LOCUS       40924_49752        4293 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast integrating plasmid with a LEU2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  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 4293)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4293)
  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..4293
                     /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"
     CDS             complement(800..1891)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1892..2299)
                     /label=LEU2 promoter
     promoter        2387..2491
                     /label=AmpR promoter
     CDS             2492..3349
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3523..4111
                     /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 tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      541 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      601 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      661 caccgtcatc accgaaacgc gcgagacgaa agggcctacc ctatgaacat attccatttt
      721 gtaatttcgt gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata
      781 aaaaaagaga atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga
      841 cttccgtggt actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct
      901 ttttaactgc atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca
      961 tcattgcagc agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag
     1021 cagaaccgtg gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca
     1081 aggacgcaga tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga
     1141 tatcaccaaa catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta
     1201 ggatcatggc ggcagaatca atcaattgat gttgaacctt caatgtagga aattcgttct
     1261 tgatggtttc ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat
     1321 ccaaggacca aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg
     1381 tgattctttg cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat
     1441 cgtcttcctt tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt
     1501 cagtaccttt agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa
     1561 agttacatgg tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt
     1621 gttcaggtct aacactacct gtaccccatt taggaccacc cacagcacct aacaaaacgg
     1681 catcaacctt cttggaggct tccagcgcct catctggaag tgggacacct gtagcatcga
     1741 tagcagcacc accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag
     1801 aaatagcttt aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg
     1861 gcaaaacgac gatcttctta ggggcagaca ttagaatggt atatccttga aatatatata
     1921 tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac
     1981 ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa
     2041 gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccgcg gctctcacct
     2101 ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat
     2161 taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc
     2221 gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata
     2281 cctgagtatt cccacagttg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2341 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2401 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2461 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2521 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2581 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2641 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2701 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2761 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2821 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2881 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2941 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3001 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3061 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3121 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3181 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3241 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3301 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3361 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3421 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3481 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3541 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3601 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3661 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3721 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3781 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3841 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3901 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3961 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4021 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4081 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4141 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4201 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4261 gcgcagcgag tcagtgagcg aggaagcgga aga
//
