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Coding

Part:BBa_K1085004

Designed by: Claudio Tiecher   Group: iGEM13_Groningen   (2013-09-10)

RBS Start-codon SpiderSilkSubunitN1 Strep-tag

This BioBrick contains the coding sequence for part of a spider silk protein (SubunitN1) with a Bacillus subtilis ribosome binding site (RBS) and the 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.

SubunitN1 codes for a novel 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.

Related BioBricks

BBa_K1085005 is the same except it has no strep-tag between Start-codon and the SilkSubunitN1 in the sequence.

BBa_K1085000 is the same except it has a SpiderSilkSubunitE1 instead of SpiderSilkSubunitN1.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 68
  • 1000
    COMPATIBLE WITH RFC[1000]


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