Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V018116 | CMV-R-GECO1.2 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
It contains a Red intensiometric genetically encoded Ca2+-indicators for optical imaging 1.2
- Vector Name:
- CMV-R-GECO1.2
- Antibiotic Resistance:
- Ampicillin, 100 μg/mL
- Length:
- 4455 bp
- Type:
- Mammalian Expression Vectors
- Source/Author:
- Wu J, Liu
- Copy Number:
- High copy number
- Promoter:
- high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
CMV-R-GECO1.2 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.
References
- Wu J, Liu L, Matsuda T, Zhao Y, Rebane A, Drobizhev M, Chang YF, Araki S, Arai Y, March K, Hughes TE, Sagou K, Miyata T, Nagai T, Li WH, Campbell RE. Improved orange and red Ca²± indicators and photophysical considerations for optogenetic applications. ACS Chem Neurosci. 2013 Jun 19;4(6):963-72. doi: 10.1021/cn400012b. Epub 2013 Mar 19. PMID: 23452507; PMCID: PMC3689190.
CMV-R-GECO1.2 vector Sequence
LOCUS Exported 4455 bp DNA circular SYN 19-SEP-2025
DEFINITION Red intensiometric genetically encoded Ca2+-indicators for optical
imaging 1.2.
ACCESSION .
VERSION .
KEYWORDS CMV-R-GECO1.2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4455)
AUTHORS Wu J, Liu L, Matsuda T, Zhao Y, Rebane A, Drobizhev M, Chang YF,
Araki S, Arai Y, March K, Hughes TE, Sagou K, Miyata T, Nagai T, Li
WH, Campbell RE
TITLE Improved Orange and Red Ca Indicators and Photophysical
Considerations for Optogenetic Applications.
JOURNAL ACS Chem Neurosci. 2013 Mar 19.
PUBMED 23452507
REFERENCE 2 (bases 1 to 4455)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Chem
Neurosci. 2013 Mar 19."
FEATURES Location/Qualifiers
source 1..4455
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 64..443
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 444..647
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 598..618
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 692..711
/label=T7
/note="T7 promoter, forward primer"
promoter 692..710
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 745..1998
/codon_start=1
/label=R-GECO1.2
/translation="MVDSSRRKWNKAGHAVRAIGRLSSPVVSERMYPEDGALKSEIKKG
LRLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQCERAEGRHST
GGMDELYKGGTGGSLVSKGEEDNRAIVKEFMRFKLHMEGSVNGHEFEIEGEGEGRPYEA
FQTAKLKVTKGGPLPFAWDILSPQLMYGSKAYIKHPADIPDYFKLSFPEGFRWERVMNF
EDGGIIHVSQDTSLQDGVFIYKVKLRGTNFPPDGPVMQKKTMGWEATRDQLTEEQIAEF
KEAFSLFDKDGDGTMTTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTFDFPEFLTM
MARKMNDTDSEEEIREAFRVFDKDGNGYIGAAELRHVMTDLGEKITDEEVDEMIRVADI
DGDGQVNYEEFVQMMTAK"
CDS 760..816
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKAGHAVRAIGRLSS"
primer_bind 1512..1531
/label=mCherry-F
/note="mCherry, forward primer"
promoter complement(2059..2077)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2060..2077)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2097..2114)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2103..2327
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(2529..3117)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2609..2628)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3288..4148)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3911..3930
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4149..4253)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4328..4347)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"