LOCUS       Exported                6274 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pXCEGFPCAH DNA, complete sequence.
ACCESSION   LC379924
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6274)
  AUTHORS   Nariya H, Miyata S, Kuwahara T, Okabe A.
  TITLE     Development and characterization of a xylose-inducible gene 
            expression system for Clostridium perfringens
  JOURNAL   Appl. Environ. Microbiol. 77 (23), 8439-8441 (2011)
  PUBMED    21965407
REFERENCE   2  (bases 1 to 6274)
  AUTHORS   Nariya H, Miyata S.
  TITLE     Development of xylose-inducible EGFP fusion Expression Vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6274)
  AUTHORS   Nariya H, Miyata S.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2018) Contact:Hirofumi Nariya Graduate School of 
            Biosphere Science, Hiroshima University, Laboratory of Food 
            Microbiology and Hygiene; 1-4-4, Kagamiyama 1-chome, 
            Higashi-Hiroshima, Hiroshima 739-8528, Japan
REFERENCE   4  (bases 1 to 6274)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6274
                     /organism="Expression vector pXCEGFPCAH"
                     /mol_type="other DNA"
                     /db_xref="taxon:2171677"
     rep_origin      266..430
                     /note="ori; derived from Clostridium perfringens plasmid 
                     pIP404 (INSD:M32882.1)"
     CDS             complement(493..1713)
                     /codon_start=1
                     /gene="rep"
                     /product="replication enzyme"
                     /label=rep
                     /note="derived from Clostridium perfringens plasmid pIP404 
                     (INSD:M32882.1)"
                     /protein_id="BBD84638.1"
                     /translation="MRNALKLNKKNYIDNIHSRSKGWITRSVIDKKGYSQWHYKYAELK
                     DLDMSDENIYITLNTFYKPCRRLENIKELNTLFIDLDYYKTGKTKDQVLMDLEKNYFNQ
                     SIPIPNYVIDSGRGMYLIWIINAVPSKALPLWKAVQEYLYNQLKYFGADRQALDATRIL
                     RVPGSINSKSKTVVNILDEYEYIYDLREIQNGFLPELKPYERKKGRPSKINYIYRERSL
                     YYGRIQDIIKLCELREYDLKGHRELILFLYRYYLCSFTEDIEKALNDVLELNSMFRQHL
                     SEREVIRATRSAERCYLDKNKQYKYKNETLIELLEITEEEQKYMTIIISKKEYKRRENI
                     RGKKNYQEQLKAKGKATKKEELNVLRKKIKALKEKGFKNKEITLMLEVPIKTLERHITY
                     MKKNGLL"
     gene            complement(493..1713)
                     /gene="rep"
                     /label=rep
     CDS             complement(2085..3245)
                     /codon_start=1
                     /gene="xylR"
                     /product="transcriptional repressor of the xylose operon"
                     /label=xylR
                     /note="derived from Clostridium difficile ATCC 9689"
                     /protein_id="BBD84639.1"
                     /translation="MVTDKYTIREMNERLVLEQIIKNGPISRASIASTIGLNKATISAI
                     TKKLIDESLVHEIGIGNSTHSGGRKPILLVFNKCAGISLSMDVGYDYIFSSLSYLDGTI
                     INSKKLTDIQVSKDNVIQLIDEIINSYNISKIDTPYKVIGLTLAIHGITCENKVLFTPY
                     YNLNEIDLYSILSKKYDFPIHIENEANLTALAENTFSTVHNSLLSLSIHSGFGSGIIIN
                     NKLYSGRNGMSGEIGHTIIMPNGKLCPCGNRGCLEQYCSEKKVFEQLSSLENIPKIDSD
                     IVKQLYYEDNQNAKKVIHEFCSYLTIAINNAITTYAPEIIYLNSQIISDIPEILQITKD
                     MLVSSFNKGINIEISSLGSEASLYGGSAVNIKSFLNIQNLTLINEI"
     gene            complement(2085..3245)
                     /gene="xylR"
                     /label=xylR
     CDS             3459..4292
                     /codon_start=1
                     /gene="CpEGFP-HA-His"
                     /product="His tagged enhanced green fluorescent protei"
                     /label=CpEGFP-HA-His
                     /note="derived from Aequorea victoria GFP gene; amino-acid 
                     substitution (p.Phe64Leu, p.Ser65Thr) and codon-optimized 
                     for Clostridium perfringens"
                     /protein_id="BBD84640.1"
                     /translation="MDVDACIDGSGTEFLQKLELESRSMSKGEELFTGVVPILVELDGD
                     VNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKR
                     HDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH
                     KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDN
                     HYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKYPYDVPDYAHHHHHH"
     gene            3459..4292
                     /gene="CpEGFP-HA-His"
                     /label=CpEGFP-HA-His
     CDS             4245..4271
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4272..4289
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4602..5225)
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /note="derived from Clostridium perfringens catP gene 
                     (INSD:M74769.1)"
                     /protein_id="BBD84641.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            complement(4602..5225)
                     /gene="catP"
                     /label=catP
     rep_origin      5499..6087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5920..6140
                     /note="oriEC; replication region from pUC18"