Reporter

Part:BBa_K2082113:Design

Designed by: Carsten Hain   Group: iGEM16_Bielefeld-CeBiTec   (2016-10-14)


Stop-GFP (K3*) under control of constitutive promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 705


Design Notes

Stop position was determined by FoldX modelling. Reversion of the stop codon to every other amino acid should result in a functional protein.

Figure 1: Extensive in silico stability scan for GFP (A) and mapping of the stability changes for a complete in silico alanine scan (B). The stability changes for mutating each position inside GFP were calculated usign FoldX. The stability changes are calculated in ΔΔG (difference of mutant folding energy to wild type folding energy, low means more stable). As overview the complete values for mutating to alanine were mapped on the GFP structure. (PDB: 2WUR)


Source

cloning by side-directed mutagensis

References