Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007549 | pcDNA3-HA2-Keap1 | 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:
- pcDNA3-HA2-Keap1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7314 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- G418
- Copy Number:
- High Copy
- Promoter:
- SV40
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7
- 3' Primer:
- SP6
pcDNA3-HA2-Keap1 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.
pcDNA3-HA2-Keap1 vector Sequence
LOCUS V007549 7314 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007549
VERSION V007549
KEYWORDS pcDNA3-HA2-Keap1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7314)
AUTHORS Furukawa M, Xiong Y
TITLE BTB protein Keap1 targets antioxidant transcription factor Nrf2 for
ubiquitination by the Cullin 3-Roc1 ligase.
JOURNAL Mol Cell Biol. 2005 Jan . 25(1):162-71.
PUBMED 15601839
REFERENCE 2 (bases 1 to 7314)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7314)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell
Biol. 2005 Jan . 25(1):162-71."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7314
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(63..80)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(234..822)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(996..1853)
/label="AmpR"
/note="beta-lactamase"
promoter complement(1854..1958)
/label="AmpR promoter"
primer_bind complement(2033..2052)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2224..2603
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2604..2807
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 2852..2870
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2886..2912
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 2922..2948
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 2970..4841
/gene="KEAP1"
/label="Kelch-like ECH-associated protein 1"
/note="Kelch-like ECH-associated protein 1 from Homo
sapiens. Accession#: Q14145"
promoter complement(4856..4874)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 4900..5124
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 5170..5598
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5612..5941
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 6008..6799
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
polyA_signal 6976..7109
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(7146..7162)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(7170..7186)
/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(7194..7224)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(7239..7260)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."