LOCUS 40924_49482 4854 bp DNA circular SYN 01-JAN-1980 DEFINITION Yeast centromeric plasmid with a TRP1 marker. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4854) AUTHORS Gietz RD, Sugino A. TITLE New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. JOURNAL Gene 1988;74:527-34. PUBMED 3073106 REFERENCE 2 (bases 1 to 4854) TITLE Direct Submission REFERENCE 3 (bases 1 to 4854) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; date: "1988"; volume: "74"; pages: "527-34" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT The sequence of this plasmid was reconstructed using the description from Gietz and Sugino. FEATURES Location/Qualifiers source 1..4854 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 106..127 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /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)." primer_bind 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature complement(233..289) /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(290..306) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature complement(1064..2227) /label=CEN/ARS /note="S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416" promoter complement(2741..2842) /label=TRP1 promoter promoter 2948..3052 /label=AmpR promoter CDS 3053..3910 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4084..4672 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg 301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt 481 gcggtatttc acaccgcata tatcgctggg ccattctcat gaagaatatc ttgaatttat 541 tgtcatatta ctagttggtg tggaagtcct aatatcggtg atcaatatag tggttgacat 601 gctggctagt caacattgag ccttttgatc atgcaaatat attacggtat tttacaatca 661 aatatcaaac ttaactattg actttataac ttatttaggt ggtaacattc ttataaaaaa 721 gaaaaaaatt actgcaaaac agtactagct tttaacttgt atcctaggtt atctatgctg 781 tctcaccata gagaatatta cctatttcag aatgtatgtc catgattcgc cgggtaaata 841 catataatac acaaatctgg cttaataaag tctataatat atctcataaa gaagtgctaa 901 attggctagt gctatatatt tttaagaaaa tttcttttga ctaagtccat atcgactttg 961 taaaagttca cattagcata catatattac acgagccaga aatagtaact tttgcctaaa 1021 tcacaaattg caaaatttaa ttgcttgcaa aaggtcacat gcttataatc aactttttta 1081 aaaatttaaa atactttttt attttttatt tttaaacata aatgaaataa tttatttatt 1141 gtttatgatt accgaaacat aaaacctgct caagaaaaag aaactgtttt gtccttggaa 1201 aaaaagcact acctaggagc ggccaaaatg ccgaggcttt catagcttaa actctttaca 1261 gaaaataggc attatagatc agttcgagtt ttcttattct tccttccggt tttatcgtca 1321 cagttttaca gtaaataagt atcacctctt agagttcgat gataagctgt caaacatgag 1381 aattaattcc cacatgttaa aatagtgaag gagcatgttc ggcacacagt ggaccgaacg 1441 tggggtaagt gcactagggt ccggttaaac ggatctcgca ttgatgaggc aacgctaatt 1501 atcaacatat agattgttat ctatctgcat gaacacgaaa tctttacttg acgacttgag 1561 gctgatggtg tttatgcaaa gaaaccactg tgtttaatat gtgtcactgt ttgatattac 1621 tgtcagcgta gaagataata gtaaaagcgg ttaataagtg tatttgagat aagtgtgata 1681 aagtttttac agcgaaaaga cgataaatac aagaaaatga ttacgaggat acggagagag 1741 gtatgtacat gtgtatttat atactaagct gccggcggtt gtttgcaaga ccgagaaaag 1801 gctagcaaga atcgggtcat tgtagcgtat gcgcctgtga acattctctt caacaagttt 1861 gattccattg cggtgaaatg gtaaaagtca accccctgcg atgtatattt tcctgtacaa 1921 tcaatcaaaa agccaaatga tttagcatta tctttacatc ttgttatttt acagatttta 1981 tgtttagatc ttttatgctt gcttttcaaa aggcttgcag gcaagtgcac aaacaatact 2041 taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt 2101 tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc 2161 catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat 2221 gtaagctctc ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa 2281 gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg 2341 aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca 2401 aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa 2461 tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa 2521 ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc 2581 ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt 2641 cagcatcgga atctagtgca cattctgcgg cctctgtgct ctgcaagccg caaactttca 2701 ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt 2761 gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc 2821 cttttctttt ttcgaccgaa ttaattcttg aagacgaaag ggcctcgtga tacgcctatt 2881 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 2941 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 3001 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 3061 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 3121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 3181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 3241 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 3301 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 3361 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 3421 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 3481 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 3541 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 3601 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 3661 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 3721 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 3781 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 3841 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 3901 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 3961 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 4021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 4081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 4141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 4201 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 4261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 4321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 4381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 4441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 4501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 4561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 4621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 4681 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 4741 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 4801 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga //