pAAV-UBC-S2-cAMPr vector (V017216)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017216 pAAV-UBC-S2-cAMPr 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:
pAAV-UBC-S2-cAMPr
Antibiotic Resistance:
Ampicillin
Length:
7742 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Adeno-associated virus
Promoter:
UbC
Growth Temperature:
37℃

pAAV-UBC-S2-cAMPr vector Map

pAAV-UBC-S2-cAMPr7742 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500UbC promoter3-methyl-2-oxobutanoate hydroxymethyltransferasecAMP-dependent protein kinase catalytic subunit alphaHAWPRESV40 poly(A) signaloriAmpRAmpR promoterf1 ori

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.

pAAV-UBC-S2-cAMPr vector Sequence

LOCUS       V017216                 7742 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017216
VERSION     V017216
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7742)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             414..1205
                     /gene="panB"
                     /label="3-methyl-2-oxobutanoate hydroxymethyltransferase"
                     /note="3-methyl-2-oxobutanoate hydroxymethyltransferase
                     from Escherichia coli (strain ATCC 8739 / DSM 1576 / NBRC
                     3972 / NCIMB 8545 / WDCM 00012 / Crooks). Accession#:
                     B1IQK6"
     CDS             1356..2408
                     /gene="Prkaca"
                     /label="cAMP-dependent protein kinase catalytic subunit
                     alpha"
                     /note="cAMP-dependent protein kinase catalytic subunit
                     alpha from Mus musculus. Accession#: P05132"
     CDS             3633..3659
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    3984..4572
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(4615..4736)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5264..5852)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6026..6883)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6884..6988)
                     /label="AmpR promoter"
     rep_origin      complement(7014..7469)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"