Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012799 | pBBR1MCS5-Tac-EGFP | 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
pBBR1MCS vectors are small in size, contain unique cloning sites within the lacZα gene, and are mobilizable and compatible with various plasmid incompatibility groups.
- Vector Name:
- pBBR1MCS5-Tac-EGFP
- Antibiotic Resistance:
- Gentamicin
- Length:
- 5542 bp
- Type:
- Expression plasmid
- Replication origin:
- pBBR1 oriV
- Copy Number:
- Low copy number
- Promoter:
- tac
- Cloning Method:
- Enzyme Cut
- 3' Primer:
- M13 rev
- Fusion Tag:
- EGFP
- Growth Strain(s):
- JM108
- Growth Temperature:
- 37℃
pBBR1MCS5-Tac-EGFP 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
- Zhang BN, Qi B, Chu WK, Song F, Li S, Dong Q, Shao Z, Zhang B, Du X, Ma X, Jhanji V, Zhou Q. Norepinephrine as the Intrinsic Contributor to Contact Lens-Induced Pseudomonas aeruginosa Keratitis. Invest Ophthalmol Vis Sci. 2023 May 1;64(5):26. doi: 10.1167/iovs.64.5.26. PMID: 37234000; PMCID: PMC10226609.
pBBR1MCS5-Tac-EGFP vector Sequence
LOCUS Exported 5542 bp DNA circular SYN 03-SEP-2024
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pBBR1MCS5-Tac-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5542)
AUTHORS NOVOPRO
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5542)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5542
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 1023..1794
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS 1795..2454
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
primer_bind 3166..3182
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3192..3210
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 3243..3259
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 3261..3289
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS 3319..4035
/label=EGFP
/note="enhanced GFP"
primer_bind complement(4064..4080)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(4110..4128)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4149..4165)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4173..4189)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4197..4227)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4242..4263)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4533..4561
/label=Pc promoter
/note="class 1 integron promoter"
CDS 4750..5280
/label=GmR
/note="gentamycin acetyltransferase"