LOCUS       sequence-257387-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control when used in combination with pAWH-largeT. 
            Expression of Gal4BD-Bait hybrid protein. Homology regions for 
            recombination with pAWH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  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 6338)
  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: pBSWHhc-Lam
FEATURES             Location/Qualifiers
     source          1..6338
                     /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"
     protein_bind    complement(1840..1864)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1903..1950
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2032..2056)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      complement(2493..3081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2573..2592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3410..3457
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3482..3501
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3689..4480)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4481..4585)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4628..4646
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4783..5064
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5065..5739
                     /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(5842..6297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5979..6000)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6191..6210
                     /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 ttatctgaag aggtggtcag ccgcgaggtg tccggcatca
     1381 aggccgccta cgaggccgag ctcggggatg cccgcaagac ccttgactca gtagccaagg
     1441 agcgcgcccg cctgcagctg gagctgagca aagtgcgtga ggagtttaag gagctgaaag
     1501 cgcgcaatac caagaaggag ggtgacctga tagctgctca ggctcggctg aaggacctgg
     1561 aggctctgct gaactccaag gaggccgcac tgagcactgc tctcagtgag aagcgcacgc
     1621 tggagggcga gctgcatgat ctgcggggcc aggtggccaa gcttgaggca gccctaggtg
     1681 aggccaagaa gcaacttcag gatgagatgc tgcggcgggt ggatgctgag aacaggctgc
     1741 agaccatgaa ggaggaactg gacttccaga agaacatcta cagtgaggag ctgcgtgaga
     1801 ccaagcgccg tcatgagacc cgactggtgg agattgacaa cccagctttc ttgtacaaag
     1861 tggtaagtaa ctaagatccg tcgacctgca gcggccgcat aactagcata accccttggg
     1921 gcctctaaac gggtcttgag gggttttttg cgcgcttgca gccaagctaa ttccgggcga
     1981 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     2041 aattttaaag tgactcttag gttttaaaac gttatcccta gtttaaacac ccataatacc
     2101 cataatagct gtttgccaat taccctgtta tccctattac ttctaaatct gaggaggaaa
     2161 atcgaatcgc taccttagcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac
     2221 aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt
     2281 tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagg
     2341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     3241 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     3301 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     3361 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     3421 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     3481 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     3541 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     3601 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     3661 tcggtcattt cgaaccccag agtcccgctt atttgccgac taccttggtg atctcgcctt
     3721 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     3781 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     3841 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc
     3901 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt
     3961 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa
     4021 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca
     4081 agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc
     4141 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt
     4201 cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg
     4261 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca
     4321 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt
     4381 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg
     4441 atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt
     4501 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4561 aaaataaaca aataggggtt ccgcgcacat ctaagaaacc attattatca tgacattaac
     4621 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ttatccctat tacttctaaa
     4681 tctgaggagg aaaatcgaat cgctacctta ggaccgttat agttactatc accatatatg
     4741 taactggtga ttctttcgct accttagagt gcaccataga tcaacgacat tactatatat
     4801 ataatatagg aagcatttaa tagaacagca tcgtaatata tgtgtacttt gcagttatga
     4861 cgccagatgg cagtagtgga agatattctt tattgaaaaa tagcttgtca ccttacgtac
     4921 aatcttgatc cggagctttt ctttttttgc cgattaagaa ttaattcggt cgaaaaaaga
     4981 aaaggagagg gccaagaggg agggcattgg tgactattga gcacgtgagt atacgtgatt
     5041 aagcacacaa aggcagcttg gagtatgtct gttattaatt tcacaggtag ttctggtcca
     5101 ttggtgaaag tttgcggctt gcagagcaca gaggccgcag aatgtgctct agattccgat
     5161 gctgacttgc tgggtattat atgtgtgccc aatagaaaga gaacaattga cccggttatt
     5221 gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata gttcaggcac tccgaaatac
     5281 ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt tggctctggt caatgattac
     5341 ggcattgata tcgtccaact gcatggagat gagtcgtggc aagaatacca agagttcctc
     5401 ggtttgccag ttattaaaag actcgtattt ccaaaagact gcaacatact actcagtgca
     5461 gcttcacaga aacctcattc gtttattccc ttgtttgatt cagaagcagg tgggacaggt
     5521 gaacttttgg attggaactc gatttctgac tgggttggaa ggcaagagag ccccgaaagc
     5581 ttacatttta tgttagctgg tggactgacg ccagaaaatg ttggtgatgc gcttagatta
     5641 aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt aaaagactct
     5701 aacaaaatag caaatttcgt caaaaatgct aagaaatagg ttattactga gtagtattta
     5761 tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg aaataccgca cagatgcgta
     5821 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5881 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5941 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6001 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6061 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6121 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6181 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     6241 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     6301 ttcgccattc aggctgcgca actgttggga agggcgat
//
