Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V011758 | pBluescript II KS(+) | In stock, 1 week for quality controls |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pBluescript II KS(+)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 2961 bp
- Type:
- I.M.A.G.E. Consortium Plasmids
- Replication origin:
- ori
- Source/Author:
- Alting-Mees MA, Short JM.
- Copy Number:
- High copy number
- Promoter:
- T3
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
pBluescript II KS(+) 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.
pBluescript II KS(+) vector Sequence
LOCUS Exported 2961 bp DNA circular SYN 30-SEP-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pBluescript II KS(+)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2961)
AUTHORS 11111111
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2961
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(839..2961,1..838)
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(2932..2961,1..2931)
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 185..206
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 221..251
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 259..275
/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)."
primer_bind 283..299
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 295..870
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
FPRQALNRGLPLGFRFSALRHLDPKKLD"
promoter 320..338
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 351..458
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 368..384
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(418..434)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 467..485
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(495..511)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 653..1108
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1134..1238
/gene="bla"
/label=AmpR promoter
CDS 1239..2099
/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"
rep_origin 2270..2858
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"