Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012786 | pHP13-P43 | 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
Promoter 43 is a constitutive promoter that constitutively expresses the P43 protein in B.subtilis. This promoter has been shown to be recognized and active during the exponential and lag phases of growth. It has been hypothesized that the ability to recognize the promoter in exponential and lag phase of growth is due to the recognition of the promoter by both sigma factor 55 (the major sigma factor A) and sigma factor 37 (the lag phase sigma factor B) 1. The P43 promoter has been previously used for constitutive expression of exogenous genes within B.subtilis vectors 2. The context with which we used the promoter P43 is as a Polymerase Per Second (PoPS) generator.
- Vector Name:
- pHP13-P43
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 5100 bp
- Type:
- B. subtilis Expression vector
- Replication origin:
- ori
- Copy Number:
- High copy number
- Promoter:
- P43
- Cloning Method:
- Enzyme Cut
pHP13-P43 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.
pHP13-P43 vector Sequence
LOCUS V012786 5100 bp DNA circular SYN 17-NOV-2021
DEFINITION Exported.
ACCESSION V012786
VERSION V012786
KEYWORDS pHP13-P43
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 5100)
AUTHORS Jakobsen OM, Benichou A, Flickinger MC, Valla S, Ellingsen TE,
Brautaset T.
TITLE Upregulated transcription of plasmid and chromosomal ribulose
monophosphate pathway genes is critical for methanol assimilation
rate and methanol tolerance in the methylotrophic bacterium Bacillus
methanolicus
JOURNAL J. Bacteriol. 188 (8), 3063-3072 (2006)
PUBMED 16585766
REFERENCE 2 (bases 1 to 5100)
AUTHORS Jakobsen OM, Benichou A, Flickinger MC, Valla S, Ellingsen TE,
Brautaset T.
TITLE Direct Submission
JOURNAL Submitted (18-NOV-2005) Department of Biotechnology, Norwegian
University of Science and Technology, Sem Saelands vei 6/8,
Trondheim 7491, Norway
REFERENCE 3 (bases 1 to 5100)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5100)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J.
Bacteriol."; date: "2006"; volume: "188"; issue: "8"; pages:
"3063-3072"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-NOV-2005) Department of Biotechnology, Norwegian University of
Science and Technology, Sem Saelands vei 6/8, Trondheim 7491,
Norway"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5100
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 565..1153
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 1441..1462
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1477..1507
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1515..1531
/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 1539..1555
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1956..1972)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2184..2831)
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
CDS complement(join(4665..5100,1..296))
/gene="ermC"
/label="rRNA adenine N-6-methyltransferase"
/note="rRNA adenine N-6-methyltransferase from
Staphylococcus aureus. Accession#: P02979"