LOCUS       V003087                 7076 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003087
VERSION     V003087
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7076)
  AUTHORS   Desai RP, Papoutsakis ET.
  TITLE     Antisense RNA strategies for metabolic engineering of Clostridium
            acetobutylicum
  JOURNAL   J. Bacteriol. 65, 936-945 (1999)
REFERENCE   2  (bases 1 to 7076)
  AUTHORS   Soucaille P, Papoutsakis ET.
  TITLE     Clostridium acetobutylicum vectors using three clostridial
            promoters: expression of synthetic acetone-pathway operons and
            antisense RNA production
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7076)
  AUTHORS   Soucaille P, Papoutsakis ET.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-NOV-2002) Laboratoire de Biotechnologie-Bioprocedes,
            UMR CNRS 5504, UR INRA 792, INSA, 135, avenue de Rangueil, Toulouse
            31077, France
REFERENCE   4  (bases 1 to 7076)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7076)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Bacteriol. 65, 936-945 (1999)"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (25-NOV-2002) Laboratoire de Biotechnologie-Bioprocedes, UMR CNRS
            5504, UR INRA 792, INSA, 135, avenue de Rangueil, Toulouse 31077,
            France"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(465..905)
                     /codon_start=1
                     /gene="repL"
                     /product="RepL"
                     /label="repL"
                     /note="Bacillus subtilis plasmid pIM13 replication protein"
                     /protein_id="AAO39977.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            complement(465..905)
                     /gene="repL"
                     /label="repL"
     CDS             complement(1356..2087)
                     /gene="ermC'"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Bacillus
                     subtilis. Accession#: P13956"
     regulatory      2450..2666
                     /label="Clostridium acetobutylicum ATCC824 adc promoter"
                     /note="Clostridium acetobutylicum ATCC824 adc promoter"
                     /regulatory_class="promoter"
     CDS             2694..3347
                     /gene="ctfA"
                     /label="Acetoacetyl-CoA:acetate/butyrate CoA transferase
                     alpha subunit"
                     /note="Acetoacetyl-CoA:acetate/butyrate CoA transferase
                     alpha subunit from Clostridium acetobutylicum (strain ATCC
                     824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787).
                     Accession#: P33752"
     CDS             3352..4014
                     /gene="ctfB"
                     /label="Acetoacetyl-CoA:acetate/butyrate CoA-transferase
                     beta subunit"
                     /note="Acetoacetyl-CoA:acetate/butyrate CoA-transferase
                     beta subunit from Clostridium acetobutylicum (strain ATCC
                     824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787).
                     Accession#: P23673"
     CDS             4050..4781
                     /gene="adc"
                     /label="Acetoacetate decarboxylase"
                     /note="Acetoacetate decarboxylase from Clostridium
                     acetobutylicum (strain ATCC 824 / DSM 792 / JCM 1419 / LMG
                     5710 / VKM B-1787). Accession#: P23670"
     primer_bind     complement(4855..4871)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4879..4895)
                     /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)."
     promoter        complement(4903..4933)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4948..4969)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5257..5845)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6019..6876)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6877..6981)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgagc ttaatcgcat
      421 ttcatagatt gacctcccaa taactacgtg gtgttattgg gaggtcaatc tatttcattt
      481 gcctcttgct caaagttccc aaattcgagt aagaggtatt tttgtttttg gtcgtcgcct
      541 ctcattagta gttcagggtt taacattaat actccagttt ttctttttat aatatttcct
      601 tcttctaaga ttttaagtgt tgttattact gtttgtagac ttgttcctgt agcttttgct
      661 atttctcttg ttgtagctat cattgtattg ttacttaagt ggacattatc taggatatag
      721 ttaacgattt taagtttttt tccgccaatc atatctaaca tacttattaa ttgcactata
      781 tatgccttta cgaagttacc agacgtttgt ttacggtata acttgtctac ctctatgact
      841 tctccacttt cttcgtctat gagcctctga gagcctttat agactgttcc atatctttct
      901 ttcatctttt tctcactcct tattttaaac tattctaact atatcataac tgttctaaaa
      961 aaaaaagaac atttgttaaa agaaattaga acaaaatgag tgaaaaatta gaacaaacaa
     1021 attccttata aaccttatca tctcaaccta tattaagatt ttacctagtt gaatcttctt
     1081 ttctatataa agcgtcggag catatcaggg ggttatctaa cgtaaatgct acccttcggc
