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_K1694044BBa_K1694044 Version 1 (Component)Pcons+B0034+Lpp-OmpA-N+scFv(Anti-EGFR)+B0030+RFP+J61048
BBa_K1694045BBa_K1694045 Version 1 (Component)Pcons+B0034+Lpp-OmpA-N+scFv(Anti-HER2)+B0030+BFP+J61048
BBa_K1694034BBa_K1694034 Version 1 (Component)Pcons+B0034+Lpp-OmpA-N+scFv(Anti-EGFR)+B0030+GFP+J61048
BBa_K1694033BBa_K1694033 Version 1 (Component)Pcons+B0034+Lpp-OmpA-N+scFv(Anti-VEGF)+B0030+GFP+J61048
BBa_K1159014BBa_K1159014 Version 1 (Component)Membrane-anchored Erythromycin Esterase Type II in RFC[25] N-Part
BBa_K1159008BBa_K1159008 Version 1 (Component)Secretory Catechol-2,3-dioxygenase (IgKappa-SigP_XylE) in RFC[25] N-Part
BBa_K1159018BBa_K1159018 Version 1 (Component)Membrane-anchored Catechol-2,3-dioxygenase (XylE) in RFC[25] N-Part
BBa_K1053120BBa_K1053120 Version 1 (Component)RNA scaffold containing HTLV-1 Rex peptide and λN peptide aptamer
RGKBBa_K1362434 Version 1 (Component)RGK (extein)
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_K2170201BBa_K2170201 Version 1 (Component)Single chain avidin with ribosomal binding site (RBS_scAvidin) in RFC[25] N-part
BBa_K1997019BBa_K1997019 Version 1 (Component)P+R->GFP10->Zif-N->RBS->Zif-C->GFP11->RBS->GFP1-9->TER
BBa_K331025BBa_K331025 Version 1 (Component)RBS with C-terminal Oligo Arginine - ECFP Fusion
BBa_K331023BBa_K331023 Version 1 (Component)RBS with C-terminal Oligo Arginine - EYFP Fusion
BBa_K1694053BBa_K1694053 Version 1 (Component)Pcons+B0034+Lpp-OmpA-N+scFv(Anti-VEGF)+B0034+amilCP+B0015
BBa_K1694035BBa_K1694035 Version 1 (Component)Pcons+B0034+Lpp-OmpA-N+scFv(Anti-HER2)+B0030+GFP+J61048
BBa_K1362300BBa_K1362300 Version 1 (Component)NpuDnaE N-Intein RFC[105] assembly construct (with His6)
BBa_K1491027BBa_K1491027 Version 1 (Component)T7 RNA Polymerase with Dead ER Intein
BBa_J70554BBa_J70554 Version 1 (Component)Split GFP C-terminal domain
C-IgAPBBa_K195410 Version 1 (Component)C-terminal of IgA protease
RFC105 NCBBa_K1362417 Version 1 (Component)C-terminal GG-linker overhang GG<u>TGGT</u>=Gly+Gly RFC[105] overhang NC
BBa_K1159019BBa_K1159019 Version 1 (Component)Membrane-anchored Human Protein Phosphatase 1 (PP1) in RFC[25] N-Part
BBa_K1159005BBa_K1159005 Version 1 (Component)Secretory Erythromycin Esterase Type II (IgKappa-SigP_EreB) in RFC[25] N-Part
BBa_K1159108BBa_K1159108 Version 1 (Component)N-TEV-Protease_36AALinker_PIF6 (Second part for a light-switchable TEV-Protease) in RFC[25]
BBa_K1159107BBa_K1159107 Version 1 (Component)N-TEV-Protease_36AALinker_PIF3 (Second part for a light-switchable TEV-Protease) in RFC[25]
BBa_K2170141BBa_K2170141 Version 1 (Component)Tetracycline inducible Outer membrane protein A signal peptide (TetR_TetO_OmpA) in RFC[25] N-part
BBa_K1510929BBa_K1510929 Version 1 (Component)Circuit injected in Streptococcus mutans that allows S.m. yield N-Acyl homoserine lactone.
BBa_K1362055BBa_K1362055 Version 1 (Component)MinD MTS as RFC[105] C-terminal insert
eYFP_SH3 gBBa_K1602041 Version 1 (Component)Inducible generator of eYFP with C-terminal SH3 ligand
BBa_K331031BBa_K331031 Version 1 (Component)Tet Repressible C-terminal Arg Tagged EYFP (no terminator)
CPXBBa_I728500 Version 1 (Component)CPX Terminal Surface Display Protein with Polystyrene-Binding Peptide
BBa_K2017000BBa_K2017000 Version 1 (Component)C-split Cas9 + DnaE C-intein
BBa_K1362052BBa_K1362052 Version 1 (Component)Carrier for RFC[105] C-terminal inserts
BBa_K1316007BBa_K1316007 Version 1 (Component)yqjF promoter coupled to mKate2 reporter gene and Rhamnose promoter coupled to N-genes
BBa_K1316008BBa_K1316008 Version 1 (Component)ybiJ promoter coupled to mKate2 reporter gene and Rhamnose promoter coupled to N-genes
BBa_K1362172BBa_K1362172 Version 1 (Component)Split sfGFP reconstitution C-terminal non-splicing control half
BBa_K249005BBa_K249005 Version 1 (Component)C-terminal Arginine Fusion Vector
BBa_K1362051BBa_K1362051 Version 1 (Component)Intein protease with arabinose inducible regulatory promoter/ repressor araC-Pbad-T7RBS-His-SspDnaBC
BBa_K1638036BBa_K1638036 Version 1 (Component)Sce VMA Intein with promoter
BBa_K1362054BBa_K1362054 Version 1 (Component)SsrA degradation tag as RFC[105] C-terminal insert
mCherryCBBa_K1093017 Version 1 (Component)mCherry C-terminal
BBa_K1333002BBa_K1333002 Version 1 (Component)M13 bacteriophage g3p with start codon of "ATG" and a FLAG tag on C-terminal
BBa_I712078BBa_I712078 Version 1 (Component)TEV protease (C-terminal half)
BBa_J70549BBa_J70549 Version 1 (Component)GFP C-Terminal fragment