BBa_K1362111BBa_K1362111 Version 1 (Component)RBS + SspDnaX C-intein RFC[105] assembly construct (with His6)
BBa_K1321120BBa_K1321120 Version 1 (Component)SmtA metallothionein fused to linker-dCBD
BBa_J133572BBa_J133572 Version 1 (Component)Protein Linker SSGSSG Library
BBa_K608404BBa_K608404 Version 1 (Component)IPTG-inducible Promoter with plastic binding domain-tagged GFP
BBa_J133580BBa_J133580 Version 1 (Component)Protein Linker SSGSSG Library
Gp41-1(C)BBa_K1362413 Version 1 (Component)Gp41-1 C-Intein cloning piece
BBa_K782062BBa_K782062 Version 1 (Component)ΔCBD Alginate lyase (mature variant)
Strep-tagBBa_M0031 Version 1 (Component)Strep-tag II affinity tag
BBa_M36009BBa_M36009 Version 1 (Component)5' Bicistronic UTR (strong), does not include ATG start
BBa_K1491025BBa_K1491025 Version 1 (Component)T7 RNA Polymerase with ER Intein
BBa_K1399006BBa_K1399006 Version 1 (Component)GFP (mut3b) with LAA-ssrA degradation tag
BBa_K1399008BBa_K1399008 Version 1 (Component)GFP (mut3b) with DAS-ssrA degradation tag
BBa_K1362444BBa_K1362444 Version 1 (Component)GGG linker with RGK (extein)
BBa_K1435000BBa_K1435000 Version 1 (Component)4-HT inducible intein
BBa_K1123012BBa_K1123012 Version 1 (Component)His-Tag and Thrombine Cleavage Site
BBa_J70628BBa_J70628 Version 1 (Component)RFC12 Sce VMA Intein Internal Domain
BBa_K180000BBa_K180000 Version 1 (Component)Hybrid promoter (trp & lac regulated -- tac pR)
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.
SV40 NLSBBa_J63008 Version 1 (Component)SV40 nuclear localization sequence from SV40; yeast codon optimized
BBa_K1362160BBa_K1362160 Version 1 (Component)RBS + Gp41-1 N-intein RFC[105] assembly construct (with His6)
RGKBBa_K1362434 Version 1 (Component)RGK (extein)
BBa_K1582029BBa_K1582029 Version 1 (Component)Protein Extraction Tag: Linker+sJanus
BBa_K1679022BBa_K1679022 Version 1 (Component)KIG, kcoil+intein+gfp
BBa_K1679027BBa_K1679027 Version 1 (Component)VMA, a thermosensitive intein
BBa_M36084BBa_M36084 Version 1 (Component)Histidine Tag and Stop Codon
BBa_K1679037BBa_K1679037 Version 1 (Component)VMA Dead, an inactivated intein
BBa_I13770BBa_I13770 Version 1 (Component)Codon CBA
Tat PROMBBa_K895000 Version 1 (Component)Eco_Ptat2
BBa_K212001BBa_K212001 Version 1 (Component)Tar-Envz (Taz)
BBa_K364308BBa_K364308 Version 1 (Component)Human PXR LBD
BBa_K2027001BBa_K2027001 Version 1 (Component)AE Blue-CBD-FlagLumioHisTag Fusion Protein
IBMc318IBMc318 Version 1 (Component)pSB1A3-BbsI-G-ER-LBD-S-SapI
BBa_K1491027BBa_K1491027 Version 1 (Component)T7 RNA Polymerase with Dead ER Intein
BBa_K1453000BBa_K1453000 Version 1 (Component)ssDsbA-mRFP-Lgt-TAL USB-His Tag
BBa_K404112BBa_K404112 Version 1 (Component)Cytosine deaminase (CD)
BBa_K1332002BBa_K1332002 Version 1 (Component)Histidine tag (8 AA) and RFP (without stop codon)
BBa_K1321348BBa_K1321348 Version 1 (Component)sfGFP fused to dCBD with linker in RFC 25
SspDnaB(N*BBa_K1362142 Version 1 (Component)RBS + SspDnaB N-intein RFC[105] non-splicing control construct (with His6)
BBa_K1332001BBa_K1332001 Version 1 (Component)Histidine tag (8 AA) and RFP
BBa_K1321345BBa_K1321345 Version 1 (Component)NiBP fused to dCBD with linker in RFC 25
eYFP_GBDliBBa_K1602037 Version 1 (Component)eYFP with C-terminal GBD ligand and RBS BBa_B0034
BBa_K654058BBa_K654058 Version 1 (Component)Thioesterase (TesA from E. coli) with 8-His Tag
csbDBO_3025 Version 1 (Component) cddBO_2718 Version 1 (Component) λOtagBBa_K365008 Version 1 (Component)Lambda tag
BBa_K1453407BBa_K1453407 Version 1 (Component)Enzyme USB(AarI)-TAL USB-His Tag
BBa_K1453406BBa_K1453406 Version 1 (Component)Enzyme USB(BsmAI)-TAL USB-His Tag