LOCUS       Exported                8005 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pBJ2, complete sequence.
ACCESSION   HM441078
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8005)
  AUTHORS   Equbal MJ, Agarwal GP.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUN-2010) Department of Biochemical Engineering & 
            Biotechnology, Indian Institute of Technology - Delhi, Hauz Khas, 
            New Delhi, Delhi 110016, India
REFERENCE   2  (bases 1 to 8005)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8005
                     /organism="Shuttle expression vector pBJ2"
                     /mol_type="other DNA"
                     /note="Corynebacterium glutamicum - Escherichia coli 
                     shuttle expression vector"
                     /db_xref="taxon:903396"
     regulatory      complement(26..73)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    complement(140..157)
                     /label=hexa-histidine tag
                     /note="hexa-histidine tag"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    complement(158..203)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    complement(207..299)
                     /label=N-terminal tag
                     /note="N-terminal tag"
     misc_feature    complement(207..239)
                     /label=T7 tag
                     /note="T7 tag"
     CDS             complement(207..239)
                     /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"
     misc_feature    complement(243..260)
                     /label=thrombin cleavage site
                     /note="thrombin cleavage site"
     CDS             complement(243..260)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    complement(270..287)
                     /label=hexa-histidine tag
                     /note="hexa-histidine tag"
     CDS             complement(270..287)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             306..328
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(307..313)
                     /regulatory_class="ribosome_binding_site"
     protein_bind    343..367
                     /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(368..386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(370..386)
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     CDS             461..1120
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="confers resistance to chloramphenicol"
                     /protein_id="ADO00978.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            461..1120
                     /gene="cat"
                     /label=cat
     promoter        1672..1749
                     /gene="lacI"
                     /label=lacI promoter
     misc_feature    2701..5451
                     /note="derived from BL1 replicon of Corynebacterium 
                     glutamicum plasmid pBL1 INSD accession X03987; involved in 
                     plasmid pBJ1 replication in Corynebacterium glutamicum"
     misc_feature    5593..5735
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5921..6509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5922
                     /label=derived from colE1 origin from pBR322
                     /note="derived from colE1 origin from pBR322"
     CDS             6631..7446
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aph
                     /note="confers resistance to kanamycin"
                     /protein_id="ADO00979.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            6631..7446
                     /gene="aph"
                     /label=aph
     rep_origin      complement(7539..7994)
                     /direction=LEFT
                     /label=phage f1 replication origin
                     /note="phage f1 replication origin"