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PurposeHomology-directed repair (HDR) donor plasmid with homology arms to ATP1A1 to introduce the Q118R and N129D (RD) mutations conferring cellular resistance to ouabain
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 86551 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepUC19
- Total vector size (bp) 3217
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Vector typeMammalian Expression, CRISPR, TALEN ; Co-selection via HDR using ouabain
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Selectable markersOuabain
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 nameATP1A1
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Alt nameATPase Na+/K+ transporting subunit alpha 1
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Alt nameGene ID: 476
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SpeciesH. sapiens (human)
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MutationQ118R, N129D
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GenBank ID476
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Entrez GeneATP1A1 (a.k.a. CMT2DD, HOMGSMR2)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Acc65I (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer cagggttattgtctcatgagcgg
- 3′ sequencing primer tgagcgaggaagcggaagag (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Article Citing this Plasmid
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.
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:
ATP1A1_plasmid_donor_RD was a gift from Yannick Doyon (Addgene plasmid # 86551 ; http://n2t.net/addgene:86551 ; RRID:Addgene_86551) -
For your References section:
Marker-free coselection for CRISPR-driven genome editing in human cells. Agudelo D, Duringer A, Bozoyan L, Huard CC, Carter S, Loehr J, Synodinou D, Drouin M, Salsman J, Dellaire G, Laganiere J, Doyon Y. Nat Methods. 2017 Apr 17. doi: 10.1038/nmeth.4265. 10.1038/nmeth.4265 PubMed 28417998