Coding

Part:BBa_K368000:Design

Designed by: Emanuel Stiegeler   Group: iGEM10_LMU-Munich   (2010-07-31)
Revision as of 18:07, 3 August 2010 by Emanuel stiegeler (Talk | contribs) (→‎Design Notes)

Pbad::pduD-aurF


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal NheI site found at 1328
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 1470
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1986
    Illegal NgoMIV site found at 2783
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1774
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1847
    Illegal BsaI.rc site found at 2486
    Illegal SapI site found at 961
    Illegal SapI.rc site found at 1907


Design Notes

PduD contained one PstI restriction side. Therefore position 1345 was mutated via two step PCR from t to a, resulting in a silent mutation. File:Muh.jpg

Source

pduD was amplified from gDNA of C.freundii (DSMZ strain number: DSM 30039). aurF was amplified from gDNA of S. thioluteus (DSMZ strain number: 40027)

References

Parsons, J. B., S. Frank, D. Bhella, M. Liang, M. B. Prentice, D. P. Mulvihill & M. J. Warren, (2010) Synthesis of empty bacterial microcompartments, directed organelle protein incorporation, and evidence of filament-associated organelle movement. Mol Cell 38: 305-315.

Simurdiak, M., J. Lee & H. Zhao, (2006) A new class of arylamine oxygenases: evidence that p-aminobenzoate N-oxygenase (AurF) is a di-iron enzyme and further mechanistic studies. Chembiochem 7: 1169-1172.