Part:BBa_K1085037

FliZ Strep-tag SpiderSilkSubunitE1 SpiderSilkSubunitE2

This biobrick contains signal sequence FliZ attached to silk subunit N1 with an N-terminal strep tag and silk subunit N2.

The FliZ codes for the signal sequence FliZ. FliZ might be involved in the anchoring of the rod to the membrane [1].

The Strep-tag, fused at the N-terminal, is used both as a binding tag to use to indicate its production and as a binding component to Streptavidin.

Subunit N1 codes for a spider silk protein that is optimized for maximal expression in Bacillus subtilis 168. The primary goal of the algorithm was to optimize the codon selection based on their availability scores. The secondary goal was to prevent the formation of secondary RNA structures in close proximity to the RBS, and the tertiary goal was to minimize the number of restriction sites.

The amino acid sequence is based on the adapted MaSp2 sequences described in the paper of Brooks et. al. 2008.

[1] http://www.uniprot.org/uniprot/P35536, visited 4-10-2013.

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 82
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]