CaV2.1 (pcDNA6) vector (V017341)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017341 CaV2.1 (pcDNA6) 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:
CaV2.1 (pcDNA6)
Antibiotic Resistance:
Ampicillin
Length:
12495 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Selection Marker:
Blast
Promoter:
SV40
5' Primer:
CMV-F
Growth Temperature:
37℃

CaV2.1 (pcDNA6) vector Map

CaV2.1 (pcDNA6)12495 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000CMV enhancerCMV promoterT7 promoterVoltage-dependent P/Q-type calcium channel subunit alpha-1AV5 tag6xHisbGH poly(A) signalf1 oriSV40 promoterEM7 promoterBSDlac 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.

CaV2.1 (pcDNA6) vector Sequence

LOCUS       V017341                12495 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017341
VERSION     V017341
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12495)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        629..647
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             755..7273
                     /gene="Cacna1a"
                     /label="Voltage-dependent P/Q-type calcium channel subunit
                     alpha-1A"
                     /note="Voltage-dependent P/Q-type calcium channel subunit
                     alpha-1A from Rattus norvegicus. Accession#: P54282"
     CDS             8126..8167
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             8177..8194
                     /label="6xHis"
                     /note="6xHis affinity tag"
     polyA_signal    8223..8447
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8493..8921
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8935..9264
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        9312..9359
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             9378..9773
                     /label="BSD"
                     /note="blasticidin S deaminase"
     promoter        complement(10152..10182)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(10197..10218)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(10506..11094)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(11268..12125)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(12126..12230)
                     /label="AmpR promoter"