Coding

Part:BBa_K2168001:Design

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


pBAD/araC - (N-6xHis) - Ag43 - sfGFP


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
    Illegal SapI.rc site found at 4414


Design Notes

The location for the 6xHis tag is the same as that for BBa_K2168000(Ag43 tagged with 6xHis on N-terminal under pBAD/araC promoter) and uses the same design principles. For independent folding, a flexible GGGGSGGGGS linker was added between Ag43 and sfGFP, and the linker was choosen to be not very hydrophilic or hydrophobic, since Ag43 lies very close to lipid membranes.


Source

This part was made from Gibson assemblies from fragments of two previous parts - BBa_K1352000 and BBa_I746908, which code for Ag43 and sfGFP respectively.

References