Coding

Part:BBa_K1085000

Designed by: Claudio Tiecher   Group: iGEM13_Groningen   (2013-08-30)

RBS Start-codon Strep-tag SpiderSilkSubunitE1

This BioBrick contains the coding sequence for part of the MaSp2 silk protein (SubunitE1) with a Bacillus subtilis ribosome binding site (RBS) and a Strep-tag in front of it.

The RBS is there to allow the ribosomes to bind and start to translate the DNA.

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 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.


Related BioBricks

BBa_K1085001 is the same except it has no strep-tag between Start-codon and the SilkSubunit in the sequence.

BBa_K1085004 is the same except it has a SpiderSilkSubunitN1 instead of SpiderSilkSubunitE1.


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds
//chassis/prokaryote/bsubtilis
//collections/silk
Parameters
n/aRBS strep-tag SpiderSilkSubunitE1