LOCUS       sequence-349521-        7551 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 7551)
  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 7551)
  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: pTL249
FEATURES             Location/Qualifiers
     source          1..7551
                     /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      1586..1633
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1660..1847
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1715..1739)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1871..1887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1871..1887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1884..1906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1895..1911
                     /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(1919..1925,1926..1943,1944..1949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1964..1985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2102..2119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2273..2861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2353..2372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3190..3237
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3262..3281
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3469..4263)
                     /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(3469..4263)
                     /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(3580..3599)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4190..4209
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4264..4368)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4395..5737
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5358..5405)
                     /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     5778..5796
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5834..5856)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5956..5975
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5996..6277
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6278..6952
                     /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(7055..7510)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7192..7213)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7404..7423
                     /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 tcaaggtgaa catggccagc gacaaatgcc gcagcaaagc catggccctg gtggcaagca
     1381 ccagcggtgt ggatagcgtg gccctggcag gtgatggcaa agatcaggtg gtggtggtgg
     1441 gcgatggtgt ggacagcatc aaactgacca ccgccctgcg caaaaaagtg ggtcatgcca
     1501 ccctgatgac cgtgggcgaa gtgaaaaaag aagagaaaaa accggaacat gcctaaggat
     1561 ccgtcgacct gcagcggccg cataactagc ataacccctt ggggcctcta aacgggtctt
     1621 gaggggtttt ttgcgcgctt gcagccaagc taattccggg cgaatttctt atgatttatg
     1681 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
     1741 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
     1801 tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat gcaagcttgg
     1861 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1921 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1981 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2041 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2101 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2161 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2221 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2281 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2341 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2401 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2461 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2521 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2581 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2641 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2701 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2761 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2821 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2881 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2941 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3001 aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga
     3061 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
     3121 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
     3181 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
     3241 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
     3301 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     3361 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
     3421 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
     3481 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     3541 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     3601 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     3661 gccattttcc accatgatat tcggcaagca ggcatcgcca tgagtcacga cgagatcctc
     3721 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     3781 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     3841 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     3901 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     3961 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     4021 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     4081 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     4141 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     4201 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     4261 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4321 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4381 aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     4441 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     4501 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     4561 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     4621 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
     4681 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
     4741 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
     4801 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
     4861 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
     4921 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
     4981 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
     5041 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
     5101 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
     5161 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
     5221 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
     5281 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
     5341 gttttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
     5401 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     5461 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     5521 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     5581 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     5641 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     5701 catccttcaa tgctatcatt tcctttgata ttggatcata ctaagaaacc attattatca
     5761 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     5821 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5881 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5941 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat agatcaacga
     6001 cattactata tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac
     6061 tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg
     6121 tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc
     6181 ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg
     6241 agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg
     6301 tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc
     6361 tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat
     6421 tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg
     6481 cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct
     6541 ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata
     6601 ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat
     6661 actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc
     6721 aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga
     6781 gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga
     6841 tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg
     6901 tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac
     6961 tgagtagtat ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc
     7021 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     7081 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     7141 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     7201 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     7261 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     7321 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     7381 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     7441 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     7501 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga t
//
