LOCUS       40924_49462        6103 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast centromeric plasmid with a LEU2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6103)
  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 6103)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6103)
  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..6103
                     /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"
     CDS             complement(2592..3683)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3684..4091)
                     /label=LEU2 promoter
     promoter        4197..4301
                     /label=AmpR promoter
     CDS             4302..5159
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5333..5921
                     /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 acgatcgatg
     2461 ataagctgtc aaacatgaga attaattcta ccctatgaac atattccatt ttgtaatttc
     2521 gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca taaaaaaaga
     2581 gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc gacttccgtg
     2641 gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac ctttttaact
     2701 gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa catcattgca
     2761 gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg agcagaaccg
     2821 tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc caaggacgca
     2881 gatggcaaca aacccaagga acctgggata acggaggctt catcggagat gatatcacca
     2941 aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac taggatcatg
     3001 gcggcagaat caatcaattg atgttgaacc ttcaatgtag gaaattcgtt cttgatggtt
     3061 tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt atccaaggac
     3121 caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct tgtgattctt
     3181 tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc atcgtcttcc
     3241 tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa gtcagtacct
     3301 ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc aaagttacat
     3361 ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc ttgttcaggt
     3421 ctaacactac ctgtacccca tttaggacca cccacagcac ctaacaaaac ggcatcaacc
     3481 ttcttggagg cttccagcgc ctcatctgga agtgggacac ctgtagcatc gatagcagca
     3541 ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc agaaatagct
     3601 ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc tggcaaaacg
     3661 acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata tatatatatt
     3721 gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc acctattgga
     3781 aaaaacaata ggtccttaaa taatattgtc aacttcaagt attgtgatgc aagcatttag
     3841 tcatgaacgc ttctctattc tatatgaaaa gccggttccg cggctctcac ctttcctttt
     3901 tctcccaatt tttcagttga aaaaggtata tgcgtcaggc gacctctgaa attaacaaaa
     3961 aatttccagt catcgaattt gattctgtgc gatagcgccc ctgtgtgttc tcgttatgtt
     4021 gaggaaaaaa ataatggttg ctaagagatt cgaactcttg catcttacga tacctgagta
     4081 ttcccacagt taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4141 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4201 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4261 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4321 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4381 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4441 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4501 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4561 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4621 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4681 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4741 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4801 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4861 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4921 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4981 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5041 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5101 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5161 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5221 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5281 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5341 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5401 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5461 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5521 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5581 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5641 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5701 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5761 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5821 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5881 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5941 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6001 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6061 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aga
//
