LOCUS       sequence-349983-        7527 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 7527)
  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 7527)
  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: pTL372
FEATURES             Location/Qualifiers
     source          1..7527
                     /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      1562..1609
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1636..1823
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1691..1715)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1847..1863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1847..1863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1860..1882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1871..1887
                     /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(1895..1901,1902..1919,1920..1925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1940..1961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2078..2095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2249..2837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2329..2348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3166..3213
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3238..3257
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3445..4239)
                     /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(3445..4239)
                     /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(3556..3575)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4166..4185
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4240..4344)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4371..5713
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5334..5381)
                     /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     5754..5772
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5810..5832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5932..5951
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5972..6253
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6254..6928
                     /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(7031..7486)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7168..7189)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7380..7399
                     /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 attcgtgcag aaaattgtga
     1321 ttaaagtgca tatgagcagc gataaatgcc gccgcaaagc aatggccctg gcagccagca
     1381 ccggtggtgt tgtgagcgtg gaactggccg gtgatgatcg cagcaaagtg gtggtggtgg
     1441 gcgatgtgga tagcattggt ctgaccaatg ccctgcgccg taaagtggat ggtagcgccg
     1501 aactggtgga agtgagcgat gccagcaaaa aaggatccgt cgacctgcag cggccgcata
     1561 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc gcgcttgcag
     1621 ccaagctaat tccgggcgaa tttcttatga tttatgattt ttattattaa ataagttata
     1681 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     1741 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct
     1801 cttattgacc acacctctac cggcatgcaa gcttggcgta atcatggtca tagctgtttc
     1861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg
     3001 ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg
     3061 gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg
     3121 gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     3181 caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg
     3241 tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat
     3301 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     3361 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     3421 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     3481 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     3541 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     3601 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     3661 caagcaggca tcgccatgag tcacgacgag atcctcgccg tcgggcatgc tcgccttgag
     3721 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     3781 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     3841 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     3901 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     3961 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     4021 cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga
     4081 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     4141 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     4201 ggccggagaa cctgcgtgca atccatcttg ttcaatcata ctcttccttt ttcaatatta
     4261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aacgaagcat
     4381 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     4441 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     4501 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     4561 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     4621 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     4681 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     4741 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     4801 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     4861 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     4921 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     4981 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     5041 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     5101 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     5161 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     5221 caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt
     5281 ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc
     5341 ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
     5401 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
     5461 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
     5521 cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct
     5581 gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct
     5641 gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct
     5701 ttgatattgg atcatactaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5761 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5821 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5881 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     5941 agcagattgt actgagagtg caccatagat caacgacatt actatatata taatatagga
     6001 agcatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac gccagatggc
     6061 agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca atcttgatcc
     6121 ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa aaggagaggg
     6181 ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa
     6241 ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt
     6301 ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg ctgacttgct
     6361 gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat
     6421 ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt
     6481 gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat
     6541 cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg gtttgccagt
     6601 tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa
     6661 acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg aacttttgga
     6721 ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct tacattttat
     6781 gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa atggcgttat
     6841 tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta acaaaatagc
     6901 aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat ttaagtattg
     6961 tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7021 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     7081 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     7141 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     7201 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7261 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7321 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7381 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7441 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     7501 ggctgcgcaa ctgttgggaa gggcgat
//
