Translational_Unit

Part:BBa_K515004:Design

Designed by: Atipat Patharagulpong   Group: iGEM11_Imperial_College_London   (2011-09-07)
Revision as of 15:11, 18 September 2011 by Cs3109 (Talk | contribs)

T4 Anti-Holin


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 241


Design Notes

This sequence has been taken from BBa_K112808 and has been modified through the addition of an insulator sequence between our newly created RBS and the coding sequence. The insulator sequence has been specially developed to allow the easy switching of our parts between promoters while maintaining little homology with the vector making recombination events between the insulator sequence and the vector unlikely. We have also added a double TAA terminator sequence instead of a single TGA sequence. By making this part available on its own we have increased its modularity and therefore its possible applications. We have also improved the double terminator and with the insulator sequence we have made this part comply with our new standard. The insulator sequence will allow for easy primer designing and will one day allow for the better characterization of promoters since they will be isolated from the RBS and will therefore not influence the strength of the RBS.

The chosen RBS is very strong to ensure that the amount of Anti-Holin transcribed from the genome will be able to negate the amount of Holin transcribed from the high copy plasmid. The values used to make the RBS and the promoter chosen have all been influenced by our modelling.

Source

The coding sequence has been extracted through PCR with Phusion from BBa_K112808. The gene originates from the enterobacteria phage T4.

References