Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V003154 | pSKI074 | 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
- Vector Name:
- pSKI074
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10450 bp
- Type:
- Activation tagging vector
- Replication origin:
- oriV
- Source/Author:
- Weigel D, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK, Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Wang Z, Xia Y, Dixon RA, Harrison MJ, Lamb CJ, Yanofs
- Promoter:
- MAS
pSKI074 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.
pSKI074 vector Sequence
LOCUS 40924_40587 10450 bp DNA circular SYN 18-DEC-2018
DEFINITION Activation tagging vector pSKI074, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10450)
AUTHORS Weigel D, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK,
Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM,
Nguyen JT, Sato S, Wang Z, Xia Y, Dixon RA, Harrison MJ, Lamb CJ,
Yanofsky MF, Chory J.
TITLE Activation tagging in Arabidopsis
JOURNAL Plant Physiol. 122 (4), 1003-1013 (2000)
PUBMED 10759496
REFERENCE 2 (bases 1 to 10450)
AUTHORS Weigel D, Blazquez MA, Borevitz J, Christensen SK, Fankhauser C,
Ferrandiz C, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Xia Y,
Wang Z, Dixon RA, Harrison MJ, Lamb CJ, Yanofsky MF, Chory J.
TITLE Direct Submission
JOURNAL Submitted (22-DEC-1999) Plant Biology Laboratory, The Salk Institute
for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA
92037, USA
REFERENCE 3 (bases 1 to 10450)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10450)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Physiol."; date: "2000"; volume: "122"; issue: "4"; pages:
"1003-1013"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-DEC-1999) Plant Biology Laboratory, The Salk Institute for
Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10450
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(13..31)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(52..68)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(76..92)
/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)."
promoter complement(100..130)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(145..166)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(454..1042)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1216..2073)
/label=AmpR
/note="beta-lactamase"
promoter complement(2074..2178)
/label=AmpR promoter
rep_origin complement(2207..2662)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
misc_feature 2663..2829
/label=derived from pUC19
/note="derived from pUC19"
primer_bind 2807..2823
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2830..2848
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 2881..2897
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
regulatory 2894..4178
/label=from cauliflower mosaic virus 35S gene promoter
/note="from cauliflower mosaic virus 35S gene promoter"
/regulatory_class="enhancer"
misc_feature 4270..4294
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
oriT complement(5497..5606)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin 6401..7113
/label=oriV
/note="incP origin of replication"
misc_feature 7700..7724
/label=LB T-DNA repeat
/note="left border repeat from octopine Ach5 T-DNA"
primer_bind complement(8045..8061)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 8362..8741
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
CDS 8771..9562
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
regulatory 9746..10430
/label=from mannopine synthase
/note="from mannopine synthase"
/regulatory_class="terminator"