LOCUS       sequence-349652-        7581 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 7581)
  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 7581)
  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: pTL318
FEATURES             Location/Qualifiers
     source          1..7581
                     /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      1616..1663
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1690..1877
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1745..1769)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1901..1917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1901..1917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1914..1936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1925..1941
                     /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(1949..1955,1956..1973,1974..1979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1994..2015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2132..2149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2303..2891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2383..2402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3220..3267
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3292..3311
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3499..4293)
                     /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(3499..4293)
                     /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(3610..3629)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4220..4239
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4294..4398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4425..5767
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5388..5435)
                     /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     5808..5826
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5864..5886)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5986..6005
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6026..6307
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6308..6982
                     /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(7085..7540)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7222..7243)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7434..7453
                     /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 attcatgaaa attgtgctga
     1321 aagtgccgat tacctgcaaa aaatgcaaaa gctgcattct gcagattgtg agccgcaaca
     1381 aaggcgtgaa gagcctgacc ttcgacgacg aaaaaagcac cctgaccgtg attggtgagg
     1441 tggatgtggt ggtgatcgtg gataaactgc gccatccgaa gaagggcaaa gagaaacgcg
     1501 agggctatat ggtggaagtg atggccgtga gcgatgagaa aaaggaagcc gaggagaaaa
     1561 agaaaaaaga tgaagaagaa aagaaaggat ccgtcgacct gcagcggccg cataactagc
     1621 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgcgcgctt gcagccaagc
     1681 taattccggg cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     1741 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     1801 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt
     1861 gaccacacct ctaccggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1921 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1981 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2041 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2101 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2161 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2221 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2281 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2701 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3001 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     3061 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     3121 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     3181 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     3241 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     3301 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     3361 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     3421 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     3481 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3541 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     3601 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     3661 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     3721 ggcatcgcca tgagtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     3781 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     3841 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     3901 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     3961 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4021 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4081 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     4141 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     4201 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     4261 agaacctgcg tgcaatccat cttgttcaat catactcttc ctttttcaat attattgaag
     4321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa gcatctgtgc
     4441 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     4501 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     4561 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     4621 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     4681 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     4741 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     4801 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     4861 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     4921 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     4981 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     5041 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     5101 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     5161 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     5221 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     5281 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     5341 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     5401 tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt
     5461 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg
     5521 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa
     5581 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata
     5641 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta
     5701 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata
     5761 ttggatcata ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5821 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5881 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5941 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6001 ttgtactgag agtgcaccat agatcaacga cattactata tatataatat aggaagcatt
     6061 taatagaaca gcatcgtaat atatgtgtac tttgcagtta tgacgccaga tggcagtagt
     6121 ggaagatatt ctttattgaa aaatagcttg tcaccttacg tacaatcttg atccggagct
     6181 tttctttttt tgccgattaa gaattaattc ggtcgaaaaa agaaaaggag agggccaaga
     6241 gggagggcat tggtgactat tgagcacgtg agtatacgtg attaagcaca caaaggcagc
     6301 ttggagtatg tctgttatta atttcacagg tagttctggt ccattggtga aagtttgcgg
     6361 cttgcagagc acagaggccg cagaatgtgc tctagattcc gatgctgact tgctgggtat
     6421 tatatgtgtg cccaatagaa agagaacaat tgacccggtt attgcaagga aaatttcaag
     6481 tcttgtaaaa gcatataaaa atagttcagg cactccgaaa tacttggttg gcgtgtttcg
     6541 taatcaacct aaggaggatg ttttggctct ggtcaatgat tacggcattg atatcgtcca
     6601 actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc cagttattaa
     6661 aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac agaaacctca
     6721 ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt tggattggaa
     6781 ctcgatttct gactgggttg gaaggcaaga gagccccgaa agcttacatt ttatgttagc
     6841 tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg ttattggtgt
     6901 tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt
     6961 cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt attgtttgtg
     7021 cacttgccga tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     7081 caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     7141 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     7201 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     7261 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     7321 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     7381 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     7441 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     7501 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc
     7561 gcaactgttg ggaagggcga t
//
