Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V011924 | ptdTomato-C1 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- ptdTomato-C1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5442 bp
- Type:
- Fluorescent Protein Genes & Plasmids
- Replication origin:
- ori
- Source/Author:
- Clontech
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
ptdTomato-C1 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.
ptdTomato-C1 vector Sequence
LOCUS ptdTomato-C1. 5442 bp DNA circular SYN 01-JAN-1980 DEFINITION Vector for fusing tdTomato to the N-terminus of a partner protein. ACCESSION . VERSION . KEYWORDS ptdTomato-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5442) AUTHORS Clontech TITLE Direct Submission REFERENCE 2 (bases 1 to 5442) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5442 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 613..2040 /label=tdTomato /note="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" misc_feature 2041..2106 /label=MCS /note="multiple cloning site" misc_feature 2115..2125 /label=stop codons /note="stop codons" /note="stop codons in all three reading frames" polyA_signal 2230..2351 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(2358..2813) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2840..2944 /label=AmpR promoter promoter 2946..3303 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3338..4129 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" polyA_signal 4364..4411 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4740..5328 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"