LOCUS       Exported                5286 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDS132, complete sequence.
ACCESSION   AY489048
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5286)
  AUTHORS   Philippe N, Alcaraz JP, Coursange E, Geiselmann J, Schneider D.
  TITLE     Improvement of pCVD442, a suicide plasmid for gene allele exchange 
            in bacteria
  JOURNAL   Plasmid 51 (3), 246-255 (2004)
  PUBMED    15109831
REFERENCE   2  (bases 1 to 5286)
  AUTHORS   Philippe N, Alcaraz JP, Coursange E, Geiselmann J, Schneider D.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2003) LAPM, UMR 5163, BP53 CERMO, Grenoble 38041, 
            France
REFERENCE   3  (bases 1 to 5286)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5286
                     /organism="Cloning vector pDS132"
                     /mol_type="genomic DNA"
                     /note="derived from pCVD442 without IS1 sequences and bla 
                     gene replaced by cat gene"
                     /db_xref="taxon:267428"
     rep_origin      1..392
                     /note="R6K ori; similar to Escherichia coli sequence 
                     deposited in GenBank Accession Number V00623"
     rep_origin      4..392
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    418..2477
                     /note="mob RP4; similar to Escherichia coli sequence 
                     deposited in GenBank Accession Number X54459"
     CDS             complement(1010..1381)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1414..1523
                     /note="incP origin of transfer"
     CDS             complement(2489..3148)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /EC_number="2.3.1.28"
                     /label=cat
                     /note="similar to Escherichia coli cat gene deposited in 
                     GenBank Accession Number V00623 and to HCM1.206 deposited 
                     in GenBank Accession Number AL513383"
                     /protein_id="AAS67528.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(2489..3148)
                     /gene="cat"
                     /label=cat
     CDS             complement(3246..4667)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase precursor"
                     /EC_number="2.4.1.10"
                     /label=sacB
                     /note="similar to Bacillus subtilis BSU34450 deposited in 
                     GenBank Accession Number AL009126"
                     /protein_id="AAS67529.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(3246..4667)
                     /gene="sacB"
                     /label=sacB
     mat_peptide     complement(3249..4580)
                     /gene="sacB"
                     /product="levansucrase"
     sig_peptide     complement(4581..4667)
                     /gene="sacB"
     promoter        complement(4668..5113)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    5135..5208
                     /note="multiple cloning site 
                     (XbaI/SalI/PstI/SalI/PstI/SalI/XbaI/SphI)"
     protein_bind    5239..5255
                     /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)."