LOCUS       V003088                 6995 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003088
VERSION     V003088
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6995)
  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 6995)
  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  JOURNAL   Unpublished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 6995)
  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 6995)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6995)
  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            "Unpublished"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..6995
                     /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="AAO39989.1"
                     /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..2485
                     /label="Clostridium acetobutylicum ATCC824 thl promoter"
                     /note="Clostridium acetobutylicum ATCC824 thl promoter"
                     /regulatory_class="promoter"
     CDS             2613..3266
                     /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             3271..3933
                     /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             3969..4700
                     /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(4774..4790)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4798..4814)
                     /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(4822..4852)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4867..4888)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5176..5764)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5938..6795)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6796..6900)
                     /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 cgacttttta
     2461 acaaaatata ttgataaaaa taataatagt gggtataatt aagttgttag agaaaacgta
     2521 taaattaggg ataaactatg gaacttatga aatagattga aatggtttat ctgttacccc
     2581 gtaggatcca gaatttaaaa ggagggatta aaatgaactc taaaataatt agatttgaaa
     2641 atttaaggtc attctttaaa gatgggatga caattatgat tggaggtttt ttaaactgtg
     2701 gcactccaac caaattaatt gattttttag ttaatttaaa tataaagaat ttaacgatta
     2761 taagtaatga tacatgttat cctaatacag gtattggtaa gttaatatca aataatcaag
     2821 taaaaaagct tattgcttca tatataggca gcaacccaga tactggcaaa aaacttttta
     2881 ataatgaact tgaagtagag ctctctcccc aaggaactct agtggaaaga atacgtgcag
     2941 gcggatctgg cttaggtggt gtactaacta aaacaggttt aggaactttg attgaaaaag
     3001 gaaagaaaaa aatatctata aatggaacgg aatatttgtt agagctacct cttacagccg
     3061 atgtagcatt aattaaaggt agtattgtag atgaggccgg aaacaccttc tataaaggta
     3121 ctactaaaaa ctttaatccc tatatggcaa tggcagctaa aaccgtaata gttgaagctg
     3181 aaaatttagt tagctgtgaa aaactagaaa aggaaaaagc aatgaccccc ggagttctta
     3241 taaattatat agtaaaggag cctgcataaa atgattaatg ataaaaacct agcgaaagaa
     3301 ataatagcca aaagagttgc aagagaatta aaaaatggtc aacttgtaaa cttaggtgta
     3361 ggtcttccta ccatggttgc agattatata ccaaaaaatt tcaaaattac tttccaatca
     3421 gaaaacggaa tagttggaat gggcgctagt cctaaaataa atgaggcaga taaagatgta
     3481 gtaaatgcag gaggagacta tacaacagta cttcctgacg gcacattttt cgatagctca
     3541 gtttcgtttt cactaatccg tggtggtcac gtagatgtta ctgttttagg ggctctccag
     3601 gtagatgaaa agggtaatat agccaattgg attgttcctg gaaaaatgct ctctggtatg
     3661 ggtggagcta tggatttagt aaatggagct aagaaagtaa taattgcaat gagacataca
     3721 aataaaggtc aacctaaaat tttaaaaaaa tgtacacttc ccctcacggc aaagtctcaa
     3781 gcaaatctaa ttgtaacaga acttggagta attgaggtta ttaatgatgg tttacttctc
     3841 actgaaatta ataaaaacac aaccattgat gaaataaggt ctttaactgc tgcagattta
     3901 ctcatatcca atgaacttag acccatggct gtttagaaag aattcttgat atcaggaagg
     3961 tgacttttat gttaaaggat gaagtaatta aacaaattag cacgccatta acttcgcctg
     4021 catttcctag aggaccctat aaatttcata atcgtgagta ttttaacatt gtatatcgta
     4081 cagatatgga tgctcttcgt aaagttgtgc cagagccttt agaaattgat gagcccttag
     4141 tcaggtttga aattatggca atgcatgata cgagtggact tggttgttat acagaaagcg
     4201 gacaggctat tcccgtaagc tgtaatggag ttaagggaga ttatcttcat atgatgtatt
     4261 tagataatga gcctgcaatt gcagtaggaa gggaattaag tgcatatcct aaaaagctcg
     4321 ggtatccaaa gctttttgtg gattcagata ctttagtagg aactttagac tatggaaaac
     4381 ttagagttgc gacagctaca atggggtaca aacataaagc cttagatgct aatgaagcaa
     4441 aggatcaaat ttgtcgccct aattatatgt tgaaaataat acccaattat gatggaagcc
     4501 ctaggatatg tgagcttata aatgcgaaaa tcacagatgt taccgtacat gaagcttgga
     4561 caggaccaac tcgactgcag ttatttgatc acgctatggc gccacttaat gatttgccag
     4621 taaaagagat tgtttctagc tctcacattc ttgcagatat aatattgcct agagctgaag
     4681 ttatatatga ttatcttaag taataaaaat aagagttacc ttaaatggta actcttattt
     4741 ttttaatgtc gactcataga attcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4801 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4861 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4921 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca aagctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6541 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6901 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6961 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
