Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011647 | pFastBacHT A | 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:
- pFastBacHT A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4856 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- Promoter:
- polyhedrin
pFastBacHT A vector 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.
pFastBacHT A vector Sequence
LOCUS pFastBacHT_A. 4856 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector for expressing 6xHis-TEV-tagged proteins in insect cells
using the Bac-to-Bac(R) baculovirus system. For other reading
frames, use pFastBac(TM)HT B or pFastBac(TM)HT C.
ACCESSION .
VERSION .
KEYWORDS pFastBacHT A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4856)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4856)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Transform into DH10Bac(TM) E. coli cells.
FEATURES Location/Qualifiers
source 1..4856
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 2..457
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 484..588
/label=AmpR promoter
CDS 589..1446
/label=AmpR
/note="beta-lactamase"
rep_origin 1620..2208
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
mobile_element complement(2511..2735)
/label=Tn7R
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(2805..3335)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(3524..3552)
/label=Pc promoter
/note="class 1 integron promoter"
promoter 3904..3995
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
CDS 4050..4052
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 4062..4079
/label=6xHis
/note="6xHis affinity tag"
CDS 4101..4121
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
misc_feature 4119..4222
/label=MCS
/note="MCS"
/note="multiple cloning site"
polyA_signal 4346..4480
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
mobile_element complement(4509..4674)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"