Reporter

Part:BBa_K199035:Design

Designed by: Alyndria Thompson   Group: iGEM09_MoWestern_Davidson   (2009-07-13)


Tet Resistance with CGGUC Addition


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 149
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 295
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 321
    Illegal NgoMIV site found at 689
    Illegal NgoMIV site found at 849
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We positioned the start codon, ATG, before the 5-bp codon and continued with the rest of the Tet gene. This way, the RBS wouldn't translate the reporter gene without reading over the 5-base codon first. In order to ligate our edited 5' sequence to the unchanged downstream portion of the Tet gene, we utilized the restriction enzyme BamHI that cuts at a single site 290-bp into BBa_J31007.


Source

The template of the gene comes from Part BBa_J31007. We designed a forward primer that included ATG, the 5-bp addition CGGTC and the first 20 nucleotides on the 5' end of Tet after ATG. This made the edited 5' sequence for our Tet.

References