BBa_K180005BBa_K180005 Version 1 (Component)GoL - Primary plasmid (part 1)/RPS - Paper primary plasmid (part 1) [LuxR generator]
pSBBs0KBBa_K823026 Version 1 (Component)pSB<sub>Bs</sub>0K-P<sub>spac</sub> (replicative Bacillus subtilis expression vector; IPTG inducible
GG100BBa_K2145125 Version 1 (Component)This part contains 2 fluorescent protein coding sites (RFP and GFP) with a spacer
GG98BBa_K2145123 Version 1 (Component)This part contains 2 fluorescent protein coding sites (RFP and GFP) with a spacer
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.