pET-44a(+) vector (V010964)

LOCUS       pET-44a(+).        7311 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing 6xHis-tagged NusA fusion proteins. 
            For other reading frames, use pET-44b(+) or pET-44c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-44a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7311)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7311)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      complement(141..168)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(260..346)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(512..529)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(536..568)
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             complement(681..695)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(714..731)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(747..791)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(801..818)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(834..2318)
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
     CDS             complement(2325..2342)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(2352..2354)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(2362..2384)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2399..2423)
                     /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)."
     promoter        complement(2424..2442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2755..2832
                     /label=lacI promoter
     CDS             2833..3912
                     /label=lacI
                     /note="lac repressor"
     protein_bind    3928..3949
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4487..4675
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4780..4922
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5108..5696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5777..5869
                     /label=AmpR promoter
     CDS             5870..6727
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(6845..7300)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"