p2CT-His-MBP-Lbu_C2c2_R472A_H477A
(Plasmid
#83483)
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PurposeBacterial expression of L. buccalis C2c2 CRISPR effector (HEPN nuclease 1 inactive mutant).
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 83483 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonep2CT
- Backbone size w/o insert (bp) 5960
- Total vector size (bp) 9437
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Vector typeBacterial 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)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameLbu_C2c2_R472A_H477A
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Alt nameLbuCas13a
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SpeciesLeptotrichia buccalis
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Insert Size (bp)3477
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MutationR472A, H477A
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GenBank IDGI_506250229
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Tag
/ Fusion Protein
- His6-MBP (N terminal on insert)
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer MBP forward (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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
Codon Optimized for E. Coli expression. His-MBP fusion tag can be removed by TEV protease
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:
p2CT-His-MBP-Lbu_C2c2_R472A_H477A was a gift from Jennifer Doudna (Addgene plasmid # 83483 ; http://n2t.net/addgene:83483 ; RRID:Addgene_83483) -
For your References section:
Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. East-Seletsky A, O'Connell MR, Knight SC, Burstein D, Cate JH, Tjian R, Doudna JA. Nature. 2016 Sep 26. doi: 10.1038/nature19802. 10.1038/nature19802 PubMed 27669025