Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011649 | pFastBac Dual | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pFastBac Dual
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5238 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- Promoter:
- p10
pFastBac Dual 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.
pFastBac Dual vector Sequence
LOCUS pFastBac_Dual. 5238 bp DNA circular SYN 01-JAN-1980
DEFINITION Dual-promoter vector for co-expression of two proteins in insect
cells using the Bac-to-Bac(R) baculovirus system.
ACCESSION .
VERSION .
KEYWORDS pFastBac Dual
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5238)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5238)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Transform into DH10Bac(TM) E. coli cells.
FEATURES Location/Qualifiers
source 1..5238
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 102..557
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 584..688
/label=AmpR promoter
CDS 689..1546
/label=AmpR
/note="beta-lactamase"
rep_origin 1720..2308
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
mobile_element complement(2611..2835)
/label=Tn7R
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(2905..3435)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(3624..3652)
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal complement(4161..4209)
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
misc_feature 4274..4337
/label=MCS
/note="MCS"
/note="multiple cloning site"
promoter complement(4345..4454)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 4478..4569
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
misc_feature 4606..4704
/label=MCS
/note="MCS"
/note="multiple cloning site"
polyA_signal 4828..4962
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
mobile_element complement(4991..5156)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"