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pCDNA3-HA-FLAG-RBP2 vector (V016638)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016638 pCDNA3-HA-FLAG-RBP2 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:
pCDNA3-HA-FLAG-RBP2
Antibiotic Resistance:
Ampicillin
Length:
11774 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Selection Marker:
Neo/G418
Promoter:
SP6
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pCDNA3-HA-FLAG-RBP2 vector Map

Copy Sequence View Map
pCDNA3-HA-FLAG-RBP211774 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500CMV enhancerCMV promoterT7 promoterHAFLAGLysine-specific demethylase 5ASP6 promoterbGH poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

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-HA-FLAG-RBP2 vector Sequence

LOCUS       V016638                11774 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016638
VERSION     V016638
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11774)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..933
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             952..975
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1021..5886
                     /gene="KDM5A"
                     /label="Lysine-specific demethylase 5A"
                     /note="Lysine-specific demethylase 5A from Homo sapiens.
                     Accession#: P29375"
     promoter        complement(7326..7344)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    7370..7594
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7640..8068
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8082..8412
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             8479..9270
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    9447..9580
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(9665..9695)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9710..9731)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(10019..10607)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10781..11638)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(11639..11743)
                     /label="AmpR promoter"