Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004990 | pLL29 | 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:
- pLL29
- Antibiotic Resistance:
- Streptomycin
- Length:
- 7029 bp
- Type:
- Single-copy integtation vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Luong TT, Lee CY.
pLL29 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.
pLL29 vector Sequence
LOCUS V004990 7029 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004990
VERSION V004990
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7029)
AUTHORS Luong TT, Lee CY.
TITLE Improved single-copy integration vectors for Staphylococcus aureus
JOURNAL J. Microbiol. Methods 70 (1), 186-190 (2007)
PUBMED 17512993
REFERENCE 2 (bases 1 to 7029)
AUTHORS Luong TT, Lee CY.
TITLE Direct Submission
JOURNAL Submitted (06-APR-2007) Microbiology and Immunology, Univ of
Arkansas for Medical Sciences, 4301 W. Markham St. Mail Slot 511,
Little Rock, AR 72205, USA
REFERENCE 3 (bases 1 to 7029)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7029)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J.
Microbiol. Methods"; date: "2007"; volume: "70"; issue: "1"; pages:
"186-190"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-APR-2007) Microbiology and Immunology, Univ of Arkansas for
Medical Sciences, 4301 W. Markham St. Mail Slot 511, Little Rock, AR
72205, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7029
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..57
/label="MCS"
/note="pUC18/19 multiple cloning site"
CDS complement(715..2091)
/gene="tet"
/label="Tetracycline resistance protein"
/note="Tetracycline resistance protein from Staphylococcus
aureus. Accession#: P02983"
CDS complement(3288..4076)
/label="SmR"
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
CDS complement(5331..6278)
/label="Rep101"
/note="RepA protein needed for replication with the pSC101
origin"
rep_origin complement(6326..6548)
/direction=LEFT
/label="pSC101 ori"
/note="low-copy replication origin that requires the Rep101
protein"