Composite
Part:BBa_K2483004:Design
Designed by: Bryan Nowack Group: iGEM17_Potsdam (2017-09-16)
regulated dCas9 with sgRNAs and IAA enzymes fused to MS2 and PP7
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 3004
Illegal NheI site found at 9656
Illegal NheI site found at 9930 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 4412
Illegal BglII site found at 6221
Illegal BglII site found at 9644
Illegal BglII site found at 9918
Illegal BamHI site found at 725
Illegal BamHI site found at 7166 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 5928
Illegal NgoMIV site found at 8872 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The LacI-dCas9 construct was put in reverse complement to the other parts to utilize the temrinators on both ends of the plasmid backbone, thereby reducing additional planning.
Source
For the sources for dCas9 and the IAA fusion proteins, see their respective parts (BBa_K2483002 and BBa_K2483000). The sgRNA design was taken from iGEM Warwick and modified to include the target sites we were using (BBa_K1994017 and BBa_K1994016).