Part:BBa_K2805008:Design

PhiReX -> mRuby

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 6966
  Illegal XbaI site found at 4492
  Illegal SpeI site found at 4888
  Illegal PstI site found at 5348
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 6966
  Illegal SpeI site found at 4888
  Illegal PstI site found at 5348
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal suffix found in sequence at 3021
  Illegal EcoRI site found at 6966
  Illegal BglII site found at 885
  Illegal BglII site found at 1380
  Illegal BglII site found at 4312
  Illegal BglII site found at 4827
  Illegal BglII site found at 7446
  Illegal BamHI site found at 5911
  Illegal BamHI site found at 7032
  Illegal BamHI site found at 8163
  Illegal XhoI site found at 1413
  Illegal XhoI site found at 1432
  Illegal XhoI site found at 6154
  Illegal XhoI site found at 7440
  Illegal XhoI site found at 8172
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 6966
  Illegal XbaI site found at 4492
  Illegal SpeI site found at 4888
  Illegal PstI site found at 5348
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 6966
  Illegal XbaI site found at 4492
  Illegal SpeI site found at 4888
  Illegal PstI site found at 5348
  Illegal NgoMIV site found at 8483
  Illegal AgeI site found at 2847
  Illegal AgeI site found at 7056
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1629
  

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

Designed by Lena Hochrein et al. (Nucleic Acids Research, 2017, doi: 10.1093/nar/gkx610). PIF3 and PhyBNT originate in Arabidopsis thaliana.

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. Hochrein L, Machens F, Messerschmidt K, Mueller-Roeber B. PhiReX: A programmable and red light-regulated protein expression switch for yeast. Nucleic Acids Res. 2017;45(15):9193-9205. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5587811/. doi: 10.1093/nar/gkx610.

3. 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.