May

Competent E. coli KRX and BL21(D3) cells

Glycerol stocks

Plasmid preparation with 'Jugend Forscht'

June

PCR of pGGA select

IDT gene fragment resuspesion and PCR

Golden Gate of lacZ, mScarlet, sfGFP and NanoLuc constructs

Transformation into BL21(D3) and DH5⍺

Cloning of constructs into DH5⍺

Sequencing of mScarlet, sfGFP, lacZ and NanoLuc

Repeat Golden Gate of mScarlet, sfGFP, lacZ and NanoLuc

July

Colony PCR of mScarlet, sfGFP, lacZ and NanoLuc in DH5⍺

Golden Gate of lacZ

Transformation of mScarlet, sfGFP, lacZ and NanoLuc into KRX

Colony PCR of constructs in KRX

Sequencing of lacZ and NanoLuc

Liquid culture assay with lacZ constructs

Sequencing of R2 and R3 mScarlet constructs

Design & Order of R5 construct

Golden Gate of R2 sfGFP, R3 lacZ and all R5 constructs

Liquid culture assay with R2 and R4 lacZ ribotswitch

Sequencing of R2 sfGFP, R5 sfGFP, and R5 NanoLuc

August

Testing all sfGFP constructs in plate reader

Testing all mScarlet constructs in plate reader

Testing all NanoLuc constructs in plate reader

Sequencing of R5 mScarlet, R3 lacZ and R5 lacZ

Transformation of R1, R2, R3 and R4 in lacZ

Testing all lacZ constructs with ONPG assay

September

Testing lacZ in BL21(D3) and KRX with X-Gal

Testing sfGFP, mScarlet and NanoLuc constructs in plate reader

Testing cell-free kit