LOCUS       sequence-348915-        7563 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 7563)
  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 7563)
  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: pTL82
FEATURES             Location/Qualifiers
     source          1..7563
                     /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      1598..1645
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1672..1859
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1727..1751)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1883..1899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1883..1899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1907..1923
                     /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(1931..1937,1938..1955,1956..1961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1976..1997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2114..2131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2285..2873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2365..2384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3202..3249
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3274..3293
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3481..4275)
                     /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(3481..4275)
                     /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(3592..3611)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4202..4221
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4276..4380)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4407..5749
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5370..5417)
                     /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     5790..5808
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5846..5868)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5968..5987
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6008..6289
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6290..6964
                     /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(7067..7522)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7204..7225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7416..7435
                     /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 attccgcaaa gaaattatta
     1321 ttcgcatgca cgtgaaaagc gacaaatgcc aggccaaagc catgaaagtg gccgccgcag
     1381 tgagcggtgt ggaaagcgtt agcctggccg gtggtgataa aagcctgctg ctggtgattg
     1441 gtaccggcgt ggatagcaac aaactggtga agaaactgaa gaagaaggtg ggcgaagccg
     1501 aaattgtgga actgcgcacc cacgatacct ttgaaccggc agcactgcct ctgccgggta
     1561 ccaaataagg atccgtcgac ctgcagcggc cgcataacta gcataacccc ttggggcctc
     1621 taaacgggtc ttgaggggtt ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc
     1681 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     1741 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     1801 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
     1861 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1921 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3001 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     3061 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     3121 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     3181 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     3241 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     3301 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     3361 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     3421 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     3481 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     3541 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     3601 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     3661 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgagtcac
     3721 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     3781 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     3841 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     3901 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     3961 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     4021 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     4081 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     4141 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     4201 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     4261 atcttgttca atcatactct tcctttttca atattattga agcatttatc agggttattg
     4321 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4381 cacatttccc cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa
     4441 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     4501 agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     4561 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4621 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     4681 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     4741 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     4801 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     4861 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     4921 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     4981 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     5041 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     5101 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     5161 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     5221 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     5281 caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc
     5341 agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac
     5401 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
     5461 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
     5521 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
     5581 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
     5641 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
     5701 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca tactaagaaa
     5761 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5821 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5881 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5941 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6001 atagatcaac gacattacta tatatataat ataggaagca tttaatagaa cagcatcgta
     6061 atatatgtgt actttgcagt tatgacgcca gatggcagta gtggaagata ttctttattg
     6121 aaaaatagct tgtcacctta cgtacaatct tgatccggag cttttctttt tttgccgatt
     6181 aagaattaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact
     6241 attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat
     6301 taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc
     6361 cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag
     6421 aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa
     6481 aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga
     6541 tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc
     6601 gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa
     6661 agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt
     6721 tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt
     6781 tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga
     6841 aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt
     6901 ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa
     6961 ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc gatctatgcg
     7021 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     7081 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     7141 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     7201 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     7261 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     7321 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     7381 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     7441 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     7501 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     7561 gat
//
