LOCUS       Exported                6995 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pSOS95, complete sequence.
ACCESSION   AY187686
VERSION     .
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6995
                     /organism="Shuttle vector pSOS95"
                     /mol_type="genomic DNA"
                     /note="Escherichia coli-Clostridium acetobutylicum shuttle 
                     vector"
                     /db_xref="taxon:220089"
     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(1353..2087)
                     /codon_start=1
                     /gene="mlsR"
                     /product="MlsR"
                     /label=mlsR
                     /note="Bacillus subtilis plasmid pIM13 
                     macrolide-lincosamide-streptogramin B resistance protein"
                     /protein_id="AAO39990.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVQRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIFGNIPY
                     NISTDIIRKIVFDSIADEIYLIVEYGFAKRLLNTKRSLALFLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLNRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     gene            complement(1353..2087)
                     /gene="mlsR"
                     /label=mlsR
     regulatory      2450..2485
                     /regulatory_class="promoter"
                     /note="Clostridium acetobutylicum ATCC824 thl promoter"
     CDS             2613..3269
                     /codon_start=1
                     /gene="ctfA"
                     /product="CtfA"
                     /label=ctfA
                     /note="acetoacetyl-CoA:acetate/butyrate:CoA transferase 
                     subunit A"
                     /protein_id="AAO39991.1"
                     /translation="MNSKIIRFENLRSFFKDGMTIMIGGFLNCGTPTKLIDFLVNLNIK
                     NLTIISNDTCYPNTGIGKLISNNQVKKLIASYIGSNPDTGKKLFNNELEVELSPQGTLV
                     ERIRAGGSGLGGVLTKTGLGTLIEKGKKKISINGTEYLLELPLTADVALIKGSIVDEAG
                     NTFYKGTTKNFNPYMAMAAKTVIVEAENLVSCEKLEKEKAMTPGVLINYIVKEPA"
     gene            2613..3269
                     /gene="ctfA"
                     /label=ctfA
     CDS             3271..3936
                     /codon_start=1
                     /gene="ctfB"
                     /product="CtfB"
                     /label=ctfB
                     /note="acetoacetyl-CoA:acetate/butyrate:CoA transferase 
                     subunit B"
                     /protein_id="AAO39992.1"
                     /translation="MINDKNLAKEIIAKRVARELKNGQLVNLGVGLPTMVADYIPKNFK
                     ITFQSENGIVGMGASPKINEADKDVVNAGGDYTTVLPDGTFFDSSVSFSLIRGGHVDVT
                     VLGALQVDEKGNIANWIVPGKMLSGMGGAMDLVNGAKKVIIAMRHTNKGQPKILKKCTL
                     PLTAKSQANLIVTELGVIEVINDGLLLTEINKNTTIDEIRSLTAADLLISNELRPMAV"
     gene            3271..3936
                     /gene="ctfB"
                     /label=ctfB
     gene            3969..4706
                     /gene="adc"
                     /label=adc
     CDS             3969..4703
                     /codon_start=1
                     /gene="adc"
                     /product="Adc"
                     /label=adc
                     /note="acetoacetate decarboxylase"
                     /protein_id="AAO39993.1"
                     /translation="MLKDEVIKQISTPLTSPAFPRGPYKFHNREYFNIVYRTDMDALRK
                     VVPEPLEIDEPLVRFEIMAMHDTSGLGCYTESGQAIPVSCNGVKGDYLHMMYLDNEPAI
                     AVGRELSAYPKKLGYPKLFVDSDTLVGTLDYGKLRVATATMGYKHKALDANEAKDQICR
                     PNYMLKIIPNYDGSPRICELINAKITDVTVHEAWTGPTRLQLFDHAMAPLNDLPVKEIV
                     SSSHILADIILPRAEVIYDYLK"
     primer_bind     complement(4774..4790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4798..4814
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    4867..4888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(5935..6795)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="AAO39994.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5935..6795)
                     /gene="bla"
                     /label=bla
     promoter        complement(6796..6900)
                     /gene="bla"
                     /label=AmpR promoter