Reporter

Part:BBa_K199006:Design

Designed by: Shamita Punjabi   Group: iGEM09_MoWestern_Davidson   (2009-06-26)


Tet Resistance with CCACU 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 ATG before the 5-base codon and continued with the rest of the Tet resistance gene. This way, the RBS wouldn't read the reporter gene without reading over the 5-base codon first. In order to ligate our 5-base pair addition at the desired position in the reporter gene, we utilized the restriction enzyme BamHI which cut at a single site 290 bp into the gene.


Source

The template of the gene comes from BioBrick part J31007. We designed a forward primer that included the first 20 nucleotides on the 5' end, and a reverse primer that included the last 20 nucleotides on the 3' end of the Tet resistance gene.


References