Part:BBa_K2805006:Design

PIF3-NLS-VP16

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 1065
  Illegal SpeI site found at 1461
  Illegal PstI site found at 1921
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 1461
  Illegal PstI site found at 1921
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 885
  Illegal BglII site found at 1400
  Illegal BamHI site found at 2484
  Illegal XhoI site found at 2727
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 1065
  Illegal SpeI site found at 1461
  Illegal PstI site found at 1921
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1065
  Illegal SpeI site found at 1461
  Illegal PstI site found at 1921
  
• 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

This is one part of the PhiReX system, designed by Hochrein et al. (Nucleic Acids Research, 2017, doi: 10.1093/nar/gkx610). PhyBNT and PIF3 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.