Coding

Part:BBa_K2168000:Design

Designed by: Arunavo Chakraborty   Group: iGEM16_IISc_Bangalore   (2016-10-13)


pBAD/araC - N(6xHis) Ag43


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1963
    Illegal NgoMIV site found at 3187
    Illegal NgoMIV site found at 3697
    Illegal NgoMIV site found at 3718
    Illegal AgeI site found at 979
    Illegal AgeI site found at 2713
    Illegal AgeI site found at 3457
    Illegal AgeI site found at 3871
    Illegal AgeI site found at 4113
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

Ag43 is an outer membrane E coli protein, so it has a N-terminal 52 amino acid long signal peptide sequence, which is cleaved after Ag43 pro-protein is transported into the periplasmic space. Since strong dependence has been demonstrated on the signal peptide cleavage on the signal peptide-protein junction, the 6xHis tag should be added after Ala53, the residue immediately after the signal peptide sequence.


Source

This comes from Gibson assembly of fragments from a previous biobrick, BBa_K1352000 by Team:Aberdeen_Scotland, 2014.

References