Part:BBa_K1085019
RBS Start-codon EstA Strep-tag SpiderSilkSubunitE1 SpiderSilkSubunitE2 SilkTail Stop-codon
This BioBrick contains the coding sequence for two parts of the MaSp2 spider silk proteins (SubunitE1 and SubunitE2) with a Bacillus subtilis ribosome binding site (RBS) EstA and strep-tag infront of it and a Silktail at the end.
The RBS is there to allow the ribosomes to bind and start to translate the DNA.
The estA codes for the EstA precursor protein which therefore facilitates the secretion of the produced protein.
The Strep-tag, fused at the N terminal, is used both as a binding tag to use to inidicate its production and as a binding component to Streptavidin.
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 amino acid sequences of the Subunits are based on the adapted MaSp2 sequences discribed in the paper of Brooks et. al. 2008.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 650
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 262
- 1000COMPATIBLE WITH RFC[1000]
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