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
//
