LOCUS Exported 8218 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A dual reporter vector modified from Sharon et al (Nature Biotech, 2012) containing Ura3 and Nat1 selectable markers, a constant background RFP (TEF2::mCherry), and a variable YFP (pTpA+MCS::yEVenus).. ACCESSION . VERSION . KEYWORDS yeast_DualReporter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8218) AUTHORS Carl G. de Boer, Eeshit Dhaval Vaishnav, Ronen Sadeh, Esteban Luis Abeyta, Nir Friedman, Aviv Regev TITLE Deciphering eukaryotic cis-regulatory logic with 100 million random promoters JOURNAL BioRxiv REFERENCE 2 (bases 1 to 8218) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8218 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(103..963) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 726..745 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(964..1068) /gene="bla" /label=AmpR promoter terminator 1133..1320 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" CDS complement(1347..2057) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1625..1644) /label=mCherry-F /note="mCherry, forward primer" primer_bind 1894..1912 /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(2674..2696) /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 2896..2914 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2896..2913 /label=SP6 /note="SP6 promoter, forward primer" promoter 2988..3203 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 3204..4007 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" primer_bind complement(4275..4296) /label=F1ori-F /note="F1 origin, forward primer" CDS 4541..5257 /codon_start=1 /product="yeast-enhanced green fluorescent protein" /label=yeGFP /note="codons optimized for translation in S. cerevisiae and C. albicans" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 5278..5465 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" gene 5540..6659 /label=natMX6 /note="yeast selectable marker conferring nourseothricin resistance" promoter 5540..5883 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 5884..6456 /codon_start=1 /gene="Streptomyces noursei nat1" /product="nourseothricin acetyltransferase" /label=NrsR /note="confers resistance to nourseothricin" /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT LCGLDTALYDGTASDGEQALYMSMPCP" terminator 6462..6659 /label=TEF terminator /note="Ashbya gossypii TEF terminator" misc_feature 6726..7229 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind complement(7285..7304) /label=T7 /note="T7 promoter, forward primer" promoter complement(7286..7304) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7391..7408) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7562..8150) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7642..7661) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 61 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 241 gcgagaccca cgttcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 1021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 1081 aaaagtgcca cctgacgtca tctatattac cctgttatcc ctagcggatc tgccggtaga 1141 ggtgtggtca ataagagcga cctcatacta tacctgagaa agcaacctga cctacaggaa 1201 agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt aaaatttgta 1261 tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata agaaattcgc 1321 ttatttagaa gtggcgcgcc ggtccgttac ttgtacagct cgtccatgcc gccggtggag 1381 tggcggccct cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg 1441 atgtccaact tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg 1501 taggtggtct tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg 1561 atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc 1621 atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc 1681 ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg 1741 tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag 1801 tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac 1861 aggatgtccc aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg 1921 gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg 1981 gagccctcca tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc 2041 tcgcccttgc tcaccatggt actagtgttt agttaattat agttcgttga ccgtatattc 2101 taaaaacaag tactccttaa aaaaaaacct tgaagggaat aaacaagtag aatagataga 2161 gagaaaaata gaaaatgcaa gagaatttat atattagaaa gagagaaaga aaaatggaaa 2221 aaaaaaaata ggaaaagcca gaaatagcac tagaaggagc gacaccagaa aagaaggtga 2281 tggaaccaat ttagctatat atagttaact accggctcga tcatctctgc ctccagcata 2341 gtcgaagaag aatttttttt ttcttgaggc ttctgtcagc aactcgtatt ttttctttct 2401 tttttggtga gcctaaaaag ttcccacgtt ctcttgtacg acgccgtcac aaacaacctt 2461 atgggtaatt tgtcgcggtc tgggtgtata aatgtgtggg tgcaacatga atgtacggag 2521 gtagtttgct gattggcggt ctatagatac cttggttatg gcgccctcac agccggcagg 2581 ggaagcgcct acgcttgaca tctactatat gtaagtatac ggccccatat ataggccctt 2641 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 2701 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 2761 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 2821 agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 2881 tatcatacac atacgattta ggtgacacta tagaacgcgg ccgccagctg aagctttaac 2941 tatgcggcat cagagcagat tgtactgaga gtgcaccata ccaccttttc aattcatcat 3001 ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct 3061 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat 3121 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa 3181 cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc 3241 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt 3301 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc 3361 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca 3421 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa 3481 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag 3541 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt 3601 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat 3661 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga 3721 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg 3781 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat 3841 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg 3901 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa 3961 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat 4021 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat 4081 taccctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 4141 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 4201 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 4261 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 4321 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 4381 caagtgctag caggaatgat gcaaaaggtt cccgattcga actgcatttt tttcacatct 4441 cgaggttacg gctgtttctt aattaaaaaa agatagaaaa cattaggagt gtaacacaag 4501 actttcggat cctgagcagg caagataaac gaaggcaaag atgtctaaag gtgaagaatt 4561 attcactggt gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt 4621 ttctgtctcc ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaattgat 4681 ttgtactact ggtaaattgc cagttccatg gccaacctta gtcactactt taggttatgg 4741 tttgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt tcaagtctgc 4801 catgccagaa ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa 4861 gaccagagct gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg 4921 tattgatttt aaagaagatg gtaacatttt aggtcacaaa ttggaataca actataactc 4981 tcacaatgtt tacatcactg ctgacaaaca aaagaatggt atcaaagcta acttcaaaat 5041 tagacacaac attgaagatg gtggtgttca attagctgac cattatcaac aaaatactcc 5101 aattggtgat ggtccagtct tgttaccaga caaccattac ttatcctatc aatctgcctt 5161 atccaaagat ccaaacgaaa agagagacca catggtcttg ttagaatttg ttactgctgc 5221 tggtattacc catggtatgg atgaattgta caaataaggc gcgccacttc taaataagcg 5281 aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac 5341 aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc 5401 ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct 5461 accggcagat ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc 5521 gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt 5581 gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt 5641 ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc 5701 ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca 5761 cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca 5821 tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca 5881 accatgggta ccactcttga cgacacggct taccggtacc gcaccagtgt cccgggggac 5941 gccgaggcca tcgaggcact ggatgggtcc ttcaccaccg acaccgtctt ccgcgtcacc 6001 gccaccgggg acggcttcac cctgcgggag gtgccggtgg acccgcccct gaccaaggtg 6061 ttccccgacg acgaatcgga cgacgaatcg gacgacgggg aggacggcga cccggactcc 6121 cggacgttcg tcgcgtacgg ggacgacggc gacctggcgg gcttcgtggt cgtctcgtac 6181 tccggctgga accgccggct gaccgtcgag gacatcgagg tcgccccgga gcaccggggg 6241 cacggggtcg ggcgcgcgtt gatggggctc gcgacggagt tcgcccgcga gcggggcgcc 6301 gggcacctct ggctggaggt caccaacgtc aacgcaccgg cgatccacgc gtaccggcgg 6361 atggggttca ccctctgcgg cctggacacc gccctgtacg acggcaccgc ctcggacggc 6421 gagcaggcgc tctacatgag catgccctgc ccctaatcag tactgacaat aaaaagattc 6481 ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat 6541 caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt 6601 taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc 6661 tgtcgattcg atactaacgc cgccatccag tgtcgaaaac gagctcgaat tcctgggtcc 6721 ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa atatataaat 6781 taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt 6841 aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca 6901 ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt 6961 gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc 7021 taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt 7081 tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa 7141 aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg 7201 aggcgcgtgt aagttacagg caagcgatcc gtccgatatc atcagatcca ctagtggcct 7261 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg 7321 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 8041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggatttt //