Basic Vector Information
CMV-ER-LAR-GECO1 vector Vector Map
Plasmid Resuspension 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 ℃.
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"
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