LOCUS       sequence-349514-        7545 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pGBKT7 yeast two-hybrid plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7545)
  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 7545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pTL246
FEATURES             Location/Qualifiers
     source          1..7545
                     /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      1580..1627
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1654..1841
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1709..1733)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1865..1881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1865..1881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1878..1900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1889..1905
                     /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(1913..1919,1920..1937,1938..1943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1958..1979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2096..2113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2267..2855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2347..2366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3184..3231
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3256..3275
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3463..4257)
                     /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(3463..4257)
                     /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(3574..3593)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4184..4203
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4258..4362)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4389..5731
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5352..5399)
                     /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     5772..5790
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5828..5850)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5950..5969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5990..6271
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6272..6946
                     /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(7049..7504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7186..7207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7398..7417
                     /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 tcaaagtgag catgccgtgc gagaaaagcc gcagcaaagc catggcactg gtggcacgtg
     1381 ccagcggtgt gaatagcatg gaagtgaccg gcgatggcaa agatcgcctg caggtggtgg
     1441 gtgatggtgt tgatcctgtt tgcctggtgg cctgtctgcg tcgcaaaatc ggctatgccg
     1501 aaatcgtgca ggtggaagaa gttaaggaca aaaaaccgga agaaaaataa ggatccgtcg
     1561 acctgcagcg gccgcataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1621 ttttttgcgc gcttgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt
     1681 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     1741 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     1801 gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgcaagc ttggcgtaat
     1861 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1921 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3001 tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg
     3061 ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc
     3121 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     3181 tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     3241 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga
     3301 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     3361 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     3421 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     3481 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3541 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3601 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3661 ttccaccatg atattcggca agcaggcatc gccatgagtc acgacgagat cctcgccgtc
     3721 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     3781 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     3841 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     3901 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     3961 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4021 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     4081 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4141 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4201 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatact
     4261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4381 gccacctgaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc
     4441 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag
     4501 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag
     4561 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     4621 gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc
     4681 ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct
     4741 ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc
     4801 tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact
     4861 gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct
     4921 ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca
     4981 ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa
     5041 tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt
     5101 tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa
     5161 gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag
     5221 caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg
     5281 ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc
     5341 aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca
     5401 aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac atacagctca
     5461 ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat gagaagaacg
     5521 gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg taggatgaaa
     5581 ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt atgcttcctt
     5641 cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta gtctcatcct
     5701 tcaatgctat catttccttt gatattggat catactaaga aaccattatt atcatgacat
     5761 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     5821 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5881 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     5941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagatca acgacattac
     6001 tatatatata atataggaag catttaatag aacagcatcg taatatatgt gtactttgca
     6061 gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct
     6121 tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta attcggtcga
     6181 aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca cgtgagtata
     6241 cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca caggtagttc
     6301 tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat gtgctctaga
     6361 ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa caattgaccc
     6421 ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt caggcactcc
     6481 gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg ctctggtcaa
     6541 tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag aataccaaga
     6601 gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca acatactact
     6661 cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag aagcaggtgg
     6721 gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc aagagagccc
     6781 cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg gtgatgcgct
     6841 tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa
     6901 agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta ttactgagta
     6961 gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa taccgcacag
     7021 atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc
     7081 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     7141 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     7201 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     7261 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     7321 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     7381 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     7441 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     7501 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgat
//
