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This page is a work in progress.
The screening plasmid is designed to enable fast characterization of PoPS-based parts and devices. It consists of 4 components: (1) Tunable input (2) Input measurement (3) Part/Device insertion site (4) Output measurement.
Screening Plasmid 1.0
Terminator Characterization via Screening Plasmid 1.0
Inverter Characterization via Screening Plasmid 1.0
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. An early list of possible improvements:
- Brighter FPs (in particular RFP is very dim)
- Emerald & tdTomato look pretty promising
- Lower copy plasmid
- New Inducible Expression System
- Propionate-inducible induction system, has a larger dynamic range than pBAD and doesn't require a specialized strain. (i.e. no knockouts of native systems needed.)
- It looks like MC4100 is missing the propionate metabolism genes and so wouldn't work with this system, these are the two genes in K12 are homologous to the prpBCDE operon:
- 349236..350405, 350439..351890 Unfortunately, there is a 98kb deletion from the MC4100 (a K12 derivative) genome from 274723-371962, so it's missing these genes.
- However, I'm not tied to MC4100, would rather get this working in K12 and then we could maybe make some progress on doing directed deletions of K12 as planned in the Biobricks Standard Strain.
 Khlebnikov et al,Modulation of gene expression from the arabinose-inducible araBAD promoter. J Ind Microbiol Biotechnol. 2002 Jul;29(1):34-7.
 Effect of gene location, mRNA secondary structures, and RNase sites on expression of two genes in an engineered operon. Biotechnol Bioeng. 2002 Dec 30;80(7):762-76.