Part:BBa_K1085024

RBS Start-codon EstA Strep-tag SpiderSilkSubunitN1 SpiderSilkSubunitN2

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.

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.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 650
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 707
  Illegal AgeI site found at 262
  
• 1000
  COMPATIBLE WITH RFC[1000]