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  • pAD-GAL4-2.1

    價格:1500元

    聯系方式:I47-825O-882O

    買家導航

    載體基本信息

    出品公司: Stratagene(Agilent)
    載體名稱: pAD-GAL4-2.1
    質粒類型: 酵母雙雜交載體;酵母噬粒載體
    克隆方法: 多克隆位點,限制性內切酶
    啟動子: ADH1
    載體大小: 7653 bp
    5' 測序引物及序列: --
    3' 測序引物及序列: --
    載體標簽:
    載體抗性: 氨芐青霉素Ampicillin
    真核篩選標記: LEU2
    克隆菌株: XL1-Blue MRF'
    表達菌株: YRG-2
    備注: 含a beta-gal報告基因。
    配套載體為pBD-GAL4-Cam。
    產品目錄號: #838401-13
    組成型/誘導型: 組成型

    載體質粒圖譜和多克隆位點信息

    pAD-GAL4-2.1載體圖譜
    pAD-GAL4-2.1 多克隆位點

    pAD-GAL4-2.1 載體特征

    載體簡介

    酵母雙雜交系統
    
    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). 
    
    雙雜交載體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). 
    

    載體序列

    pAD-GAL4-2.1, 7653 bp                         version 122001
    
    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 AATGATGGAA GACACTAAAG  
     101  GAAAAAATTA ACGACAAAGA CAGCACCAAC AGATGTCGTT GTTCCAGAGC  
     151  TGATGAGGGG TATCTTCGAA CACACGAAAC TTTTTCCTTC CTTCATTCAC  
     201  GCACACTACT CTCTAATGAG CAACGGTATA CGGCCTTCCT TCCAGTTACT  
     251  TGAATTTGAA ATAAAAAAAG TTTGCCGCTT TGCTATCAAG TATAAATAGA  
     301  CCTGCAATTA TTAATCTTTT GTTTCCTCGT CATTGTTCTC GTTCCCTTTC  
     351  TTCCTTGTTT CTTTTTCTGC ACAATATTTC AAGCTATACC AAGCATACAA  
     401  TCAACTCCAA GCTTTGCAAA GATGGATAAA GCGGAATTAA TTCCCGAGCC  
     451  TCCAAAAAAG AAGAGAAAGG TCGAATTGGG TACCGCCGCC AATTTTAATC  
     501  AAAGTGGGAA TATTGCTGAT AGCTCATTGT CCTTCACTTT CACTAACAGT  
     551  AGCAACGGTC CGAACCTCAT AACAACTCAA ACAAATTCTC AAGCGCTTTC  
     601  ACAACCAATT GCCTCCTCTA ACGTTCATGA TAACTTCATG AATAATGAAA  
     651  TCACGGCTAG TAAAATTGAT GATGGTAATA ATTCAAAACC ACTGTCACCT  
     701  GGTTGGACGG ACCAAACTGC GTATAACGCG TTTGGAATCA CTACAGGGAT  
     751  GTTTAATACC ACTACAATGG ATGATGTATA TAACTATCTA TTCGATGATG  
     801  AAGATACCCC ACCAAACCCA AAAAAAGAGA TCGAATTAGG ATCCTCTGCT  
     851  AGCAGAGAAT TCAATTCTCT AATGCTTCTC GAGAGTATTA GTCGACTCTA  
     901  GAGCCCTATA GTGAGTCGTA TTACTGCAGA GATCTATGAA TCGTAGATAC  
     951  TGAAAAACCC CGCAAGTTCA CTTCAACTGT GCATCGTGCA CCATCTCAAT  
    1001  TTCTTTCATT TATACATCGT TTTGCCTTCT TTTATGTAAC TATACTCCTC  
    1051  TAAGTTTCAA TCTTGGCCAT GTAACCTCTG ATCTATAGAA TTTTTTAAAT  
    1101  GACTAGAATT AATGCCCATC TTTTTTTTGG ACCTAAATTC TTCATGAAAA  
    1151  TATATTACGA GGGCTTATTC AGAAGCTTTG GACTTCTTCG CCAGAGGTTT  
    1201  GGTCAAGTCT CCAATCAAGG TTGTCGGCTT GTCTACCTTG CCAGAAATTT  
    1251  ACGAAAAGAT GGAAAAGGGT CAAATCGTTG GTAGATACGT TGTTGACACT  
    1301  TCTAAATAAG CGAATTTCTT ATGATTTATG ATTTTTATTA TTAAATAAGT  
    1351  TATAAAAAAA ATAAGTGTAT ACAAATTTTA AAGTGACTCT TAGGTTTTAA  
    1401  AACGAAAATT CTTGTTCTTG AGTAACTCTT TCCTGTAGGT CAGGTTGCTT  
    1451  TCTCAGGTAT AGCATGAGGT CGCTCTTATT GACCACACCT CTACCGGCAT  
    1501  GCCCGAAATT CCCCTACCCT ATGAACATAT TCCATTTTGT AATTTCGTGT  
