Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000976 | pSMT3-M | 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:
- pSMT3-M
- Length:
- 5708 bp
- Type:
- Shuttle vector for mycobacteria
- Replication origin:
- ori
- Selection Marker:
- Hygromycin
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
pSMT3-M 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.
pSMT3-M vector Sequence
LOCUS V000976 5708 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000976
VERSION V000976
KEYWORDS pSMT3-M
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 5708)
AUTHORS Carroll P, Schreuder LJ, Muwanguzi-Karugaba J, Wiles S, Robertson
BD, Ripoll J, Ward TH, Bancroft GJ, Schaible UE, Parish T
TITLE Sensitive detection of gene expression in mycobacteria under
replicating and non-replicating conditions using optimized far-red
reporters.
JOURNAL PLoS One. 2010 . 5(3):e9823.
PUBMED 20352111
REFERENCE 2 (bases 1 to 5708)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5708)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.
2010 . 5(3):e9823."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5708
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(436..452)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3307..4302)
/gene="hyg"
/label="Hygromycin-B 7''-O-kinase"
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
rep_origin 4672..5260
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5414..5431
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 5548..5569
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5584..5614
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5622..5638
/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 5646..5662
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"