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Part:BBa_K4850008:Design
GFP-sacB Negative Selection Marker
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 729
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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