LOCUS       sequence-257390-        7498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 7498)
  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 7498)
  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
FEATURES             Location/Qualifiers
     source          1..7498
                     /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..1441
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1466..1471,1472..1489,1490..1496)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1550..2209
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1616..1635)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2551..2856
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2735..2754
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2897..3021)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3060..3107
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3189..3213)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      complement(3650..4238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3730..3749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4567..4614
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4639..4658
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4846..5640)
                     /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(4846..5640)
                     /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(4957..4976)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5567..5586
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5641..5745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5788..5806
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5943..6224
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6225..6899
                     /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(7002..7457)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7139..7160)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7351..7370
                     /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 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     1381 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     1441 ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
     1501 gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1561 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1621 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1681 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1741 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
     1801 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1861 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1921 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1981 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2041 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2101 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2161 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc
     2221 cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag
     2281 aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt
     2341 attacagtga cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata
     2401 tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct
     2461 ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct
     2521 cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga
     2581 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
     2641 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     2701 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     2761 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     2821 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc
     2881 cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg
     2941 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     3001 agctttcttg tacaaagtgg taagtaacta agatccgtcg acctgcagcg gccgcataac
     3061 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcgc gcttgcagcc
     3121 aagctaattc cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     3181 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgtt atccctagtt
     3241 taaacaccca taatacccat aatagctgtt tgccaattac cctgttatcc ctattacttc
     3301 taaatctgag gaggaaaatc gaatcgctac cttagcgcct gtttcaggct gtctatgtgt
     3361 gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt
     3421 ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc
     3481 tgttcatggt gaacaggcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4381 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     4441 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     4501 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     4561 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     4621 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     4681 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     4741 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     4801 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     4861 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     4921 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     4981 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     5041 attttccacc atgatattcg gcaagcaggc atcgccatga gtcacgacga gatcctcgcc
     5101 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     5161 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     5221 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     5281 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     5341 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     5401 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     5461 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     5521 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     5581 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     5641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ctaagaaacc
     5761 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
     5821 ttatccctat tacttctaaa tctgaggagg aaaatcgaat cgctacctta ggaccgttat
     5881 agttactatc accatatatg taactggtga ttctttcgct accttagagt gcaccataga
     5941 tcaacgacat tactatatat ataatatagg aagcatttaa tagaacagca tcgtaatata
     6001 tgtgtacttt gcagttatga cgccagatgg cagtagtgga agatattctt tattgaaaaa
     6061 tagcttgtca ccttacgtac aatcttgatc cggagctttt ctttttttgc cgattaagaa
     6121 ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg agggcattgg tgactattga
     6181 gcacgtgagt atacgtgatt aagcacacaa aggcagcttg gagtatgtct gttattaatt
     6241 tcacaggtag ttctggtcca ttggtgaaag tttgcggctt gcagagcaca gaggccgcag
     6301 aatgtgctct agattccgat gctgacttgc tgggtattat atgtgtgccc aatagaaaga
     6361 gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata
     6421 gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt
     6481 tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat gagtcgtggc
     6541 aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact
     6601 gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt
     6661 cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa
     6721 ggcaagagag ccccgaaagc ttacatttta tgttagctgg tggactgacg ccagaaaatg
     6781 ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga
     6841 caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg
     6901 ttattactga gtagtattta tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg
     6961 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     7021 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     7081 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     7141 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     7201 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     7261 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     7321 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     7381 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     7441 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgat
//
