pET-44c(+) vector (V010962)

Basic Vector Information

Vector Name:
pET-44c(+)
Antibiotic Resistance:
Ampicillin
Length:
7309 bp
Type:
pET & Duet Vectors (Novagen)
Replication origin:
ori
Source/Author:
Novagen (EMD Millipore)
Copy Number:
High copy number

pET-44c(+) vector Map

pET-44c(+)7309 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200T7 terminatorrrnB T2 terminatorrrnB T1 terminator6xHisHSV tagenterokinase sitethrombin siteS-Tag6xHisNusA6xHisATGRBSlac operatorT7 promoterlacI promoterlacICAP binding siteropbomoriAmpR promoterAmpRf1 ori

pET-44c(+) vector Sequence

LOCUS       pET-44c(+).        7309 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing 6xHis-tagged NusA fusion proteins. 
            For other reading frames, use pET-44a(+) or pET-44b(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-44c(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7309)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7309)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7309
                     /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(679..693)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(712..729)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(745..789)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(799..816)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(832..2316)
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
     CDS             complement(2323..2340)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(2350..2352)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(2360..2382)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2397..2421)
                     /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(2422..2440)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2753..2830
                     /label=lacI promoter
     CDS             2831..3910
                     /label=lacI
                     /note="lac repressor"
     protein_bind    3926..3947
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4485..4673
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4778..4920
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5106..5694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5775..5867
                     /label=AmpR promoter
     CDS             5868..6725
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(6843..7298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"

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