contain the 2μ origin for replication. For selection in yeast, the pAD-GAL4-2.1 phagemid vector contains the LEU2 gene and the pBD-GAL4 Cam phagemid vector contains the TRP1 gene. The hybrid protein is expressed by the alcohol dehydrogenase 1 (ADH1) promoter (P ADH1) and is terminated by the ADH1 terminator (T ADH1). 
    

    载体序列

    pBD-GAL4-Cam, 6491 bp                          version 045002
    
    NOTE: The following sequence has been verified for accuracy
    at the junctions. The remainder of the sequence has been 
    obtained from existing data. 
    
    
        1  GCTTGCATGC AACTTCTTTT CTTTTTTTTT CTTTTCTCTC TCCCCCGTTG
       51  TTGTCTCACC ATATCCGCAA TGACAAAAAA ATGATGGAAG ACACTAAAGG
      101  AAAAAATTAA CGACAAAGAC AGCACCAACA GATGTCGTTG TTCCAGAGCT
      151  GATGAGGGGT ATCTCGAAGC ACACGAAACT TTTTCCTTCC TTCATTCACG
      201  CACACTACTC TCTAATGAGC AACGGTATAC GGCCTTCCTT CCAGTTACTT
      251  GAATTTGAAA TAAAAAAAAG TTTGCTGTCT TGCTATCAAG TATAAATAGA
      301  CCTGCAATTA TTAATCTTTT GTTTCCTCGT CATTGTTCTC GTTCCCTTTC
      351  TTCCTTGTTT CTTTTTCTGC ACAATATTTC AAGCTATACC AAGCATACAA
      401  TCAACTCCAA GCTTGAAGCA AGCCTCCTGA AAGATGAAGC TACTGTCTTC
      451  TATCGAACAA GCATGCGATA TTTGCCGACT TAAAAAGCTC AAGTGCTCCA
      501  AAGAAAAACC GAAGTGCGCC AAGTGTCTGA AGAACAACTG GGAGTGTCGC
      551  TACTCTCCCA AAACCAAAAG GTCTCCGCTG ACTAGGGCAC ATCTGACAGA
      601  AGTGGAATCA AGGCTAGAAA GACTGGAACA GCTATTTCTA CTGATTTTTC
      651  CTCGAGAAGA CCTTGACATG ATTTTGAAAA TGGATTCTTT ACAGGATATA
      701  AAAGCATTGT TAACAGGATT ATTTGTACAA GATAATGTGA ATAAAGATGC
      751  CGTCACAGAT AGATTGGCTT CAGTGGAGAC TGATATGCCT CTAACATTGA
      801  GACAGCATAG AATAAGTGCG ACATCATCAT CGGAAGAGAG TAGTAACAAA
      851  GGTCAAAGAC AGTTGACTGT ATCGCCGGAA TTCGCCCGGG CCTCGAGCCC
      901  GGGTCGACTC TAGAGCCCTA TAGTGAGTCG TATTACTGCA GCCAAGCTAA
      951  TTCCGGGCGA ATTTCTTATG ATTTATGATT TTTATTATTA AATAAGTTAT
     1001  AAAAAAAATA AGTGTATACA AATTTTAAAG TGACTCTTAG GTTTTAAAAC
     1051  GAAAATTCTT GTTCTTGAGT AACTCTTTCC TGTAGGTCAG GTTGCTTTCT
     1101  CAGGTATAGC ATGAGGTCGC TCTTATTGAC CACACCTCTA CCGGCATGCC
     1151  GGCAAGTGCA CAAACAATAC TTAAATAAAT ACTACTCAGT AATAACCTAT
     1201  TTCTTAGCAT TTTTGACGAA ATTTGCTATT TTGTTAGAGT CTTTTACACC
     1251  ATTTGTCTCC ACACCTCCGC TTACATCAAC ACCAATAACG CCATTTAATC
     1301  TAAGCGCATC ACCAACATTT TCTGGCGTCA GTCCACCAGC TAACATAAAA
     1351  TGTAAGCTTT CGGGGCTCTC TTGCCTTCCA ACCCAGTCAG AAATCGAGTT
     1401  CCAATCCAAA AGTTCACCTG TCCCACCTGC TTCTGAATCA AACAAGGGAA
     1451  TAAACGAATG AGGTTTCTGT GAAGCTGCAC TGAGTAGTAT GTTGCAGTCT
     1501  TTTGGAAATA CGAGTCTTTT AATAACTGGC AAACCGAGGA ACTCTTGGTA
     1551  TTCTTGCCAC GACTCATCTC CATGCAGTTG GACGATATCA ATGCCGTAAT
     1601  CATTGACCAG AGCCAAAACA TCCTCCTTAG GTTGATTACG AAACACGCCA
     1651  ACCAAGTATT TCGGAGTGCC TGAACTATTT TTATATGCTT TTACAAGACT
     1701  TGAAATTTTC CTTGCAATAA CCGGGTCAAT TGTTCTCTTT CTATTGGGCA
     1751  CACATATAAT ACCCAGCAAG TCAGCATCGG AATCTAGAGC ACATTCTGCG
     1801  GCCTCTGTGC TCTGCAAGCC GCAAACTTTC ACCAATGGAC CAGAACTACC
     1851  TGTGAAATTA ATAACAGACA TACTCCAAGC TGCCTTTGTG TGCTTAATCA
     1901  CGTATACTCA CGTGCTCAAT AGTCACCAAT GCCCTCCCTC TTGGCCCTCT
