LOCUS       sequence-349992-        7548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGBKT7 yeast two-hybrid plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7548)
  AUTHORS   Bentham AR, Petit-Houdenot Y, Win J, Chuma I, Terauchi R, Banfield 
            MJ, Kamoun S, Langner T
  TITLE     A single amino acid polymorphism in a conserved effector of the 
            multihost blast fungus pathogen expands host-target binding 
            spectrum.
  JOURNAL   PLoS Pathog. 2021 Nov 10;17(11):e1009957. doi: 
            10.1371/journal.ppat.1009957. eCollection 2021 Nov.
  PUBMED    34758051
REFERENCE   2  (bases 1 to 7548)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2021-11-10"; volume: "17(11):e1009957. doi"; pages:
            " 10.1371/journal.ppat.1009957. eCollection 2021 Nov"
COMMENT     Sequence Label: pTL95
FEATURES             Location/Qualifiers
     source          1..7548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     1213..1232
                     /label=T7
                     /note="T7 promoter, forward primer"
     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      1583..1630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1657..1844
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1712..1736)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1868..1884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1868..1884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1881..1903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1892..1908
                     /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(join(1916..1922,1923..1940,1941..1946))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1961..1982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2099..2116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2270..2858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2350..2369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3187..3234
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3259..3278
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3466..4260)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3466..4260)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3577..3596)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4187..4206
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4261..4365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4392..5734
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5355..5402)
                     /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)."
     primer_bind     5775..5793
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5831..5853)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5953..5972
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5993..6274
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6275..6949
                     /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(7052..7507)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7189..7210)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7401..7420
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
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 attcaaacag aaaattgtga
     1321 tcaaagccac catgagcaac gccaaaagcc gtgcccaggc aatggtgctg gccagcaaag
     1381 ccaatggtgt gggtagcgtg ggcattaccg gcgatctgaa agatcagctg gaagtggtgg
     1441 gtgtgggcat tgatattgcc tgcctggtgc gctgcctgcg caaaaaactg cgctatgccg
     1501 agatcgtgaa ggtggaagag gtgaaggaca aaccggagga agagaaaaaa taaggatccg
     1561 tcgacctgca gcggccgcat aactagcata accccttggg gcctctaaac gggtcttgag
     1621 gggttttttg cgcgcttgca gccaagctaa ttccgggcga atttcttatg atttatgatt
     1681 tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag
     1741 gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct
     1801 caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgca agcttggcgt
     1861 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1921 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1981 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2041 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2101 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2161 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2221 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2281 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2341 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2401 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2461 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2521 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2581 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2641 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2701 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2761 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2821 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2881 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2941 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3001 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     3061 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     3121 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     3181 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     3241 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     3301 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     3361 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     3421 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     3481 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     3541 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     3601 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     3661 attttccacc atgatattcg gcaagcaggc atcgccatga gtcacgacga gatcctcgcc
     3721 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     3781 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     3841 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     3901 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     3961 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     4021 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     4081 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     4141 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     4201 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     4261 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4321 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4381 agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     4441 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     4501 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     4561 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     4621 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     4681 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     4741 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     4801 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     4861 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     4921 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     4981 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     5041 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     5101 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     5161 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     5221 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     5281 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt
     5341 ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
     5401 tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc
     5461 tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga
     5521 acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg
     5581 aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc
     5641 cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat
     5701 ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt attatcatga
     5761 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5821 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5881 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     5941 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataga tcaacgacat
     6001 tactatatat ataatatagg aagcatttaa tagaacagca tcgtaatata tgtgtacttt
     6061 gcagttatga cgccagatgg cagtagtgga agatattctt tattgaaaaa tagcttgtca
     6121 ccttacgtac aatcttgatc cggagctttt ctttttttgc cgattaagaa ttaattcggt
     6181 cgaaaaaaga aaaggagagg gccaagaggg agggcattgg tgactattga gcacgtgagt
     6241 atacgtgatt aagcacacaa aggcagcttg gagtatgtct gttattaatt tcacaggtag
     6301 ttctggtcca ttggtgaaag tttgcggctt gcagagcaca gaggccgcag aatgtgctct
     6361 agattccgat gctgacttgc tgggtattat atgtgtgccc aatagaaaga gaacaattga
     6421 cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata gttcaggcac
     6481 tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt tggctctggt
     6541 caatgattac ggcattgata tcgtccaact gcatggagat gagtcgtggc aagaatacca
     6601 agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact gcaacatact
     6661 actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt cagaagcagg
     6721 tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa ggcaagagag
     6781 ccccgaaagc ttacatttta tgttagctgg tggactgacg ccagaaaatg ttggtgatgc
     6841 gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt
     6901 aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg ttattactga
     6961 gtagtattta tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg aaataccgca
     7021 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     7081 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     7141 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     7201 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     7261 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     7321 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     7381 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     7441 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     7501 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgat
//
