Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V000089 | pBV220 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pBV220 (Cat No.: V000089) is a high-performance E. coli expression vector engineered around the λ phage-derived PL and PR promoters, offering efficient and cost-effective gene expression control through its core features: it is equipped with λPL and λPR promoters that exhibit strong transcriptional initiation capability, regulated by a temperature-sensitive λ repressor (ts) which eliminates the need for chemical inducers, with temperature-dependent regulation where the active repressor inhibits PL/PR promoters at 30℃ and shifting the culture temperature to 42℃ inactivates the repressor to trigger target gene expression; the vector offers cloning and expression advantages such as a multi-cloning site (MCS) downstream of the SD sequence to facilitate insertion of foreign genes with an initiation ATG for expressing non-fusion proteins, and contains strong rrnB T1 and T2 transcription terminators to prevent read-through and enhance system stability.
- Vector Name:
- pBV220
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3665 bp
- Type:
- E. coli Expression Vectors
- Replication origin:
- ori
- Copy Number:
- High copy number
- Promoter:
- pR/pL
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- pBV220-F: 5′-AAGAAGGGCAGCATTCAAAG-3‘
- 3' Primer:
- pBV220-R: 5′-CTGCGTTCTGATTTAATCTG-3‘
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 30℃
pBV220 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
- Ma X, Liang X, Li Y, Feng Q, Cheng K, Ma N, Zhu F, Guo X, Yue Y, Liu G, Zhang T, Liang J, Ren L, Zhao X, Nie G. Modular-designed engineered bacteria for precision tumor immunotherapy via spatiotemporal manipulation by magnetic field. Nat Commun. 2023 Mar 23;14(1):1606.
pBV220 vector Sequence
LOCUS Exported 3665 bp DNA circular SYN 30-SEP-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3665)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3665)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 3665)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3665)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 3665)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3665
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(253..3665,1..252)
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 80..130
/label=pL promoter
/note="lambda PL"
terminator 490..576
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 668..695
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 714..805
/label=AmpR promoter
CDS 806..1663
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1837..2425
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2780..3490)
/codon_start=1
/label=lambda repressor (ts)
/note="temperature-sensitive variant of the phage lambda
repressor"
/translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
VGKVIASQWPEETFG"
promoter 3531..3580
/label=pR promoter
/note="lambda PR"