LOCUS       sequence-349970-        7605 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 7605)
  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 7605)
  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: pTL376
FEATURES             Location/Qualifiers
     source          1..7605
                     /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      1640..1687
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1714..1901
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1769..1793)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1925..1941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1925..1941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1938..1960)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1949..1965
                     /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(1973..1979,1980..1997,1998..2003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2018..2039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2156..2173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2327..2915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2407..2426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3244..3291
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3316..3335
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3523..4317)
                     /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(3523..4317)
                     /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(3634..3653)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4244..4263
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4318..4422)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4449..5791
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5412..5459)
                     /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     5832..5850
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5888..5910)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6010..6029
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6050..6331
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6332..7006
                     /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(7109..7564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7246..7267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7458..7477
                     /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 attcgccgat ccggtggtgc
     1321 tgcgcatgga actgcattgc gccggctgcg cccagaaagt gaaaaagagc atcaaacatc
     1381 tggccggcgt ggaaagcgtg gccgcagatg tggccaccaa taccgttgtg gttgccggta
     1441 ccgccgaagc cgcagcactg aaagcccgca tcgaagccaa gaccaaaaaa ccggtggagg
     1501 tggttagcgc aggtggtggc ggtgcagcag ctaaaaaacc ggccgccgaa ccgaaagccg
     1561 tgaaagatga cggcggcgag aaaaaagacg cacaggccaa agaagaaaaa ggatccgtcg
     1621 acctgcagcg gccgcataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1681 ttttttgcgc gcttgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt
     1741 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     1801 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     1861 gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgcaagc ttggcgtaat
     1921 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1981 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2041 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2101 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2161 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2221 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2281 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2341 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2401 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2461 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2521 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2581 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2641 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2701 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2761 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2821 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2881 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2941 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3001 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3061 tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg
     3121 ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc
     3181 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     3241 tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     3301 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga
     3361 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     3421 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     3481 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     3541 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3601 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3661 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3721 ttccaccatg atattcggca agcaggcatc gccatgagtc acgacgagat cctcgccgtc
     3781 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     3841 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     3901 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     3961 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     4021 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4081 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     4141 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4201 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4261 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatact
     4321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4441 gccacctgaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc
     4501 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag
     4561 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag
     4621 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     4681 gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc
     4741 ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct
     4801 ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc
     4861 tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact
     4921 gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct
     4981 ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca
     5041 ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa
     5101 tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt
     5161 tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa
     5221 gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag
     5281 caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg
     5341 ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc
     5401 aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca
     5461 aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac atacagctca
     5521 ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat gagaagaacg
     5581 gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg taggatgaaa
     5641 ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt atgcttcctt
     5701 cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta gtctcatcct
     5761 tcaatgctat catttccttt gatattggat catactaaga aaccattatt atcatgacat
     5821 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     5881 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5941 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     6001 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagatca acgacattac
     6061 tatatatata atataggaag catttaatag aacagcatcg taatatatgt gtactttgca
     6121 gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct
     6181 tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta attcggtcga
     6241 aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca cgtgagtata
     6301 cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca caggtagttc
     6361 tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat gtgctctaga
     6421 ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa caattgaccc
     6481 ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt caggcactcc
     6541 gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg ctctggtcaa
     6601 tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag aataccaaga
     6661 gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca acatactact
     6721 cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag aagcaggtgg
     6781 gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc aagagagccc
     6841 cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg gtgatgcgct
     6901 tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa
     6961 agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta ttactgagta
     7021 gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa taccgcacag
     7081 atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc
     7141 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     7201 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     7261 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     7321 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     7381 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     7441 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     7501 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     7561 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgat
//
