LOCUS       sequence-349655-        7629 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 7629)
  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 7629)
  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: pTL319
FEATURES             Location/Qualifiers
     source          1..7629
                     /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      1664..1711
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1738..1925
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1793..1817)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1949..1965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1949..1965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1962..1984)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1973..1989
                     /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(1997..2003,2004..2021,2022..2027))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2042..2063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2180..2197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2351..2939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2431..2450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3268..3315
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3340..3359
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3547..4341)
                     /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(3547..4341)
                     /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(3658..3677)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4268..4287
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4342..4446)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4473..5815
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5436..5483)
                     /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     5856..5874
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5912..5934)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6034..6053
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6074..6355
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6356..7030
                     /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(7133..7588)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7270..7291)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7482..7501
                     /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 attcatgggc atcctggtga
     1321 ttagcgccga tctgaaatgc tgccgctgca aagagaagct gagcaagatc ctgtgcagcc
     1381 tgcgcgataa atatggcatc gaaaaaactg agtacgagga caaggatgat cgcgtgattg
     1441 tgcgcggcaa ttttccgacc gataaactgc gcagcgtgat ttggtgcaaa gccggccgca
     1501 aactgattcg cgatatcgcc gtggttgatg tgtggccgcc gccgaaaaaa ccggaaaccg
     1561 caggtggtac cacccctgca ccggcagttg caaatccgcc gccggcagca acagttgcag
     1621 caaccccgcc gaaaggatcc gtcgacctgc agcggccgca taactagcat aaccccttgg
     1681 ggcctctaaa cgggtcttga ggggtttttt gcgcgcttgc agccaagcta attccgggcg
     1741 aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac
     1801 aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc
     1861 ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga ccacacctct
     1921 accggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1981 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2041 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2101 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2161 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2221 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2281 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2521 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2761 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3061 gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg
     3121 ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa
     3181 aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc
     3241 agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct
     3301 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     3361 ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3421 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     3481 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     3541 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     3601 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     3661 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     3721 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     3781 agtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     3841 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     3901 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     3961 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     4021 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     4081 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     4141 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     4201 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     4261 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     4321 caatccatct tgttcaatca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4381 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4441 tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag
     4501 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4561 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     4621 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     4681 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     4741 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     4801 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     4861 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     4921 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     4981 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     5041 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5101 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5161 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     5221 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     5281 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     5341 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     5401 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     5461 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     5521 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     5581 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     5641 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     5701 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     5761 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatact
     5821 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5881 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5941 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6001 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6061 tgcaccatag atcaacgaca ttactatata tataatatag gaagcattta atagaacagc
     6121 atcgtaatat atgtgtactt tgcagttatg acgccagatg gcagtagtgg aagatattct
     6181 ttattgaaaa atagcttgtc accttacgta caatcttgat ccggagcttt tctttttttg
     6241 ccgattaaga attaattcgg tcgaaaaaag aaaaggagag ggccaagagg gagggcattg
     6301 gtgactattg agcacgtgag tatacgtgat taagcacaca aaggcagctt ggagtatgtc
     6361 tgttattaat ttcacaggta gttctggtcc attggtgaaa gtttgcggct tgcagagcac
     6421 agaggccgca gaatgtgctc tagattccga tgctgacttg ctgggtatta tatgtgtgcc
     6481 caatagaaag agaacaattg acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc
     6541 atataaaaat agttcaggca ctccgaaata cttggttggc gtgtttcgta atcaacctaa
     6601 ggaggatgtt ttggctctgg tcaatgatta cggcattgat atcgtccaac tgcatggaga
     6661 tgagtcgtgg caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt
     6721 tccaaaagac tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc
     6781 cttgtttgat tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga
     6841 ctgggttgga aggcaagaga gccccgaaag cttacatttt atgttagctg gtggactgac
     6901 gccagaaaat gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg
     6961 aggtgtggag acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc
     7021 taagaaatag gttattactg agtagtattt atttaagtat tgtttgtgca cttgccgatc
     7081 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     7141 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     7201 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     7261 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     7321 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     7381 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     7441 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     7501 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     7561 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     7621 aagggcgat
//