    1551  CGTTTCTATT ATGAATTTCA TTTATAAAGT TTATGTACAA ATATCATAAA  
    1601  AAAAGAGAAT CTTTTTAAGC AAGGATTTTC TTAACTTCTT CGGCGACAGC  
    1651  ATCACCGACT TCGGTGGTAC TGTTGGAACC ACCTAAATCA CCAGTTCTGA  
    1701  TACCTGCATC CAAAACCTTT TTAACTGCAT CTTCAATGGC CTTACCTTCT  
    1751  TCAGGCAAGT TCAATGACAA TTTCAACATC ATTGCAGCAG ACAAGATAGT  
    1801  GGCGATAGGG TCAACCTTAT TCTTTGGCAA ATCTGGAGCA GAACCGTGGC  
    1851  ATGGTTCGTA CAAACCAAAT GCGGTGTTCT TGTCTGGCAA AGAGGCCAAG  
    1901  GACGCAGATG GCAACAAACC CAAGGAACCT GGGATAACGG AGGCTTCATC  
    1951  GGAGATGATA TCACCAAACA TGTTGCTGGT GATTATAATA CCATTTAGGT  
    2001  GGGTTGGGTT CTTAACTAGG ATCATGGCGG CAGAATCAAT CAATTGATGT  
    2051  TGAACCTTCA ATGTAGGAAA TTCGTTCTTG ATGGTTTCCT CCACAGTTTT  
    2101  TCTCCATAAT CTTGAAGAGG CCAAAACATT AGCTTTATCC AAGGACCAAA  
    2151  TAGGCAATGG TGGCTCATGT TGTAGGGCCA TGAAAGCGGC CATTCTTGTG  
    2201  ATTCTTTGCA CTTCTGGAAC GGTGTATTGT TCACTATCCC AAGCGACACC  
    2251  ATCACCATCG TCTTCCTTTC TCTTACCAAA GTAAATACCT CCCACTAATT  
    2301  CTCTGACAAC AACGAAGTCA GTACCTTTAG CAAATTGTGG CTTGATTGGA  
    2351  GATAAGTCTA AAAGAGAGTC GGATGCAAAG TTACATGGTC TTAAGTTGGC  
    2401  GTACAATTGA AGTTCTTTAC GGATTTTTAG TAAACCTTGT TCAGGTCTAA  
    2451  CACTACCTGT ACCCCATTTA GGACCACCCA CAGCACCTAA CAAAACGGCA  
    2501  TCAACCTTCT TGGAGGCTTC CAGCGCCTCA TCTGGAAGTG GGACACCTGT  
    2551  AGCATCGATA GCAGCACCAC CAATTAAATG ATTTTCGAAA TCGAACTTGA  
    2601  CATTGGAACG AACATCAGAA ATAGCTTTAA GAACCTTAAT GGCTTCGGCT  
    2651  GTGATTTCTT GACCAACGTG GTCACCTGGC AAAACGACGA TCTTCTTAGG  
    2701  GGCAGACATT AGAATGGTAT ATCCTTGAAA TATATATATA TATTGCTGAA  
    2751  ATGTAAAAGG TAAGAAAAGT TAGAAAGTAA GACGATTGCT AACCACCTAT  
    2801  TGGAAAAAAC AATAGGTCCT TAAATAATAT TGTCAACTTC AAGTATTGTG  
    2851  ATGCAAGCAT TTAGTCATGA ACGCTTCTCT ATTCTATATG AAAAGCCGGT  
    2901  TCCGGCCTCT CACCTTTCCT TTTTCTCCCA ATTTTTCAGT TGAAAAAGGT  
    2951  ATATGCGTCA GGCGACCTCT GAAATTAACA AAAAATTTCC AGTCATCGAA  
    3001  TTTGATTCTG TGCGATAGCG CCCCTGTGTG TTCTCGTTAT GTTGAGGAAA  
    3051  AAAATAATGG TTGCTAAGAG ATTCGAACTC TTGCATCTTA CGATACCTGA  
    3101  GTATTCCCAC AGTTGGGGAT CTCGACTCTA GCTAGAGGAT CAATTCGTAA  
    3151  TCATGGTCAT AGCTGTTTCC TGTGTGAAAT TGTTATCCGC TCACAATTCC  
    3201  ACACAACATA CGAGCCGGAA GCATAAAGTG TAAAGCCTGG GGTGCCTAAT  
    3251  GAGTGAGGTA ACTCACATTA ATTGCGTTGC GCTCACTGCC CGCTTTCCAG  
    3301  TCGGGAAACC TGTCGTGCCA GTTATATGCG GCCGCATAAT TTAGGCGCGT  
    3351  AAATTGTAAG CGTTAATATT TTGTTAAAAT TCGCGTTAAA TTTTTGTTAA  
    3401  ATCAGCTCAT TTTTTAACCA ATAGGCCGAA ATCGGCAAAA TCCCTTATAA  
    3451  ATCAAAAGAA TAGACCGAGA TAGGGTTGAG TGTTGTTCCA GTTTGGAACA  
    3501  AGAGTCCACT ATTAAAGAAC GTGGACTCCA ACGTCAAAGG GCGAAAAACC  
    3551  GTCTATCAGG GCGATGGCCC ACTACGTGAA CCATCACCCT AATCAAGTTT  
    3601  TTTGGGGTCG AGGTGCCGTA AAGCACTAAA TCGGAACCCT AAAGGGAGCC  
