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PurposeExpresses human NKCC1 with an N-terminal 3xFlag-YFP-(chloride-sensing) tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites. Native hNKCC1 amino acid sequence
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 49060 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3.1 neo
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Backbone manufacturerLife Technologies
- Backbone size w/o insert (bp) 5316
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namehuman NKCC1 (synthetic)
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Alt nameSlc12A2 (synthetic)
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Alt nameNa-K-Cl cotransporter
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SpeciesH. sapiens (human), Synthetic
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Insert Size (bp)4479
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MutationCl-sensitive YFP (EYFP/ V163S A206K) in hNKCC1 in NT17
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Entrez GeneSLC12A2 (a.k.a. BSC, BSC-2, BSC2, CCC1, KILQS, NKCC1, PPP1R141)
- Promoter CMV
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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
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Academic/Nonprofit Terms
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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.
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.
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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