Coding

Part:BBa_K2787005:Design

Designed by: Weiyi Tang   Group: iGEM18_ShanghaiTech   (2018-10-09)


RFP with LVA tag under innate RBS, GFP with LVA tag under C9 orthogonal RBS with orthogonal 16S rRNA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 949
    Illegal NheI site found at 972
    Illegal NheI site found at 1879
    Illegal NheI site found at 1902
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2741
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 616
    Illegal AgeI site found at 728
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1652


Design Notes

Though most of the open database will show the 16s RNA only contains 1542bp, it is still necessary to obtain about 300 bp more since it will help to form the 16S rRNA precursor, otherwise, it may not function appropriately.


Source

The part is cloned from the Escherichia coli genome and is mutated in Shine-Dalgarno sequence and 722 to 723 bp which will form a bulge proximal to the minor groove of the SD helix formed between the ribosome and mRNA. The orthogonal 16S rRNA sequence was published by Rackham, Oliver and Chin, Jason W. in their Nature Chemical Biology paper A network of orthogonal ribosome·mRNA pairs(doi:10.1038/nchembio719).


References