LOCUS 40924_49737 4303 bp DNA circular SYN 01-JAN-1980 DEFINITION Yeast/E. coli integrating shuttle vector with a URA3 marker. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4303) AUTHORS Hill JE, Myers AM, Koerner TJ, Tzagoloff A. TITLE Yeast/E. coli shuttle vectors with multiple unique restriction sites. JOURNAL Yeast 1986;2:163-7. PUBMED 3333305 REFERENCE 2 (bases 1 to 4303) TITLE Direct Submission REFERENCE 3 (bases 1 to 4303) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; date: "1986"; volume: "2"; pages: "163-7" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..4303 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 15..71 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(75..91) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(367..1167) /codon_start=1 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(1168..1378) /label=URA3 promoter promoter 2182..2286 /label=AmpR promoter CDS 2287..3144 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3318..3906 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 4194..4215 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 4230..4260 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4268..4284 /label=lac operator /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)." ORIGIN 1 atgaccatga ttacgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg 61 catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt 121 acccaactta atcgccttgc agcacatccc cccttcgcca gctggcgtaa tagcgaagag 181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg 241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata 301 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt 361 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg 421 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat 481 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc 541 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc 601 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc 661 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa 721 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac 781 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta 841 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc 901 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat 961 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc 1021 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt 1081 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc 1141 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcggttttt gttctgtgca 1201 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata 1261 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa 1321 aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa 1381 gctcttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag 1441 atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat 1501 gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat 1561 ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttattttag 1621 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt 1681 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa 1741 taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca 1801 catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca 1861 tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt tatggtgcac tctcagtaca 1921 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 1981 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 2041 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 2101 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 2161 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 2221 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 2281 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 2341 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 2401 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 2461 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 2521 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 2581 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 2641 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 2701 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 2761 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 2821 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 2881 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 2941 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 3001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 3061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 3121 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 3181 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 3241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 3301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 3361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 3421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 3481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 3541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 3601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 3661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc 3721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 3781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 3841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 3901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 3961 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4021 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4081 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatcc 4141 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 4201 agttacctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 4261 tgtggaattg tgagcggata acaatttcac acaggaaaca gct //