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Design of gene circuits used in this protocol (A) DNA sequence of B0034-MCS1-pSB1A2 between <t>RFC10</t> prefix and suffix. The solid green arrow marks the start codon of the open reading frame. (B) Schematic diagrams of synthetic gene circuits constructed and characterized in this study. Self-activation circuits include non–phase-separating controls (OP152 and OP177) and phase-separating circuits incorporating intrinsically disordered regions (OP153 and OP203). (C) Colocalization circuits include CT149, which contains 12× lacO operator sites adjacent to the target promoter, and IC41, a matched control lacking lacO sites. All circuits are based on AraC-mediated positive feedback and are expressed in E. coli.
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Design of gene circuits used in this protocol (A) DNA sequence of B0034-MCS1-pSB1A2 between <t>RFC10</t> prefix and suffix. The solid green arrow marks the start codon of the open reading frame. (B) Schematic diagrams of synthetic gene circuits constructed and characterized in this study. Self-activation circuits include non–phase-separating controls (OP152 and OP177) and phase-separating circuits incorporating intrinsically disordered regions (OP153 and OP203). (C) Colocalization circuits include CT149, which contains 12× lacO operator sites adjacent to the target promoter, and IC41, a matched control lacking lacO sites. All circuits are based on AraC-mediated positive feedback and are expressed in E. coli.
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Design of gene circuits used in this protocol (A) DNA sequence of B0034-MCS1-pSB1A2 between <t>RFC10</t> prefix and suffix. The solid green arrow marks the start codon of the open reading frame. (B) Schematic diagrams of synthetic gene circuits constructed and characterized in this study. Self-activation circuits include non–phase-separating controls (OP152 and OP177) and phase-separating circuits incorporating intrinsically disordered regions (OP153 and OP203). (C) Colocalization circuits include CT149, which contains 12× lacO operator sites adjacent to the target promoter, and IC41, a matched control lacking lacO sites. All circuits are based on AraC-mediated positive feedback and are expressed in E. coli.
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Design of gene circuits used in this protocol (A) DNA sequence of B0034-MCS1-pSB1A2 between RFC10 prefix and suffix. The solid green arrow marks the start codon of the open reading frame. (B) Schematic diagrams of synthetic gene circuits constructed and characterized in this study. Self-activation circuits include non–phase-separating controls (OP152 and OP177) and phase-separating circuits incorporating intrinsically disordered regions (OP153 and OP203). (C) Colocalization circuits include CT149, which contains 12× lacO operator sites adjacent to the target promoter, and IC41, a matched control lacking lacO sites. All circuits are based on AraC-mediated positive feedback and are expressed in E. coli.

Journal: STAR Protocols

Article Title: Protocol for condensate-based stabilization of gene circuit dynamics under growth-mediated dilution in E. coli

doi: 10.1016/j.xpro.2026.104536

Figure Lengend Snippet: Design of gene circuits used in this protocol (A) DNA sequence of B0034-MCS1-pSB1A2 between RFC10 prefix and suffix. The solid green arrow marks the start codon of the open reading frame. (B) Schematic diagrams of synthetic gene circuits constructed and characterized in this study. Self-activation circuits include non–phase-separating controls (OP152 and OP177) and phase-separating circuits incorporating intrinsically disordered regions (OP153 and OP203). (C) Colocalization circuits include CT149, which contains 12× lacO operator sites adjacent to the target promoter, and IC41, a matched control lacking lacO sites. All circuits are based on AraC-mediated positive feedback and are expressed in E. coli.

Article Snippet: BglII-RFP- RFC10 surfix , E1010 , RFP-BglII-F , VR , 880 bp.

Techniques: Sequencing, Construct, Activation Assay, Control