CaV1.2 vector (V000220)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000220 CaV1.2 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:
CaV1.2
Antibiotic Resistance:
Ampicillin
Length:
11709 bp
Type:
Mammalian Expression
Replication origin:
ori
Promoter:
CMV
Cloning Method:
Restriction Enzyme
5' Primer:
T7
3' Primer:
BGH-rev

CaV1.2 vector Map

CaV1.211709 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500CAP binding sitelac promoterIn lacZ geneSV40 poly(A) signalBSDEM7 promoterSV40 promoterpBABE 3'f1 oribGH poly(A) signal6xHisV5 tagVoltage-dependent L-type calcium channel subunitalpha-1CT7 promoterCMV promoterCMV enhancerpRS-markerAmpR promoterAmpRoriL4440

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.

CaV1.2 vector Sequence

Copy Sequence

Download GenBank File(.gb)

LOCUS       V000220                11709 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000220
VERSION     V000220
KEYWORDS    CaV1.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11709)
  AUTHORS   Helton TD, Xu W, Lipscombe D
  TITLE     Neuronal L-type calcium channels open quickly and are inhibited
            slowly.
  JOURNAL   J Neurosci. 2005 Nov 2. 25(44):10247-51.
   PUBMED   16267232
REFERENCE   2  (bases 1 to 11709)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 11709)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Neurosci.
            2005 Nov 2. 25(44):10247-51."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label="M13/pUC Reverse"
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     205..221
                     /label="M13 Reverse"
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    complement(258..391)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             complement(552..947)
                     /label="BSD"
                     /note="blasticidin S deaminase"
     promoter        complement(966..1013)
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     promoter        complement(1069..1279)
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     primer_bind     1303..1323
                     /label="pBABE 3'"
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1332..1760)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1442..1463)
                     /label="F1ori-F"
                     /note="F1 origin, forward primer"
     primer_bind     1654..1673
                     /label="F1ori-R"
                     /note="F1 origin, reverse primer"
     polyA_signal    complement(1806..2030)
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2059..2076)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(2086..2127)
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             complement(2332..8748)
                     /gene="Cacna1c"
                     /label="Voltage-dependent L-type calcium channel subunit
                     alpha-1C"
                     /note="Voltage-dependent L-type calcium channel subunit
                     alpha-1C from Mus musculus. Accession#: Q01815"
     promoter        complement(8892..8910)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8955..9158)
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(9159..9538)
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9710..9729
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     promoter        9804..9908
                     /label="AmpR promoter"
     CDS             9909..10766
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      10940..11528
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     11682..11699
                     /label="L4440"
                     /note="L4440 vector, forward primer"