LOCUS       sequence-349778-        7512 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 7512)
  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 7512)
  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: pTL325
FEATURES             Location/Qualifiers
     source          1..7512
                     /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      1547..1594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1621..1808
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1676..1700)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1832..1848)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1832..1848)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1845..1867)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1856..1872
                     /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(1880..1886,1887..1904,1905..1910))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1925..1946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2063..2080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2234..2822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2314..2333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3151..3198
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3223..3242
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3430..4224)
                     /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(3430..4224)
                     /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(3541..3560)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4151..4170
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4225..4329)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4356..5698
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5319..5366)
                     /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     5739..5757
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5795..5817)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5917..5936
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5957..6238
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6239..6913
                     /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(7016..7471)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7153..7174)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7365..7384
                     /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 attcgtggaa gtggtggtgc
     1321 tgcgcgtgag cctgcattgc aaaggctgcg ccggcaaagt gaagaaacat atcagcaaaa
     1381 tggaaggcgt gaccagcctg gacatcgata tcgccaccaa aaaggtgacc gtggtgggtc
     1441 atgtgacccc gctgagcgtt ctgaccgccg tgagcaagat taaaccggcc cagttttggc
     1501 cgattagcag cagcagcgga tccgtcgacc tgcagcggcc gcataactag cataacccct
     1561 tggggcctct aaacgggtct tgaggggttt tttgcgcgct tgcagccaag ctaattccgg
     1621 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     1681 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     1741 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     1801 tctaccggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1861 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1921 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1981 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2041 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2641 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2941 aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg
     3001 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     3061 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     3121 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     3181 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     3241 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga
     3301 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     3361 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     3421 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     3481 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     3541 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     3601 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     3661 atgagtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc
     3721 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     3781 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     3841 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     3901 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     3961 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     4021 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac
     4081 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     4141 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     4201 gtgcaatcca tcttgttcaa tcatactctt cctttttcaa tattattgaa gcatttatca
     4261 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4321 ggttccgcgc acatttcccc gaaaagtgcc acctgaacga agcatctgtg cttcattttg
     4381 tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     4441 ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat
     4501 ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     4561 catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac
     4621 ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta
     4681 gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact
     4741 gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa
     4801 agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca
     4861 agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca
     4921 gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt
     4981 ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc
     5041 tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa
     5101 aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat
     5161 atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag
     5221 cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag
     5281 tgcgtcttca gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag
     5341 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     5401 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     5461 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     5521 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     5581 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     5641 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat
     5701 actaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5761 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5821 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5881 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5941 gagtgcacca tagatcaacg acattactat atatataata taggaagcat ttaatagaac
     6001 agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag tggaagatat
     6061 tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc ttttcttttt
     6121 ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca
     6181 ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat
     6241 gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag
     6301 cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt
     6361 gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa
     6421 agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc
     6481 taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg
     6541 agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt
     6601 atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat
     6661 tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc
     6721 tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact
     6781 gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag
     6841 cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa
     6901 tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgccg
     6961 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     7021 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7081 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7141 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7201 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7261 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7321 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7381 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7441 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     7501 gggaagggcg at
//