     1951  CCTTTTCTTT TTTCGACCGA ATTAATTCGT AATCATGGTC ATAGCTGTTT
     2001  CCTGTGTGAA ATTGTTATCC GCTCACAATT CCACACAACA TACGAGCCGG
     2051  AAGCATAAAG TGTAAAGCCT GGGGTGCCTA ATGAGTGAGG TAACTCACAT
     2101  TAATTGCGTT GCGCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC
     2151  CAGCTGGTTC TTTCCGCCTC AGGACTCTTC CTTTTTCAAT ATTATTGAAG
     2201  CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT
     2251  AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG AAAAGTGCCA
     2301  CCTGACGCGC CCTGTAGCGG CGCATTAAGC GCGGCGGGTG TGGTGGTTAC
     2351  GCGCAGCGTG ACCGCTACAC TTGCCAGCGC CCTAGCGCCC GCTCCTTTCG
     2401  CTTTCTTCCC TTCCTTTCTC GCCACGTTCG CCGGCTTTCC CCGTCAAGCT
     2451  CTAAATCGGG GGCTCCCTTT AGGGTTCCGA TTTAGTGCTT TACGGCACCT
     2501  CGACCCCAAA AAACTTGATT AGGGTGATGG TTCACGTAGT GGGCCATCGC
     2551  CCTGATAGAC GGTTTTTCGC CCTTTGACGT TGGAGTCCAC GTTCTTTAAT
     2601  AGTGGACTCT TGTTCCAAAC TGGAACAACA CTCAACCCTA TCTCGGTCTA
     2651  TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT TGGTTAAAAA
     2701  ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG
     2751  CTTACAATTT ACGCGCCTAA ATTATGCGGC CGCATATAAC TGGATTAATG
     2801  AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCTCTTCCG
     2851  CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT GCGGCGAGCG
     2901  GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA
     2951  TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC
     3001  GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC
     3051  GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA ACCCGACAGG
     3101  ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC
     3151  CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT TCTCCCTTCG
     3201  GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC TCAGTTCGGT
     3251  GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC
     3301  CCGACCGCTG CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA
     3351  AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA GGATTAGCAG
     3401  AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGG TGGCCTAACT
     3451  ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT GCTGAAGCCA
     3501  GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC
     3551  CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT ACGCGCAGAA
     3601  AAAAAGGATC TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT
     3651  CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA GATTATCAAA
     3701  AAGGATCTTC ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA
     3751  TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA ATGCTTAATC
     3801  AGTGAGGCAC CTATCTCAGC GATCTGTCTA TTTCGTTCAT CCATAGTTGC
     3851  CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC TTACCATCTG
     3901  GCCCCAGTGC TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT
     3951  TTATCAGCAA TAAACCAGCC AGCCGCCTGT GACGGAAGAT CACTTCGCAG
     4001  AATAAATAAA TCCTGGTGTC CCTGTTGATA CCGGGAAGCC CTGGGCCAAC
     4051  TTTTGGCGAA AATGAGACGT TGATCGGCAC GTAAGAGGTT CCAACTTTCA
     4101  CCATAATGAA ATAAGATCAC TACCGGGCGT ATTTTTTGAG TTGTCGAGAT
