LOCUS       Exported                4861 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCtrlki-kanMX6 DNA, complete sequence.
ACCESSION   AB623233
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  AUTHORS   Takeda J, Taoka M, Isobe T.
  TITLE     Cloning vector pCtrlki-kanMX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4861)
  AUTHORS   Takeda J, Taoka M, Isobe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-APR-2011) Contact:Jun Takeda Tokyo Metropolitan 
            University, Graduate School of Science and Engineering; 1-1 
            Minami-osawa, Hachioji, Tokyo 192-0397, Japan
REFERENCE   3  (bases 1 to 4861)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4861
                     /organism="Cloning vector pCtrlki-kanMX6"
                     /lab_host="Escherichia coli DH5-alpha"
                     /mol_type="other DNA"
                     /db_xref="taxon:1004559"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    231..266
                     /label=Multiple cloning site 1
                     /note="Multiple cloning site 1"
     regulatory      278..510
                     /regulatory_class="terminator"
                     /note="SV40 poly(A) signal sequence"
     polyA_signal    391..472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      517..899
                     /regulatory_class="promoter"
                     /note="Ptef"
     gene            556..1912
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        556..899
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             900..1709
                     /codon_start=1
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aminoglycoside 3-phosphotransferase
                     /note="Kanamycin resistance gene"
                     /protein_id="BAL27687.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      1710..1946
                     /regulatory_class="terminator"
                     /note="Ttef"
     terminator      1715..1912
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1947..1976
                     /label=Multiple cloning site 2
                     /note="Multiple cloning site 2"
     primer_bind     complement(1990..2006)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2219..2674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2956..3060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3061..3921
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4092..4680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"