LOCUS Exported 3846 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Position 6 module- URA3. ACCESSION . VERSION . KEYWORDS pMV10 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3846) AUTHORS Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G TITLE COSPLAY: An expandable toolbox for combinatorial and swift generation of expression plasmids in yeast. JOURNAL PLoS One. 2019 Aug 28;14(8):e0220694. doi: 10.1371/journal.pone.0220694. eCollection 2019. PUBMED 31461461 REFERENCE 2 (bases 1 to 3846) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3846 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 35..57 /label=M13/pUC Forward /note="In lacZ gene" promoter 139..354 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 355..1158 /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(1296..1312) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1296..1312) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1309..1331) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1320..1336 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1344..1374) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1389..1410 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1527..1544) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1698..2286) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1778..1797) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2457..3317) /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 3080..3099 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3318..3422) /gene="bla" /label=AmpR promoter primer_bind 3490..3508 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3546..3568) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3668..3687 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg 61 accagtgaat tcgagctcgg tacctcgcga atgcatctag atatcggatc ccggtctcga 121 gcaaccacag cttttcaatt caattcatca tttttttttt attctttttt ttgatttcgg 181 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag 241 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca 301 gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg 361 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt 421 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag 481 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg 541 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc 601 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa 661 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac 721 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag 781 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga 841 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt 901 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat catgacaccc 961 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat 1021 gtggtgtcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg 1081 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga 1141 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa 1201 attagagctt caatttaatt atatcagtta ttaccctcca tagagaccgg gcccgtcgac 1261 tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 1321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 1381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 1441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 1501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 1561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 1621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 1681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 1741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 1801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 1861 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 1921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 1981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 2041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 2101 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 2161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 2221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 2281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 2341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 2401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 2461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 2521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 2581 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag 2641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 2701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 2761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 2821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 2881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 2941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 3001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 3061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 3121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 3181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 3241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 3301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 3361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 3421 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta 3481 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt 3541 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc 3601 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt 3661 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg 3721 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac 3781 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga 3841 tgtgct //