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Beta-arrestin2 GFP WT vector (V000378)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000378 Beta-arrestin2 GFP WT In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
Beta-arrestin2 GFP WT
Antibiotic Resistance:
Kanamycin
Length:
5954 bp
Type:
Mammalian Expression
Replication origin:
ori
Copy Number:
High Copy
Promoter:
CMV
Cloning Method:
Restriction Enzyme
3' Primer:
EGFP-N

Beta-arrestin2 GFP WT vector Map

Copy Sequence View Map
Beta-arrestin2 GFP WT5954 bp60012001800240030003600420048005400CMV enhancerCMV promoterBeta-arrestin-2EGFPSV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRTK-pA-RHSV TK poly(A) signaloriCMV

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.

Beta-arrestin2 GFP WT vector Sequence

LOCUS       V000378                 5954 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000378
VERSION     V000378
KEYWORDS    Beta-arrestin2 GFP WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5954)
  AUTHORS   Shenoy SK, Lefkowitz RJ
  TITLE     Receptor-specific ubiquitination of beta-arrestin directs assembly
            and targeting of seven-transmembrane receptor signalosomes.
  JOURNAL   J Biol Chem. 2005 Apr 15;280(15):15315-24. Epub 2005 Feb 7.
   PUBMED   15699045
REFERENCE   2  (bases 1 to 5954)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5954)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
            Chem."; date: "2005-04-15"; volume: "280(15)"; pages: "15315-24.
            Epub 2005 Feb 7"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             583..1812
                     /gene="Arrb2"
                     /label="Beta-arrestin-2"
                     /note="Beta-arrestin-2 from Rattus norvegicus. Accession#:
                     P29067"
     CDS             1840..2556
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    2682..2803
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2810..3265)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3292..3396
                     /label="AmpR promoter"
     promoter        3398..3755
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3790..4581
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     primer_bind     complement(4772..4791)
                     /label="TK-pA-R"
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4816..4863
                     /label="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5192..5780
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5895..5954
                     /label="CMV"
                     /note="Human cytomegalovirus immediate early
                     enhancer/promoter"