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Mouse Grin1 ORF clone (NM_008169) (Cat. No.: V032194)

pCMV-Neo-Grin1(mouse)-DsRed27490 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200CMV enhancerCMV promoterGrin1(NM_008169)DsRed2SV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRHSV TK poly(A) signalori
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Basic Information
Name:
pCMV-Neo-Grin1(mouse)-DsRed2
Accession ID:
NM_008169.3
Antibiotic Resistance:
Kanamycin
Length:
7490 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Selection Marker:
Neo/G418;DsRed2
Promoter:
CMV
5' Primer:
CMV-F
Fusion Tag:
DsRed2
Growth Strain(s):
DH5a
Growth Temperature:
37℃
Expression Method:
Transient
Gene Synonyms:
GluN1; GluRdelta1; GluRzeta1; M100174; NMD-R1; Nmdar; NMDAR1; NR1; Rgsc174
Transcript Definition:
Mus musculus glutamate receptor, ionotropic, NMDA1 (zeta 1) (Grin1), transcript variant 1, mRNA
Cat. No.: V032194 pCMV-Neo-Grin1(mouse)-DsRed2
$ 298.8
In stock, 1-2 weeks for quality controls
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Two tubes of transfection grade plasmid, each tube is about 100 µg (lyophilized).

Resources

Transcript ID
NM_008169.3
Fusion Tag
DsRed2
Selection Marker
Neo/G418;DsRed2
Expression Method
Transient

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.

Mouse Grin1 ORF clone (NM_008169) (Cat. No.: V032194) Sequence

LOCUS       V032194                 7490 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032194
VERSION     V032194
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             624..3437
                     /label="Grin1(NM_008169)"
                     /note="Grin1(NM_008169)"
                     /gene="Grin1"
     CDS             3480..4157
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="improved tetrameric variant of DsRed fluorescent
                     protein"
                     /transl_table=1
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL*"
                     /label="DsRed2"
     polyA_signal    4278..4399
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4406..4861)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4888..4992
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4994..5351
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5386..6180
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6412..6459
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6788..7376
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"