pcDNA3.1 Flag YFP hNKCC1 (NT17)
(Plasmid #49085)

• Purpose: Expresses human NKCC1 with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites. Native hNKCC1 amino acid sequence
• Depositing Lab: Biff Forbush
• Publication: Somasekharan et al PLoS One. 2013 Dec 5;8(12):e82060. doi: 10.1371/journal.pone.0082060.

Backbone

• Vector backbone: pcDNA3.1 neo
• Backbone manufacturer: Life Technologies
• Backbone size w/o insert (bp): 5316
• 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: human NKCC1 (synthetic)
• Alt name: Slc12A2 (synthetic)
• Alt name: Na-K-Cl cotransporter
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 4479
• Mutation: 262 silent mutations from native hNKCC1
• Entrez Gene: SLC12A2 (a.k.a. BSC, BSC-2, BSC2, CCC1, KILQS, NKCC1, PPP1R141)
• Promoter: CMV
• Tags / Fusion Proteins:
  • 3x Flag (N terminal on insert)
  • mVenus (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: PmeI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Synthetic cDNA encoding full length human NKCC1, with 29 silent restriction sites in NKCC1 coding region

This plasmid has also been described in Monette et al., J Biol Chem. 2014 Jan 22 (https://www.ncbi.nlm.nih.gov/pubmed/24451383)