Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010143 | CMV-ER-LAR-GECO1 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- CMV-ER-LAR-GECO1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4517 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- TAATACGACTCACTATAGGG
- 3' Primer:
- TAGAAGGCACAGTCGAGG
CMV-ER-LAR-GECO1 vector Map
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.
CMV-ER-LAR-GECO1 vector Sequence
LOCUS 40924_500 4517 bp DNA circular SYN 13-MAY-2021
DEFINITION Expresses LAR-GECO1 in the endoplasmic reticulum in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4517)
AUTHORS Wu J, Prole DL, Shen Y, Lin Z, Gnanasekaran A, Liu Y, Chen L, Zhou
H, Chen SR, Usachev YM, Taylor CW, Campbell RE
TITLE Red fluorescent genetically encoded Ca2+ indicators for use in
mitochondria and endoplasmic reticulum.
JOURNAL Biochem J. 2014 Nov 15;464(1):13-22. doi: 10.1042/BJ20140931.
PUBMED 25164254
REFERENCE 2 (bases 1 to 4517)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4517)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName:
"Biochem J"; date: "2014-11-15- 15"; volume: "464"; issue: "1";
pages: "13-22"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4517
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal complement(42..266)
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 292..310
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS complement(386..1633)
/codon_start=1
/label=CAR-GECO1
/note="CAR-GECO1 is a basic (constitutively fluorescent)
red fluorescent protein. It has high acid sensitivity."
/translation="VDSSRRKWNKAGHAWRAIGRLSSPVVSERMYPEDGALKSEIKKGL
RLKDGGHYAAEVKTTYKAKKPVQLPGAYVVDIKLDIVSHNEDYTIVEQCERAEGRHSTG
GMDELYKGGTGGSLVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAF
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYIKHPADIPDYFKLSFPEGFRWERVMNFE
DGGIIHVNQDSSLQDGVFIYKVKLRGTNFPPDGPVMQKKTMGWEATRDQLTEEQIAEFK
EAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMISEVDADGDGTFDFPEFLTMM
ARKMNYTDSEEEIREAFRVADKDGNGYIGAAELRHAMTDIGEKLTDEEVDEMIRVADID
GDGQVNYEEFVQMMTAK"
promoter complement(1722..1740)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1785..1988)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(1989..2368)
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 2540..2559
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2634..2738
/label=AmpR promoter
CDS 2739..3596
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3770..4358
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"