LOCUS       sequence-266985-        6806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive control when used in combination with pAWH-largeT. 
            Expression of Gal4BD-p53 hybrid protein. Homology regions for 
            recombination with pAWH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  AUTHORS   Yang JS, Garriga-Canut M, Link N, Carolis C, Broadbent K, 
            Beltran-Sastre V, Serrano L, Maurer SP
  TITLE     rec-YnH enables simultaneous many-by-many detection of direct 
            protein-protein and protein-RNA interactions.
  JOURNAL   Nat Commun. 2018 Sep 14;9(1):3747. doi: 10.1038/s41467-018-06128-x.
  PUBMED    30217970
REFERENCE   2  (bases 1 to 6806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-09-14"; volume: "9(1):3747. doi"; pages: " 
            10.1038/s41467-018-06128-x"
COMMENT     Sequence Label: pBKWH-p53
FEATURES             Location/Qualifiers
     source          1..6806
                     /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"
     protein_bind    1317..1341
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1503..1514)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2305..2329)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2368..2415
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2497..2521)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      complement(2958..3546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3038..3057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3875..3922
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3947..3966
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4154..4948)
                     /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(4154..4948)
                     /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(4265..4284)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4875..4894
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4949..5053)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5096..5114
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5251..5532
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5533..6207
                     /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(6310..6765)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6447..6468)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6659..6678
                     /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 attcccgggg atctccacaa
     1321 gtttgtacaa aaaagcaggc ttacctgtca ccgagacccc tgggccagtg gcccctgccc
     1381 cagccactcc atggcccctg tcatcttttg tcccttctca aaaaacttac cagggcaact
     1441 atggcttcca cctgggcttc ctgcagtctg ggacagccaa gtctgttatg tgcacgtact
     1501 ctcctcccct caataagcta ttctgccagc tggcgaagac gtgccctgtg cagttgtggg
     1561 tcagcgccac acctccagct gggagccgtg tccgcgccat ggccatctac aagaagtcac
     1621 agcacatgac ggaggtcgtg agacgctgcc cccaccatga gcgctgctcc gatggtgatg
     1681 gcctggctcc tccccagcat cttatccggg tggaaggaaa tttgtatccc gagtatctgg
     1741 aagacaggca gacttttcgc cacagcgtgg tggtacctta tgagccaccc gaggccggct
     1801 ctgagtatac caccatccac tacaagtaca tgtgtaatag ctcctgcatg gggggcatga
     1861 accgccgacc tatccttacc atcatcacac tggaagactc cagtgggaac cttctgggac
     1921 gggacagctt tgaggttcgt gtttgtgcct gccctgggag agaccgccgt acagaagaag
     1981 aaaatttccg caaaaaggaa gtcctttgcc ctgaactgcc cccagggagc gcaaagagag
     2041 cgctgcccac ctgcacaagc gcctctcccc cgcaaaagaa aaaaccactt gatggagagt
     2101 atttcaccct caagatccgc gggcgtaaac gcttcgagat gttccgggag ctgaatgagg
     2161 ccttagagtt aaaggatgcc catgctacag aggagtctgg agacagcagg gctcactcca
     2221 gctacctgaa gaccaagaag ggccagtcta cttcccgcca taaaaaaaca atggtcaaga
     2281 aagtggggcc tgactcagac tgaaacccag ctttcttgta caaagtggta agtaactaag
     2341 atccgtcgac ctgcagcggc cgcataacta gcataacccc ttggggcctc taaacgggtc
     2401 ttgaggggtt ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc ttatgattta
     2461 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     2521 cttaggtttt aaaacgttat ccctagttta aacacccata atacccataa tagctgtttg
     2581 ccaattaccc tgttatccct attacttcta aatctgagga ggaaaatcga atcgctacct
     2641 tagcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg
     2701 ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat taggtgttac
     2761 atgctgttca tctgttacat tgtcgatctg ttcatggtga acaggcgctc ggtcgttcgg
     2821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg
     3721 cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt
     3781 ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca
     3841 gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt
     3901 ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt
     3961 gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg
     4021 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     4081 gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac
     4141 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     4201 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     4261 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     4321 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     4381 cgccatgagt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca
     4441 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     4501 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     4561 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     4621 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     4681 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     4741 gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct
     4801 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     4861 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4921 ctgcgtgcaa tccatcttgt tcaatcatac tcttcctttt tcaatattat tgaagcattt
     4981 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5041 taggggttcc gcgcacatct aagaaaccat tattatcatg acattaacct ataaaaatag
     5101 gcgtatcacg aggccctttc gtcttcactt atccctatta cttctaaatc tgaggaggaa
     5161 aatcgaatcg ctaccttagg accgttatag ttactatcac catatatgta actggtgatt
     5221 ctttcgctac cttagagtgc accatagatc aacgacatta ctatatatat aatataggaa
     5281 gcatttaata gaacagcatc gtaatatatg tgtactttgc agttatgacg ccagatggca
     5341 gtagtggaag atattcttta ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg
     5401 gagcttttct ttttttgccg attaagaatt aattcggtcg aaaaaagaaa aggagagggc
     5461 caagagggag ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag
     5521 gcagcttgga gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt
     5581 tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg
     5641 ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt
     5701 tcaagtcttg taaaagcata taaaaatagt tcaggcactc cgaaatactt ggttggcgtg
     5761 tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc
     5821 gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt
     5881 attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa
     5941 cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat
     6001 tggaactcga tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg
     6061 ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt
     6121 ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca
     6181 aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt
     6241 ttgtgcactt gccgatctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     6301 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     6361 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     6421 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     6481 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6541 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6601 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6661 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6721 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     6781 gctgcgcaac tgttgggaag ggcgat
//
