Part:BBa_K1723025

CYC_3 promoter

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

Usage and Biology

S. cerevisiae