BBa_K1218014

BBa_K1218014 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1218014
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Sophia Liang
Date created: 2013-08-28 11:00:00
Date modified: 2015-05-08 01:09:42

dCas9-ω
Activator



Types
DnaRegion

Roles
Coding

CDS

Sequences BBa_K1218014_sequence (Version 1)

Description

CRISPR-Cas is a bacterial immune system that remembers and targets foreign viral DNA by storing DNA sequences, or spacers, between clustered regularly interspaced short palindromic repeats (CRISPRs). RNA transcripts of the spacers are then used to sense homologous DNA, which is cleaved by CRISPR-associated (Cas) proteins.

This part codes for the tracrRNA, a mutated Cas9 protein, and minimal CRISPR array of a type II CRISPR-Cas system. The CRISPR array includes two CRISPR repeats separated by a spacer with two BsaI sites. Digestion with BsaI allows for insertion of a new spacer, thus changing the sequence targeted by Cas9. However, the Cas9 protein is mutated or "dead" and is fused to a RNAP omega ("w") subunit. Binding of the Cas9-RNAP-subunit fusion activates transcription of the targeted gene.

Notes

Mutagenesis to remove internal EcoRI site

Source

CRISPR-Cas system from streptococcus pyogenes; cloned from plasmids obtained from Bikard et al.

igem#experience
None
 
igem#sampleStatus
In stock
igem#status
Available
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1218014/1