BBa_I759041BBa_I759041 Version 1 (Component)cis5-repressed, tet-regulated YFP
BBa_I759035BBa_I759035 Version 1 (Component)cis2-repressed, tet-regulated YFP
BBa_K315023BBa_K315023 Version 1 (Component)pTet-loxBri(F)-RBS-RFP-loxBri(F)
BBa_S03736BBa_S03736 Version 1 (Component)pLac-lox-RBS-Tet (in pSB1A2)
BBa_K323164BBa_K323164 Version 1 (Component)VioA and VioB enzymes fused with zinc fingers under pBAD promoter in pSB4K5
BBa_K1140006BBa_K1140006 Version 1 (Component)pTet + 37 oC RNA thermometer + mCherry (LVA)
BBa_K208013BBa_K208013 Version 1 (Component)Tet Repressible Promoter (BBa_R0040) and RBS (BBa_B0034)
BBa_S03766BBa_S03766 Version 1 (Component)RBS-Kan-RBS-Tet-RBS-RFP (pSB1A7)
BBa_K315045BBa_K315045 Version 1 (Component)pTet+loxP forward+RBS+RFP+loxN forward
BBa_K323163BBa_K323163 Version 1 (Component)VioC, VioD and VioE enzymes fused with zinc fingers under pBAD promoter in pSB4C5
BBa_K194000BBa_K194000 Version 1 (Component)cln2 PEST destabilization domain for rapid protein turnover
cln2BBa_K105014 Version 1 (Component)cln2 PEST destabilization domain for rapid protein turnover
BBa_K091119BBa_K091119 Version 1 (Component)LacI protein generator with a pTet promoter
BBa_K876016BBa_K876016 Version 1 (Component)plac-lacO-RBS-GFP-T-pTet-LacI
BBa_M36305BBa_M36305 Version 1 (Component)Transcription terminator pT-T7 from bacteriophage
BBa_K2123115BBa_K2123115 Version 1 (Component)Universal promoter (Tac + JK26) for both growth phase with downstream mer operator + K081014
BBa_K2123114BBa_K2123114 Version 1 (Component)Stationary phase promoter in tandem (3 repetition) with downstream mer operator + RFP (K081014)
BBa_K516222BBa_K516222 Version 1 (Component)AiiA protein generator with Ptet and RBS B0032
BBa_K2123117BBa_K2123117 Version 1 (Component)Novel RFP device regulated by mercury: MerR (regulatory protein) + Stationary phase with mer operato
BBa_K2123116BBa_K2123116 Version 1 (Component)Universal promoter for both phase of growth in tandem with downstram mer operator + RFP (K081014)
BBa_S03737BBa_S03737 Version 1 (Component)pLac-lox-RFP(reverse)-TT-lox-RBS-Tet (psB1A2)
BBa_K2066500BBa_K2066500 Version 1 (Component)UNS 2 Sequence, from Torella et al., 2013
SEGASEGA_collection Version 1 (Collection)In the Standardized Genome Architecture (SEGA), genomic integration of DNA fragments is enabled by λ-Red recombineering and so-called landing pads that are a common concept in synthetic biology and typically contain features that i) enable insertion of additional genetic elements and ii) provide well-characterized functional parts such as promoters and genes, and iii) provides insulation against genome context-dependent effects. The SEGA landing pads allow for reusable homology regions and time-efficient construction of parallel genetic designs with a minimal number of reagents and handling steps. SEGA bricks, typically synthetic DNA or PCR fragments, are integrated on the genome simply by combining the two reagents (i.e. competent cells and DNA), followed by incubation steps, and successful recombinants are identified by visual inspection on agar plates. The design of the SEGA standard was heavily influenced by the Standard European Vector Architecture (SEVA). SEGA landing pads typically hosts two major genetic “control elements” that influence gene expression on the transcriptional (C1), and translational (C2) level. Furthermore, landing pads contain gadgets such as selection and counterselection markers.
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.