Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012134 | 2XMyc-LRRK2-WT | 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:
- 2XMyc-LRRK2-WT
- Antibiotic Resistance:
- Kanamycin
- Length:
- 11985 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- SV40pA-R
2XMyc-LRRK2-WT 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.
2XMyc-LRRK2-WT vector Sequence
LOCUS V012134 11985 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012134
VERSION V012134
KEYWORDS 2XMyc-LRRK2-WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11985)
TITLE The Parkinson Disease-associated Leucine-rich Repeat Kinase 2
(LRRK2) Is a Dimer That Undergoes Intramolecular Autophosphorylation
JOURNAL J Biol Chem. 2008 Jun 13;283(24):16906-14.
PUBMED 18397888
REFERENCE 2 (bases 1 to 11985)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11985)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem."; date: "2008-06-13"; volume: "283(24)"; pages: "16906-14"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11985
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 68..371
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 621..639
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 671..680
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 680..709
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 725..754
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
protein_bind 773..797
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 812..8392
/gene="LRRK2"
/label="Leucine-rich repeat serine/threonine-protein kinase
2"
/note="Leucine-rich repeat serine/threonine-protein kinase
2 from Homo sapiens. Accession#: Q5S007"
protein_bind complement(8414..8438)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(8488..8506)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 8780..8901
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(8908..9363)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 9390..9492
/label="AmpR promoter"
promoter 9496..9853
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 9888..10679
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
primer_bind complement(10870..10889)
/label="TK-pA-R"
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 10914..10961
/label="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 11290..11878
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"