Data repository for work contributing to the paper "Sculpting DNA-based synthetic cells through phase separation and phase-targeted activity".

Confocal micrographs are presented as raw .TIFF files with corresponding metadata .xml file, which can be opened using text editing software such as Notepad. Most confocal micrographs were collected using a Leica TCS SP5 confocal microscope (objectives used: HC PL FLUOTAR 20 × 0.50 N.A. dry objective and HCX PL APO CS 20 × 0.70 N.A. immersion objective (Leica) using type F immersion liquid (Leica)). Other confocal micrographs were obtained using a Leica STELLARIS 8 Inverted Confocal Microscope (HC PL APO 20 × 0.75 N.A. CS2 dry objective (Leica)).

Widefield epifluorescence micrographs are presented as .nd2 files which can be read using the free Nikon Elements Viewer software. Metadata is embedded into these files. These micrographs were collected using a Nikon Eclipse Ti2-E inverted microscope (Plan Fluor 20 × 0.75 N.A and Plan Fluor 40 × 0.9 5 N.A dry objectives (Nikon)).

SAXS data was collected at the I22 beamline at the Diamond Light Source in Oxfordshire, using a radiation wavelength of 1 angstrom, beam size ∼300 µm wide by 100 µm high, and an exposure time of 100 ms. Data is presented as .nsx files, which can be read using the DAWN software or custom code.

NB: some data, particularly confocal and SAXS, are labelled according to an internal labelling scheme and do not match the sample nomenclature used in the manuscript. Where necessary, we have provided a .txt file matching internal labelling to that used in the manuscript.

UVvis data was collected using an Agilent Cary 3500 UV-vis spectrophotometer, data is presented in .xlsx format. DLS data was collected using a Malvern Panalytical Zetasizer, data is presented in .xlsx format.

Bulk fluorescence measurements were acquired using a BMG CLARIOstar Plus plate reader, data is presented in .csv format.

See the main manuscript and SI (all open access) for further details on samples and data collection methods.