LOCUS       sequence-349602-        7539 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 7539)
  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 7539)
  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: pTL312
FEATURES             Location/Qualifiers
     source          1..7539
                     /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      1574..1621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1648..1835
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1703..1727)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1859..1875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1859..1875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1872..1894)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1883..1899
                     /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(1907..1913,1914..1931,1932..1937))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1952..1973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2090..2107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2261..2849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2341..2360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3178..3225
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3250..3269
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3457..4251)
                     /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(3457..4251)
                     /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(3568..3587)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4178..4197
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4252..4356)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4383..5725
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5346..5393)
                     /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     5766..5784
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5822..5844)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5944..5963
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5984..6265
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6266..6940
                     /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(7043..7498)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7180..7201)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7392..7411
                     /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 attccagcag aaagtgatcc
     1321 tgaaggtgag cagcatgagc gataccaaga tgaagcagaa agccatggaa accgtggccg
     1381 atatctacgg catcgatagc attgccgccg atcacaaaga ccagaagatg accgtgatcg
     1441 gtgaggtgga caccgtggaa atcgccaaaa aactgaaaaa atttggcaaa gtggatatta
     1501 ttagcgtggg cccggccaaa gaagagaaaa aagatgataa aaaaggatcc gtcgacctgc
     1561 agcggccgca taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1621 gcgcgcttgc agccaagcta attccgggcg aatttcttat gatttatgat ttttattatt
     1681 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     1741 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     1801 catgaggtcg ctcttattga ccacacctct accggcatgc aagcttggcg taatcatggt
     1861 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1921 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1981 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2041 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2101 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2161 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2221 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2281 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2341 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2401 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2461 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2521 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2581 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2641 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2701 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2761 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2821 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2881 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2941 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3001 agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt
     3061 cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat
     3121 ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa
     3181 caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc
     3241 cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact
     3301 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     3361 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     3421 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3481 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3541 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3601 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3661 catgatattc ggcaagcagg catcgccatg agtcacgacg agatcctcgc cgtcgggcat
     3721 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3781 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3841 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3901 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     3961 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     4021 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     4081 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4141 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4201 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tactcttcct
     4261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4381 tgaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt
     4441 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat
     4501 tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct
     4561 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     4621 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     4681 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     4741 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     4801 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     4861 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     4921 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     4981 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     5041 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     5101 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     5161 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     5221 gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag
     5281 tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc
     5341 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac ttcaaagcgt
     5401 ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc
     5461 acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag aacggcatag
     5521 tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat gaaaggtagt
     5581 ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt ccttcagcac
     5641 taccctttag ctgttctata tgctgccact cctcaattgg attagtctca tccttcaatg
     5701 ctatcatttc ctttgatatt ggatcatact aagaaaccat tattatcatg acattaacct
     5761 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5821 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5881 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     5941 atgcggcatc agagcagatt gtactgagag tgcaccatag atcaacgaca ttactatata
     6001 tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt tgcagttatg
     6061 acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc accttacgta
     6121 caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg tcgaaaaaag
     6181 aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag tatacgtgat
     6241 taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta gttctggtcc
     6301 attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc tagattccga
     6361 tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg acccggttat
     6421 tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca ctccgaaata
     6481 cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg tcaatgatta
     6541 cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc aagagttcct
     6601 cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac tactcagtgc
     6661 agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag gtgggacagg
     6721 tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga gccccgaaag
     6781 cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg cgcttagatt
     6841 aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg taaaagactc
     6901 taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg agtagtattt
     6961 atttaagtat tgtttgtgca cttgccgatc tatgcggtgt gaaataccgc acagatgcgt
     7021 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     7081 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7141 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7201 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7261 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     7321 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7381 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     7441 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     7501 attcgccatt caggctgcgc aactgttggg aagggcgat
//
