pX330-Flag-dHeFSpCas9
(Plasmid
#92116)
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PurposeExpression plasmid for human codon-optimized dead/inactive increased fidelity HeFSpCas9 (without U6-sgRNA coding sequence)
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Depositing Lab
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Depositing OrganizationHUN-REN Biological Research Centre Szeged (HUN-REN Szegedi Biológiai Kutatóközpont)
Located in Hungary -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 92116 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 92116-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 92116-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepX330-like (without U6-sgRNA coding sequence)
- Total vector size (bp) 8074
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Vector typeMammalian Expression, CRISPR
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)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namedead/inactive HeFSpCas9 with FLAG tag
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SpeciesS. pyogenes
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Insert Size (bp)4272
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MutationD10A, N497A, R661A, Q695A, H840A, K848A, Q926A, K1003A, R1060A
- Promoter Cbh
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Tags
/ Fusion Proteins
- 3xFLAG (N terminal on insert)
- NLS (N terminal on insert)
- NLS (C terminal on insert)
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer gggtggagtatttacggtaaactgcccacttgg
- 3′ sequencing primer CGGCGTACTGGTCGCCGATC
- (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
Alternate plasmid name: pCbh-3xFLAG-NLS-dHeFSpCas9-NLS (without U6-sgRNA coding sequence).
For detailed information and plasmid usage, please see the associated publication.
DNA (Catalog # 92116-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
-
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.
DNA (Catalog # 92116-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:
pX330-Flag-dHeFSpCas9 was a gift from Ervin Welker (Addgene plasmid # 92116 ; http://n2t.net/addgene:92116 ; RRID:Addgene_92116) -
For your References section:
Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Kulcsar PI, Talas A, Huszar K, Ligeti Z, Toth E, Weinhardt N, Fodor E, Welker E. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8. 10.1186/s13059-017-1318-8 [pii] PubMed 28985763
Map uploaded by the depositor.
