Part:BBa_K4850008:Design

GFP-sacB Negative Selection Marker

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 729
  
• 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 644
  

Design Notes

The protein linker linking the GFP gene and sacB gene should be a rigid protein linker.

Source

The genetic information of both GFP sequence and sacB sequence were obtained from the lab. The sequence of the protein linker A(EAAAK)₂A protein linker was obtained from the Arai et al. (2001) paper.

References

[1] Arai, R., Ueda, H., Kitayama, A., Kamiya, N., & Nagamune, T. (2001). Design of the linkers which effectively separate domains of a bifunctional fusion protein. Protein Engineering Design & Selection, 14(8), 529–532.https://doi.org/10.1093/protein/14.8.529

[2] Chen, X., Zaro, J. L., & Shen, W. (2013). Fusion protein linkers: Property, design and functionality. Advanced Drug Delivery Reviews, 65(10), 1357–1369. https://doi.org/10.1016/j.addr.2012.09.039