pGBT9

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  • Model: PVT4036
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pGBT9

PVT4036      2ug


pGBT9 Plasmid information

Bacterial Resistance:
Growth Strain:
Expression: Yeast
Use: Yeast expression

pGBT9 Plasmid description

pGBT9 generates a hybrid protein that contains the sequences for the GAL4 DNA-binding domain (DNA-BD; a.a. 1–147). For the construction of a hybrid protein, the gene encoding the protein of interest is ligated into the MCS in the correct orientation and in the correct reading frame such that a fusion protein is generated. The fusion protein is expressed in yeast host cells from the constitutive ADH1 promoter; transcription is terminated at the ADH1 transcription termination signal. The hybrid protein is targeted to the yeast nucleus by nuclear localization sequences that are an intrinsic part of the GAL4 DNA-BD . pGBT9 is a shuttle vector that replicates autonomously in both E. coli and S. cerevisiae. It carries the bla gene (for ampicillin resistance in E. coli) and the TRP1 nutritional marker that allow yeast auxotrophs carrying pGBT9 to grow on limiting synthetic medium lacking Trp.
pGBT9-plasmid

pGBT9 Plasmid Sequence

LOCUS       Exported                5524 bp ds-DNA     circular SYN 10-SEP-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 20

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 5524)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4

FEATURES             Location/Qualifiers

     source          1..5524

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        5..406

                     /gene="S. cerevisiae ADH1"

                     /note="ADH1 promoter"

                     /note="promoter for alcohol dehydrogenase 1"

     CDS             434..874

                     /codon_start=1

                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"

                     /product="DNA binding domain of the GAL4 transcriptional 

                     activator"

                     /note="GAL4 DNA binding domain"

                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK

                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK

                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"

     terminator      921..1108

                     /gene="S. cerevisiae ADH1"

                     /note="ADH1 terminator"

                     /note="transcription terminator for the S. cerevisiae 

                     alcohol dehydrogenase 1 (ADH1) gene"

     CDS             complement(1161..1835)

                     /codon_start=1

                     /gene="S. cerevisiae TRP1"

                     /product="phosphoribosylanthranilate isomerase, required 

                     for tryptophan biosynthesis"

                     /note="TRP1"

                     /note="yeast auxotrophic marker"

                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK

                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES

                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW

                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA

                     KK"

     promoter        complement(1836..1937)

                     /gene="S. cerevisiae TRP1"

                     /note="TRP1 promoter"

     primer_bind     complement(1952..1968)

                     /note="M13 rev"

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

                     variants"

     protein_bind    1976..1992

                     /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(2000..2030)

                     /note="lac promoter"

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

     rep_origin      complement(2354..2942)

                     /direction=LEFT

                     /note="ori"

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

                     replication"

     CDS             complement(3113..3973)

                     /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(3974..4078)

                     /gene="bla"

                     /note="AmpR promoter"

     rep_origin      4360..5524

                     /note="2u ori"

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

ORIGIN

        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc

       61 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga

      121 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac

      181 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct

      241 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga

      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt

      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca

      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact

      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg

      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga

      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga

      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt

      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac

      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag

      841 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcccgggga tccgtcgacc

      901 tgcagccaag ctaattccgg gcgaatttct tatgatttat gatttttatt attaaataag

      961 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat

     1021 tcttgttctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg

     1081 tcgctcttat tgaccacacc tctaccggca tgccggcaag tgcacaaaca atacttaaat

     1141 aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta

     1201 gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt

     1261 aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag

     1321 ctttcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca

     1381 cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct

     1441 gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg

     1501 aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat atcaatgccg

     1561 taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac gccaaccaag

     1621 tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat tttccttgca

     1681 ataaccgggt caattgttct ctttctattg ggcacacata taatacccag caagtcagca

     1741 tcggaatcta gagcacattc tgcggcctct gtgctctgca agccgcaaac tttcaccaat

     1801 ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt tgtgtgctta

     1861 atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc ctctcctttt

     1921 cttttttcga ccgaattaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt

     1981 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg

     2041 cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg

     2101 gaaacctgtc gtgccagctg gattaatgaa tcggccaacg cgcggggaga ggcggtttgc

     2161 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc

     2221 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata

     2281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg

     2341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct

     2401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa

     2461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc

     2521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt

     2581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg

     2641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg

     2701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct

     2761 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc

     2821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg

     2881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc

     2941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt

     3001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa

     3061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat

     3121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct

     3181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg

     3241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag

     3301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta

     3361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg

     3421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg

     3481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct

     3541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta

     3601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg

     3661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc

     3721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg

     3781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga

     3841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg

     3901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat

     3961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc

     4021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca

     4081 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct

     4141 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa

     4201 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga

     4261 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact

     4321 atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg

     4381 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac

     4441 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg

     4501 aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt

     4561 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc

     4621 accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa

     4681 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt

     4741 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt

     4801 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg

     4861 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa

     4921 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat

     4981 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct

     5041 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata

     5101 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag

     5161 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa

     5221 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa

     5281 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga

     5341 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt

     5401 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg

     5461 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt

     5521 cgtt

//

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Product is for research use only!

 

 

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