Coding

Part:BBa_K1012001:Design

Designed by: John Allan   Group: iGEM13_Dundee   (2013-08-28)


Protein Phosphatase 1 with HA tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 527
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The natural PstI site within the human PP1 gene was silently removed by site-directed mutagenesis

Source

We thank Dr James Hastie (Division of Signal Transduction Therapy, College of Life Sciences, University of Dundee) for providing the pGEX6P-1 PP1 CAT gamma vector (Kelsall et al. 2011) encoding a GST fusion to PP1. This vector served as a PCR template to allow cloning of the PP1 coding sequence into pSB1C3, remove illegal restriction sites, and submit to the Registry under RFC[10] standard.

References

Kelsall IR, Voss M, Munro S, Cuthbertson DJ, Cohen PT. (2011) R3F, a novel membrane associated glycogen targeting subunit of protein phosphatase 1 regulates glycogen synthase in astrocytoma cells in response to glucose and extracellular signals. Journal of Neurochemistry 118:596-610.