LOCUS       Exported                6099 bp DNA     circular SYN 18-JUL-2025
DEFINITION  Yeast centromeric plasmid with a LEU2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  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 6099)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6099)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6099)
  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     SGRef: number: 3; type: "Journal Article"
COMMENT     The sequence of this plasmid was reconstructed using the description
            from Gietz and Sugino.
FEATURES             Location/Qualifiers
     source          1..6099
                     /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"
     misc_feature    complement(1063..2225)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     CDS             complement(2587..3681)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3682..4087)
                     /label=LEU2 promoter
     promoter        4193..4297
                     /label=AmpR promoter
     CDS             4298..5158
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5329..5917
                     /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 atcgctgggc cattctcatg aagaatatct tgaatttatt
      541 gtcatattac tagttggtgt ggaagtccta atatcggtga tcaatatagt ggttgacatg
      601 ctggctagtc aacattgagc cttttgatca tgcaaatata ttacggtatt ttacaatcaa
      661 atatcaaact taactattga ctttataact tatttaggtg gtaacattct tataaaaaag
      721 aaaaaaatta ctgcaaaaca gtactagctt ttaacttgta tcctaggtta tctatgctgt
      781 ctcaccatag agaatattac ctatttcaga atgtatgtcc atgattcgcc gggtaaatac
      841 atataataca caaatctggc ttaataaagt ctataatata tctcataaag aagtgctaaa
      901 ttggctagtg ctatatattt ttaagaaaat ttcttttgac taagtccata tcgactttgt
      961 aaaagttcac attagcatac atatattaca cgagccagaa atagtaactt ttgcctaaat
     1021 cacaaattgc aaaatttaat tgcttgcaaa aggtcacatg cttataatca acttttttaa
     1081 aaatttaaaa tactttttta ttttttattt ttaaacataa atgaaataat ttatttattg
     1141 tttatgatta ccgaaacata aaacctgctc aagaaaaaga aactgttttg tccttggaaa
     1201 aaaagcacta cctaggagcg gccaaaatgc cgaggctttc atagcttaaa ctctttacag
     1261 aaaataggca ttatagatca gttcgagttt tcttattctt ccttccggtt ttatcgtcac
     1321 agttttacag taaataagta tcacctctta gagttcgatg ataagctgtc aaacatgaga
     1381 attaattcca catgttaaaa tagtgaagga gcatgttcgg cacacagtgg accgaacgtg
     1441 gggtaagtgc actagggtcc ggttaaacgg atctcgcatt gatgaggcaa cgctaattat
     1501 caacatatag attgttatct atctgcatga acacgaaatc tttacttgac gacttgaggc
     1561 tgatggtgtt tatgcaaaga aaccactgtg tttaatatgt gtcactgttt gatattactg
     1621 tcagcgtaga agataatagt aaaagcggtt aataagtgta tttgagataa gtgtgataaa
     1681 gtttttacag cgaaaagacg ataaatacaa gaaaatgatt acgaggatac ggagagaggt
     1741 atgtacatgt gtatttatat actaagctgc cggcggttgt ttgcaagacc gagaaaaggc
     1801 tagcaagaat cgggtcattg tagcgtatgc gcctgtgaac attctcttca acaagtttga
     1861 ttccattgcg gtgaaatggt aaaagtcaac cccctgcgat gtatattttc ctgtacaatc
     1921 aatcaaaaag ccaaatgatt tagcattatc tttacatctt gttattttac agattttatg
     1981 tttagatctt ttatgcttgc ttttcaaaag gcttgcaggc aagtgcacaa acaatactta
     2041 aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg
     2101 ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca
     2161 tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt
     2221 aagctctcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt
     2281 tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa
     2341 gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa
     2401 ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac gatcgatgat
     2461 aagctgtcaa acatgagaat taattctacc ctatgaacat attccatttt gtaatttcgt
     2521 gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga
     2581 atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttcggtggt
     2641 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc
     2701 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc
     2761 agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag cagaaccgtg
     2821 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga
     2881 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa
     2941 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc
     3001 ggcagaatca atcaattgat gttgaacctt caatgtagga aattcgttct tgatggtttc
     3061 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca
     3121 aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg
     3181 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt
     3241 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt
     3301 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg
     3361 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct
     3421 aacactacct gtaccccatt taggaccacc cacagcacct aacaaaacgg catcagcctt
     3481 cttggaggct tccagcgcct catctggaag tggaacacct gtagcatcga tagcagcacc
     3541 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt
     3601 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac
     3661 gatcttctta ggggcagaca ttagaatggt atatccttga aatatatata tatattgctg
     3721 aaatgtaaaa ggtaagaaaa gttagaaagt aagacgattg ctaaccacct attggaaaaa
     3781 acaataggtc cttaaataat attgtcaact tcaagtattg tgatgcaagc atttagtcat
     3841 gaacgcttct ctattctata tgaaaagccg gttccggcgc tctcaccttt cctttttctc
     3901 ccaatttttc agttgaaaaa ggtatatgcg tcaggcgacc tctgaaatta acaaaaaatt
     3961 tccagtcatc gaatttgatt ctgtgcgata gcgcccctgt gtgttctcgt tatgttgagg
     4021 aaaaaaataa tggttgctaa gagattcgaa ctcttgcatc ttacgatacc tgagtattcc
     4081 cacagttaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4141 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4201 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4261 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4321 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4381 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4441 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4501 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4561 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4621 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4681 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4741 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4801 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4861 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4921 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4981 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5041 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5101 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5161 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5221 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5281 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5341 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5401 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5461 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5521 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5581 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5641 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5701 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5761 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5821 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5881 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5941 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6001 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6061 gaccgagcgc agcgagtcag tgagcgagga agcggaaga
//
