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Public
BBa_K1413045
BBa_K1413045 Version 1 (Component)
A fusion of Universal Transposon Plasmid and pSB1C3
Public
AraC_TEV-F
BBa_K627008 Version 1 (Component)
Fusion part of arabinose-inducible induction system and the TEV protease
Public
ssTorA_CS-
BBa_K627012 Version 1 (Component)
Fusion of TorA sig-seq, TEV protease cleavage site and b-lactamase
Public
BBa_K371054
BBa_K371054 Version 1 (Component)
MPF(meta-prefix)+[GFP+10*GS+A] fusion protein+MSF(meta-suffix))
Public
BBa_K774102
BBa_K774102 Version 1 (Component)
Multi sensor - for calculation of specific concentrations of nitrates nitrites and nitric oxide usin
Public
Intein_assisted_Bisection_Mapping
Intein_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.
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