LOCUS       40924_49487        5603 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast centromeric plasmid with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5603)
  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 5603)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5603)
  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..5603
                     /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(2565..3365)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3366..3581)
                     /label=URA3 promoter
     promoter        3697..3801
                     /label=AmpR promoter
     CDS             3802..4659
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4833..5421
                     /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 attgggtaat aactgatata attaaattga agctctaatt
     2521 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     2581 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     2641 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     2701 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     2761 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     2821 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     2881 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     2941 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     3001 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     3061 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     3121 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     3181 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     3241 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     3301 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     3361 gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt taagaatact
     3421 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     3481 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac
     3541 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga attgaaaagc taattcttga
     3601 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3661 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3721 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3781 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3841 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3901 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3961 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4021 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4081 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4141 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4201 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4261 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4321 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4381 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4441 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4501 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4561 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4621 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4681 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4741 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4801 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4861 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4921 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4981 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5041 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5101 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5161 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5221 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5281 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5341 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5401 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5461 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5521 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5581 tcagtgagcg aggaagcgga aga
//
