ATP1A1_T804N_hPGK1_EBFP_Donor
(Plasmid
#187455)
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PurposeHDR donor plasmid to introduce the T804N mutation conferring cellular resistance to ouabain and the EBFP cassette to ATP1A1 intron 17.
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Depositing Lab
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Depositing OrganizationUniversite Laval
Located in Canada -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 187455 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 187455-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 187455-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepUC19
- Total vector size (bp) 5382
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Vector typeMammalian Expression
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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-T804N EBFP donor
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SpeciesH. sapiens (human)
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Insert Size (bp)1584
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Entrez GeneATP1A1 (a.k.a. CMT2DD, HOMGSMR2)
- Promoter hPGK1
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site NheI (not destroyed)
- 5′ sequencing primer TGAGCGAGGAAGCGGAAGAG
- 3′ sequencing primer CAGGGTTATTGTCTCATGAGCGG
- (Common Sequencing Primers)
Resource Information
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
This plasmid can be used in combination with pX330_ATP1A1_G7 or eSpCas9(1.1)_No_FLAG_ATP1A1_G7 to target EBFP to ATP1A1 intron 17. A user-specified transgene of interest can also be cloned using restriction cloning sites upstream (KpnI, AflII, NcoI) and downstream (SbfI) of the EBFP gene cassette.
Please visit https://doi.org/10.1101/2021.11.02.464583 for bioRxiv preprint.
DNA (Catalog # 187455-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 187455-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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_T804N_hPGK1_EBFP_Donor was a gift from Yannick Doyon (Addgene plasmid # 187455 ; http://n2t.net/addgene:187455 ; RRID:Addgene_187455) -
For your References section:
Marker-free co-selection for successive rounds of prime editing in human cells. Levesque S, Mayorga D, Fiset JP, Goupil C, Duringer A, Loiselle A, Bouchard E, Agudelo D, Doyon Y. Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z. 10.1038/s41467-022-33669-z PubMed 36207338