Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007025 | DP6 | 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:
- DP6
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 8639 bp
- Type:
- Bacterial Expression
- Replication origin:
- CloDF13 ori
- Copy Number:
- High Copy
- Promoter:
- araBAD
DP6 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.
DP6 vector Sequence
LOCUS V007025 8639 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007025
VERSION V007025
KEYWORDS DP6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8639)
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 8639)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8639)
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..8639
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..876
/label="araC"
/note="L-arabinose regulatory protein"
primer_bind complement(929..946)
/label="pBAD Reverse"
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 1099..1185
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1277..1304
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 1468..1486
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 1556..2827
/label="M13 gene III"
/note="pIII"
rep_origin 2987..3725
/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."
CDS 3878..4534
/label="CmR"
/note="chloramphenicol acetyltransferase"
terminator complement(4553..4584)
/label="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS complement(4614..5237)
/label="PmCDA1"
/note="cytidine deaminase from the sea lamprey Petromyzon
marinus (Rogozin et al., 2007)"
CDS complement(5270..5521)
/label="UGI"
/note="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
CDS complement(5553..6080)
/gene="mprA"
/label="Transcriptional repressor MprA"
/note="Transcriptional repressor MprA from Escherichia coli
(strain K12). Accession#: P0ACR9"
CDS complement(6112..6654)
/gene="seqA"
/label="Negative modulator of initiation of replication"
/note="Negative modulator of initiation of replication from
Escherichia coli (strain K12). Accession#: P0AFY8"
CDS complement(6686..7519)
/gene="dam"
/label="DNA adenine methylase"
/note="DNA adenine methylase from Escherichia coli (strain
K12). Accession#: P0AEE8"
CDS complement(7551..8279)
/gene="dnaQ"
/label="DNA polymerase III subunit epsilon"
/note="DNA polymerase III subunit epsilon from Escherichia
coli (strain K12). Accession#: P03007"
promoter complement(8329..8613)
/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)"