CMV-ER-LAR-GECO1 vector (V010143)

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 promoterCAR-GECO1T7 promoterCMV 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       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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