Coding
RGE

Part:BBa_K1442102:Design

Designed by: Becky Seeley   Group: iGEM14_Warwick   (2014-10-07)

Reverse GFP for E.coli


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 NgoMIV site found at 742
    Illegal NgoMIV site found at 770
    Illegal NgoMIV site found at 799
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 763


Design Notes

There are two restriction sites surrounding the 3’UTR AflII and BsiW1 which can be used to change the RDRP promotor.

The GFP has been codon optimised for E.coli IDTs codon optimising software.

The GFP and RBS are in the system in the reverse complement as they are transcribed to forwards sense in the incidence of RdRp functioning correctly.

Two stop codons are added after the T7 promoter to ensure no translation occurs here.

Source

The sequence for GFP was obtained from the NCBI database and was codon optimised.

References

https://www.idtdna.com/CodonOpt