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Addgene

pcDNA3.1 Flag Cl-sensitive YFP hNKCC1 WT (NT13)
(Plasmid #49060)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 49060 Standard format: Plasmid sent in bacteria as agar stab 1 $85

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
    Cl-sensitive YFP (EYFP/ V163S A206K) in hNKCC1 in NT17
  • 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)
    • YFP (chloride-sensitive) (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
  • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Plasmid constructed from pcDNA3.1 Flag YFP hNKCC1 WT (NT17). See Addgene plasmid# 49085 (http://www.addgene.org/49085/) for plasmid map of wild-type construct.

How to cite this plasmid ( Back to top)

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:

    pcDNA3.1 Flag Cl-sensitive YFP hNKCC1 WT (NT13) was a gift from Biff Forbush (Addgene plasmid # 49060 ; http://n2t.net/addgene:49060 ; RRID:Addgene_49060)
  • For your References section:

    Functional Expression of Human NKCC1 from a Synthetic Cassette-Based cDNA: Introduction of Extracellular Epitope Tags and Removal of Cysteines. Somasekharan S, Monette MY, Forbush B. PLoS One. 2013 Dec 5;8(12):e82060. doi: 10.1371/journal.pone.0082060. PubMed 24339991