Coding
mNeon

Part:BBa_K5121012:Design

Designed by: Carlo Famularo   Group: iGEM24_Sydney-Australia   (2024-09-04)


GGG-mNeon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 124
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We were supplied with a pCDFDuet plasmid containing mNeonGreen with an N-terminal polyhistidine tag. We engineered this plasmid to add a TEV protease cleavage site (ENLYFQ/G) and tri-glycine sequence downstream of the his-tag. The TEV cleavage site serves two purposes: 1) it allows selective elution of mNeonGreen from Ni-NTA columns by performing on-bead cleavage; and 2) cleavage leaves a poly-glycine sequence on the N-terminus of mNeonGreen. This makes mNetTEVonGreen suitable for sortase-mediated conjugation reactions, which we performed on modified R-bodies we engineered (BBa_K5121027, BBa_K5121028).

Source

E. coli