LOCUS       Exported                5295 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pK4RP DNA, complete sequence.
ACCESSION   LC363920
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  AUTHORS   Pavlova M, Klvana M, Jesenska A, Prokop Z, Konecna H, Sato T, Tsuda 
            M, Nagata Y, Damborsky J.
  TITLE     The identification of catalytic pentad in the haloalkane 
            dehalogenase DhmA from Mycobacterium avium N85: reaction mechanism 
            and molecular evolution
  JOURNAL   J. Struct. Biol. 157 (2), 384-392 (2007)
  PUBMED    17084094
REFERENCE   2  (bases 1 to 5295)
  AUTHORS   Kishida K, Ogawa N, Ichihashi E, Kato H, Nagata Y, Ohtsubo Y, Tsuda 
            M.
  TITLE     Establishment of Plasmid Vector and Allelic Exchange Mutagenesis 
            Systems in a Mycobacterial Strain that is Able to Degrade Polycyclic
            Aromatic Hydrocarbons
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5295)
  AUTHORS   Kishida K, Tsuda M.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JAN-2018) Contact:Kouhei Kishida Graduate School of 
            Life Sciences, Tohoku University, Department of Environmental Life 
            Science; 2-1-1 Katahira, Sendai, Miyagi 980-8577, Japan
REFERENCE   4  (bases 1 to 5295)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5295
                     /organism="Cloning vector pK4RP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2077117"
     CDS             complement(250..531)
                     /codon_start=1
                     /gene="repB"
                     /product="DNA-binding replication protein"
                     /label=repB
                     /note="putative;similar to putative DNA-binding replication
                     protein from Rhodococcus erythropolis pFAJ2600"
                     /protein_id="BBC62170.1"
                     /translation="MGAETPARRTRTAREVAERIGASPRTVRRIIAEPRASYEARAAER
                     RKQVLELRASGMKLREIAAEVGMSVGGVGTILHHARKTEQSKAEGAMA"
     gene            complement(250..531)
                     /gene="repB"
                     /label=repB
     CDS             complement(521..1441)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label=repA
                     /note="putative;similar to putative theta replicase from 
                     Rhodococcus rhodochrous pKA22 and RepA proteins from 
                     Mycobacterium fortuitum pAL5000 and related plasmids."
                     /protein_id="BBC62171.1"
                     /translation="MDMSGGSLSGDWEQLWLPLWPLATDDLLLGVYRMPRQDALDRRYL
                     EANPQALSNLLVVDVDHPDAALRALSAAGNHPLPNAIVENPRNGHAHAVWALTEPFTRT
                     EYARRKPLAYAAAVNEGLRRAVDGDAAYSGLMTKNPTHSAWDTHWIHAETRSLADLEHD
                     LGKHMPPPRWRQSKRRREDPVGLGRNCMLFETARTWAYRELRCHWGDPEGLGKAIQVEA
                     ADLNAAFSEPLPVSEVRAIAASIHRWIVTKSRMWADGPAVYEATFVAIQSARGRKMTEK
                     KREANRRRATKYDRDLVRKEATDGS"
     gene            complement(521..1441)
                     /gene="repA"
                     /label=repA
     primer_bind     complement(2615..2631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2639..2655
                     /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(2663..2693)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2708..2729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3186..3805
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      complement(3201..3789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4171..4986
                     /codon_start=1
                     /gene="KanR"
                     /product="kanamycin resistance protein"
                     /label=KanR
                     /protein_id="BBC62172.1"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            4171..4986
                     /gene="KanR"
                     /label=KanR
     primer_bind     5205..5221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      5231..5259
                     /regulatory_class="promoter"
                     /note="putative rrn promoter"