pBD-GAL4-Cam

价格:1500元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Stratagene(Agilent)
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: #838401-13
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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载体应用举例

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