Composite

Part:BBa_K2683031:Design

Designed by: Kristi Turton   Group: iGEM18_Lethbridge   (2018-10-13)


P22 Capsid


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 177
    Illegal AgeI site found at 783
    Illegal AgeI site found at 1005
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Due to the Size of this construct and its need for the scaffolding protein to encapsulate finding surface modifications had to be compatible with the outer surface f the capsid. The DNA construct was built with primer extension. Current illegal cut sites within the sequence will be removed by site directed mutagenesis.

Source

P22 is a capsid protein originally from the P22 bacteriophage. P22 was a originally in a pBad plasmid donated in kind by the Wiedenheft lab in the University of Montana. Biobricking was done by primer extension.

References

[1]O’Neil, A., Reichhardt, C., Johnson, B., Prevelige, P., and Douglas, T. (2011) Genetically programmed in vivo packaging of protein cargo and its controlled release from bacteriophage P22. Angewandte Chemie International edition. 50, 7425-7428


[2]O’Neil, A., Prevelidge, P., Basu, G., and Douglas, T. (2012) Coconfinement of fluorescent proteins: spatially enforced communication of GFP and mCherry encapsulated within the P22 capsid. 13, 3902-3907


[3]Qazi, S., Miettinen, H., Wilkinson, R., McCoy, K., Douglas, T., and Wiedenheft, B. (2016)Programmed self-assembly of an active P22-Cas9 Nanocarrier System. Molecular Pharmaceutics. 13, 1191-1196