CMV-ER-LAR-GECO1 vector (V010143#)

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

CMV-ER-LAR-GECO14517 bp600120018002400300036004200bGH poly(A) signalSP6 promotermCherry-FT7CMV promoterCMV enhancerpRS-markerAmpR promoterAmpRori

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

Copy Sequence

Download GeneBank File(.gb)

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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