pcDNA3-ΔN-RNMT
(Plasmid #112708)

• Purpose: Expresses FLAG-NES-RNMT 121-476 D203A ("ΔN-RNMT") in mammalian cells. Overexpression inhibits cytoplasmic cap methylation.
• Depositing Lab: Daniel Schoenberg
• Publication: Trotman et al Nucleic Acids Res. 2017 Oct 13;45(18):10726-10739. doi: 10.1093/nar/gkx801.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5446
• Total vector size (bp): 6603
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: RNMT
• Alt name: RNA guanine-7 methyltransferase
• Alt name: Cap methyltransferase
• Species: H. sapiens (human)
• Insert Size (bp): 1167
• Mutation: deleted amino acids 2-120, mutated active-site D203 to A
• Entrez Gene: RNMT (a.k.a. CMT1, CMT1c, MET, Met, N7-MTase, RG7MT1, cm1p, hCMT1, hMet)
• Promoter: CMV
• Tags / Fusion Proteins:
  • FLAG (N terminal on insert)
  • HIV Rev nuclear export signal (NES)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (destroyed during cloning)
• 3′ cloning site: KpnI (destroyed during cloning)
• 5′ sequencing primer: T7 promoter (TAATACGACTCACTATAGG)
• 3′ sequencing primer: SP6 promoter (ATTTAGGTGACACTATAG)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Source of RNMT sequence: pET16b-His-RNMT from Stuart Shuman, Sloan Kettering
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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