Reporter

Part:BBa_K1943020:Design

Designed by: Shixin Lu, Shiqiang Tang   Group: iGEM16_SUSTech_Shenzhen   (2016-10-13)


R-GECO


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE 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.

References

  1. Parts Registry Assembly Help
  2. Parts Registry Transformation Guideline