Intermediate

Part:BBa_K260016:Design

Designed by: Kaj Bernhardt   Group: iGEM09_BIOTEC_Dresden   (2009-10-18)
Revision as of 15:56, 18 October 2009 by Kaj (Talk | contribs)

TT_FRT_GFP. (double terminator, FRT site, optimised GFP)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 144
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 144
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 144
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 809


Design Notes

A strong promoter was necessary and we use BBa_J23100. An appropriate reading frame for the FRT site had to be found to prevent premature Stop-codons and hydrophobic amino acid residues. Further, the gene encoding GFP (mut3b) was codon optimised for expression in both E. coli and H. sapiens. For potential use in H. sapiens, the CDS were also made free of polyA sites, exon junctions and CpG dinucleotides where possible.


Source

This part was synthesised by "Mr Gene" (Geneart) and is available in the original pMA-BsdR-@CC2FosMB plasmid backbone (BBa_K260010), but also pCC2FosMB BBa_K260001, which is already half-way towards constructing the full P_FRT_dhfr_TT_FRT_GFP reporter.

References