Team:Stanford/Part Collection

Stanford iGEM Team Wiki

Parts Collection

Parts Collection

Part Name
1 BBa_K3258001 M13 pIV
2 BBa_K3258003 M13 pII
3 BBa_K3258004 M13 pIX
4 BBa_K3258005 M13 pVII
5 BBa_K3258006 Infection Reporter (Design Using BBa_K2832003)
6 BBa_K3258007 M13 pIII Neg
7 BBa_K3258008 M13 Helper Plasmid for PREDCEL+
8 BBa_K3258009 Selection Plasmid PREDCEL+
9 BBa_K3258010 T7 to T3 Selection Schema PREDCEL+
10 BBa_K3258011 lacI-derived biosensor selection schema (positive selection) PREDCEL+
11 BBa_K3258012 lacI-derived biosensor selection schema (negative selection) PREDCEL+
12 BBa_K3258013 Selection Plasmid T7 to T3 PREDCEL+
13 BBa_K3258014 Selection Plasmid T7 to T3 PREDCEL+ (Alternate Version)
14 BBa_K3258019 M13 pVI

Best Parts Collection

We submit PREDCEL+ SHOP as our best new parts collection. SHOP stands for Selection Helper and Other Plasmids, and is our collection of components to assist in the implementation of selection schema and directed evolution using our improved PREDCEL+ directed evolution technique. To make PREDCEL more accessible to iGEM teams, we created a standard set of three plasmids to implement user-defined selection schema. It improves PREDCEL in three ways. First, our selection is based off pVI instead of pIII expression - significant because pIII renders cells immune to infection. Second, our plasmid architecture enables ‘infection reporter’ use, which provides a visual indicator of the number of cells infected in a given E. coli population. This feedback is used to modulate selection stringency, driving evolution forward. Finally, pIII expression is triggered post-infection, preventing multiple phage phenotypes from infecting a cell. This stops “cheater” phenotypes from proliferating as described by Brodel et al. (2016). To document this collection, we present all M13 phage genes not previously documented alongside composites we used for evolving T7 polymerase to recognize a T3 promoter and new lacI based biosensors.

Team Pic
Generalized and simplified selection schema for directed protein evolution with PREDCEL+ technique.