Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Huang Xingxu
Date created: 2013-09-12 11:00:00
Date modified: 2015-05-08 01:09:32
coding sequence of Cas9 from CRISPR system type II
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_K1160000_sequence (Version 1) |
Description
CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system provide protection against mobile genetic element by guided specific DNA cleavage. Cas9 (CRISPR-associated protein 9) belongs to type II CRISPR system and contains HNH motif and three RuvC-like motifs, homologous to HNH and RuvC endonuclease, respectively. Cas9 has strong DNA cleavage activity, thus it showed potential use in gene editing.This part contains the coding sequence of cas9 from Streptococcus pyogenes (UniProtKB AC number:Q99ZW2). Some "redundant" sequence was added to ensure the Cas9's normal function or just for the convenience to detect it in cell.
The cas9 protein must be guided by two RNAs, crRNA(CRISPR RNA) and tracrRNA(trans-activating crRNA), to complete its specific DNA cleavage function. We build another part-a chimeic RNA-that combines the feature two RNAs. For more details, refer to part BBa_K1160001.
With this part, you can
1)directly use it to express cas9 protein in cell or
2)use T7 RNA polymerase to get cas9's mRNA, which can be further modified to prepare authentic mRNA that can be used for translation in vivo.
Notes
to be addedSource
Streptococcus pyogenes| Sequence Annotation | Location | Component / Role(s) |
| Ca9 coding sequence CMV promoter CMV enhancer bGH poly(A) signal f1 ori BSD ori AmpR | 904,5010 615,818 235,614 5120,5344 5390,5818 6276,6644 7404,7992 8163,9023 | feature/protein CDS feature/promoter promoter sequence_feature feature/misc feature/tag tag sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc CDS feature/protein |