Part:BBa_K3927001

C120-CYC

This part encodes for a truncated CYCp core promoter with C120 sequence replacing the native upstream activating sequence.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 104
  Illegal SpeI site found at 79
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 86
  Illegal SpeI site found at 79
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 104
  Illegal SpeI site found at 79
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 104
  Illegal SpeI site found at 79
  
• 1000
  COMPATIBLE WITH RFC[1000]

Description

Usage

NLS-VP16-EL222 was expressed constitutively using the native promoter ACT1p in S.cerevisiae strain BY4741 from an episomal plasmid alongside C120-CYC upstream of the fluorescent protein mKO (Figure 1). BY4741 containing this construct was exposed to blue light as well as darkness for 6 hours, and compared to BY4741 without the construct present (Figure 2).

Design

Blue light induced activation of this promoter is dependent on simultaneous expression of an EL222 based transcription factor containing a nuclear localization sequence, and an activation domain.

References

1. Motta-Mena, L. B., Reade, A., Mallory, M. J., Glantz, S., Weiner, O. D., Lynch, K. W., & Gardner, K. H. (2014). An optogenetic gene expression system with rapid activation and deactivation kinetics. Nature chemical biology, 10(3), 196–202. https://doi.org/10.1038/nchembio.1430

2. Benzinger D, Khammash M. Pulsatile inputs achieve tunable attenuation of gene expression variability and graded multi-gene regulation. Nat Commun. 2018 Aug 30;9(1):3521. doi: 10.1038/s41467-018-05882-2. PMID: 30166548; PMCID: PMC6117348