pGBKT7- GW/ pGBKT7- GW质粒/ pGBKT7- GW Plasmid

pGBKT7-GW

Catalog No. PVT11255
Packing 2ug

 

pGBKT7-GW Informaiton
Function yeast Hybrid plasmid

Promoter: ADH1 promoter

Plasmid size: 8981bp

Prokaryotic resistance: kanamycin Kan

Screening markers: TRP1

Cloned strain: DH5 alpha Escherichia coli

Culture conditions: 37 centigrade, aerobic, LB

 

pGBKT7-GW Description

PGBKT7-GW plasmid is a yeast two hybrid plasmid cloned by GateWay.

 

pGBKT7-GW Sequence

LOCUS       Exported                8981 bp ds-DNA     circular SYN 11-OCT-2017

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8981)

  AUTHORS   .

  TITLE     Direct Submission

           

FEATURES             Location/Qualifiers

     source          1..8981

                     /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 oncogene) epitope tag"

                     /label=Myc

                     /translation="EQKLISEEDL"

     protein_bind    1287..1411

                     /gene="mutant version of attR"

                     /label=attR1

                     /bound_moiety="LR Clonase(TM)"

                     /note="recombination site for the Gateway(R) LR reaction"

     promoter        1436..1466

                     /label=lac UV5 promoter

                     /note="E. coli lac promoter with an ""up"" mutation"

     CDS             1520..2179

                     /codon_start=1

                     /gene="cat"

                     /product="chloramphenicol acetyltransferase"

                     /label=CmR

                     /note="confers resistance to chloramphenicol"

                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL

                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS

                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM

                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"

     CDS             2521..2826

                     /codon_start=1

                     /gene="ccdB"

                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"

                     /label=ccdB

                     /note="Plasmids containing the ccdB gene cannot be 

                     propagated in standard E. coli strains."

                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK

                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"

     protein_bind    complement(2867..2991)

                     /gene="mutant version of attR"

                     /label=attR2

                     /bound_moiety="LR Clonase(TM)"

                     /note="recombination site for the Gateway(R) LR reaction"

     terminator      3015..3062

                     /label=T7 terminator

                     /note="transcription terminator for bacteriophage T7 RNA 

                     polymerase"

     terminator      3089..3276

                     /gene="S. cerevisiae ADH1"

                     /label=ADH1 terminator

                     /note="transcription terminator for the S. cerevisiae 

                     alcohol dehydrogenase 1 (ADH1) gene"

     primer_bind     complement(3300..3316)

                     /label=M13 rev

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     protein_bind    3324..3340

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /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(3348..3378)

                     /label=lac promoter

                     /note="promoter for the E. coli lac operon"

     protein_bind    3393..3414

                     /label=CAP binding site

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(3702..4290)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     polyA_signal    4619..4666

                     /label=HSV TK poly(A) signal

                     /note="herpes simplex virus thymidine kinase 

                     polyadenylation signal (Cole and Stacy, 1985)"

     CDS             complement(4899..5693)

                     /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

                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     promoter        complement(5694..5798)

                     /gene="bla"

                     /label=AmpR promoter

     rep_origin      5825..7167

                     /label=2u ori

                     /note="yeast 2u plasmid origin of replication"

     protein_bind    complement(6788..6835)

                     /label=FRT

                     /bound_moiety="FLP recombinase from the Saccharomyces 

                     cerevisiae 2u plasmid"

                     /note="FLP-mediated recombination occurs in the 8-bp core 

                     sequence TCTAGAAA (Turan and Bode, 2011)."

     promoter        7426..7707

                     /gene="S. cerevisiae TRP1"

                     /label=TRP1 promoter

     CDS             7708..8382

                     /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(8485..8940)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

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 acaaaaaaaa tgatggaaga

      421 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg

      481 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct

      541 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat 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 catatgacaa gtttgtacaa aaaagctgaa cgagaaacgt

     1321 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat

     1381 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac

     1441 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc

     1501 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc

     1561 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa

     1621 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa

     1681 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg

     1741 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt

     1801 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg

     1861 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt

     1921 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac

     1981 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg

     2041 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc

     2101 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga

     2161 gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc

     2221 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat

     2281 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat

     2341 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga

     2401 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg

     2461 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa

     2521 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta

     2581 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt

     2641 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc

     2701 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg

     2761 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga

     2821 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat

     2881 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt

     2941 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg tctcgacctg

     3001 cagcggccgc ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt

     3061 tgcgcgcttg cagccaagct aattccgggc gaatttctta tgatttatga tttttattat

     3121 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa

     3181 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata

     3241 gcatgaggtc gctcttattg accacacctc taccggcatg caagcttggc gtaatcatgg

     3301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc

     3361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg

     3421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc

     3481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact

     3541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta

     3601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag

     3661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc

     3721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta

     3781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg

     3841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc

     3901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac

     3961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac

     4021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg

     4081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga

     4141 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt

     4201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag

     4261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct

     4321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg

     4381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat

     4441 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct

     4501 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa

     4561 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga

     4621 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt

     4681 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac

     4741 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg

     4801 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc

     4861 gtcgcttggt cggttcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg

     4921 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg

     4981 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga

     5041 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc

     5101 accatgatat tcggcaagca ggcatcgtca tgggtcacga cgagatcctc gccgtcgggc

     5161 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc

     5221 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt

     5281 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca

     5341 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc

     5401 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct

     5461 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca

     5521 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc

     5581 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc

     5641 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catactcttc

     5701 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt

     5761 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca

     5821 cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat

     5881 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct

     5941 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg

     6001 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga

     6061 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga

     6121 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat

     6181 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact

     6241 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt

     6301 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac

     6361 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg

     6421 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt

     6481 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc

     6541 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc

     6601 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa

     6661 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac

     6721 agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa

     6781 gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc