pGADT7 AD; pGBKT7 Vector；pGADT7-T；pGBKT7-Lam；SD/-Trp；Clontech；Yeast two-hybrid (Y2H)酵母双杂交；
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货号
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产品名称 |
规格 |
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| MF3604-2UG |
pGADT7 AD Vector 酵母双杂交克隆载体 |
2μg |
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基本信息
| 载体类型： |
酵母双杂交系统载体 |
| 载体大小： |
7987bp |
| 原核抗性： |
氨苄青霉素（Ampicillin, Amp） |
| 克隆菌株： |
DH5α |
| 载体标签 |
HA |
| 筛选标记： |
LEU2 |
| 启动子： |
ADH1 |
| 复制子： |
2μ |
| 表达宿主： |
酵母细胞 |
| 5’测序引物： |
T7: TAATACGACTCACTATAGGG |
| 3’测序引物： |
根据序列设计引物 |
质粒图谱
描述：pGADT7 AD是一种酵母双杂交表达载体，特别设计用来表达介于GAL4 AD（a.a. 768-881）和目的蛋白之间的融合蛋白。在ADH1全长启动子的驱动下高水平表达融合蛋白。融合蛋白含一个HA表位标签。为了便于体外转录/翻译，在GAL4 AD和HA表位标签之间又引入T7启动子。
保存与运输方法
保存：-20℃保存，至少1年有效。
运输：冰袋运输。
注意事项
1） 我司提供的载体/质粒/菌株都经过严格的质量控制。请收到货后先进行转化，再挑取单克隆扩增。产品使用中若遇到质量问题，请和我司及时沟通。
2） 为了您的安全和健康，请穿实验服并戴一次性手套操作。
质粒序列
LOCUS Exported 7987 bp ds-DNA circular SYN 09-SEP-2012
DEFINITION Yeast two-hybrid “”prey”” vector for expressing proteins fused to
the GAL4 activation domain.
KEYWORDS pGADT7 AD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7987)
AUTHORS Clontech
TITLE Direct Submission
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7987
/organism=”synthetic DNA construct”
/lab_host=”Saccharomyces cerevisiae”
/mol_type=”other DNA”
promoter 771..1475
/gene=”S. cerevisiae ADH1″
/note=”ADH1 promoter”
/note=”promoter for alcohol dehydrogenase 1″
CDS 1491..1493
/codon_start=1
/product=”start codon for expression in yeast”
/note=”ATG”
/translation=”M”
CDS 1521..1541
/codon_start=1
/product=”nuclear localization signal of SV40 large T
antigen”
/note=”SV40 NLS”
/translation=”PKKKRKV”
CDS 1557..1898
/codon_start=1
/gene=”Saccharomyces cerevisiae GAL4 (truncated)”
/product=”activation domain of the GAL4 transcriptional
activator”
/note=”GAL4 activation domain”
/translation=”ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE”
promoter 1904..1922
/note=”T7 promoter”
/note=”promoter for bacteriophage T7 RNA polymerase”
CDS 1935..1937
/codon_start=1
/product=”start codon for in vitro
transcription/translation”
/note=”ATG”
/translation=”M”
CDS 1941..1967
/codon_start=1
/product=”HA (human influenza hemagglutinin) epitope tag”
/note=”HA”
/translation=”YPYDVPDYA”
misc_feature 1968..2035
/label=MCS
/note=”MCS”
/note=”multiple cloning site”
terminator 2415..2602
/gene=”S. cerevisiae ADH1″
/note=”ADH1 terminator”
/note=”transcription terminator for alcohol dehydrogenase
1″
CDS complement(2719..3813)
/codon_start=1
/gene=”S. cerevisiae LEU2″
/product=”3-isopropylmalate dehydrogenase, required for
leucine biosynthesis”
/note=”LEU2″
/note=”yeast auxotrophic marker”
/translation=”MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA”
promoter complement(3814..4219)
/gene=”S. cerevisiae LEU2″
/note=”LEU2 promoter”
primer_bind complement(4261..4277)
/note=”M13 rev”
/note=”common sequencing primer, one of multiple similar
variants”
protein_bind 4285..4301
/bound_moiety=”lac repressor encoded by lacI”
/note=”lac operator”
/note=”The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG).”
