Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V006635 | MP6 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- MP6
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6681 bp
- Type:
- Bacterial Expression
- Replication origin:
- CloDF13 ori
- Copy Number:
- High Copy
- Promoter:
- araBAD
- Cloning Method:
- Ligation Independent Cloning
- 5' Primer:
- pBAD-F
- 3' Primer:
- CAT-F
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
MP6 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.
MP6 vector Sequence
LOCUS V006635 6681 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006635
VERSION V006635
KEYWORDS MP6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6681)
AUTHORS Badran AH, Liu DR
TITLE Development of potent in vivo mutagenesis plasmids with broad
mutational spectra.
JOURNAL Nat Commun. 2015 Oct 7;6:8425. doi: 10.1038/ncomms9425.
PUBMED 26443021
REFERENCE 2 (bases 1 to 6681)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6681)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2015-10-7"; pages: "
10.1038/ncomms9425"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6681
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..285
/label="araBAD promoter"
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 335..1063
/gene="dnaQ"
/label="DNA polymerase III subunit epsilon"
/note="DNA polymerase III subunit epsilon from Escherichia
coli (strain K12). Accession#: P03007"
CDS 1095..1928
/gene="dam"
/label="DNA adenine methylase"
/note="DNA adenine methylase from Escherichia coli (strain
K12). Accession#: P0AEE8"
CDS 1960..2502
/gene="seqA"
/label="Negative modulator of initiation of replication"
/note="Negative modulator of initiation of replication from
Escherichia coli (strain K12). Accession#: P0AFY8"
CDS 2534..3061
/gene="mprA"
/label="Transcriptional repressor MprA"
/note="Transcriptional repressor MprA from Escherichia coli
(strain K12). Accession#: P0ACR9"
CDS 3093..3344
/label="UGI"
/note="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
CDS 3377..4000
/label="PmCDA1"
/note="cytidine deaminase from the sea lamprey Petromyzon
marinus (Rogozin et al., 2007)"
terminator complement(4030..4061)
/label="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS complement(4080..4736)
/label="CmR"
/note="chloramphenicol acetyltransferase"
rep_origin complement(4889..5627)
/direction=LEFT
/label="CloDF13 ori"
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
terminator complement(5720..5763)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(5780..6655)
/label="araC"
/note="L-arabinose regulatory protein"