Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V011661 | pBacPAK8 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pBacPAK8 is a 5538 bp baculovirus transfer vector designed for high-level recombinant protein expression in insect cells. It utilizes the strong polyhedrin promoter, has an ampicillin resistance gene for selection in E. coli, and contains a multiple cloning site for gene insertion.
- Vector Name:
- pBacPAK8
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5538 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- polyhedrin
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pBacPAK8 vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Krammer F, Schinko T, Palmberger D, Tauer C, Messner P, Grabherr R. Trichoplusia ni cells (High Five) are highly efficient for the production of influenza A virus-like particles: a comparison of two insect cell lines as production platforms for influenza vaccines. Mol Biotechnol. 2010 Jul;45(3):226-34. doi: 10.1007/s12033-010-9268-3. PMID: 20300881; PMCID: PMC4388404.
pBacPAK8 vector Sequence
LOCUS 40924_5679 5538 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBacPAK8, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5538)
AUTHORS Kitts PA.
TITLE CLONTECH Vectors On Disc version 1.3
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5538)
AUTHORS Kitts PA.
TITLE Direct Submission
JOURNAL Submitted (07-OCT-1993) Paul A. Kitts, CLONTECH Laboratories, Inc.,
1020 East Meadow Circle, Palo Alto, CA 94303, USA
REFERENCE 3 (bases 1 to 5538)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5538)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(07-OCT-1993) Paul A. Kitts, CLONTECH Laboratories, Inc., 1020 East
Meadow Circle, Palo Alto, CA 94303, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT This vector can be obtained from CLONTECH Laboratories, Inc., 1020
East Meadow Circle, Palo Alto, CA 94303, USA. To place an order call
(415) 424-8222 or (800) 662-2566, extension 1. International
customers, please contact your local distributor. For technical
information, call (415) 424- 8222 or (800) 662-2566, extension 3.
This sequence has been compiled from information in the sequence
databases, published literature and other sources, together with
partial sequences obtained by CLONTECH. If you suspect there is an
error in this sequence, please contact CLONTECH's Technical
Technical Service Department at (415) 424-8222 or (800) 662-2566,
extension 3 or E-mail TECH@CLONTECH.COM.
FEATURES Location/Qualifiers
source 1..5538
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 327..1154
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
promoter 1158..1249
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
misc_recomb 1337..2694
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
rep_origin 2841..3296
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3578..3682
/label=AmpR promoter
CDS 3683..4540
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4714..5302
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"