BBa_K1723000

BBa_K1723000 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1723000
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Gregoire Thouvenin
Date created: 2015-08-09 11:00:00
Date modified: 2015-08-10 10:13:34

dCas9-ω



Types
DnaRegion

Roles
CDS

Coding

Sequences BBa_K1723000_sequence (Version 1)

Description

This part codes for a enzymatically inactive Cas9 protein (dCas9) that is fused to rpoZ, the omega subunit of E.Coli RNA Polymerase (RNAP).

Cas9 (CRISPR associated protein 9) is an RNA-guided DNA endonuclease that targets and cleaves any DNA sequence complementary to its guide RNA (gRNA). Catalytically ???dead??? Cas9 (dCas9) lack the ability to cleave DNA. When fused to a RNAP recruiting element (in this case the omega subunit of RNAP), chimeric dCas9 can act as a programmable transcription activator. dCas9 may also act as a DNA transcription inhibitor: depending on its gRNA-determined binding site, it has been shown in yeasts to sterically hinder RNAP recruitment to promoter sequences.

Notes

The original dCas9-ω given to us by the Bikard lab contains an EcoRI restriction site. We performed a site-directed mutagenesis to remove it.

Source

A plasmid containing dCas9_ω (pdCas9) was given to our iGEM team by David Bikard.

Sequence Annotation Location Component / Role(s)
linker
rpoZ (omega subunit)
dCas9
4105,4113
4114,4383
1,4104
feature/misc sequence_feature
feature/binding non_covalent_binding_site
feature/protein CDS
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1723000/1