LOCUS       Exported                3964 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pCM3 DNA, complete sequence.
ACCESSION   AB562893
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3964)
  AUTHORS   Tanaka H, Nariya H, Suzuki M, Houchi H, Tamai E, Miyata S, Okabe A.
  TITLE     High-level production and purification of clostripain expressed in a
            virulence-attenuated strain of Clostridium perfringens
  JOURNAL   Protein Expr. Purif. 76 (1), 83-89 (2011)
  PUBMED    20940055
REFERENCE   2  (bases 1 to 3964)
  AUTHORS   Suzuki M, Okabe A.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAY-2010) Contact:Motoo Suzuki Kagawa University, 
            Department of Microbiology, Faculty of Medicine; 1750-1 Ikenobe, 
            Miki-cho, Kita-gun, Kagawa 761-0793, Japan
REFERENCE   3  (bases 1 to 3964)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3964
                     /organism="Shuttle vector pCM3"
                     /mol_type="other DNA"
                     /note="Derivative of pJIR418, reconstruction of the vector 
                     and significantly expansion of multiple cloning site; 
                     Shuttle vector using Clostridium perfringens and 
                     Escherichia coli"
                     /db_xref="taxon:768197"
     misc_feature    1..1985
                     /note="derived from pJIR418(kms); Our laboratory collection
                     of pJIR418, originally provided by J. Rood; Complete 
                     sequence of pJIR418 has been reported as Sloan,J. et al, 
                     Plasmid 27, 207-219, 1992; accession number M77169"
     misc_difference 252
                     /compare="M77169.1"
                     /note="insertion of A confirmed by sequencing analysis of 
                     pCM3 and pJIR418(kms); corresponding to position between 
                     1176 and 1177 of published sequence of pJIR418"
     rep_origin      266..430
                     /note="Derived from pJIR418(kms); the 64 and 1 bp-deletions
                     were found in pJIR418(kms), corresponding to region 1246 to
                     1309 and 1229 of pJIR418, respectively; Originally derived 
                     from complete sequence of pIP404 (Garnier & Cole, Plasmid 
                     19, 134-150, 1988; accession number M32882); C.perfringens 
                     pIP404 replication origin; ori"
     misc_difference 304..305
                     /note="deletion of the 1 base pair corresponding to the 
                     position 1229 of pJIR418, confirmed by sequencing analysis 
                     of pCM3 and pJIR418(kms)"
     misc_difference 320..321
                     /note="deletion of the 64 base pairs corresponding to the 
                     position from 1246 to 1309 of pJIR418, confirmed by 
                     sequencing analysis of pCM3 and pJIR418(kms)"
     CDS             complement(493..1713)
                     /codon_start=1
                     /gene="rep"
                     /product="replication enzyme"
                     /label=rep
                     /note="derived from complete sequence of pIP404 (Garnier & 
                     Cole, Plasmid 19, 134-150, 1988; accession number M32882); 
                     derived from pJIR418(kms); the mutation of G to C was found
                     in pJIR418(kms), corresponding to the position at 2223 of 
                     pJIR418"
                     /protein_id="BAJ34716.1"
                     /translation="MRNALKLNKKNYIDNIHSRSKGWITRSVIDKKGYSQWHYKYAELK
                     DLDMSDENIYITLNTFYKPCRRLENIKELNTLFIDLDYYKTGKTKDQVLMDLEKNYFNQ
                     SIPIPNYVIDSGRGMYLIWIINAVPSKALPLWKAVQEYLYNQLKYFGADRQALDATRIL
                     RVPGSINSKSKTVVNILDEYEYIYDLREIQNGFLPELKPYERKKGRPSKINYIYRERSL
                     YYGRIQDIIKLCELREYDLKGHRELILFLYRYYLCSFTEDIEKALNDVLELNSMFRQHL
                     SEREVIRATRSAERCYLDKNKQYKYKNETLIELLEITEEEQKYMTIIISKKEYKRRENI
                     RGKKNYQEQLKAKGKATKKEELNVLRKKIKALKEKGFKNKEITLMLEVPIKTLERHITY
                     MKKNGLL"
     gene            complement(493..1713)
                     /gene="rep"
                     /label=rep
     misc_difference 1108
                     /compare="M77169.1"
                     /note="mutation by PCR to abolish a NdeI site in 
                     pJIR418(kms), when constructed pCM3; silent mutation; Pro 
                     (CCA) in rep gene of pJIR418(kms) to Pro (CCT) of pCM3"
     misc_difference 1234
                     /compare="M77169.1"
                     /note="The position is C confirmed by sequencing analysis 
                     of pCM3 and pJIR418(kms); although C is G in originally 
                     published sequence of pJIR418 (position at 2223)"
     misc_feature    1986..2099
                     /label=Artificial multiple cloning site (ScaI-SmaI)
                     /note="Artificial multiple cloning site (ScaI-SmaI)"
     misc_feature    2100..3964
                     /note="derived from pJIR418(kms); Our laboratory collection
                     of pJIR418, originally provided by J. Rood; Complete 
                     sequence of pJIR418 has been reported as Sloan,J. et al, 
                     Plasmid 27, 207-219, 1992; accession number M77169"
     CDS             complement(2292..2915)
                     /codon_start=1
                     /gene="catP"
                     /gene_synonym="CAT"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /note="derived from C. perfringens catP (T. Bannam & J. 
                     Rood, unpublished results; accession number M74769)"
                     /protein_id="BAJ34717.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            complement(2292..2915)
                     /gene="catP"
                     /gene_synonym="CAT"
                     /label=catP
     rep_origin      3189..3777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_difference 3336
                     /compare="M77169.1"
                     /note="The position is T confirmed by sequencing analysis 
                     of pCM3 and pJIR418(kms); although T is C in originally 
                     published sequence of pJIR418 (position at 6506)"
     rep_origin      3610..3830
                     /note="oriEC; replication region from pUC18"