LOCUS       sequence-348916-        7590 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 7590)
  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 7590)
  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: pTL83
FEATURES             Location/Qualifiers
     source          1..7590
                     /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"
     regulatory      1398..1407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      1625..1672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1699..1886
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1754..1778)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1910..1926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1910..1926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1923..1945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1934..1950
                     /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(1958..1964,1965..1982,1983..1988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2003..2024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2141..2158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2312..2900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2392..2411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3229..3276
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3301..3320
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3508..4302)
                     /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(3508..4302)
                     /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(3619..3638)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4229..4248
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4303..4407)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4434..5776
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5397..5444)
                     /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     5817..5835
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5873..5895)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5995..6014
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6035..6316
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6317..6991
                     /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(7094..7549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7231..7252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7443..7462
                     /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 aaaatcgtgc
     1321 tgaaactgcc gctggatgac gagcgcaaaa aacgcaaagc cttcaaagcc gccgtgggta
     1381 tgaatggtgt gaccagcgcc accatggagg gcgataaaat cattgtggtg ggcgatggcg
     1441 ttgatcctat taccctgacc accatgctgc gccgtagcct gggctatgcc gaactgctga
     1501 gcgtgagcag cggtgatgac aagaagaaag gcgacggtta cggctatggc tatggtggcg
     1561 tgatgatgta tggcggcggc aaagaaggca aataaggatc cgtcgacctg cagcggccgc
     1621 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgcgcgcttg
     1681 cagccaagct aattccgggc gaatttctta tgatttatga tttttattat taaataagtt
     1741 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     1801 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc
     1861 gctcttattg accacacctc taccggcatg caagcttggc gtaatcatgg tcatagctgt
     1921 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1981 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2101 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2161 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2221 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     3061 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     3121 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     3181 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     3241 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     3301 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     3361 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     3421 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     3481 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     3541 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     3601 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     3661 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     3721 cggcaagcag gcatcgccat gagtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     3781 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     3841 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     3901 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     3961 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     4021 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     4081 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     4141 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     4201 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     4261 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atactcttcc tttttcaata
     4321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4381 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaacgaag
     4441 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     4501 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     4561 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     4621 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     4681 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     4741 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     4801 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     4861 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     4921 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     4981 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     5041 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     5101 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     5161 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     5221 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     5281 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     5341 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     5401 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa
     5461 cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt cacgtcgcac
     5521 ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata gtgcgtgttt
     5581 atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag tctagtacct
     5641 cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta
     5701 gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt
     5761 cctttgatat tggatcatac taagaaacca ttattatcat gacattaacc tataaaaata
     5821 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5881 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5941 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     6001 cagagcagat tgtactgaga gtgcaccata gatcaacgac attactatat atataatata
     6061 ggaagcattt aatagaacag catcgtaata tatgtgtact ttgcagttat gacgccagat
     6121 ggcagtagtg gaagatattc tttattgaaa aatagcttgt caccttacgt acaatcttga
     6181 tccggagctt ttcttttttt gccgattaag aattaattcg gtcgaaaaaa gaaaaggaga
     6241 gggccaagag ggagggcatt ggtgactatt gagcacgtga gtatacgtga ttaagcacac
     6301 aaaggcagct tggagtatgt ctgttattaa tttcacaggt agttctggtc cattggtgaa
     6361 agtttgcggc ttgcagagca cagaggccgc agaatgtgct ctagattccg atgctgactt
     6421 gctgggtatt atatgtgtgc ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa
     6481 aatttcaagt cttgtaaaag catataaaaa tagttcaggc actccgaaat acttggttgg
     6541 cgtgtttcgt aatcaaccta aggaggatgt tttggctctg gtcaatgatt acggcattga
     6601 tatcgtccaa ctgcatggag atgagtcgtg gcaagaatac caagagttcc tcggtttgcc
     6661 agttattaaa agactcgtat ttccaaaaga ctgcaacata ctactcagtg cagcttcaca
     6721 gaaacctcat tcgtttattc ccttgtttga ttcagaagca ggtgggacag gtgaactttt
     6781 ggattggaac tcgatttctg actgggttgg aaggcaagag agccccgaaa gcttacattt
     6841 tatgttagct ggtggactga cgccagaaaa tgttggtgat gcgcttagat taaatggcgt
     6901 tattggtgtt gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat
     6961 agcaaatttc gtcaaaaatg ctaagaaata ggttattact gagtagtatt tatttaagta
     7021 ttgtttgtgc acttgccgat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7081 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     7141 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     7201 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     7261 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     7321 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     7381 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     7441 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     7501 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     7561 tcaggctgcg caactgttgg gaagggcgat
//
