TAGZyme pQE-2 vector (V010759)

LOCUS       TAGZyme_pQE-2.        4758 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a lacIq repressor gene for bacterial expression of 
            6xHis-tagged proteins that can be processed by TAGZyme.
ACCESSION   .
VERSION     .
KEYWORDS    TAGZyme pQE-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  AUTHORS   Qiagen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4758)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For TAGZyme cleavage, a glutamine should be inserted at an uneven 
            amino acid position, followed by the first amino acid of the 
            expressed protein.
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    62..78
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     RBS             101..106
                     /note="ribosome binding site"
     CDS             115..117
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             121..141
                     /label=7xHis
                     /note="6xHis affinity tag"
     misc_feature    139..199
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    199..209
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     terminator      215..309
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             353..1009
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      1077..1163
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(1222..1243)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1259..2338)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(2339..2416)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    2608..2748
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2934..3522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3696..4553)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4554..4658)
                     /label=AmpR promoter