     1141 tcgctttcgc tcggcattga cgtcagatac tgcaccccct gaacccccat gctccaacag
     1201 caaaaaggaa actttttgct gcttttccga cgcttattcg cttcgctcat atttatatag
     1261 aaaagaagtg aatgcgcaaa agacataatc gattcacaaa aaataggtac acgaaaaaca
     1321 agttaaggga tgcagtttat gcatccctta acttacttat taaataattt atagctattg
     1381 aaaagagata agaattgttc aaagctaata ttgtttaaat cgtcaattcc tgcatgtttt
     1441 aaggaattgt taaattgatt ttttgtaaat attttcttgt attctttgtt aacccatttc
     1501 ataacgaaat aattatactt ctgtttatct ttgtgtgata ttcttgattt ttttctattt
     1561 aatctgataa gtgagctatt cactttaggt ttaggatgaa aatattctct tggaaccata
     1621 cttaatatag aaatatcaac ttctgccatt aaaaataatg ccaatgagcg ttttgtattt
     1681 aataatcttt tagcaaaccc gtattccacg attaaataaa tctcatcagc tatactatca
     1741 aaaacaattt tgcgtattat atccgtactt atgttataag gtatattacc aaatatttta
     1801 taggattggt ttttaggaaa tttaaactgc aatatatcct tgtttaaaac ttggaaatta
     1861 tcgtgatcaa caagtttatt ttctgtagtt ttgcataatt tatggtctat ttcaatggca
     1921 gttacgaaat tacacctctg tactaattca agggtaaaat gcccttttcc tgagccgatt
     1981 tcaaagatat tatcatgttc atttaatctt atatttgtca ttattttatc tatattatgt
     2041 tttgaagtaa taaagttttg actgtgtttt atatttttct cgttcattat aaccctcttt
     2101 attttttcct ccttataaaa ttagtataat tatagcacga gctctgataa atatgaacat
     2161 gatgagtgat cgttaaattt atactgcaat ctgatgcgat tattgaataa aagatatgag
     2221 agatttatct agtttctttt tttacaagaa aaaagaaagt tcttaaaggt tttatacttt
     2281 tggtcgtaga gcacacggtt taacgactta attacgaagt aaataagtct agtgtgttag
     2341 actttaatgt ttttttaagg cattagtgca tttaagcgtc agagcatggc tttatgccga
     2401 gaaaactatt ggttggaatg gcgtgtgtgt tagccaaagc tcctgcaggt cgacaaaaca
     2461 cctccacata agtttatata aatctattat gcagaatttt ttaggaagtg tacttttatt
     2521 ttcgaaagct agtagaaaaa attaatatat ggtgtaaaaa tttacttaaa aaaacaatat
     2581 gtgttataat gtaaatataa ataaatagga ctagaggcga tttataatgt aagataaagt
     2641 atgttagaaa agctaaacat tattggatcc agaatttaaa aggagggatt aaaatgaact
     2701 ctaaaataat tagatttgaa aatttaaggt cattctttaa agatgggatg acaattatga
     2761 ttggaggttt tttaaactgt ggcactccaa ccaaattaat tgatttttta gttaatttaa
     2821 atataaagaa tttaacgatt ataagtaatg atacatgtta tcctaataca ggtattggta
     2881 agttaatatc aaataatcaa gtaaaaaagc ttattgcttc atatataggc agcaacccag
     2941 atactggcaa aaaacttttt aataatgaac ttgaagtaga gctctctccc caaggaactc
     3001 tagtggaaag aatacgtgca ggcggatctg gcttaggtgg tgtactaact aaaacaggtt
     3061 taggaacttt gattgaaaaa ggaaagaaaa aaatatctat aaatggaacg gaatatttgt
     3121 tagagctacc tcttacagcc gatgtagcat taattaaagg tagtattgta gatgaggccg
     3181 gaaacacctt ctataaaggt actactaaaa actttaatcc ctatatggca atggcagcta
     3241 aaaccgtaat agttgaagct gaaaatttag ttagctgtga aaaactagaa aaggaaaaag
     3301 caatgacccc cggagttctt ataaattata tagtaaagga gcctgcataa aatgattaat
     3361 gataaaaacc tagcgaaaga aataatagcc aaaagagttg caagagaatt aaaaaatggt
     3421 caacttgtaa acttaggtgt aggtcttcct accatggttg cagattatat accaaaaaat
     3481 ttcaaaatta ctttccaatc agaaaacgga atagttggaa tgggcgctag tcctaaaata
     3541 aatgaggcag ataaagatgt agtaaatgca ggaggagact atacaacagt acttcctgac
     3601 ggcacatttt tcgatagctc agtttcgttt tcactaatcc gtggtggtca cgtagatgtt
     3661 actgttttag gggctctcca ggtagatgaa aagggtaata tagccaattg gattgttcct
     3721 ggaaaaatgc tctctggtat gggtggagct atggatttag taaatggagc taagaaagta
     3781 ataattgcaa tgagacatac aaataaaggt caacctaaaa ttttaaaaaa atgtacactt
     3841 cccctcacgg caaagtctca agcaaatcta attgtaacag aacttggagt aattgaggtt
     3901 attaatgatg gtttacttct cactgaaatt aataaaaaca caaccattga tgaaataagg
     3961 tctttaactg ctgcagattt actcatatcc aatgaactta gacccatggc tgtttagaaa
     4021 gaattcttga tatcaggaag gtgactttta tgttaaagga tgaagtaatt aaacaaatta
     4081 gcacgccatt aacttcgcct gcatttccta gaggacccta taaatttcat aatcgtgagt
     4141 attttaacat tgtatatcgt acagatatgg atgctcttcg taaagttgtg ccagagcctt
     4201 tagaaattga tgagccctta gtcaggtttg aaattatggc aatgcatgat acgagtggac
     4261 ttggttgtta tacagaaagc ggacaggcta ttcccgtaag ctgtaatgga gttaagggag
     4321 attatcttca tatgatgtat ttagataatg agcctgcaat tgcagtagga agggaattaa
     4381 gtgcatatcc taaaaagctc gggtatccaa agctttttgt ggattcagat actttagtag
     4441 gaactttaga ctatggaaaa cttagagttg cgacagctac aatggggtac aaacataaag
     4501 ccttagatgc taatgaagca aaggatcaaa tttgtcgccc taattatatg ttgaaaataa
     4561 tacccaatta tgatggaagc cctaggatat gtgagcttat aaatgcgaaa atcacagatg
     4621 ttaccgtaca tgaagcttgg acaggaccaa ctcgactgca gttatttgat cacgctatgg
     4681 cgccacttaa tgatttgcca gtaaaagaga ttgtttctag ctctcacatt cttgcagata
     4741 taatattgcc tagagctgaa gttatatatg attatcttaa gtaataaaaa taagagttac
     4801 cttaaatggt aactcttatt tttttaatgt cgactcatag aattcgtaat catggtcata
     4861 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4921 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aaagctcacg
     5461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5761 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7021 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
