BBa_K1723024
1
BBa_K1723024
CYC_2 promoter
2015-09-17T11:00:00Z
2015-09-18T01:06:40Z
Synthesized as a G-Block
CYC_2 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_2 gRNA (produced by BBa_K1723011): 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_2 gRNA (produced by BBa_K1723015): 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_2 gRNA (produced by BBa_K1723019). 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
false
false
_2143_
25334
25334
9
false
This promoter was specifically designed to be controlled by complexes constituted of dCas9_VP64 and gRNAs c3_2, c6_2 and c7_2. The design of the promoter is based on promoter CYC_0 (BBa_K1723022). It shows specific mutations that make c3_2, c6_2 and c7_2 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 boxes of CYC_0 intact.
false
Cyril Pulver
annotation2469844
1
c3_2
range2469844
1
39
58
annotation2469847
1
c7_2
range2469847
1
58
77
annotation2469849
1
C6_2
range2469849
1
126
145
BBa_K1723024_sequence
1
aagcttgatatcgaattcctgcagcccgggtactgtatatatacccctgcataccctatggtcggaagatctagaatatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattgcctgaaccgccgcataaattactatacttctatagacacacaaacacaaatacacacactaatctagatattaaa
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z