    3651  CCCGATTTAG AGCTTGACGG GGAAAGCCGG CGAACGTGGC GAGAAAGGAA  
    3701  GGGAAGAAAG CGAAAGGAGC GGGCGCTAGG GCGCTGGCAA GTGTAGCGGT  
    3751  CACGCTGCGC GTAACCACCA CACCCGCCGC GCTTAATGCG CCGCTACAGG  
    3801  GCGCGTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC CCCTATTTGT  
    3851  TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA GACAATAACC  
    3901  CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTCCT GAGGCGGAAA  
    3951  GAACCAGCTG TGGAATGTGT GTCAGTTAGG GTGTGGAAAG TCCCCAGGCT  
    4001  CCCCAGCAGG CAGAAGTATG CAAAGCATGC ATCTCAATTA GTCAGCAACC  
    4051  AGGTGTGGAA AGTCCCCAGG CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT  
    4101  GCATCTCAAT TAGTCAGCAA CCATAGTCCC GCCCCTAACT CCGCCCATCC  
    4151  CGCCCCTAAC TCCGCCCAGT TCCGCCCATT CTCCGCCCCA TGGCTGACTA  
    4201  ATTTTTTTTA TTTATGCAGA GGCCGAGGCC GCCTCGGCCA TTACAGCTGG  
    4251  ATTAATGAAT CGGCCAACGC GCGGGGAGAG GCGGTTTGCG TATTGGGCGC  
    4301  TCTTCCGCTT CCTCGCTCAC TGACTCGCTG CGCTCGGTCG TTCGGCTGCG  
    4351  GCGAGCGGTA TCAGCTCACT CAAAGGCGGT AATACGGTTA TCCACAGAAT  
    4401  CAGGGGATAA CGCAGGAAAG AACATGTGAG CAAAAGGCCA GCAAAAGGCC  
    4451  AGGAACCGTA AAAAGGCCGC GTTGCTGGCG TTTTTCCATA GGCTCCGCCC  
    4501  CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC  
    4551  CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG  
    4601  CGCTCTCCTG TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT  
    4651  CCCTTCGGGA AGCGTGGCGC TTTCTCATAG CTCACGCTGT AGGTATCTCA  
    4701  GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA CGAACCCCCC  
    4751  GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC TTGAGTCCAA  
    4801  CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA  
    4851  TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG  
    4901  CCTAACTACG GCTACACTAG AAGGACAGTA TTTGGTATCT GCGCTCTGCT  
    4951  GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC  
    5001  AAACCACCGC TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA GCAGATTACG  
    5051  CGCAGAAAAA AAGGATCTCA AGAAGATCCT TTGATCTTTT CTACGGGGTC  
    5101  TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTG GTCATGAGAT  
    5151  TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTT  
    5201  AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG  
    5251  CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA  
    5301  TAGTTGCCTG ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA  
    5351  CCATCTGGCC CCAGTGCTGC AATGATACCG CGAGACCCAC GCTCACCGGC  
    5401  TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCC GAGCGCAGAA  
    5451  GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGG  
    5501  GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC  
    5551  CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT  
    5601  TCAGCTCCGG TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG  
