Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Cyril Pulver
Date created: 2015-09-17 11:00:00
Date modified: 2015-09-18 01:17:34
CYC_3 promoter
| Types | DnaRegion |
| Roles | promoter Regulatory |
| Sequences | BBa_K1723025_sequence (Version 1) |
Description
CYC_3 is a mutated version of the pCYC Yeast Promoter (BBa_K124000) preceded by a PAM sequence rich region (NGG) that allows binding of dCas9_VP64 together with the appropriate guide RNA (gRNA) to 20 base pair specificity determinant sequences flanked by a PAM sequence. It is activated by binding of the complex formed of dCas9_VP64 (BBa_K1723021) and c3_3 gRNA (produced by BBa_K1723012): recruitment of RNA polymerase II by the VP64 subunit on the promoter will express a downstream gene. It is inhibited by binding of the complex formed of dCas9_VP64 and c6_3 gRNA (produced by BBa_K1723016): the complex will sterically prevent RNA polymerase II from binding to the promoter. It is inhibited through the same mechanism by binding of the complex formed of dCas9_VP64 and c7_3 gRNA (produced by BBa_K1723020). If any combination of those complexes is expressed, the promoter will be inhibited. A similar mechanism has been proven to work with CYC_0 (BBa_K1723022) and gRNAs c3_0 (produced by BBa_K1723009), c6_0 (produced by BBa_K1723013) and c7_0 (produced by BBa_K172017) [1].To learn more about how this part was used specific to our project, please follow this link: http://2015.igem.org/Team:EPF_Lausanne/Part_Collection
Notes
This promoter was specifically designed to be controlled by complexes constituted of dCas9_VP64 and gRNAs c3_1, c6_1 and c7_1. The design of the promoter is based on promoter CYC_0 (BBa_K1723022). It shows specific mutations that make c3_3, c6_3 and c7_3 sites differ from c3_0, c6_0 and c7_0 sites which are present on CYC_0. Since it has been demonstrated with CYC_0 that inhibition prevails over activation when combining the co-expression of dCas9_VP64 and those gRNAs [1], this promoter can be used as a basic cellular computation tool.Mutations were carefully designed to keep both TATA sequences [2] of CYC_0 intact.