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Purpose(Empty Backbone) Used for expression of sgRNA in mammalian cells
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Depositing Lab
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Publication
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Sequence Information
Ordering
This material is available to academics and nonprofits only.
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 118055 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Backbone
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Vector backbonepSpCas9(BB)-2A-Puro (PX459) V2.0 (Addgene#62988)
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Vector typeMammalian Expression
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Selectable markersBlasticidin
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
Cloning Information
- 5′ sequencing primer AGCTGGGAGGCGACAAAAGG (Common Sequencing Primers)
Resource Information
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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.
Depositor Comments
pSpCas9(BB)-2A-Puro (PX459) V2.0 was digested with EcoRI to remove the T2A-Puro cassette, and a P2A-Blasticidin cassette was amplified by PCR, digested with EcoRI, and ligated.
The BbsI site (GAAGAC) in the blasticidin-resistance gene was mutated to GAGGAC with no change in amino acids.
gRNA inserts can be sequenced using pLKO.1 5’ primer, 5’-GACTATCATATGCTTACCGT-3’.
How to cite this plasmid
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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:
pSpCas9(BB)-2A-Blast was a gift from Ken-Ichi Takemaru (Addgene plasmid # 118055 ; http://n2t.net/addgene:118055 ; RRID:Addgene_118055)