    5651  TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT CCGATCGTTG TCAGAAGTAA  
    5701  GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG CATAATTCTC  
    5751  TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCA  
    5801  ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC  
    5851  GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC  
    5901  TCATCATTGG AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG  
    5951  CTGTTGAGAT CCAGTTCGAT GTAACCCACT CGTGCACCCA ACTGATCTTC  
    6001  AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA ACAGGAAGGC  
    6051  AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTC  
    6101  ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT  
    6151  CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG  
    6201  TTCCGCGCAC ATTTCCCCGA AAAGTGCCAC CTGACGTCTA AGAAACCATT  
    6251  ATTATCATGA CATTAACCTA TAAAAATAGG CGTATCACGA GGCCCTTTCG  
    6301  TCTCGCGCGT TTCGGTGATG ACGGTGAAAA CCTCTGACAC ATGCAGCTCC  
    6351  CGGAGACGGT CACAGCTTGT CTGTAAGCGG ATGCCGGGAG CAGACAAGCC  
    6401  CGTCAGGGCG CGTCAGCGGG TGTTGGCGGG TGTCGGGGCT GGCTTAACTA  
    6451  TGCGGCATCA GAGCAGATTG TACTGAGAGT GCACCATAAC GCATTTAAGC  
    6501  ATAAACACGC ACTATGCCGT TCTTCTCATG TATATATATA TACAGGCAAC  
    6551  ACGCAGATAT AGGTGCGACG TGAACAGTGA GCTGTATGTG CGCAGCTCGC  
    6601  GTTGCATTTT CGGAAGCGCT CGTTTTCGGA AACGCTTTGA AGTTCCTATT  
    6651  CCGAAGTTCC TATTCTCTAG CTAGAAAGTA TAGGAACTTC AGAGCGCTTT  
    6701  TGAAAACCAA AAGCGCTCTG AAGACGCACT TTCAAAAAAC CAAAAACGCA  
    6751  CCGGACTGTA ACGAGCTACT AAAATATTGC GAATACCGCT TCCACAAACA  
    6801  TTGCTCAAAA GTATCTCTTT GCTATATATC TCTGTGCTAT ATCCCTATAT  
    6851  AACCTACCCA TCCACCTTTC GCTCCTTGAA CTTGCATCTA AACTCGACCT  
    6901  CTACATTTTT TATGTTTATC TCTAGTATTA CTCTTTAGAC AAAAAAATTG  
    6951  TAGTAAGAAC TATTCATAGA GTGAATCGAA AACAATACGA AAATGTAAAC  
    7001  ATTTCCTATA CGTAGTATAT AGAGACAAAA TAGAAGAAAC CGTTCATAAT  
    7051  TTTCTGACCA ATGAAGAATC ATCAACGCTA TCACTTTCTG TTCACAAAGT  
    7101  ATGCGCAATC CACATCGGTA TAGAATATAA TCGGGGATGC CTTTATCTTG  
    7151  AAAAAATGCA CCCGCAGCTT CGCTAGTAAT CAGTAAACGC GGGAAGTGGA  
    7201  GTCAGGCTTT TTTTATGGAA GAGAAAATAG ACACCAAAGT AGCCTTCTTC  
    7251  TAACCTTAAC GGACCTACAG TGCAAAAAGT TATCAAGAGA CTGCATTATA  
    7301  GAGCGCACAA AGGAGAAAAA AAGTAATCTA AGATGCTTTG TTAGAAAAAT  
    7351  AGCGCTCTCG GGATGCATTT TTGTAGAACA AAAAAGAAGT ATAGATTCTT  
    7401  TGTTGGTAAA ATAGCGCTCT CGCGTTGCAT TTCTGTTCTG TAAAAATGCA  
    7451  GCTCAGATTC TTTGTTTGAA AAATTAGCGC TCTCGCGTTG CATTTTTGTT  
    7501  TTACAAAAAT GAAGCACAGA TTCTTCGTTG GTAAAATAGC GCTTTCGCGT  
    7551  TGCATTTCTG TTCTGTAAAA ATGCAGCTCA GATTCTTTGT TTGAAAAATT  
    7601  AGCGCTCTCG CGTTGCATTT TTGTTCTACA AAATGAAGCA CAGATGCTTC  
    7651  GTT
    
    

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