Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V002370 | pUCP20T-mCherry | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pUCP20T-mCherry
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4878 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Barbier M, Damron FH.
- Growth Strain(s):
- Stbl3
pUCP20T-mCherry 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.
pUCP20T-mCherry vector Sequence
LOCUS 40924_45323 4878 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUCP20T-mCherry, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4878)
AUTHORS Barbier M, Damron FH.
TITLE Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling
JOURNAL PLoS ONE 11 (3), E0146827 (2016)
PUBMED 26937640
REFERENCE 2 (bases 1 to 4878)
AUTHORS Barbier M, Damron FH.
TITLE Direct Submission
JOURNAL Submitted (06-OCT-2015) Microbiology, Immunology and Cell Biology,
West Virginia University, School of Medicine, One Medical Center
Drive, Morgantown, WV 26506, USA
REFERENCE 3 (bases 1 to 4878)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4878)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2016"; volume: "11"; issue: "3"; pages: "E0146827"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-OCT-2015) Microbiology, Immunology and Cell Biology, West
Virginia University, School of Medicine, One Medical Center Drive,
Morgantown, WV 26506, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4878
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..200
/label=AmpR promoter
CDS 201..1058
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1232..1820
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
oriT complement(1892..2000)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
protein_bind 2384..2405
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2420..2450
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2458..2474
/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)."
primer_bind 2482..2498
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 2522..3229
/codon_start=1
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(3272..3288)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3452..4282)
/codon_start=1
/label=pRO1600 Rep
/note="replication protein for the broad-host-range plasmid
pRO1600 from Pseudomonas aeruginosa"
/translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
rep_origin 4296..4647
/label=pRO1600 oriV
/note="broad-host-range origin of replication from
Pseudomonas aeruginosa plasmid pRO1600; requires the
pRO1600 Rep protein for replication (West et al., 1994)"