pAD-GAL4-2.1

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: pAD-GAL4-2.1
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: 7653 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: 氨苄青霉素Ampicillin
真核筛选标记: LEU2
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pBD-GAL4-Cam。
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

pAD-GAL4-2.1载体图谱
pAD-GAL4-2.1 多克隆位点

pAD-GAL4-2.1 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

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

载体质粒图谱pdf版和相关资料下载

载体应用举例

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