LOCUS       sequence-349800-        7632 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 7632)
  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 7632)
  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: pTL342
FEATURES             Location/Qualifiers
     source          1..7632
                     /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"
     CDS             complement(1546..1563)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1609..1632)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      1667..1714
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1741..1928
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1796..1820)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1952..1968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1952..1968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1965..1987)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1976..1992
                     /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(2000..2006,2007..2024,2025..2030))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2045..2066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2183..2200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2354..2942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2434..2453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3271..3318
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3343..3362
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3550..4344)
                     /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(3550..4344)
                     /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(3661..3680)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4271..4290
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4345..4449)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4476..5818
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5439..5486)
                     /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     5859..5877
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5915..5937)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6037..6056
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6077..6358
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6359..7033
                     /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(7136..7591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7273..7294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7485..7504
                     /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 attcatgagc accattgtga
     1321 tgatggtgga cctggaatgc gatcgctgct atcgcaaaat tcgccgcgtg ctgtgcaaac
     1381 tgcaggacaa agccagcatc aaagccatca gctatgacga gaaaaataat accgtgaccg
     1441 tggccggtcc gtttgacgca gatgaagtga gcgaccgtct gtgtagcagc gccggcaaag
     1501 tgattaccga tattcgcgtg gtgggtggtg ccaaaccgat gccgggtggt ggtggtggtg
     1561 gtgcaaaagc ccatgcaaac aaaccggccg gcaaagatgg tagcgcaggt ggtggtggtg
     1621 gtggtggtgg tggtaaagga tccgtcgacc tgcagcggcc gcataactag cataacccct
     1681 tggggcctct aaacgggtct tgaggggttt tttgcgcgct tgcagccaag ctaattccgg
     1741 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     1801 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     1861 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     1921 tctaccggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1981 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2041 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2101 gaaacctgtc gtgccagctg cattaatgaa 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 gaagaacagt 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 atgagtaacc tgaggctatg gcagggcctg
     3121 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     3181 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     3241 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     3301 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     3361 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga
     3421 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     3481 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     3541 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     3601 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     3661 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     3721 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     3781 atgagtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc
     3841 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     3901 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     3961 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     4021 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     4081 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     4141 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac
     4201 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     4261 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     4321 gtgcaatcca tcttgttcaa tcatactctt cctttttcaa tattattgaa gcatttatca
     4381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4441 ggttccgcgc acatttcccc gaaaagtgcc acctgaacga agcatctgtg cttcattttg
     4501 tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     4561 ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat
     4621 ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     4681 catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac
     4741 ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta
     4801 gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact
     4861 gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa
     4921 agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca
     4981 agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca
     5041 gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt
     5101 ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc
     5161 tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa
     5221 aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat
     5281 atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag
     5341 cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag
     5401 tgcgtcttca gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag
     5461 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     5521 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     5581 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     5641 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     5701 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     5761 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat
     5821 actaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5881 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5941 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6001 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6061 gagtgcacca tagatcaacg acattactat atatataata taggaagcat ttaatagaac
     6121 agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag tggaagatat
     6181 tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc ttttcttttt
     6241 ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca
     6301 ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat
     6361 gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag
     6421 cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt
     6481 gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa
     6541 agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc
     6601 taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg
     6661 agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt
     6721 atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat
     6781 tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc
     6841 tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact
     6901 gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag
     6961 cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa
     7021 tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgccg
     7081 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     7141 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7201 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7261 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7321 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7381 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7441 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7501 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7561 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     7621 gggaagggcg at
//
