Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010549 | pLI50 | 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:
- pLI50
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5505 bp
- Type:
- S. aureus-E. coli shuttle vector
- Replication origin:
- ori
- Selection Marker:
- Chloramphenicol in S. aureus at 37C
- Cloning Method:
- Restriction Enzyme
pLI50 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.
pLI50 vector Sequence
LOCUS V010549 5505 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V010549
VERSION V010549
KEYWORDS pLI50
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 5505)
AUTHORS Lee CY, Buranen SL, Ye ZH
TITLE Construction of single-copy integration vectors for Staphylococcus
aureus.
JOURNAL Gene. 1991 Jul 15. 103(1):101-5.
PUBMED 1652539
REFERENCE 2 (bases 1 to 5505)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5505)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene. 1991
Jul 15. 103(1):101-5."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5505
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..57
/label="MCS"
/note="pUC18/19 multiple cloning site"
CDS complement(585..1232)
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
CDS 1737..2738
/label="repB"
/note="RepB replication protein"
primer_bind complement(3247..3269)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
misc_feature 3355..3495
/label="bom"
/note="basis of mobility region from pBR322"
primer_bind complement(3510..3527)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(3681..4269)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4443..5300)
/label="AmpR"
/note="beta-lactamase"
promoter complement(5301..5405)
/label="AmpR promoter"
primer_bind 5473..5491
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"