LOCUS       sequence-349522-        7560 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 7560)
  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 7560)
  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: pTL250
FEATURES             Location/Qualifiers
     source          1..7560
                     /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      1595..1642
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1669..1856
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1724..1748)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1880..1896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1880..1896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1893..1915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1904..1920
                     /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(1928..1934,1935..1952,1953..1958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1973..1994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2111..2128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2282..2870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2362..2381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3199..3246
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3271..3290
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3478..4272)
                     /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(3478..4272)
                     /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(3589..3608)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4199..4218
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4273..4377)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4404..5746
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5367..5414)
                     /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     5787..5805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5843..5865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5965..5984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6005..6286
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6287..6961
                     /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(7064..7519)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7201..7222)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7413..7432
                     /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 aaaattgtga
     1321 tcaaagtgtg cgccccgtgc gaaggttgcc gtgcaaaagc actggaagtg gccgcacgtg
     1381 ccgccgatgg tgtgattagc ctggccatta ccggcgatga tcgcgataaa ctggaagtgg
     1441 tgggcgtggg tgtggatgtg acccgcctgg tgatttgcct gcgcaaaaag gtgtgctacg
     1501 ccgagattct gctggtggaa gaggagaaag aagaagaaga aaagaaagaa ccggaatgca
     1561 aataaggatc cgtcgacctg cagcggccgc ataactagca taaccccttg gggcctctaa
     1621 acgggtcttg aggggttttt tgcgcgcttg cagccaagct aattccgggc gaatttctta
     1681 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     1741 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     1801 aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg
     1861 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1921 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1981 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2041 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3001 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
     3061 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
     3121 cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
     3181 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
     3241 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
     3301 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     3361 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     3421 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     3481 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     3541 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     3601 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     3661 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gagtcacgac
     3721 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
     3781 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     3841 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     3901 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     3961 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     4021 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     4081 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     4141 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     4201 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     4261 ttgttcaatc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4321 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4381 atttccccga aaagtgccac ctgaacgaag catctgtgct tcattttgta gaacaaaaat
     4441 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     4501 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     4561 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     4621 acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt
     4681 ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt
     4741 tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta
     4801 aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca
     4861 cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat
     4921 ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc
     4981 gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat
     5041 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
     5101 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
     5161 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
     5221 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
     5281 tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga
     5341 gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc
     5401 ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg
     5461 cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata
     5521 tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat
     5581 ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt
     5641 atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg
     5701 gattagtctc atccttcaat gctatcattt cctttgatat tggatcatac taagaaacca
     5761 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5821 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5881 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5941 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6001 gatcaacgac attactatat atataatata ggaagcattt aatagaacag catcgtaata
     6061 tatgtgtact ttgcagttat gacgccagat ggcagtagtg gaagatattc tttattgaaa
     6121 aatagcttgt caccttacgt acaatcttga tccggagctt ttcttttttt gccgattaag
     6181 aattaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt ggtgactatt
     6241 gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt ctgttattaa
     6301 tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca cagaggccgc
     6361 agaatgtgct ctagattccg atgctgactt gctgggtatt atatgtgtgc ccaatagaaa
     6421 gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag catataaaaa
     6481 tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta aggaggatgt
     6541 tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag atgagtcgtg
     6601 gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat ttccaaaaga
     6661 ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc ccttgtttga
     6721 ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg actgggttgg
     6781 aaggcaagag agccccgaaa gcttacattt tatgttagct ggtggactga cgccagaaaa
     6841 tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg gaggtgtgga
     6901 gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg ctaagaaata
     6961 ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgccgat ctatgcggtg
     7021 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     7081 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     7141 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     7201 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7261 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     7321 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7381 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7441 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7501 gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat
//
