LOCUS       sequence-349786-        7515 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 7515)
  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 7515)
  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: pTL331
FEATURES             Location/Qualifiers
     source          1..7515
                     /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      1550..1597
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1624..1811
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1679..1703)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1835..1851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1835..1851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1848..1870)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1859..1875
                     /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(1883..1889,1890..1907,1908..1913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1928..1949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2066..2083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2237..2825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2317..2336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3154..3201
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3226..3245
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3433..4227)
                     /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(3433..4227)
                     /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(3544..3563)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4154..4173
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4228..4332)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4359..5701
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5322..5369)
                     /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     5742..5760
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5798..5820)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5920..5939
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5960..6241
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6242..6916
                     /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(7019..7474)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7156..7177)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7368..7387
                     /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 attcccgaaa accgtggaac
     1321 tgcgcgtgag catgcattgc tatggctgcg ccaaaaaggt gcagaagcac atcagcaaaa
     1381 tggatggcgt gaccagcttc gaagtggacc tggaaagcaa gaaagtggtg gtgatcggcg
     1441 acattacccc gtatgaagtg ctggccagcg tgagcaaagt gatgaaattc gccgagctgt
     1501 gggtggcccc gaatagccag ggatccgtcg acctgcagcg gccgcataac tagcataacc
     1561 ccttggggcc tctaaacggg tcttgagggg ttttttgcgc gcttgcagcc aagctaattc
     1621 cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     1681 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     1741 tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac
     1801 acctctaccg gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1861 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1921 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1981 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2041 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2101 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2161 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2221 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2281 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2341 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2401 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2461 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2521 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2581 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2641 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2701 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2761 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2821 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2881 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2941 aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc
     3001 ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg
     3061 gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag
     3121 agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt
     3181 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     3241 tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga
     3301 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     3361 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     3421 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     3481 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     3541 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     3601 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     3661 gccatgagtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     3721 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     3781 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     3841 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     3901 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     3961 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     4021 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     4081 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     4141 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     4201 tgcgtgcaat ccatcttgtt caatcatact cttccttttt caatattatt gaagcattta
     4261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4321 aggggttccg cgcacatttc cccgaaaagt gccacctgaa cgaagcatct gtgcttcatt
     4381 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     4441 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     4501 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     4561 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     4621 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     4681 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     4741 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     4801 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     4861 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     4921 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     4981 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     5041 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     5101 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     5161 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     5221 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     5281 aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     5341 gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa
     5401 atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt
     5461 gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta
     5521 cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
     5581 cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
     5641 gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
     5701 catactaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5761 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     5821 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     5881 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     5941 tgagagtgca ccatagatca acgacattac tatatatata atataggaag catttaatag
     6001 aacagcatcg taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga
     6061 tattctttat tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt
     6121 tttttgccga ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg
     6181 gcattggtga ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag
     6241 tatgtctgtt attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca
     6301 gagcacagag gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg
     6361 tgtgcccaat agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt
     6421 aaaagcatat aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca
     6481 acctaaggag gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca
     6541 tggagatgag tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact
     6601 cgtatttcca aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt
     6661 tattcccttg tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat
     6721 ttctgactgg gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg
     6781 actgacgcca gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt
     6841 aagcggaggt gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa
     6901 aaatgctaag aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg
     6961 ccgatctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
     7021 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     7081 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     7141 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     7201 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     7261 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     7321 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     7381 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     7441 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
     7501 gttgggaagg gcgat
//
