DOI: 10.5061/dryad.rxwdbrvh9
title: DNA sequences for: Synthetic control of actin polymerization and symmetry breaking in active protocells

description: Non-linear biomolecular interactions on membranes drive membrane
 remodeling crucial for biological processes including chemotaxis,
 cytokinesis, and endocytosis. The complexity of biomolecular interactions,
 their redundancy, and the importance of spatiotemporal context in membrane
 organization impede understanding of the physical principles governing
 membrane mechanics. Developing a minimal in vitro system that mimics
 molecular signaling and mem- brane remodeling while maintaining
 physiological fidelity poses a significant challenge. Inspired by
 chemotaxis, we reconstructed chemically regulated actin polymerization
 inside vesicles, guiding membrane self-organization. An external,
 undirected chemical input induced directed actin polymerization and
 membrane deformation uncorrelated with upstream biochemical cues,
 suggesting symmetry breaking. A biophysical model incorporating actin
 dynamics and membrane mechanics proposes that uneven actin distributions
 cause non-linear membrane deformations, consistent with experimental
 findings. This protocellular system illuminates the interplay between
 actin dynamics and membrane shape during symmetry breaking, offering
 insights into chemotaxis and other cell biological processes.
description: Sequencing Files: The sequences provided were generated using
 standard cloning methods and were verified via Sanger sequencing for
 accuracy and completeness.
description: # DNA sequence files Supplementary DNA Sequences for 'Synthetic
 Control of Actin Polymerization and Symmetry Breaking in Active
 Protocells' Manuscript Title: Synthetic Control of Actin
 Polymerization and Symmetry Breaking in Active Protocells Authors: Shiva
 Razavi, Felix Wong, Bedri Abubaker-Sharif, Hideaki T. Matsubayashi, Hideki
 Nakamura, Nhung Thi Hong Nguyen, Douglas N. Robinson, Baoyu Chen, Pablo A.
 Iglesias, and Takanari Inoue. Access this dataset on Dryad:
 [https://doi.org/10.5061/dryad.rxwdbrvh9](https://doi.org/10.5061/dryad.rxwdbrvh9) ## Description of the data and file structure This dataset comprises the DNA sequences corresponding to the proteins that were purified and utilized in this study. The files are provided in Genebank (.gbk) format and include the following sequences: \- 2xStrep-mActA(1-184)-FRB-CFP: Contains the sequence of ActA(1-184) codon-optimized for mammalian expression and tagged with a Strep tag for purification. The FRB module enables the translocation of ActA via chemically inducible dimerization. The CFP tag facilitates visualization using standard microscopy techniques. \- mActA(1-584)-FKBP-CFP: Contains the sequence of ActA(1-584) codon-optimized for mammalian expression and tagged with a 6xHis tag for purification. \- Cap_32 and Cap_34: These are the dictyostelium capping protein sequences used in this study. \- YFP-FKBP and CFP-FRB: Contain chemically inducible dimerization modules that dimerize with rapamycin, resulting in a FRET signal. \- mCherry-FKBP-MARCKS: Contains FKBP tagged with MARCKS for lipid tethering. \- mCherry-FKBP: Contains FKBP tagged with a 6xHis tag for tethering to nickel-conjugated lipid. ## Sharing/Access information To received a vial of the DNA sequences please contact the corresponding authors: [srazavi@mit.edu](mailto:srazavi@mit.edu) and [jctinoue@jhmi.edu](mailto:jctinoue@jhmi.edu)