LOCUS       sequence-349987-        7530 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 7530)
  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 7530)
  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: pTL362
FEATURES             Location/Qualifiers
     source          1..7530
                     /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      1565..1612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1639..1826
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1694..1718)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1850..1866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1850..1866)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1863..1885)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1874..1890
                     /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(1898..1904,1905..1922,1923..1928))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1943..1964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2081..2098)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2252..2840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2332..2351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3169..3216
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3241..3260
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3448..4242)
                     /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(3448..4242)
                     /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(3559..3578)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4169..4188
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4243..4347)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4374..5716
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5337..5384)
                     /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     5757..5775
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5813..5835)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5935..5954
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5975..6256
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6257..6931
                     /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(7034..7489)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7171..7192)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7383..7402
                     /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 attcatgccg accctgatta
     1321 ttaccgtgga tctgcagtgc tgccgctgca acatcaagat ccagaaggtg ctgtgctgca
     1381 tgcaggaacg cggcgagctg gagatcgaaa aaattgtgta tgaaaaagat accgtggtgg
     1441 tgagcggtcc gagtacaccg cgtagcagtc cggcaagttg tggtgcacgt ccggcccgta
     1501 gcagtcgtac cagccgtagc agccgccgca aacgcggatc cgtcgacctg cagcggccgc
     1561 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgcgcgcttg
     1621 cagccaagct aattccgggc gaatttctta tgatttatga tttttattat taaataagtt
     1681 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     1741 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc
     1801 gctcttattg accacacctc taccggcatg caagcttggc gtaatcatgg tcatagctgt
     1861 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1921 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1981 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2041 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2101 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2161 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2221 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2281 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2341 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2401 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2461 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2521 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2581 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2641 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2701 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2761 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2821 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2881 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2941 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     3001 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     3061 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     3121 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     3181 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     3241 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     3301 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     3361 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     3421 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     3481 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     3541 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     3601 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     3661 cggcaagcag gcatcgccat gagtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     3721 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     3781 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     3841 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     3901 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     3961 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     4021 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     4081 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     4141 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     4201 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atactcttcc tttttcaata
     4261 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4321 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaacgaag
     4381 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     4441 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     4501 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     4561 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     4621 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     4681 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     4741 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     4801 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     4861 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     4921 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     4981 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     5041 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     5101 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     5161 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     5221 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     5281 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     5341 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa
     5401 cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt cacgtcgcac
     5461 ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata gtgcgtgttt
     5521 atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag tctagtacct
     5581 cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta
     5641 gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt
     5701 cctttgatat tggatcatac taagaaacca ttattatcat gacattaacc tataaaaata
     5761 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5821 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5881 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5941 cagagcagat tgtactgaga gtgcaccata gatcaacgac attactatat atataatata
     6001 ggaagcattt aatagaacag catcgtaata tatgtgtact ttgcagttat gacgccagat
     6061 ggcagtagtg gaagatattc tttattgaaa aatagcttgt caccttacgt acaatcttga
     6121 tccggagctt ttcttttttt gccgattaag aattaattcg gtcgaaaaaa gaaaaggaga
     6181 gggccaagag ggagggcatt ggtgactatt gagcacgtga gtatacgtga ttaagcacac
     6241 aaaggcagct tggagtatgt ctgttattaa tttcacaggt agttctggtc cattggtgaa
     6301 agtttgcggc ttgcagagca cagaggccgc agaatgtgct ctagattccg atgctgactt
     6361 gctgggtatt atatgtgtgc ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa
     6421 aatttcaagt cttgtaaaag catataaaaa tagttcaggc actccgaaat acttggttgg
     6481 cgtgtttcgt aatcaaccta aggaggatgt tttggctctg gtcaatgatt acggcattga
     6541 tatcgtccaa ctgcatggag atgagtcgtg gcaagaatac caagagttcc tcggtttgcc
     6601 agttattaaa agactcgtat ttccaaaaga ctgcaacata ctactcagtg cagcttcaca
     6661 gaaacctcat tcgtttattc ccttgtttga ttcagaagca ggtgggacag gtgaactttt
     6721 ggattggaac tcgatttctg actgggttgg aaggcaagag agccccgaaa gcttacattt
     6781 tatgttagct ggtggactga cgccagaaaa tgttggtgat gcgcttagat taaatggcgt
     6841 tattggtgtt gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat
     6901 agcaaatttc gtcaaaaatg ctaagaaata ggttattact gagtagtatt tatttaagta
     6961 ttgtttgtgc acttgccgat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7021 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     7081 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     7141 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     7201 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     7261 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     7321 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     7381 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     7441 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     7501 tcaggctgcg caactgttgg gaagggcgat
//
