LOCUS       sequence-349823-        7584 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 7584)
  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 7584)
  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: pTL367
FEATURES             Location/Qualifiers
     source          1..7584
                     /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      1619..1666
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1693..1880
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1748..1772)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1904..1920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1904..1920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1917..1939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1928..1944
                     /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(1952..1958,1959..1976,1977..1982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1997..2018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2135..2152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2306..2894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2386..2405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3223..3270
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3295..3314
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3502..4296)
                     /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(3502..4296)
                     /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(3613..3632)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4223..4242
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4297..4401)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4428..5770
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5391..5438)
                     /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     5811..5829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5867..5889)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5989..6008
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6029..6310
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6311..6985
                     /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(7088..7543)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7225..7246)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7437..7456
                     /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 attcaaacgc accatcatcg
     1321 ttaaagtgga cctggagtgc gaacgctgct atgccaaaat tgatcgcgtg ctgacccgca
     1381 ttcgcgataa aggcgaattt gtgattgatg atattgaatt tgatgtgaaa cataacaaag
     1441 tgatcgtgag cggcccgttt gatccggata aactggccga caagctgtgc tgcaaagcct
     1501 gcaaaattat taaagaaatt gaaattgtgg atctgccgcc tccgccccct ccccctgcac
     1561 ctgagccgga accggagccg ccgaaaaaag gatccgtcga cctgcagcgg ccgcataact
     1621 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcgcg cttgcagcca
     1681 agctaattcc gggcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     1741 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     1801 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     1861 attgaccaca cctctaccgg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     1921 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1981 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2041 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2101 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2161 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2221 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta
     3061 tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg
     3121 ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg
     3181 tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac
     3241 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct
     3301 ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc
     3361 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
     3421 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca
     3481 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     3541 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     3601 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     3661 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     3721 gcaggcatcg ccatgagtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct
     3781 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     3841 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     3901 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     3961 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     4021 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     4081 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag
     4141 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     4201 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     4261 cggagaacct gcgtgcaatc catcttgttc aatcatactc ttcctttttc aatattattg
     4321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgaac gaagcatctg
     4441 tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     4501 tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa
     4561 tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     4621 gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca
     4681 aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa
     4741 caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata
     4801 actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc
     4861 ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg
     4921 tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat
     4981 actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg
     5041 gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt
     5101 ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag taatactaga
     5161 gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga
     5221 tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa
     5281 tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg
     5341 gttttttgaa agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta
     5401 tactttctag agaataggaa cttcggaata ggaacttcaa agcgtttccg aaaacgagcg
     5461 cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc gcacctatat
     5521 ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt gtttatgctt
     5581 aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt acctcctgtg
     5641 atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc tttagctgtt
     5701 ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc atttcctttg
     5761 atattggatc atactaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5821 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     5881 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     5941 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6001 agattgtact gagagtgcac catagatcaa cgacattact atatatataa tataggaagc
     6061 atttaataga acagcatcgt aatatatgtg tactttgcag ttatgacgcc agatggcagt
     6121 agtggaagat attctttatt gaaaaatagc ttgtcacctt acgtacaatc ttgatccgga
     6181 gcttttcttt ttttgccgat taagaattaa ttcggtcgaa aaaagaaaag gagagggcca
     6241 agagggaggg cattggtgac tattgagcac gtgagtatac gtgattaagc acacaaaggc
     6301 agcttggagt atgtctgtta ttaatttcac aggtagttct ggtccattgg tgaaagtttg
     6361 cggcttgcag agcacagagg ccgcagaatg tgctctagat tccgatgctg acttgctggg
     6421 tattatatgt gtgcccaata gaaagagaac aattgacccg gttattgcaa ggaaaatttc
     6481 aagtcttgta aaagcatata aaaatagttc aggcactccg aaatacttgg ttggcgtgtt
     6541 tcgtaatcaa cctaaggagg atgttttggc tctggtcaat gattacggca ttgatatcgt
     6601 ccaactgcat ggagatgagt cgtggcaaga ataccaagag ttcctcggtt tgccagttat
     6661 taaaagactc gtatttccaa aagactgcaa catactactc agtgcagctt cacagaaacc
     6721 tcattcgttt attcccttgt ttgattcaga agcaggtggg acaggtgaac ttttggattg
     6781 gaactcgatt tctgactggg ttggaaggca agagagcccc gaaagcttac attttatgtt
     6841 agctggtgga ctgacgccag aaaatgttgg tgatgcgctt agattaaatg gcgttattgg
     6901 tgttgatgta agcggaggtg tggagacaaa tggtgtaaaa gactctaaca aaatagcaaa
     6961 tttcgtcaaa aatgctaaga aataggttat tactgagtag tatttattta agtattgttt
     7021 gtgcacttgc cgatctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7081 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     7141 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     7201 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     7261 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     7321 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     7381 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     7441 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     7501 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     7561 tgcgcaactg ttgggaaggg cgat
//
