Difference between revisions of "Part:BBa K368000:Design"
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===Design Notes=== | ===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. | PduD contained one PstI restriction side. Therefore position 1345 was mutated via two step PCR from t to a, resulting in a silent mutation. | ||
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===Source=== | ===Source=== | ||
Latest revision as of 18:09, 3 August 2010
Pbad::pduD-aurF
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
Illegal NheI site found at 1328 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 1470 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.
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.