Part:BBa_K1896015

mGFPuv2-mSA2 generator

This part constitutively expresses a fusion protein comprising monomeric variants of GFPuv (mGFPuv2) and Streptavidin (mSA2). When the crude lysate of a culture of E. coli that contains this part is spread over solid biotinylated poly lactic acid (PLA), the strong affinity of mSA2 to biotin causes the fusion protein to adhere to the PLA. The biotin-mSA2 interaction is strong enough to resist multiple washes with saline.

PLA is a material often used for 3D printing and biotin can be incorporated either in the raw filament itself, or coated on the printed structure [http://2016.igem.org/Team:UGent_Belgium/Filament|http://2016.igem.org/Team:UGent_Belgium/Filament]. This part was created to demonstrate that mSA2 can be used to attach proteins to 3D printed structures. The combination of Synthetic Biology and 3D printing could greatly broaden the applicability of many biological parts.

Glass slides coated with biotinylated PLA, dipped in crude cell lysate and then washed with saline. Left: the mGFPuv2-mSA2 fusion protein adheres to the PLA, right: the mGFPuv2 control easily washes off.

Sequence and Features

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 872
  Illegal AgeI site found at 932
  
• 1000
  COMPATIBLE WITH RFC[1000]