Reporter
Part:BBa_K1943020:Design
Designed by: Shixin Lu, Shiqiang Tang Group: iGEM16_SUSTech_Shenzhen (2016-10-13)
R-GECO
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 914
Illegal SapI.rc site found at 111
Illegal SapI.rc site found at 820
Design Notes
This year, R-GECO is one of core protein of our project. We find its sequence and synthesis it and ligate it to the plasmid that we transfect to mammalian cells. So we design primers and do PCR of this R-GECO coding sequence and ligate it to pSB1C3 backbone. Besides, there is a R-GECO in biobrick registry, BBa_K881000. However, there are two PstI enzyme cutting sites in BBa_K881000. Thus, we do two site-directed mutagenesis of the two PstI enzyme cutting sites by the degeneracy of condons to make it RCF10 compatible.
Source
The plasmid was from our instructor, Huangwei's lab. And we design primers to copy down by PCR.