Composite

Part:BBa_K2787001:Design

Designed by: Haoer Shi   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

Given the orthogonal RBS sequence, we intentionally added a few base pairs between RBS and the GFP start codon to ensure that the RBS function appropriately.

Source

The part was built by inserting the o-RBS and promoter before the GFP coding sequencing provided by 2018 Distribution Kit. The orthogonal RBS 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