LOCUS       40924_49737        4303 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast/E. coli integrating shuttle vector with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4303)
  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 4303)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4303)
  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..4303
                     /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
     promoter        2182..2286
                     /label=AmpR promoter
     CDS             2287..3144
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3318..3906
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4194..4215
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4230..4260
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4268..4284
                     /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 gttattttag
     1621 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt
     1681 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa
     1741 taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca
     1801 catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca
     1861 tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt tatggtgcac tctcagtaca
     1921 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     1981 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2041 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2101 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2161 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2221 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2281 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2341 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2401 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2461 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2521 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2581 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2641 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2701 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2761 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2821 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2881 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2941 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3121 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3181 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     3721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3961 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4021 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4081 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatcc
     4141 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4201 agttacctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4261 tgtggaattg tgagcggata acaatttcac acaggaaaca gct
//
