Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012859 | pHB201 | 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
10μg/ml Chloramphenicol in E.coli
- Vector Name:
- pHB201
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6603 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Bacillus subtilis
- Growth Strain(s):
- DH5B
- Growth Temperature:
- 37℃
pHB201 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.
pHB201 vector Sequence
LOCUS Exported 6603 bp DNA circular SYN 01-NOV-2024
DEFINITION Exported.
ACCESSION V012859
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6603)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6603)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6603
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1545..1563
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 1576..1683
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(1692..1710)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1720..1736)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 2082..2168
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2260..2287
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(2729..3376)
/gene="cat"
/label=Chloramphenicol acetyltransferase
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
CDS complement(5338..6069)
/gene="ermC"
/label=rRNA adenine N-6-methyltransferase
/note="rRNA adenine N-6-methyltransferase from
Staphylococcus aureus. Accession#: P02979"
rep_origin 6338..6603
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"