LOCUS       Exported                3223 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETBK, complete sequence.
ACCESSION   GQ421431
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3223)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Bacterial expression with pETBK
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3223)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2009) BioDynamics Laboratory Inc., 9-7 Hongo 
            2-Chome, Bunkyo-Ku, Tokyo 113-0033, Japan
REFERENCE   3  (bases 1 to 3223)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3223
                     /organism="Expression vector pETBK"
                     /mol_type="other DNA"
                     /db_xref="taxon:670877"
     promoter        171..189
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      171..187
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     RBS             221..243
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    263..280
                     /label=histidine tag
                     /note="histidine tag"
     CDS             263..280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             284..316
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             320..334
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    334..380
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      456..503
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      456..503
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             703..1518
                     /codon_start=1
                     /gene="Km"
                     /product="aminoglycoside phosphotransferase"
                     /label=Km
                     /note="confers kanamycin resistance"
                     /protein_id="ACV53165.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            703..1518
                     /gene="Km"
                     /label=Km
     rep_origin      1660..2279
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      1676..2264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2450..2592
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2694..2885)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"