Difference between revisions of "Part:BBa K3634011:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | The ccdB sequence was taken from BBa_K145151 and found to have an illegal BsaI restriction site at 222bp. The appropriate alteration was made to make the part RFC[1000] compatible: c219g (GTC (Val) to got to go (Val)). As native to E.coli, no codon optimisation step was required. | + | The ccdB sequence was taken from BBa_K145151 and found to have an illegal BsaI restriction site at 222bp. The appropriate alteration was made to make the part RFC[1000] compatible: c219g (GTC (Val) to got to go (Val)). As native to E.coli, no codon optimisation step was required. |
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===Source=== | ===Source=== | ||
Revision as of 10:53, 6 August 2020
ccdB (BsaI Removed)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The ccdB sequence was taken from BBa_K145151 and found to have an illegal BsaI restriction site at 222bp. The appropriate alteration was made to make the part RFC[1000] compatible: c219g (GTC (Val) to got to go (Val)). As native to E.coli, no codon optimisation step was required.
Source
The ccdB toxin is found natively in E.coli and the sequence specified in BBa_K145151 by KULeuven iGEM (2008). This sequence was confirmed against pAG415 GPD-EGFP and pCMV SPORT6ccdB plasmids and in silico site-directed mutagenesis removed a BsaI site so that the part was RFC[1000] standard.