LOCUS       40924_49657        5741 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal plasmid with a LEU2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5741)
  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 5741)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5741)
  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..5741
                     /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(2230..3321)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3322..3729)
                     /label=LEU2 promoter
     promoter        3835..3939
                     /label=AmpR promoter
     CDS             3940..4797
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4971..5559
                     /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 taattctacc ctatgaacat attccatttt gtaatttcgt
     2161 gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga
     2221 atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttccgtggt
     2281 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc
     2341 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc
     2401 agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag cagaaccgtg
     2461 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga
     2521 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa
     2581 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc
     2641 ggcagaatca atcaattgat gttgaacctt caatgtagga aattcgttct tgatggtttc
     2701 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca
     2761 aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg
     2821 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt
     2881 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt
     2941 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg
     3001 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct
     3061 aacactacct gtaccccatt taggaccacc cacagcacct aacaaaacgg catcaacctt
     3121 cttggaggct tccagcgcct catctggaag tgggacacct gtagcatcga tagcagcacc
     3181 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt
     3241 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac
     3301 gatcttctta ggggcagaca ttagaatggt atatccttga aatatatata tatatattgc
     3361 tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa
     3421 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc
     3481 atgaacgctt ctctattcta tatgaaaagc cggttccgcg gctctcacct ttcctttttc
     3541 tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa
     3601 tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga
     3661 ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata cctgagtatt
     3721 cccacagtta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5641 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5701 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a
//
