Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000861 | pMS2-GFP | 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:
- pMS2-GFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6019 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- M13_Rev
pMS2-GFP 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.
pMS2-GFP vector Sequence
LOCUS V000861 6019 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000861
VERSION V000861
KEYWORDS pMS2-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6019)
AUTHORS Fusco D, Accornero N, Lavoie B, Shenoy SM, Blanchard JM, Singer RH,
Bertrand E
TITLE Single mRNA molecules demonstrate probabilistic movement in living
mammalian cells.
JOURNAL Curr Biol. 2003 Jan 21. 13(2):161-7.
PUBMED 12546792
REFERENCE 2 (bases 1 to 6019)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6019)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr Biol.
2003 Jan 21. 13(2):161-7."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6019
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 750..776
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 777..788
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 1179..1895
/label="EGFP"
/note="enhanced GFP"
CDS 1896..1916
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label="SV40 NLS"
/translation="PKKKRKV"
intron 1975..2547
/label="beta-globin intron"
/note="intron from rabbit beta-globin gene"
CDS 2551..2673
/gene="HBB"
/label="Hemoglobin subunit beta"
/note="Hemoglobin subunit beta from Colobus guereza.
Accession#: Q9TT33"
primer_bind complement(2679..2698)
/label="Bglob-pA-R"
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2744..2799
/label="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2798..2817)
/label="rbglobpA-R"
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
promoter complement(3154..3172)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3179..3195)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3337..3792
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3818..3922
/label="AmpR promoter"
CDS 3923..4780
/label="AmpR"
/note="beta-lactamase"
rep_origin 4954..5542
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5696..5713
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 5830..5851
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5866..5896
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5904..5920
/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 5928..5944
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 5965..5983
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"