LOCUS       Exported                5344 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJTOOL-3, complete sequence.
ACCESSION   JF756693
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5344)
  AUTHORS   van Aartsen JJ, Rajakumar K.
  TITLE     An optimized method for suicide vector-based allelic exchange in 
            Klebsiella pneumoniae
  JOURNAL   J. Microbiol. Methods 86 (3), 313-319 (2011)
  PUBMED    21699924
REFERENCE   2  (bases 1 to 5344)
  AUTHORS   van Aartsen J, Rajakumar K.
  TITLE     pJTOOL vectors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5344)
  AUTHORS   van Aartsen J, Rajakumar K.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2011) Department of Infection, Immunity and 
            Inflammation, University of Leicester, United Kingdom, University 
            Road, Leicester, Leicestershire LE1 9HN, United Kingdom
REFERENCE   4  (bases 1 to 5344)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5344
                     /organism="Cloning vector pJTOOL-3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1003183"
     rep_origin      27..418
                     /label=from R6K
                     /note="from R6K"
     rep_origin      30..418
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    444..2503
                     /label=from RP4
                     /note="from RP4"
     CDS             complement(1036..1407)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1440..1549
                     /note="incP origin of transfer"
     CDS             complement(2515..3174)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AEB22074.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(2515..3174)
                     /gene="cat"
                     /label=cat
     CDS             complement(3272..4693)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase precursor"
                     /label=sacB
                     /protein_id="AEB22075.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(3272..4693)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(4694..5139)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     promoter        5159..5177
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5186..5293
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     5210..5226
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5260..5276)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5306..5324)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    5323..5339
                     /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)."