基本信息
別名: pGBKT7-BD
啟動子: ADH1
復(fù)制子: 2μ
質(zhì)粒分類: 酵母系列，酵母雙雜交載體
質(zhì)粒大小: 7304bp
原核抗性: Kan
真核抗性: TRP1
克隆菌株: DH5a
培養(yǎng)條件: 37度
表達(dá)宿主: 酵母細(xì)胞
質(zhì)粒屬性
質(zhì)粒宿主： 酵母菌
質(zhì)粒用途： 雜交
片段類型： ORF
片段物種： 空載體
原核抗性： Kan
真核抗性： TRP1
熒光標(biāo)記：
功能簡介
酵母雙雜交空載體
質(zhì)粒簡介
The pEXT03/BD (pGBKT7/BD) vector expresses proteins fused to amino acids 1–147 of the GAL4 DNA binding domain (DNA-BD) for constructing fusion proteins with a bait protein. pGBKT7 replicates autonomously in both E. coli and S.cerevisiae from the pUC and 2 μ ori, respectivel.
pGBKT7質(zhì)粒的參考文獻(xiàn)：Gene activation by a CRISPR-assisted trans enhancer
質(zhì)粒圖譜
質(zhì)粒序列
LOCUS Exported 7303 bp ds-DNA circular SYN 06-MAR-2019
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..7303
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 30..734
/gene="S. cerevisiae ADH1"
/label=ADH1 promoter
/note="promoter for alcohol dehydrogenase 1"
CDS 762..1202
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="DNA binding domain of the GAL4 transcriptional
activator"
/label=GAL4 DNA binding domain
/translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
promoter 1213..1231
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1251..1280
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
terminator 1338..1385
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1412..1599
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
rep_origin complement(2025..2613)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
polyA_signal 2942..2989
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(3221..4015)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=Kan
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(4016..4120)
/gene="bla"
/label=AmpR promoter
rep_origin 4147..5489
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 5748..6029
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
CDS 6030..6704
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
rep_origin complement(6807..7262)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
BASE COUNT 1977 a 1653 c 1737 g 1935 t
ORIGIN
1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcccgggg atccgtcgac
1321 ctgcagcggc cgcataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
1381 ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat
1441 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
1501 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
1561 atagcatgag gtcgctctta ttgaccacac ctctaccggc atgcaagctt ggcgtaatca
1621 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
1681 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
1741 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
1801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
1861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
1921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
1981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
2101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
2161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
2221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
2281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
2341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
2401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
2461 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
2521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
2581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
2641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
2701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
2761 tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg
2821 ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc
2881 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
2941 tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
3001 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag
3061 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
3121 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
3181 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
3241 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
3301 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
3361 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
3421 ccaccatgat attcggcaag caggcatcgc catgagtcac gacgagatcc tcgccgtcgg
3481 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
3541 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
3601 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
3661 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
3721 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
3781 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt
3841 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
3901 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
3961 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatactct
4021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
4081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
4141 cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
4201 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
4261 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
4321 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
4381 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
4441 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
4501 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
4561 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
4621 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
4681 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
4741 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
4801 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
4861 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
4921 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
4981 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
5041 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa
5101 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa
5161 gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact
5221 gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc
5281 atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg
5341 tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca
5401 gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc
5461 aatgctatca tttcctttga tattggatca tactaagaaa ccattattat catgacatta
5521 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
5581 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
5641 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
5701 aactatgcgg catcagagca gattgtactg agagtgcacc atagatcaac gacattacta
5761 tatatataat ataggaagca tttaatagaa cagcatcgta atatatgtgt actttgcagt
5821 tatgacgcca gatggcagta gtggaagata ttctttattg aaaaatagct tgtcacctta
5881 cgtacaatct tgatccggag cttttctttt tttgccgatt aagaattaat tcggtcgaaa
5941 aaagaaaagg agagggccaa gagggagggc attggtgact attgagcacg tgagtatacg
6001 tgattaagca cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg
6061 gtccattggt gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt
6121 ccgatgctga cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg
6181 ttattgcaag gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga
6241 aatacttggt tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg
6301 attacggcat tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt
6361 tcctcggttt gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca
6421 gtgcagcttc acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga
6481 caggtgaact tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg
6541 aaagcttaca ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta
6601 gattaaatgg cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag
6661 actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt
6721 atttatttaa gtattgtttg tgcacttgcc gatctatgcg gtgtgaaata ccgcacagat
6781 gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc
6841 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
6901 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
6961 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
7021 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
7081 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
7141 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
7201 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
7261 gtccattcgc cattcaggct gcgcaactgt tgggaagggc gat
