Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V006590 | pJL-TRBO-G | 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
Recommended culture condition: DH5α competent cells,kanamycin 50ug/ml,culture temperature 30℃(not 37℃)
- Vector Name:
- pJL-TRBO-G
- Antibiotic Resistance:
- Kanamycin
- Length:
- 11329 bp
- Type:
- Plant Expression
- Replication origin:
- oriV
- Selection Marker:
- Gentamicin ; Rifampicin
- Copy Number:
- Low Copy
pJL-TRBO-G 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.
pJL-TRBO-G vector Sequence
LOCUS V006590 11329 bp DNA circular SYN 13-MAY-2021 DEFINITION Exported. ACCESSION V006590 VERSION V006590 KEYWORDS pJL-TRBO-G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 11329) AUTHORS Lindbo JA TITLE TRBO: a high-efficiency tobacco mosaic virus RNA-based overexpression vector. JOURNAL Plant Physiol. 2007 Dec;145(4):1232-40. Epub 2007 Aug 24. PUBMED 17720752 REFERENCE 2 (bases 1 to 11329) TITLE Direct Submission REFERENCE 3 (bases 1 to 11329) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Physiol."; date: "2007-12"; volume: "145(4)"; pages: "1232-40. Epub 2007 Aug 24" SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..11329 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..46 /label="minimal CaMV 35S promoter" /note="minimal 35S promoter from cauliflower mosaic virus" misc_feature 58..113 /label="TMV Omega" /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" CDS 4949..5752 /gene="MP" /label="Movement protein" /note="Movement protein from Tobacco mosaic virus (strain vulgare). Accession#: P03583" CDS 5811..6527 /label="Cycle 3 GFP" /note="Cycle 3 GFP (Crameri et al., 1996)" misc_feature 7137..7161 /label="RB T-DNA repeat" /note="right border repeat from nopaline C58 T-DNA" CDS complement(7319..8464) /label="trfA" /note="trans-acting replication protein that binds to and activates oriV" CDS complement(8766..9557) /label="KanR" /note="aminoglycoside phosphotransferase" rep_origin complement(9766..10401) /direction=LEFT /label="oriV" /note="incP origin of replication" misc_feature 10481..10505 /label="LB T-DNA repeat" /note="left border repeat from nopaline C58 T-DNA" primer_bind 10586..10602 /label="KS primer" /note="common sequencing primer, one of multiple similar variants"