LOCUS       40924_49662        5241 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal plasmid with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5241)
  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 5241)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5241)
  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..5241
                     /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"
     rep_origin      complement(743..2089)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2203..3003)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3004..3219)
                     /label=URA3 promoter
     promoter        3335..3439
                     /label=AmpR promoter
     CDS             3440..4297
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4471..5059
                     /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 tatcggatcg tacttgttac ccatcattga attttgaaca
      541 tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat
      601 agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc
      661 tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc
      721 acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg
      781 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
      841 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
      901 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
      961 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     1021 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     1081 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     1141 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     1201 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     1261 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     1321 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     1381 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     1441 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     1501 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     1561 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     1621 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag
     1681 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     1741 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
     1801 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
     1861 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
     1921 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
     1981 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
     2041 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat
     2101 aagctgtcaa acatgagaat tgggtaataa ctgatataat taaattgaag ctctaatttg
     2161 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     2221 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     2281 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     2341 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     2401 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     2461 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     2521 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     2581 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     2641 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     2701 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     2761 ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac
     2821 atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt
     2881 ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa
     2941 tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga
     3001 catgatttat cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta agaatactgg
     3061 gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc
     3121 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg
     3181 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagcta attcttgaag
     3241 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3301 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3361 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3421 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3481 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3541 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3601 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3661 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3721 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3781 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3841 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3901 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3961 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4021 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4081 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4141 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4201 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4261 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4321 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4381 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4441 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4501 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4561 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4621 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4681 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4741 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4801 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4861 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4921 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4981 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5041 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5101 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5161 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5221 agtgagcgag gaagcggaag a
//
