TEV cut siBBa_K128002 Version 1 (Component)TEV tobacco etch virus protease cleavage site
BBa_K2172009BBa_K2172009 Version 1 (Component)Tac Promoter-RBS-GST-Thrombin Protease-GFP-Terminator
AraC_TEV-FBBa_K627010 Version 1 (Component)Fusion of AraC induction system and TEV protease 3
AraC_TEV-FBBa_K627008 Version 1 (Component)Fusion part of arabinose-inducible induction system and the TEV protease
ssTorA_CS-BBa_K627012 Version 1 (Component)Fusion of TorA sig-seq, TEV protease cleavage site and b-lactamase
Bacillus subtilis Collectionbsu_collection Version 1 (Collection)This collection includes information about promoters, operators, CDSs and proteins from Bacillus subtilis. Functional interactions such as transcriptional activation and repression, protein production and various protein-protein interactions are also included.
PrtDEFBBa_K258007 Version 1 (Component)Export of recombinant proteins in Escherichia coli using ABC transporter of Erwinia chrysanthemi
BBa_K079051BBa_K079051 Version 1 (Component)LacI repressor and GFP reporter proteins controlled by the J23118 promoter and Lac 1 operator
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.