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Purpose(Empty Backbone) Vector for tandem expression of two sgRNAs from two independent U6 promoters. Cas9 is expressed by Cbh promoter.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 64073 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Cloning Grade DNA | 64073-DNA.cg | 2 µg of cloning grade DNA in Tris buffer | 1 | $105 |
Backbone
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Vector backbonepX333
- Backbone size (bp) 9000
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Vector typeMammalian Expression
- Promoter CBh; U6
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Tag
/ Fusion Protein
- 3XFLAG-Cas9 (N terminal on backbone)
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 numberUnknown
Cloning Information
- Cloning method Restriction Enzyme
Resource Information
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Addgene Notes
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A portion of this plasmid was derived from a plasmid made byCas9 from pX330 (Zhang lab)
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Articles 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
pX333 for cloning a tandem of sgRNAs (BbsI and BsaI)
Information for Cloning Grade DNA (Catalog # 64073-DNA.cg) ( Back to top)
Purpose
Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.
Delivery
- Amount 2 µg
- Guaranteed Concentration 100 ng/µl +/- 5 ng/µl
- Pricing $105 USD
- Storage DNA can be stored at 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
Addgene has verified this plasmid using Next Generation Sequencing. Results are available here
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:
pX333 was a gift from Andrea Ventura (Addgene plasmid # 64073 ; http://n2t.net/addgene:64073 ; RRID:Addgene_64073) -
For your References section:
In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system. Maddalo D, Manchado E, Concepcion CP, Bonetti C, Vidigal JA, Han YC, Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A. Nature. 2014 Dec 18;516(7531):423-7. doi: 10.1038/nature13902. Epub 2014 Oct 22. 10.1038/nature13902 PubMed 25337876