pET-58-DEST vector (V010943)

LOCUS       pET-58-DEST.        8395 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) Nova bacterial destination vector for expressing proteins
            tagged at the N-terminus with NusA and at the C-terminus with S-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-58-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8395)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(585..1442)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1443..1535)
                     /label=AmpR promoter
     rep_origin      1616..2204
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2390..2532)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2637..2825)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3363..3384)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3400..4479)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4480..4557)
                     /label=lacI promoter
     promoter        4870..4888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4889..4913
                     /label=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             4928..4950
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4957..4959
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             4969..4986
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             5002..6483
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
     CDS             6493..6510
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     protein_bind    6514..6638
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6663..6693
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6747..7403
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             7748..8050
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(8094..8218)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             8223..8267
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      8323..8370
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"