lacZa-ccdBBBa_K176003 Version 1 (Component)lacZalpha-ccdB coding sequence
1NpuE(C*)BBa_K1362103 Version 1 (Component)RBS + NpuDnaE C-intein RFC[105] non-slicing control construct (with His6)
BBa_K1896006BBa_K1896006 Version 1 (Component)mGFPuv2 (N-part)
RFP1BBa_I715022 Version 1 (Component)Amino Portion of RFP
CreBBa_J61047 Version 1 (Component)Cre DNA recombinase
YFPBBa_K592101 Version 1 (Component)Yellow Fluorescent Protein (YFP)
BBa_K1491025BBa_K1491025 Version 1 (Component)T7 RNA Polymerase with ER Intein
BBa_K1362444BBa_K1362444 Version 1 (Component)GGG linker with RGK (extein)
BBa_K1916011BBa_K1916011 Version 1 (Component)CBCR2 + intein-CBD tag
BBa_K1916022BBa_K1916022 Version 1 (Component)CBCR11 + intein-CBD tag
BBa_K1916005BBa_K1916005 Version 1 (Component)Ga0100955_11928g3 + intein-CBD tag
BBa_K1916008BBa_K1916008 Version 1 (Component)Osc7112_5903g3 + intein-CBD tag
BBa_K1916010BBa_K1916010 Version 1 (Component)CBCR2 + intein-CBD tag
BBa_K1916023BBa_K1916023 Version 1 (Component)CBCR11 + intein-CBD tag
BBa_K1916017BBa_K1916017 Version 1 (Component)CBCR6 + intein-CBD tag
BBa_K1916015BBa_K1916015 Version 1 (Component)CBCR4 + intein-CBD tag
BBa_K1916020BBa_K1916020 Version 1 (Component)CBCR9 + intein-CBD tag
BBa_K1916019BBa_K1916019 Version 1 (Component)CBCR8 + intein-CBD tag
BBa_K1916009BBa_K1916009 Version 1 (Component)Osc7112_5903g3 + intein-CBD tag
BBa_K1916001BBa_K1916001 Version 1 (Component)NpF2164g5 + intein-CBD tag
BBa_K1916018BBa_K1916018 Version 1 (Component)CBCR8 + intein-CBD tag
BBa_K1916013BBa_K1916013 Version 1 (Component)CBCR3 + intein-CBD tag
BBa_K1916007BBa_K1916007 Version 1 (Component)Ga0039498_104087g2 + intein-CBD tag
BBa_K1916014BBa_K1916014 Version 1 (Component)CBCR4 + intein-CBD tag
BBa_K1916000BBa_K1916000 Version 1 (Component)NpF2164g5 + intein-CBD tag
BBa_K1916003BBa_K1916003 Version 1 (Component)Cha6605_4344g2 + intein-CBD tag
BBa_K1916006BBa_K1916006 Version 1 (Component)Ga0039498_104087g2 + intein-CBD tag
BBa_K1435000BBa_K1435000 Version 1 (Component)4-HT inducible intein
BBa_K1916002BBa_K1916002 Version 1 (Component)Cyan7427_1390g3 + intein-CBD tag
BBa_K1916021BBa_K1916021 Version 1 (Component)CBCR9 + intein-CBD tag
BBa_K1916004BBa_K1916004 Version 1 (Component)Ga0100955_11928g3 + intein-CBD tag
BBa_K1916016BBa_K1916016 Version 1 (Component)CBCR6 + intein-CBD tag
BBa_K1916012BBa_K1916012 Version 1 (Component)CBCR3 + intein-CBD tag
BBa_K1159995BBa_K1159995 Version 1 (Component)10/25 N-Part Assembly Scar
BBa_J70628BBa_J70628 Version 1 (Component)RFC12 Sce VMA Intein Internal Domain
cinIBBa_C0076 Version 1 (Component)autoinducer synthetase
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_K1362106BBa_K1362106 Version 1 (Component)RBS + NpuDnaE N-intein RFC[105] assembly construct with additional C-teminal insertion site
H6-sfGFP(NBBa_K1362471 Version 1 (Component)split sfGFPn half
RGKBBa_K1362434 Version 1 (Component)RGK (extein)
BBa_K1362107BBa_K1362107 Version 1 (Component)RBS + NpuDnaE C-intein RFC[105] assembly construct with additional N-teminal insertion site
BBa_K1679022BBa_K1679022 Version 1 (Component)KIG, kcoil+intein+gfp
BBa_K1679027BBa_K1679027 Version 1 (Component)VMA, a thermosensitive intein
BBa_K1679037BBa_K1679037 Version 1 (Component)VMA Dead, an inactivated intein
BBa_K1362901BBa_K1362901 Version 1 (Component)C-terminal tripleSWAGtag (C-intein)
BBa_K1491027BBa_K1491027 Version 1 (Component)T7 RNA Polymerase with Dead ER Intein
BBa_K208001BBa_K208001 Version 1 (Component)Phasin (PhaP)
BBa_K1151006BBa_K1151006 Version 1 (Component)HpNikR coding device
BBa_K1450002BBa_K1450002 Version 1 (Component)T7 ssDNA Binding Protein