Part:BBa_K2805003:Design

LOV2: NLS-VP16-EL222

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal suffix found in sequence at 1782
  Illegal BglII site found at 885
  Illegal XhoI site found at 1617
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1187
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The primary concern was Dueber MoClo and GoldenGate compatibility. These sequences were designed to be robustly assembled and integrated into the S. cerevisiae genome.

Source

LOV2 was identified and engineered from E. litoralis by Laura B. Motta-Mena et al. (Nature Chemical Biology 2014, doi: 10.1038/nchembio.1430).

References

1. Benzinger D, Khammash M. Pulsatile inputs achieve tunable attenuation of gene expression variability and graded multi-gene regulation. Nature Communications. 2018;9(1):3521. https://doi.org/10.1038/s41467-018-05882-2. doi: 10.1038/s41467-018-05882-2.

2. Motta-Mena L, Reade A, Mallory MJ, et al. An optogenetic gene expression system with rapid activation and deactivation kinetics. Nature Chemical Biology. 2014;10:196. http://dx.doi.org/10.1038/nchembio.1430.

3. Zhao EM, Zhang Y, Mehl J, et al. Optogenetic regulation of engineered cellular metabolism for microbial chemical production. Nature. 2018;555:683. http://dx.doi.org/10.1038/nature26141.

4. Lee ME, DeLoache WC, Cervantes B, Dueber JE. A highly characterized yeast toolkit for modular, multipart assembly. ACS Synth Biol. 2015;4(9):975-986. https://doi.org/10.1021/sb500366v. doi: 10.1021/sb500366v.