Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000803 | pAX01 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
For protein expression in Bacillus subtilis with a xylose-inducible promoter (xylA).
- Vector Name:
- pAX01
- Antibiotic Resistance:
- Ampicillin, Erythromycin
- Length:
- 7781 bp
- Type:
- Bacillus subtilis expression vectors
- Replication origin:
- ori
- Selection Marker:
- Erythromycin
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pAX01 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.
References
- Härtl B, Wehrl W, Wiegert T, Homuth G, Schumann W. 2001. Development of a New Integration Site within the Bacillus subtilis Chromosome and Construction of Compatible Expression Cassettes. J Bacteriol 183:.https://doi.org/10.1128/jb.183.8.2696-2699.2001
pAX01 vector Sequence
LOCUS V000803 7781 bp DNA circular SYN 13-JAN-2022
DEFINITION Exported.
ACCESSION V000803
VERSION V000803
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7781)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7781)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7781
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 546..580
/note="lambda t0 terminator"
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
terminator complement(660..687)
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(706..792)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(965..1699)
/gene="ermBP"
/label="rRNA adenine N-6-methyltransferase"
/note="rRNA adenine N-6-methyltransferase from Enterococcus
faecalis. Accession#: P0A4D5"
promoter 2078..2182
/label="AmpR promoter"
terminator complement(3674..3708)
/label="lambda t0 terminator"
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
promoter 4352..4456
/gene="bla"
/label="bla promoter"
/note="AmpR promoter"
CDS 4457..5314
/label="AmpR"
/note="beta-lactamase"
rep_origin 5488..6076
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(6262..6402)
/label="bom"
/note="basis of mobility region from pBR322"
CDS complement(6507..6695)
/label="rop"
/note="Rop protein, which maintains plasmids at low copy
number"