BBa_K091230BBa_K091230 Version 1 (Component)cI promoter+RBS+RFP+TT
BBa_K092500BBa_K092500 Version 1 (Component)TetR cds with RBS and Terminator + pTetR, RFP with RBS
BBa_K318501BBa_K318501 Version 1 (Component)lacI pL + RBS + RcsB + TT
[OriTR]+[RBBa_K1439002 Version 1 (Component)This part contains a reporter gene BBa_J04450, combined with OriTR. Used to test plasmid mobility.
BBa_I13006BBa_I13006 Version 1 (Component)Promoter R0040 w/ YFP (-LVA) TT
BBa_K318509BBa_K318509 Version 1 (Component)lacI pL + RBS + cI LVA + TT
BBa_K199165BBa_K199165 Version 1 (Component)pT7 + RBS + GFP + TT in pSB3T5
BBa_K2066500BBa_K2066500 Version 1 (Component)UNS 2 Sequence, from Torella et al., 2013
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.