Reporter

Part:BBa_K199074:Design

Designed by: Clifton Davis   Group: iGEM09_MoWestern_Davidson   (2009-09-20)

RBS RFP with AGGAC Addition


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Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 578
    Illegal AgeI site found at 690
  • 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 RFP 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 RFP gene, we utilized the restriction enzyme NcoI that cuts at a single site 419-bp into BBa_E1010.

Source

The template of the gene comes from Part BBa_E1010. We designed a forward primer that included RBS,ATG, the 5-bp addition AGGAC and the first 20 nucleotides on the 5' end of RFP after ATG. This made the edited 5' sequence for our RFP.

References