Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | CMV-ER-LAR-GECO1 | Antibiotic Resistance | Ampicillin |
| Length | 4517 bp | Type | Mammalian Expression |
| Copy Number | High Copy | Promoter | CMV |
| Cloning Method | Restriction Enzyme | 5' Primer | TAATACGACTCACTATAGGG |
| 3' Primer | TAGAAGGCACAGTCGAGG | ||
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 Exported 4517 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses LAR-GECO1 in the endoplasmic reticulum in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS CMV-ER-LAR-GECO1
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)
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"
FEATURES Location/Qualifiers
source 1..4517
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 42..266
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 255..272
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
promoter 292..310
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 292..309
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(850..869)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1721..1740)
/label=T7
/note="T7 promoter, forward primer"
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"
primer_bind complement(1814..1834)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 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
/gene="bla"
/label=AmpR promoter
CDS 2739..3599
/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 complement(2957..2976)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3770..4358
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4259..4278
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
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