Coding

Part:BBa_K1085029

Designed by: Mirjam Groenewold   Group: iGEM13_Groningen   (2013-09-17)

FliZ Strep-tag SpiderSilkSubunitE1

This biobrick contains signal peptide sequence of FliZ attached to silk subunit E1 with an N-terminal strep-tag.

FliZ codes for the signal peptide sequence of 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.

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.

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]


[edit]
Categories
//cds
//chassis/prokaryote/bsubtilis
//collections/silk
Parameters
None