|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||45482||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepcDNA4/TO (modified)
- Backbone size w/o insert (bp) 5000
- Total vector size (bp) 11000
Vector typeMammalian Expression ; Tetracycline Inducible
Growth in Bacteria
Copy numberHigh Copy
Alt nametransient receptor potential cation channel, subfamily M, member 7
SpeciesM. musculus (mouse)
Insert Size (bp)6000
GenBank IDNM_021450.2 NP_067425.2
Entrez GeneTrpm7 (a.k.a. 2310022G15Rik, 4833414K03Rik, 5033407O22Rik, CHA, CHAK, CHAK1, LTR, LTrpC-7, Lt, Ltpr7, Ltrpc7, TRP, TRPPLIK)
- Promoter Tetracycline-controlled CMV
/ Fusion Protein
- Flag (N terminal on insert)
- Cloning method Unknown
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site KpnI (unknown if destroyed)
- 5′ sequencing primer LNCX; CMV-F
- 3′ sequencing primer BGH-Rev (Common Sequencing Primers)
For the purpose of expressing LTRPC7 in eukaryotic cells, the depositing lab used PCR to produce an epitope tagged expression construct from two overlapping murine LTRPC7 clones. The LTRPC7 coding sequence was modified by removing the initiating methionine and replacing it with a sequence encoding a Kozak sequence, the FLAG tag and the additional sequence GCGGCCGCAT, and by placing a SpeI site just after the stop codon. These modifications result in an expressed protein which started with the following amino acid sequence: MGDYKDDDDKRPH followed by the murine LTRPC7 coding sequence starting at the second amino acid. This construct is expressed from the pcDNA4/TO vector which provides tetracycline-controlled expression from a CMV promotor.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pcDNA4/TO mTRPM7 was a gift from Andrew Scharenberg (Addgene plasmid # 45482 ; http://n2t.net/addgene:45482 ; RRID:Addgene_45482)
For your References section:LTRPC7 is a Mg.ATP-regulated divalent cation channel required for cell viability. Nadler MJ, Hermosura MC, Inabe K, Perraud AL, Zhu Q, Stokes AJ, Kurosaki T, Kinet JP, Penner R, Scharenberg AM, Fleig A. Nature. 2001 May 31;411(6837):590-5. 10.1038/35079092 PubMed 11385574
Map uploaded by the depositor.