LOCUS       40924_49542        5644 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast/E. coli shuttle vector with a LEU2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5644)
  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 5644)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5644)
  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..5644
                     /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(762..1853)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MFVPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWTLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEALRYPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPGFAKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1854..2258)
                     /label=LEU2 promoter
     rep_origin      complement(2259..3241)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3523..3627
                     /label=AmpR promoter
     CDS             3628..4485
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4659..5247
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5535..5556
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5571..5601
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5609..5625
                     /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 cgtagcctga atggcgaatg gcgcctgatg
      241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagatcc ggtacgtcgt
      301 taaggccgtt tctgacagag taaaattctt gagggaactt tcaccattat ggaaaatggt
      361 tcaagaaggt attgacttaa actccatcaa atggtcaggt catttgagtg ttttttattt
      421 gttgtatttt ttttttttta gagaaaatcc tccaatatca aattagggat tcgtagtttc
      481 atgattttct gttataccta actttttgtg tggtgccctc ctccttgtca atattaatgt
      541 taaagtgcaa ttctctttcc ttatcacgtt gagccattag tatcaatttg cttacctgta
      601 ttcctttagc tatcctcctt tttctccttc ttgataaatg tatgtagatt gcgtttgtct
      661 accctatgaa catattccat tttgtaattt cgtgtcgttt ctattatgaa tttcatttat
      721 aaagtttatg tacaaatatc ataaaaaaga gaatcttttt aagcaaggat tttcttaact
      781 tcttcggcga cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt
      841 ctgatacctg catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc
      901 aagttcaatg acaatttcaa catcattgca gcagacaaga tagtggcgat agggtcaacc
      961 ttattcttgg caaatccggg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg
     1021 ttcttgtctg gcaaagaggc caaggacgca gatggcaaca aacccaagga acctggataa
     1081 cgtaaggctt catcggagat gatatcacca aacatgttgc tggtgattat aataccattt
     1141 aggtgggttg ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc
     1201 ttcaatgtag ggaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa
     1261 gaggccaaaa cattagcttt atccaaggtc caaataggca atggtggctc atgttgtagg
     1321 gccatgaaag cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta
     1381 tcccaagcga caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact
     1441 aattctctga caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag
     1501 tctaaaagag agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct
     1561 ttacggattt ttagtaaacc ttgttcaggt ctaacactac cggtacccca ttttggacca
     1621 cccacagcac ctaacaaaac ggcatcaacc ttcttggagg cttccagtgc ctcatctgga
     1681 agtgggacac ctgtagcatc gatagcagca ccaccaatta aatgattttc gaaatcgaac
     1741 ttgacattgg aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatt
     1801 tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct taggaacaaa cattagaatg
     1861 gtatatcctt gaaatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa
     1921 gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa
     1981 cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc
     2041 cggttccggc ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc
     2101 gtcaggcgac ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat
     2161 agcgcccctg tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga
     2221 actcttgcat cttacgatac ctgagtattc ccacagttaa cgaagcatct gtgcttcatt
     2281 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2341 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     2401 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     2461 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     2521 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     2581 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     2641 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     2701 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     2761 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     2821 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     2881 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     2941 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     3001 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     3061 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     3121 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     3181 aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     3241 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3301 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3361 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3421 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3481 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3601 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3661 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3721 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3781 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3841 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3901 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3961 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4021 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4081 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4141 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4201 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4261 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4321 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4381 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4441 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4501 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4561 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4621 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4681 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4741 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4801 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4861 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4921 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4981 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5041 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5101 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5161 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5221 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5281 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5341 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5401 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5461 cgttggccga ttcattaatc cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5521 gagcgcaacg caattaatgt gagttacctc actcattagg caccccaggc tttacacttt
     5581 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5641 agct
//
