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Construction of engineered biological systems from collections of standard biological parts requires mechanisms for rapid and reliable characterization of parts. Additionally, the difficulty of rational part design necessitates library screening systems that can be employed in service of tuning part performance. Here we describe pSB1A10, a system for both characterizing and screening transcription-based parts based on their input / output function. We demonstrate the successful operation of this system by characterizing and tuning genetic inverters and transcriptional terminators.
Screening Plasmid 1.0
Characterization of Empty Screening Plasmid 1.0
- Endy:Screening plasmid/v1.0/Hairpin - characterize effectiveness of the hairpin to stabilize the mRFP1 transcript
- Endy:Screening plasmid/v1.0/RNAseE sites - characterize effectiveness of the RNAseE sites
To ensure our screening system was operating as expected we measured the expression levels of GFP and mRFP1 when the plasmid did not contain a part (e.g. input equals output).
Terminator Characterization via Screening Plasmid 1.0
Inverter Characterization via Screening Plasmid 1.0
Inverter library screening via Screening Plasmid 1.0
To be added.
Screening Plasmid 1.5
This version of the screening makes use of an AHL-based induction system (<bbpart>F2620</bbpart>).
Screening Plasmid 2.0
We are in the process of designing a new version of the screening plasmid to account for some of the shortcomings of the previous version. Details can be found here:
- Registry technical report (in progress)