URL: http://www.addgene.org/196244
Proper Citation: RRID:Addgene_196244
Insert Name: Cas9
Organism: Streptococcus pyogenes
Bacterial Resistance: Ampicillin
Defining Citation: PMID:37147433
Vector Backbone Description: Backbone Marker:pET-based custom vector; Backbone Size:6400; Vector Backbone:pEC-K-MBP; Vector Types:Bacterial Expression, CRISPR; Bacterial Resistance:Ampicillin
Comments: This submission's cysteine-free triNLS Cas9 construct bears an unexpected and unintentional point mutation in the Cas9 sequence, H1311N, that was not identified until after deposition. Analysis of the Cas9 structure suggests that this mutation is unlikely to impact enzyme function or properties. H1311 is located in the PAM-interacting domain (PID), a relatively rigid part of the Cas9 structure. In the Cas9 crystal structure (PDB ID 4UN3), the side chain of H1311 is solvent exposed and does not interact with other Cas9 amino acids or bound nucleic acid. Substitution of histidine with asparagine (a hydrophilic amino acid) should be well tolerated. We do not anticipate any consequences associated with this mutation. This mutation is reflected in the sequence files currently associated with this Addgene entry. Note that our Addgene submission 403489 (triNLS Cas9 with two cysteines) is very similar to this plasmid but it does not contain the H1311N mutation (it has H1311).
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No rating or validation information has been found for pRM053 His-TEV-Cas9 triNLS 0C.
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Source: Addgene (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org