Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V008759 | pCAMBIA1380 | 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
Agrobacterium binary vector for plant transformation, with hygromycin- and kanamycin-resistance genes.
- Vector Name:
- pCAMBIA1380
- Antibiotic Resistance:
- Kanamycin
- Length:
- 8860 bp
- Type:
- Binary vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Hajdukiewicz P, Svab Z, Maliga P.
- Promoter:
- CaMV 35S (enhanced)
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pCAMBIA1380 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
- Beernink BM, Holan KL, Lappe RR, Whitham SA. Direct Agroinoculation of Maize Seedlings by Injection with Recombinant Foxtail Mosaic Virus and Sugarcane Mosaic Virus Infectious Clones. J Vis Exp. 2021 Feb 27;(168).
pCAMBIA1380 vector Sequence
LOCUS 40924_8736 8860 bp DNA circular SYN 17-DEC-2018 DEFINITION Binary vector pCAMBIA-1380, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8860) AUTHORS Hajdukiewicz P, Svab Z, Maliga P. TITLE The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation JOURNAL Plant Mol. Biol. 25 (6), 989-994 (1994) PUBMED 7919218 REFERENCE 2 (bases 1 to 8860) AUTHORS Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A, Jefferson RA. TITLE A comprehensive set of modular vectors for advanced manipulations and efficient transformation of plants JOURNAL Unpublished REFERENCE 3 (bases 1 to 8860) AUTHORS Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A, Jefferson RA. TITLE Direct Submission JOURNAL Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / GPO Box 3200, Canberra, ACT 2601, Australia REFERENCE 4 (bases 1 to 8860) TITLE Direct Submission REFERENCE 5 (bases 1 to 8860) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol. Biol."; date: "1994"; volume: "25"; issue: "6"; pages: "989-994" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / GPO Box 3200, Canberra, ACT 2601, Australia" COMMENT SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..8860 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 6..23 /label=6xHis /note="6xHis affinity tag" terminator 58..310 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature 332..356 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 1656..2282 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS 2719..3783 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" rep_origin 3852..4046 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 4390..4530 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(4716..5304) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5394..6185) /label=KanR /note="aminoglycoside phosphotransferase" misc_feature 6610..6634 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal complement(6712..6886) /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(6929..7951) /label=HygR /note="aminoglycoside phosphotransferase from E. coli" promoter complement(8019..8696) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" misc_feature complement(8785..8821) /note="pUC8 MCS; polylinker"