Basic Vector Information
The pIEx/Bac vectors are dual-purpose vectors that offer unique flexibility for cloning and expression of proteins in Spodoptera-derived insect cells. These vectors contain the Autographa californica nuclear polyhedrosis virus (AcNPV) enhancer/immediate early promoter combination (hr5/ie1) for plasmid-mediated and early baculovirus expression, and the AcNPV p10 very late promoter for robust baculovirus-mediated expression. All pIEx/Bac plasmids carry AcNPV-derived virus regions to mediate recombination at the polh locus. The pIEx/Bac-3 vector contains an N-terminal His•Tag coding sequence with 10 histidines followed by an HRV 3C protease cleavage site and a multiple cloning site, with the option to include a C-terminal thrombin protease cleavage site and 8 aa Strep•Tag II coding sequence. pIEx/Bac-3 is also available as a 3C/LIC vector. The 3C/LIC vector is prepared for rapid directional cloning of PCR-amplified DNA. Using specifically designed primers for amplification, inserts can be efficiently cloned without the need for restriction enzyme digestion or ligation.
- Vector Name:
- pIEx/Bac-3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6802 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
- Promoter:
- IE1
- Cloning Method:
- Enzyme digestion and ligation
- Fusion Tag:
- C-His,N-Strep
pIEx/Bac-3 vector Vector Map
pIEx/Bac-3 vector Sequence
LOCUS pIEx_Bac-3. 6802 bp DNA circular SYN 01-JAN-1980
DEFINITION Insect cell and baculovirus vector encoding a cleavable N-terminal
10xHis tag and a cleavable C-terminal Strep-Tag(R) II.
ACCESSION .
VERSION .
KEYWORDS pIEx Bac-3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6802)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6802)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6802
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 329..1156
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
enhancer 1196..1678
/label=hr5 enhancer
/note="baculovirus early transcription enhancer"
promoter 1682..2273
/label=IE1 promoter
/note="promoter of the ie1 gene from the baculovirus
Autographa californica"
promoter 2283..2392
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
CDS 2394..2396
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 2400..2426
/label=9xHis
/note="9xHis affinity tag"
CDS 2436..2459
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
misc_feature 2457..2501
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 2505..2522
/label=thrombin site
/note="thrombin recognition and cleavage site"
CDS 2529..2552
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
terminator 2571..2877
/label=IE1 terminator
/note="terminator of the ie1 gene from the baculovirus
Autographa californica"
misc_recomb 2906..4263
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
rep_origin complement(4526..5114)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5288..6145)
/label=AmpR
/note="beta-lactamase"
promoter complement(6146..6250)
/label=AmpR promoter
rep_origin 6277..6732
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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