Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000961 | pMIND | 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:
- pMIND
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6778 bp
- Type:
- Bacterial Expression
- Replication origin:
- ori
- Selection Marker:
- Hygromycin
- Copy Number:
- Low Copy
- Cloning Method:
- Restriction Enzyme
pMIND vector 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.
pMIND vector Sequence
LOCUS V000961 6778 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000961
VERSION V000961
KEYWORDS pMIND
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6778)
AUTHORS Blokpoel MC, Murphy HN, O'Toole R, Wiles S, Runn ES, Stewart GR,
Young DB, Robertson BD
TITLE Tetracycline-inducible gene regulation in mycobacteria.
JOURNAL Nucleic Acids Res. 2005 . 33(2):e22.
PUBMED 15687380
REFERENCE 2 (bases 1 to 6778)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6778)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. 2005 . 33(2):e22."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6778
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 19..35
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 908..1903
/gene="hyg"
/label="Hygromycin-B 7''-O-kinase"
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
primer_bind complement(1916..1932)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS 4745..5536
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
rep_origin 5823..6411
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6565..6582
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 6699..6720
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6735..6765
/label="lac promoter"
/note="promoter for the E. coli lac operon"