Difference between revisions of "Part:BBa K3927001"

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===Usage===
 
===Usage===
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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===
 
===Design===

Revision as of 02:42, 18 October 2021


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.

Characterization

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