LOCUS       40924_49732        4406 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast/E. coli integrating shuttle vector with a LEU2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4406)
  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 4406)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4406)
  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..4406
                     /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
     promoter        2285..2389
                     /label=AmpR promoter
     CDS             2390..3247
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3421..4009
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4297..4318
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4333..4363
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4371..4387
                     /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 ccacagttca ggtggcactt ttcggggaaa
     2281 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2341 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2401 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2461 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2521 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2581 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2641 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2701 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2761 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2821 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2881 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2941 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3001 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3061 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3121 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3181 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3241 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3301 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3361 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3421 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3481 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3541 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3601 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3661 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3721 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3781 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     3841 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3901 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3961 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4021 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4081 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4141 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4201 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tccagctggc acgacaggtt
     4261 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttacc tcactcatta
     4321 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     4381 ataacaattt cacacaggaa acagct
//
