LOCUS       40924_49547        5181 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast/E. coli shuttle vector with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5181)
  AUTHORS   Hill JE, Myers AM, Koerner TJ, Tzagoloff A.
  TITLE     Yeast/E. coli shuttle vectors with multiple unique restriction 
            sites.
  JOURNAL   Yeast 1986;2:163-7.
  PUBMED    3333305
REFERENCE   2  (bases 1 to 5181)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5181)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "1986"; volume: "2"; pages: "163-7"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    15..71
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(75..91)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(367..1167)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1168..1378)
                     /label=URA3 promoter
     rep_origin      complement(1614..2778)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3060..3164
                     /label=AmpR promoter
     CDS             3165..4022
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4196..4784
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5072..5093
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5108..5138
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5146..5162
                     /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)."
ORIGIN
        1 atgaccatga ttacgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg
       61 catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      121 acccaactta atcgccttgc agcacatccc cccttcgcca gctggcgtaa tagcgaagag
      181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
      241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata
      301 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
      361 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
      421 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
      481 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
      541 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
      601 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
      661 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
      721 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
      781 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
      841 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
      901 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
      961 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     1021 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     1081 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     1141 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcggttttt gttctgtgca
     1201 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     1261 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     1321 aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     1381 gctcttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag
     1441 atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat
     1501 gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat
     1561 ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag
     1621 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     1681 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     1741 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     1801 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     1861 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     1921 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     1981 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     2041 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     2101 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     2161 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     2221 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     2281 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     2341 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     2401 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     2461 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     2521 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     2581 ttcctatact ttctagctag agaataggaa cttcggaata ggaacttcaa agcgtttccg
     2641 aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc
     2701 gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt
     2761 gtttatgctt aaatgcgtta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2821 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2881 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2941 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3001 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3061 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3181 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3241 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3301 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3361 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3421 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3481 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3541 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3601 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3661 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3721 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3781 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3841 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3901 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3961 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4021 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4081 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4141 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4201 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4261 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4321 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4381 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4441 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4501 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4561 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     4621 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4681 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4741 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4801 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4861 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4921 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4981 aaccgcctct ccccgcgcgt tggccgattc attaatccag ctggcacgac aggtttcccg
     5041 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac
     5101 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5161 aatttcacac aggaaacagc t
//
