LOCUS pRS415 6021 bp DNA circular 05-DEC-2007 DEFINITION Cloning vector pRS415, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pRS415 ORGANISM Cloning vector pRS415 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6021) AUTHORS Sikorski,R.S. and Hieter,P. TITLE A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae JOURNAL Genetics 122 (1), 19-27 (1989) PUBMED 2659436 REFERENCE 2 (bases 1 to 6021) AUTHORS Christianson,T.W., Sikorski,R.S., Dante,M., Shero,J.H. and Hieter,P. TITLE Multifunctional yeast high-copy-number shuttle vectors JOURNAL Gene 110 (1), 119-122 (1992) PUBMED 1544568 REFERENCE 3 (bases 1 to 6021) AUTHORS New England Biolabs. TITLE Direct Submission JOURNAL Submitted (05-DEC-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA COMMENT See also GenBank accession U03449. FEATURES Location/Qualifiers source 1..6021 /organism="Cloning vector pRS415" /mol_type="other DNA" gene complement(663..1769) /gene="LEU2" CDS complement(663..1769) /gene="LEU2" /codon_start=1 /product="beta-isopropylmalate dehydrogenase" /translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFD FENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRK ELQLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVA WDSEQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEF PTLKVQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLA SLPDKNTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVK KVLDAGIRTGDLGGSNSTTEVGDAVAEEVKKILA" rep_origin complement(2474..2929) /note="f1 origin of replication (+ -)" gene complement(2712..3290) /gene="lacZalpha" CDS complement(2712..3290) /gene="lacZalpha" /codon_start=1 /product="beta-galactosidase alpha fragment" /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD" misc_feature 3127..3234 /gene="lacZalpha" /note="multiple cloning site (KpnI-SacI)" rep_origin complement(3688..4275) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point); rop gene has been disrupted" gene complement(4447..5307) /gene="bla" CDS complement(4447..5307) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide complement(5239..5307) /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" misc_structure 5448..5566 /note="CEN6 (yeast chromosome VI centromere sequence)" rep_origin 5579..5952 /note="ARS209; histone H4 autonomously replicating sequence (ARS)" BASE COUNT 1663 a 1336 c 1339 g 1683 t ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg 2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt 2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg 2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta 2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa 2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt 2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg 2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 3001 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc 3061 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc 3121 gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc 3181 ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt 3241 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc 3301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg 3361 taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc 3421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 3481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 5401 aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat 5461 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa 5521 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga 5581 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt 5641 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt 5701 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta 5761 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta 5821 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa 5881 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac 5941 aggcaagcga tccgtcctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 6001 gtatcacgag gccctttcgt c //