LOCUS       sequence-349658-        7554 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 7554)
  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 7554)
  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: pTL321
FEATURES             Location/Qualifiers
     source          1..7554
                     /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      1589..1636
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1663..1850
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1718..1742)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1874..1890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1874..1890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1887..1909)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1898..1914
                     /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(1922..1928,1929..1946,1947..1952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1967..1988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2105..2122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2276..2864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2356..2375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3193..3240
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3265..3284
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3472..4266)
                     /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(3472..4266)
                     /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(3583..3602)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4193..4212
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4267..4371)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4398..5740
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5361..5408)
                     /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     5781..5799
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5837..5859)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5959..5978
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5999..6280
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6281..6955
                     /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(7058..7513)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7195..7216)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7407..7426
                     /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 attcattcag acccatgtgc
     1321 tgcgcgtgaa cattcattgc gacggctgca agcacaaggt gaaaaagctg ctgcagaaga
     1381 tcgaaggcgt gtatagcgtg gccctggacg tggataacca taaagtgacc gtgaccggca
     1441 acgtggatag cgataccctg attcgcaaac tgacccgcgg tggtaaacat gccgaactgt
     1501 ggagccagca gaaaggtggc agcaatcagg gccacaaagg cagcaacaat cagcagaaag
     1561 gatccgtcga cctgcagcgg ccgcataact agcataaccc cttggggcct ctaaacgggt
     1621 cttgaggggt tttttgcgcg cttgcagcca agctaattcc gggcgaattt cttatgattt
     1681 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     1741 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     1801 ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg catgcaagct
     1861 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1921 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1981 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2041 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2101 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2161 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2221 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2281 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2341 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2401 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2461 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2521 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2581 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2641 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2701 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2761 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2821 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2881 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2941 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3001 tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg
     3061 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg
     3121 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa
     3181 ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg
     3241 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag
     3301 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     3361 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     3421 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     3481 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     3541 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     3601 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     3661 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgagtca cgacgagatc
     3721 ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     3781 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     3841 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     3901 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     3961 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     4021 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     4081 gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     4141 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     4201 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     4261 aatcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4321 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4381 ccgaaaagtg ccacctgaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     4441 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     4501 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     4561 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4621 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     4681 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     4741 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     4801 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     4861 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     4921 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     4981 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     5041 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     5101 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     5161 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     5221 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     5281 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt
     5341 ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata
     5401 ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca
     5461 tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg
     5521 agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt
     5581 aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta
     5641 tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag
     5701 tctcatcctt caatgctatc atttcctttg atattggatc atactaagaa accattatta
     5761 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     5821 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5881 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5941 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catagatcaa
     6001 cgacattact atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg
     6061 tactttgcag ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc
     6121 ttgtcacctt acgtacaatc ttgatccgga gcttttcttt ttttgccgat taagaattaa
     6181 ttcggtcgaa aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac
     6241 gtgagtatac gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac
     6301 aggtagttct ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg
     6361 tgctctagat tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac
     6421 aattgacccg gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc
     6481 aggcactccg aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc
     6541 tctggtcaat gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga
     6601 ataccaagag ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa
     6661 catactactc agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga
     6721 agcaggtggg acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca
     6781 agagagcccc gaaagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg
     6841 tgatgcgctt agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa
     6901 tggtgtaaaa gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat
     6961 tactgagtag tatttattta agtattgttt gtgcacttgc cgatctatgc ggtgtgaaat
     7021 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     7081 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7141 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7201 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7261 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7321 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7381 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7441 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7501 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgat
//
