
Coding
CapD_CP
Part:BBa_K314012
Designed by: Chris Eiben Group: iGEM10_Washington (2010-10-08)
CapD_CP
A circularly permuted CapD with a Foldit-designed linker.
Usage and Biology
To test BBa _K314012, it was inserted into a pET29b+ vector using Kunkel Mutagenesis. CapD_CP was then produced and purified as described in the UW 2010 iGEM Team's Small Scale Protein Purification Protocol. The purified protein was then tested for activity. For a detailed description of the assay, please see the 2010 UW iGEM wiki. A protein gel was also run to determine physical qualities. The resulting data is shown below.

We also ran the purified protein samples through a mass spectrometer to make sure the first methionine was being removed by the natural E. coli methionine aminopeptidase and that the protein was the expected size. Our results indicate that a massive majority of the protein in our purified sample was within .02% error of the predicted CapD_CP size. Expected weight of CapD_CP without Methionine=55285Da, with Methionine=55417Da. Our mass spec detected a peak at 55274.8Da (no Methionine) well within the 0.02% error limit for our mass spec
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1489
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None |