pcDNA3.1 Flag YFP hNKCC1 N-terminus T,S mutations P676C I730C (NT95)
(Plasmid
#50913)
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PurposeExpresses human NKCC1 with mutated Ser and Thr residues in the N-terminus and mutations at P676C I730C. Contains N-terminal 3xFLAG-YFP tag for expression in mammalian cells
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 50913 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA 3.1 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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MutationS150N S155Q S170Q T177A S183R S193E T203A T205A T207Q T212A T217A T230A S242N T266A T268Q T274E T276R P676C I730C 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 (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 YFP hNKCC1 N-terminus T,S mutations P676C I730C (NT95) was a gift from Biff Forbush (Addgene plasmid # 50913 ; http://n2t.net/addgene:50913 ; RRID:Addgene_50913) -
For your References section:
Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion Transport and Transporter Activation Revealed by Internal Crosslinking between Transmembrane Domains 10 and 11/12. Monette MY, Somasekharan S, Forbush B. J Biol Chem. 2014 Jan 22. 10.1074/jbc.M113.542258 PubMed 24451383