LOCUS       sequence-349465-        7557 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 7557)
  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 7557)
  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: pTL121
FEATURES             Location/Qualifiers
     source          1..7557
                     /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      1592..1639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1666..1853
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1721..1745)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1877..1893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1877..1893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1890..1912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1901..1917
                     /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(1925..1931,1932..1949,1950..1955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1970..1991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2108..2125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2279..2867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2359..2378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3196..3243
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3268..3287
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3475..4269)
                     /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(3475..4269)
                     /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(3586..3605)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4196..4215
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4270..4374)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4401..5743
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5364..5411)
                     /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     5784..5802
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5840..5862)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5962..5981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6002..6283
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6284..6958
                     /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(7061..7516)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7198..7219)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7410..7429
                     /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 attcagcaaa cgcaccgtgc
     1321 tgcgcgtgga taccagctgc gccaaatgca aacgcaaagt gctgctggca gtgagcagcc
     1381 tgcagggcgt ggacaagatc gagatcgaca gcgaaaaagg caccatgacc gtgaccggtg
     1441 gtgttgatcc tgtggatgtg gtggaagcca cccgccgtaa agccggtaaa cgcgccgatg
     1501 tgctgaccat tggtccgccg ccgagtgcaa gcaccgcaag caaaccggaa gagaaaaaat
     1561 aaggatccgt cgacctgcag cggccgcata actagcataa ccccttgggg cctctaaacg
     1621 ggtcttgagg ggttttttgc gcgcttgcag ccaagctaat tccgggcgaa tttcttatga
     1681 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     1741 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     1801 ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgcaa
     1861 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1921 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1981 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2041 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2101 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2461 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2701 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3001 caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg
     3061 ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc
     3121 gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     3181 gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg
     3241 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     3301 tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     3361 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     3421 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     3481 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     3541 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     3601 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     3661 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatgag tcacgacgag
     3721 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     3781 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     3841 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     3901 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     3961 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     4021 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     4081 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     4141 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     4201 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     4261 ttcaatcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4381 tccccgaaaa gtgccacctg aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
     4441 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     4501 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
     4561 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     4621 gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta
     4681 caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct
     4741 cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg
     4801 ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt
     4861 cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc
     4921 cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt
     4981 gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact
     5041 acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta
     5101 ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga
     5161 gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac
     5221 agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat
     5281 tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg
     5341 cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga
     5401 ataggaactt caaagcgttt ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc
     5461 acatacagct cactgttcac gtcgcaccta tatctgcgtg ttgcctgtat atatatatac
     5521 atgagaagaa cggcatagtg cgtgtttatg cttaaatgcg tacttatatg cgtctattta
     5581 tgtaggatga aaggtagtct agtacctcct gtgatattat cccattccat gcggggtatc
     5641 gtatgcttcc ttcagcacta ccctttagct gttctatatg ctgccactcc tcaattggat
     5701 tagtctcatc cttcaatgct atcatttcct ttgatattgg atcatactaa gaaaccatta
     5761 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     5821 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5881 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5941 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatagat
     6001 caacgacatt actatatata taatatagga agcatttaat agaacagcat cgtaatatat
     6061 gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt attgaaaaat
     6121 agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc gattaagaat
     6181 taattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt gactattgag
     6241 cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgtctg ttattaattt
     6301 cacaggtagt tctggtccat tggtgaaagt ttgcggcttg cagagcacag aggccgcaga
     6361 atgtgctcta gattccgatg ctgacttgct gggtattata tgtgtgccca atagaaagag
     6421 aacaattgac ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag
     6481 ttcaggcact ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt
     6541 ggctctggtc aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggca
     6601 agaataccaa gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg
     6661 caacatacta ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc
     6721 agaagcaggt gggacaggtg aacttttgga ttggaactcg atttctgact gggttggaag
     6781 gcaagagagc cccgaaagct tacattttat gttagctggt ggactgacgc cagaaaatgt
     6841 tggtgatgcg cttagattaa atggcgttat tggtgttgat gtaagcggag gtgtggagac
     6901 aaatggtgta aaagactcta acaaaatagc aaatttcgtc aaaaatgcta agaaataggt
     6961 tattactgag tagtatttat ttaagtattg tttgtgcact tgccgatcta tgcggtgtga
     7021 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     7081 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7141 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     7201 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7261 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     7321 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7381 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     7441 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7501 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgat
//
