LOCUS Exported 6341 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Negative control. Expression of Gal4BD-Lam hybrid protein. Homology regions for recombination with pAWH. ACCESSION . VERSION . KEYWORDS pBKWH-Lam SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6341) 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 6341) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6341 /organism="synthetic DNA construct" /mol_type="other DNA" 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..4483) /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(3800..3819) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4410..4429 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(4484..4588) /gene="bla" /label=AmpR promoter primer_bind 4631..4649 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4786..5067 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 5068..5742 /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(5845..6300) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5982..6003) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6194..6213 /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 agtcccgctc agaagaactc gtcaagaagg cgatagaagg 3721 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt 3781 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg 3841 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat 3901 tcggcaagca ggcatcgcca tgagtcacga cgagatcctc gccgtcgggc atgctcgcct 3961 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct 4021 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt 4081 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga 4141 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc 4201 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa 4261 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac 4321 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg 4381 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc 4441 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catactcttc ctttttcaat 4501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 4561 agaaaaataa acaaataggg gttccgcgca catctaagaa accattatta tcatgacatt 4621 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacttatccc tattacttct 4681 aaatctgagg aggaaaatcg aatcgctacc ttaggaccgt tatagttact atcaccatat 4741 atgtaactgg tgattctttc gctaccttag agtgcaccat agatcaacga cattactata 4801 tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta 4861 tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg 4921 tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa 4981 agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg 5041 attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt 5101 ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc 5161 gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt 5221 attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa 5281 tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat 5341 tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc 5401 ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt 5461 gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca 5521 ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa 5581 agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga 5641 ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac 5701 tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat 5761 ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc 5821 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt 5881 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt 5941 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc 6001 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 6061 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac 6121 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg 6181 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag 6241 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt 6301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga t //