LOCUS       Exported                4999 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKIT303 DNA, complete sequence.
ACCESSION   LC317095
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  AUTHORS   Toda, H., Itoh, N.
  TITLE     Development of novel E. coli-Kocuria shuttle vector using cryptic 
            plasmid pKPAL3 from Kocuria palustris IPUFS-1 and utilization for 
            producing enantiopure (S)-styrene oxide
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4999)
  AUTHORS   Itoh N, Toda, H.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-AUG-2017) Contact:Nobuya Itoh Toyama Prefectural 
            University, Biotechnology Research Center; 5180 Kurokawa, Imizu, 
            Toyama 939-0398, Japan URL :http://www.pu-toyama.ac.jp/BR/itoh/
REFERENCE   3  (bases 1 to 4999)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4999
                     /organism="Cloning vector pKIT303"
                     /mol_type="other DNA"
                     /db_xref="taxon:2033403"
     protein_bind    39..60
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        75..105
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..129
                     /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)."
     primer_bind     137..153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             149..475
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIRQSNHSTRP
                     VAAH"
     misc_feature    163..219
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(223..239)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      452..907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1189..1293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1294..2154
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /protein_id="BBA31609.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1294..2154
                     /gene="AmpR"
                     /label=AmpR
     rep_origin      2325..2913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3141..3950
                     /codon_start=1
                     /gene="tsr"
                     /product="thiostrepton resistance protein"
                     /label=tsr
                     /protein_id="BBA31610.1"
                     /translation="MTELDTIANPSDPAVQRIIDVTKPSRSNIKTTLIEDVEPLMHSIA
                     AGVEFIEVYGSDSSPFPSELLDLCGRQNIPVRLIDSSIVNQLFKGERKAKTFGIARVPR
                     PARFGDIASRRGDVVVLDGVKIVGNIGAIVRTSLALGASGIILVDSDITSIADRRLQRA
                     SRGYVFSLPVVLSGREEAIAFIRDSGMQLMTLKADGDISVKELGDNPDRLALLFGSEKG
                     GPSDLFEEASSASVSIPMMSQTESLNVSVSLGIALHERIDRNLAANR"
     gene            3141..3950
                     /gene="tsr"
                     /label=tsr
     rep_origin      3996..4999
                     /label=pKPAL3-derived plasmid replication origin
                     /note="pKPAL3-derived plasmid replication origin"