pcDNA4/TO mTRPM7
(Plasmid #45482)

• Depositing Lab: Andrew Scharenberg
• Publication: Nadler et al Nature. 2001 May 31;411(6837):590-5.

Backbone

• Vector backbone: pcDNA4/TO (modified)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 11000
• Vector type: Mammalian Expression ; Tetracycline Inducible
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TRPM7
• Alt name: LTRPC7
• Alt name: CHAK1
• Alt name: transient receptor potential cation channel, subfamily M, member 7
• Species: M. musculus (mouse)
• Insert Size (bp): 6000
• GenBank ID: NM_021450.2 NP_067425.2
• Entrez Gene: Trpm7 (a.k.a. 2310022G15Rik, 4833414K03Rik, 5033407O22Rik, CHAK, CHAK1, LTrpC-7, Ltpr7, Ltrpc7, TRPPLIK)
• Promoter: Tetracycline-controlled CMV
• Tag / Fusion Protein:
  • Flag (N terminal on insert)

Cloning Information

• 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

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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.