Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V013944 | pFastBac1-6×His-MBP-MCS | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pFastBac1-6×His-MBP-MCS vector enables high-level expression of recombinant proteins in insect cells using the baculovirus expression system. Its key features include:
Expression: Utilizes the strong polyhedrin promoter for high protein yields
Solubility Enhancement: An MBP (Maltose Binding Protein) tag significantly improves solubility and stability of difficult-to-express proteins.
Purification & Detection: Contains a 6×His tag for simple, one-step IMAC purification (e.g., Ni-NTA). The MBP tag allows additional amylose affinity purification and aids detection.
- Vector Name:
- pFastBac1-6×His-MBP-MCS
- Antibiotic Resistance:
- Ampicillin, Gentamycin
- Length:
- 5915 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Insect cells
- Promoter:
- polyhedrin
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pFastBac1-6×His-MBP-MCS 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.
pFastBac1-6×His-MBP-MCS vector Sequence
LOCUS Exported 5915 bp DNA circular SYN 14-JUL-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5915)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5915)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5915
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 2..456
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 482..586
/label=AmpR promoter
CDS 587..1444
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1618..2206
/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)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
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 4041..4058
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 4059..5156
/codon_start=1
/label=MBP
/note="maltose binding protein from E. coli"
/translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPAAAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYAAGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEHAFNHGET
AMTINGPWAWSNIDTSAVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRVAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDAALAAAQT"
CDS 5160..5180
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
polyA_signal 5405..5539
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
mobile_element complement(5568..5733)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"