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_K1051105BBa_K1051105 Version 1 (Component)TOM40;targeting peptide to mitochondria outer membrane
BBa_I20249BBa_I20249 Version 1 (Component)Device used to characterize Ribosome Binding Sites through measurement of GFP expression
BBa_I20248BBa_I20248 Version 1 (Component)Device used to characterize Ribosome Binding Sites through measurement of GFP expression
BBa_I20250BBa_I20250 Version 1 (Component)Device used to characterize Ribosome Binding Sites through measurement of RFP expression
BBa_I20251BBa_I20251 Version 1 (Component)Device used to characterize Ribosome Binding Sites through measurement of RFP expression
BBa_K1510000BBa_K1510000 Version 1 (Component)constitutive promoter and the coding sequence for luxR protein
PLac-CI-LaBBa_I763019 Version 1 (Component)CI and LacY coding device switched on by IPTG with GFP as a reporter
BBa_K2018021BBa_K2018021 Version 1 (Component)Functional panK-II device