Part:BBa_K1442100:Design
RdRP
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 492
Illegal BglII site found at 1340 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 537
Design Notes
The 21 amino acid residues have been removed in the part sequence, ensuring cytoplasmic RdRp activity, this ensures no anchoring to the endoplasmic reticulum in human cells (Lai et al., 2004). This modification has no major effect on RdRp activity, which is complimented by previous analysis showing no significant loss of nucleotide polymerization activity (Vo et al., 2004).
A single point mutation has been introduced at position 2884, where a guanine has replaced a cytosine. This confers an Arginine to glycine substitution, which increases the number of transformed colonies obtained (Bartenschlager et al., 2001).
A PstI restriction site was removed in the part sequence (CTGCAG) and was altered to CTCCAG to allow bio brick compatibility. This was necessary, as this would have invalidated this part as permissible for iGEM's RFC25 standard.