pDEST51-LRRK2-R1441G vector (V016655)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V016655	pDEST51-LRRK2-R1441G	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:: pDEST51-LRRK2-R1441G

Antibiotic Resistance:: Ampicillin

Length:: 13420 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells

Selection Marker:: Neo/G418

Promoter:: EF-1α

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

pDEST51-LRRK2-R1441G vector Vector Map

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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.

pDEST51-LRRK2-R1441G vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V016655                13420 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016655
VERSION     V016655
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 13420)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        475..1653
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1670..1688
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1720..1744
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1759..9339
                     /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(9356..9380)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9406..9447
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             9457..9474
                     /label="6xHis"
                     /note="6xHis affinity tag"
     polyA_signal    9503..9727
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        10215..10545
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        10593..10640
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             10659..11054
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    11215..11348
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(11433..11463)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(11478..11499)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(11787..12375)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(12549..13406)
                     /label="AmpR"
                     /note="beta-lactamase"