LOCUS       V003089                 7000 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003089
VERSION     V003089
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7000)
  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 7000)
  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  JOURNAL   Unpublished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 7000)
  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 7000)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7000)
  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            "Unpublished"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..7000
                     /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="AAO39983.1"
                     /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..2590
                     /label="Clostridium acetobutylicum ATCC824 ptb promoter"
                     /note="Clostridium acetobutylicum ATCC824 ptb promoter"
                     /regulatory_class="promoter"
     CDS             2618..3271
                     /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             3276..3938
                     /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             3974..4705
                     /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(4779..4795)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4803..4819)
                     /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(4827..4857)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4872..4893)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5181..5769)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5943..6800)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6801..6905)
                     /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 cgactgtgga
     2461 tggagttaag tcagcagaaa gtataatgag aaaatataaa atataaataa ttttctaaaa
     2521 aacttaactt catgtgaaaa gtttgttaaa atataaatga gcacgttaat catttaacat
     2581 agataattgg atccagaatt taaaaggagg gattaaaatg aactctaaaa taattagatt
     2641 tgaaaattta aggtcattct ttaaagatgg gatgacaatt atgattggag gttttttaaa
     2701 ctgtggcact ccaaccaaat taattgattt tttagttaat ttaaatataa agaatttaac
     2761 gattataagt aatgatacat gttatcctaa tacaggtatt ggtaagttaa tatcaaataa
     2821 tcaagtaaaa aagcttattg cttcatatat aggcagcaac ccagatactg gcaaaaaact
     2881 ttttaataat gaacttgaag tagagctctc tccccaagga actctagtgg aaagaatacg
     2941 tgcaggcgga tctggcttag gtggtgtact aactaaaaca ggtttaggaa ctttgattga
     3001 aaaaggaaag aaaaaaatat ctataaatgg aacggaatat ttgttagagc tacctcttac
     3061 agccgatgta gcattaatta aaggtagtat tgtagatgag gccggaaaca ccttctataa
     3121 aggtactact aaaaacttta atccctatat ggcaatggca gctaaaaccg taatagttga
     3181 agctgaaaat ttagttagct gtgaaaaact agaaaaggaa aaagcaatga cccccggagt
     3241 tcttataaat tatatagtaa aggagcctgc ataaaatgat taatgataaa aacctagcga
     3301 aagaaataat agccaaaaga gttgcaagag aattaaaaaa tggtcaactt gtaaacttag
     3361 gtgtaggtct tcctaccatg gttgcagatt atataccaaa aaatttcaaa attactttcc
     3421 aatcagaaaa cggaatagtt ggaatgggcg ctagtcctaa aataaatgag gcagataaag
     3481 atgtagtaaa tgcaggagga gactatacaa cagtacttcc tgacggcaca tttttcgata
     3541 gctcagtttc gttttcacta atccgtggtg gtcacgtaga tgttactgtt ttaggggctc
     3601 tccaggtaga tgaaaagggt aatatagcca attggattgt tcctggaaaa atgctctctg
     3661 gtatgggtgg agctatggat ttagtaaatg gagctaagaa agtaataatt gcaatgagac
     3721 atacaaataa aggtcaacct aaaattttaa aaaaatgtac acttcccctc acggcaaagt
     3781 ctcaagcaaa tctaattgta acagaacttg gagtaattga ggttattaat gatggtttac
     3841 ttctcactga aattaataaa aacacaacca ttgatgaaat aaggtcttta actgctgcag
     3901 atttactcat atccaatgaa cttagaccca tggctgttta gaaagaattc ttgatatcag
     3961 gaaggtgact tttatgttaa aggatgaagt aattaaacaa attagcacgc cattaacttc
     4021 gcctgcattt cctagaggac cctataaatt tcataatcgt gagtatttta acattgtata
     4081 tcgtacagat atggatgctc ttcgtaaagt tgtgccagag cctttagaaa ttgatgagcc
     4141 cttagtcagg tttgaaatta tggcaatgca tgatacgagt ggacttggtt gttatacaga
     4201 aagcggacag gctattcccg taagctgtaa tggagttaag ggagattatc ttcatatgat
     4261 gtatttagat aatgagcctg caattgcagt aggaagggaa ttaagtgcat atcctaaaaa
     4321 gctcgggtat ccaaagcttt ttgtggattc agatacttta gtaggaactt tagactatgg
     4381 aaaacttaga gttgcgacag ctacaatggg gtacaaacat aaagccttag atgctaatga
     4441 agcaaaggat caaatttgtc gccctaatta tatgttgaaa ataataccca attatgatgg
     4501 aagccctagg atatgtgagc ttataaatgc gaaaatcaca gatgttaccg tacatgaagc
     4561 ttggacagga ccaactcgac tgcagttatt tgatcacgct atggcgccac ttaatgattt
     4621 gccagtaaaa gagattgttt ctagctctca cattcttgca gatataatat tgcctagagc
     4681 tgaagttata tatgattatc ttaagtaata aaaataagag ttaccttaaa tggtaactct
     4741 tattttttta atgtcgactc atagaattcg taatcatggt catagctgtt tcctgtgtga
     4801 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4861 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4921 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4981 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5041 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5101 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5161 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5221 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5281 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5341 gcctttctcc cttcgggaag cgtggcgctt tctcaaagct cacgctgtag gtatctcagt
     5401 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5461 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5521 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5581 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5641 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5701 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5761 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5821 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5881 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5941 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6001 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6061 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6121 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6181 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6241 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6301 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6361 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6421 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6481 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6541 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6601 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6661 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6721 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6781 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6841 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6901 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     6961 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
