Intein_assisted_Bisection_MappingIntein_assisted_Bisection_Mapping_collection Version 1 (Collection)Split inteins are powerful tools for seamless ligation of synthetic split proteins. Yet, their use remains limited because the already intricate split site identification problem is often complicated by the requirement of extein junction sequences. To address this, we augmented a mini-Mu transposon-based screening approach and devised the intein-assisted bisection mapping (IBM) method. IBM robustly revealed clusters of split sites on five proteins, converting them into AND or NAND logic gates. We further showed that the use of inteins expands functional sequence space for splitting a protein. We also demonstrated the utility of our approach over rational inference of split sites from secondary structure alignment of homologous proteins. Furthermore, the intein inserted at an identified site could be engineered by the transposon again to become partially chemically inducible, and to some extent enabled post-translational tuning on host protein function. Our work offers a generalizable and systematic route towards creating split protein-intein fusions and conditional inteins for protein activity control.
BBa_I733007BBa_I733007 Version 1 (Component)Weight cells either turn blue or die in response to both inputs and HSL level
BBa_I741109BBa_I741109 Version 1 (Component)Lambda Or operator region
BBa_K549004BBa_K549004 Version 1 (Component)LacI promotor fused with the iron dependent regulator fur
BBa_K1795400BBa_K1795400 Version 1 (Component)Truncated pSB1C3 with no Prefix or Suffix
BBa_K228013BBa_K228013 Version 1 (Component)(pSal PO) OR Gate - GFP
BBa_K415011BBa_K415011 Version 1 (Component)PtetR : RBS : LuxR : Term : PluxR/cI-OR : RBS : mCherry : Term : Plux/cI-OR : RBS : LuxI
BBa_K1341011BBa_K1341011 Version 1 (Component)OR LOGIC GATE IN Graph Theory (GFP OUTPUT DEVICE)
BBa_K415005BBa_K415005 Version 1 (Component)pLux/cI-OR : RBS-mCherry : Term : p(tetR) : RBS-luxR : Term