Composite

Part:BBa_K2066118:Design

Designed by: Kalen Clifton, Christine Gao, Andrew Halleran, Ethan Jones, Likhitha Kolla, Joseph Maniaci, John Marken, John Mitchell, Callan Monette, Adam Reiss   Group: iGEM16_William_and_Mary   (2016-10-14)


Synthetic Enhancer with 3X TetO cassette (52s) on UNS backbone


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
    COMPATIBLE WITH RFC[1000]


Design Notes

We made this part to be compatible with the iGEM Biobrick standard as well as flank the synthetic enhancer circuit (Amit et. al. 2011) with UNS sequences for easy cloning.

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 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