     4151  TTTCAGGAGC TAAGGAAGCT AAAATGGAGA AAAAAATCAC TGGATATACC
     4201  ACCGTTGATA TATCCCAATG GCATCGTAAA GAACATTTTG AGGCATTTCA
     4251  GTCAGTTGCT CAATGTACCT ATAACCAGAC CGTTCAGCTG GATATTACGG
     4301  CCTTTTTAAA GACCGTAAAG AAAAATAAGC ACAAGTTTTA TCCGGCCTTT
     4351  ATTCACATTC TTGCCCGCCT GATGAATGCT CATCCGGAAT TACGTATGGC
     4401  AATGAAAGAC GGTGAGCTGG TGATATGGGA TAGTGTTCAC CCTTGTTACA
     4451  CCGTTTTCCA TGAGCAAACT GAAACGTTTT CATCGCTCTG GAGTGAATAC
     4501  CACGACGATT TCCGGCAGTT TCTACACATA TATTCGCAAG ATGTGGCGTG
     4551  TTACGGTGAA AACCTGGCCT ATTTCCCTAA AGGGTTTATT GAGAATATGT
     4601  TTTTCGTCTC AGCCAATCCC TGGGTGAGTT TCACCAGTTT TGATTTAAAC
     4651  GTGGCCAATA TGGACAACTT CTTCGCCCCG TTTTCACCAT GGGCAAATAT
     4701  TATACGCAAG GCGACAAGGT GCTGATGCCG CTGGCGATTC AGGTTCATCA
     4751  TGCCGTTTGT GATGGCTTCC ATGTCGGCAG AATGCTTAAT GAATTACAAC
     4801  AGTACTGCGA TGAGTGGCAG GGCGGGGCGT AATTTTTTTA AGGCAGTTAT
     4851  TGGTGCCCTT AAACGCCTGG TTGCTACGCC TGAATAAGTG ATAATAAGCG
     4901  GATGAATGGC AGAAATTCGA AAGCAAATTC GACCCGGTCG TCGGTTCAGG
     4951  GCAGGGTCGT TAAATAGCCG CTTATGTCTA TTGCTGGTTT ACCGGTTTAT
     5001  TGACTACCGG AAGCAGTGTG ACCGTGTGCT TCTCAAATGC CTGAGGCCAG
     5051  TTTGCTCAGG CTCTCCCCGT GGAGGCTAAG AAACCATTAT TATCATGACA
     5101  TTAACCTATA AAAATAGGCG TATCACGAGG CCCTTTCGTC TCGCGCGTTT
     5151  CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA
     5201  CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG
     5251  TCAGCGGGTG TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA
     5301  GCAGATTGTA CTGAGAGTGC ACCATAACGC ATTTAAGCAT AAACACGCAC
     5351  TATGCCGTTC TTCTCATGTA TATATATATA CAGGCAACAC GCAGATATAG
     5401  GTGCGACGTG AACAGTGAGC TGTATGTGCG CAGCTCGCGT TGCATTTTCG
     5451  GAAGCGCTCG TTTTCGGAAA CGCTTTGAAG TTCCTATTCC GAAGTTCCTA
     5501  TTCTCTAGCT AGAAAGTATA GGAACTTCAG AGCGCTTTTG AAAACCAAAA
     5551  GCGCTCTGAA GACGCACTTT CAAAAAACCA AAAACGCACC GGACTGTAAC
     5601  GAGCTACTAA AATATTGCGA ATACCGCTTC CACAAACATT GCTCAAAAGT
     5651  ATCTCTTTGC TATATATCTC TGTGCTATAT CCCTATATAA CCTACCCATC
     5701  CACCTTTCGC TCCTTGAACT TGCATCTAAA CTCGACCTCT ACATTTTTTA
     5751  TGTTTATCTC TAGTATTACT CTTTAGACAA AAAAATTGTA GTAAGAACTA
     5801  TTCATAGAGT GAATCGAAAA CAATACGAAA ATGTAAACAT TTCCTATACG
     5851  TAGTATATAG AGACAAAATA GAAGAAACCG TTCATAATTT TCTGACCAAT
     5901  GAAGAATCAT CAACGCTATC ACTTTCTGTT CACAAAGTAT GCGCAATCCA
     5951  CATCGGTATA GAATATAATC GGGGATGCCT TTATCTTGAA AAAATGCACC
     6001  CGCAGCTTCG CTAGTAATCA GTAAACGCGG GAAGTGGAGT CAGGCTTTTT
     6051  TTATGGAAGA GAAAATAGAC ACCAAAGTAG CCTTCTTCTA ACCTTAACGG
     6101  ACCTACAGTG CAAAAAGTTA TCAAGAGACT GCATTATAGA GCGCACAAAG
     6151  GAGAAAAAAA GTAATCTAAG ATGCTTTGTT AGAAAAATAG CGCTCTCGGG
     6201  ATGCATTTTT GTAGAACAAA AAAGAAGTAT AGATTCTTTG TTGGTAAAAT
     6251  AGCGCTCTCG CGTTGCATTT CTGTTCTGTA AAAATGCAGC TCAGATTCTT
     6301  TGTTTGAAAA ATTAGCGCTC TCGCGTTGCA TTTTTGTTTT ACAAAAATGA
     6351  AGCACAGATT CTTCGTTGGT AAAATAGCGC TTTCGCGTTG CATTTCTGTT
     6401  CTGTAAAAAT GCAGCTCAGA TTCTTTGTTT GAAAAATTAG CGCTCTCGCG
     6451  TTGCATTTTT GTTCTACAAA ATGAAGCACA GATGCTTCGT T

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