Part:BBa_K1085028
RBS Start-codon SpiderSilkSubunitE1 SpiderSilkSubunitE2 x2 SilkTail Strep-tag Stop-codon
This BioBrick contains the coding sequence for three parts of the MaSp2 spider silk proteins (SubunitE1 and two SubunitE2s) with a Bacillus subtilis ribosome binding site (RBS) infront of it and a Silktail with a Strep-tag behind it.
The RBS is there to allow the ribosomes to bind and start to translate the DNA.
SubunitE1 codes for a spider silk protein that is optimized for maximal expression in Bacillus subtillus 168. The primary goal of the algorithm was to optimize the codon selection based on their availbility scores. The secondary goal was to prevent the formation of secondary RNA structures in close proximity to the RBS, and the tetrary goal was to minimize the number of restriction sites.
SubunitE2 codes for a spider silk protein that is optimized for maximal expression in Bacillus subtillus 168. The primary goal of the algorithm was to optimize the codon selection based on their availbility scores. The secondary goal was to minimize the number of restriction sites, and the tetrary goal was to prevent the formation of secondary RNA structures in close proximity to the RBS.
The Silktail is part of the C terminal domain of MaSp2 spider silk.
The Strep-tag, fused at the C terminal, is used both as a binding tag to use to inidicate its production and as a binding component to Streptavidin.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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