Difference between revisions of "Part:BBa K2066116:Design"

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===Design Notes===
 
===Design Notes===
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To be completed
 
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This part was made to put the synthetic enhancer suite (Amit et. al. 2011) onto the Biobrick backbone flanked by UNS for easy cloning.  Furthermore, we exchanged the reporter gene with sfGFP for more pronounced fluorescent measurements of the step wise transfer functions.
  
 
===Source===
 
===Source===

Latest revision as of 04:11, 29 October 2016


Synthetic Enhancer Project: 3x TetO Binding Cassette (52s) + sfGFP on UNS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 111
    Illegal NheI site found at 206
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 171
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 951
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1939


Design Notes

This part was made to put the synthetic enhancer suite (Amit et. al. 2011) onto the Biobrick backbone flanked by UNS for easy cloning. Furthermore, we exchanged the reporter gene with sfGFP for more pronounced fluorescent measurements of the step wise transfer functions.

Source

The enhancer, tet cassette, glnAp2 synthetic promoter, and NRI coding region sequences were derived from Amit, R., Garcia, H. G., Phillips, R. & Fraser, S. E. Building enhancers from the ground up: a synthetic biology approach. Cell146, 105–118 (2011). The sfGFP flourescent reporter design is inspired by C. Lou, B. Stanton, Y.-J. Chen, B. Munsky, C. A. Voigt, Ribozyme-based insulator parts buffer synthetic circuits from genetic context. Nat. Biotechnol. 30, 1137 (2012). doi:10.1038/nbt.2401 pmid:23034349. The UNS sequences at the ends of the insert are derived from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860. A huge thanks to all the researchers involved in its original creation!

References