promoter complement(4309..4339)
/note=”lac promoter”
/note=”promoter for the E. coli lac operon”
rep_origin complement(4814..5402)
/direction=LEFT
/note=”ori”
/note=”high-copy-number colE1/pMB1/pBR322/pUC origin of
replication”
CDS complement(5573..6433)
/codon_start=1
/gene=”bla”
/product=”beta-lactamase”
/note=”AmpR”
/note=”confers resistance to ampicillin, carbenicillin, and
related antibiotics”
/translation=”MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW”
promoter complement(6434..6538)
/gene=”bla”
/note=”AmpR promoter”
rep_origin 6820..7984
/note=”2u ori”
/note=”yeast 2u plasmid origin of replication”
ORIGIN
1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
61 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
1141 gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag
1201 atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct
1261 tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc cagttacttg
1321 aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat
1381 taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca
1441 caatatttca agctatacca agcatacaat caactccaag ctttgcaaag atggataaag
1501 cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt accgccgcca
1561 attttaatca aagtgggaat attgctgata gctcattgtc cttcactttc actaacagta
1621 gcaacggtcc gaacctcata acaactcaaa caaattctca agcgctttca caaccaattg
1681 cctcctctaa cgttcatgat aacttcatga ataatgaaat cacggctagt aaaattgatg
1741 atggtaataa ttcaaaacca ctgtcacctg gttggacgga ccaaactgcg tataacgcgt
1801 ttggaatcac tacagggatg tttaatacca ctacaatgga tgatgtatat aactatctat
1861 tcgatgatga agatacccca ccaaacccaa aaaaagagat ctttaatacg actcactata
1921 gggcgagcgc cgccatggag tacccatacg acgtaccaga ttacgctcat atggccatgg
1981 aggccagtga attccacccg ggtgggcatc gatacgggat ccatcgagct cgagctgcag
2041 atgaatcgta gatactgaaa aaccccgcaa gttcacttca actgtgcatc gtgcaccatc
2101 tcaatttctt tcatttatac atcgttttgc cttcttttat gtaactatac tcctctaagt
2161 ttcaatcttg gccatgtaac ctctgatcta tagaattttt taaatgacta gaattaatgc
2221 ccatcttttt tttggaccta aattcttcat gaaaatatat tacgagggct tattcagaag
2281 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
2341 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
2401 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
2461 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
2521 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
2581 ttattgacca cacctctacc ggccggtcga aattccccta ccctatgaac atattccatt
2641 ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca
2701 taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc
2761 gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac
2821 ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa
2881 catcattgca gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg
2941 agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc
3001 caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt catcggagat
3061 gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac
3121 taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag gaaattcgtt
3181 cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt
3241 atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct
3301 tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc
3361 atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa
3421 gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc
3481 aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc
3541 ttgttcaggt ctaacactac ctgtacccca tttaggacca cccacagcac ctaacaaaac
3601 ggcatcagcc ttcttggagg cttccagcgc ctcatctgga agtgggacac ctgtagcttc
3661 gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc
3721 agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc
3781 tggcaaaacg acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata
3841 tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac
3901 ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa
3961 gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc gctctcacct
4021 ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat
4081 taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc
4141 gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata
4201 cctgagtatt cccacagttg ggggatctcg actctagcta gaggatcaat tcgtaatcat
4261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
4321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
4381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctga taacttcgta
4441 taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag ccaagctagc
4501 ttggctgcag gtcgagcggc cgcgatccgg aacccttaat ataacttcgt ataatgtatg
4561 ctatacgaag ttatcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
4621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
4681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
4741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
4801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
4861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
4921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
4981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
5041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
5101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
5161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
5221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
5281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
5341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
5401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
5461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
5521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
5581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
5641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
5701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
5761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
5821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
5881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
5941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
6001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
6061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
6121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
6181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
6241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
6301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
6361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
6421 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
6481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
6541 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
6601 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
6661 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
6721 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
6781 atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg
6841 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac
6901 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg
6961 aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt
7021 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc
7081 accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa
7141 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt
7201 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt
7261 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg
7321 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa
7381 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat
7441 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct
7501 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata
7561 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag
7621 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa
7681 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa
7741 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
7801 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt
7861 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg
7921 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt
7981 cgttgct
