Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V018110 | pBSM | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pBSM, as an expression vector, serves primarily to carry the tRNA scaffold and target RNA sequence, driving recombinant RNA expression in Escherichia coli while facilitating subsequent purification.
- Vector Name:
- pBSM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3089 bp
- Type:
- Bacterial Expression
- Source/Author:
- Luc Ponchon
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pBSM 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
- Ponchon L, Beauvais G, Nonin-Lecomte S, Dardel F. A generic protocol for the expression and purification of recombinant RNA in Escherichia coli using a tRNA scaffold. Nat Protoc. 2009;4(6):947-59. doi: 10.1038/nprot.2009.67. Epub 2009 May 28. PMID: 19478810.
pBSM vector Sequence
LOCUS Exported 3089 bp DNA circular SYN 23-AUG-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBSM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3089)
AUTHORS Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte
S, Tisne C
TITLE Co-expression of RNA-protein complexes in Escherichia coli and
applications to RNA biology.
JOURNAL Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576.
Epub 2013 Jun 26.
PUBMED 23804766
REFERENCE 2 (bases 1 to 3089)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2013-08"; volume: "41(15):e150. doi"; pages: "
10.1093/nar/gkt576. Epub 2013 Jun 26"
FEATURES Location/Qualifiers
source 1..3089
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(279..297)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(318..334)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(318..334)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(331..353)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 342..358
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
promoter complement(366..396)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 411..432
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(549..566)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(720..1308)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(800..819)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1479..2339)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2102..2121
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2340..2444)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2470..2925)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2607..2628)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2819..2838
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3052..3074
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3066..3083
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3067..3083
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"