Enhancer Construction for Per and Tim

The enhancer of Per and Tim were synthesized as gBlocks (Integrated DNA Technologies) in 2× tandem multiplexes. Then two copies of the gBlocks were amplified by PCR and inserted into a reporter vector using In-Fusion HD Cloning (Clontech) to generate 4× Per and 4× Tim enhancers (Figure S4). The reporter vector contains fly heat shock mini promoter and an nlsGFP fused with PEST domain at the C terminus (destabilized GFP) as previously used for the construction of Notch reporter(Hunter et al., 2016 (link)).

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Parasram K., Bernardon N., Hammoud M., Chang H., He L., Perrimon N, & Karpowicz P. (2018). Intestinal Stem Cells Exhibit Conditional Circadian Clock Function. Stem Cell Reports, 11(5), 1287-1301.

Publication 2018

gBlocks In-Fusion HD Cloning

Heat shock Pest Vector

Corresponding Organization : University of Windsor

Other organizations : Harvard University

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Variable analysis

independent variables

Enhancer of Per and Tim were synthesized as gBlocks (Integrated DNA Technologies) in 2× tandem multiplexes
Two copies of the gBlocks were amplified by PCR and inserted into a reporter vector using In-Fusion HD Cloning (Clontech) to generate 4× Per and 4× Tim enhancers

dependent variables

Not explicitly mentioned

control variables

The reporter vector contains fly heat shock mini promoter and an nlsGFP fused with PEST domain at the C terminus (destabilized GFP) as previously used for the construction of Notch reporter(Hunter et al., 2016)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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