Part:BBa_K5121024:Design
Reb1: RebB C-terminal Cys
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 806
Illegal NheI site found at 868 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 748
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Although in vivo, the reb genes are transcriptionally independent, here they are encoded in polycistronic fashion. Since wild-type rebB contains no cysteines (and neither does rebA), this composite part contains a modified version of rebB with an added C-terminal cysteine (BBa_K5121016). This modification was made using an extension overlap PCR of the original reb1 plasmid (BBa_K5121011) with overlapping primers as described in the design notes for the basic part (BBa_K5121016). Since the R-body is a large protein polymer, we sought to enhance the accessibility of our introduced cysteine to reacting maleimides. We achieved this with a flexible poly-glycine linker placed in between the C-terminal cysteine and the remainder of the rebB coding region (N-SGGGC-C).
Source
E. coli