Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007450 | HA-p62 | 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:
- HA-p62
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6414 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Zeocin
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- BGH-rev
HA-p62 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.
HA-p62 vector Sequence
LOCUS V007450 6414 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007450
VERSION V007450
KEYWORDS HA-p62
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6414)
AUTHORS Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q
TITLE Keap1 facilitates p62-mediated ubiquitin aggregate clearance via
autophagy.
JOURNAL Autophagy. 2010 Jul 22. 6(5):.
PUBMED 20495340
REFERENCE 2 (bases 1 to 6414)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6414)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Autophagy.
2010 Jul 22. 6(5):."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6414
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 205..221
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal complement(258..391)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(524..895)
/label="BleoR"
/note="antibiotic-binding protein"
promoter complement(914..961)
/label="EM7 promoter"
/note="synthetic bacterial promoter"
promoter complement(1009..1338)
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin complement(1352..1780)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal complement(1826..2050)
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
CDS complement(2097..2123)
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS complement(2133..3452)
/gene="SQSTM1"
/label="Sequestosome-1"
/note="Sequestosome-1 from Homo sapiens. Accession#:
Q13501"
primer_bind complement(3585..3609)
/label="LNCX"
/note="Human CMV promoter, forward primer"
protein_bind 3619..3637
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind complement(3640..3658)
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter complement(3660..3863)
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(3864..4243)
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
primer_bind 4415..4434
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4509..4613
/label="AmpR promoter"
CDS 4614..5471
/label="AmpR"
/note="beta-lactamase"
rep_origin 5645..6233
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6387..6404
/label="L4440"
/note="L4440 vector, forward primer"