BBa_I3401BBa_I3401 Version 1 (Component)Lambda cI switch input device (B0034.C0051.B0015)
BBa_K584011BBa_K584011 Version 1 (Component)Lac-Lux hybrid promotor + CrtEBI + CI repressor + INP
BBa_K611017BBa_K611017 Version 1 (Component)cI Lambda Repressor and Promoter Wild Type Control
PLac-LacY-BBa_I763013 Version 1 (Component)LacY and cI coding device switched on by IPTG
BBa_I758601BBa_I758601 Version 1 (Component)Screen for binding affinity of mutant cI lambda to promotor sites
BBa_I758600BBa_I758600 Version 1 (Component)Screen for binding affinity of mutant cI lambda to promotor sites
BBa_K145112BBa_K145112 Version 1 (Component)cI under T7 and PR<sub>R</sub> dual promotor
BBa_K177035BBa_K177035 Version 1 (Component)cI repressor from E. coli phage lambda (+LVA) under control of RBS.3 (medium)
BBa_K584008BBa_K584008 Version 1 (Component)Lambda cI and LuxR regulated hybrid promotor + RBS + MelA + RBS + AFP + term
BBa_K415005BBa_K415005 Version 1 (Component)pLux/cI-OR : RBS-mCherry : Term : p(tetR) : RBS-luxR : Term
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.
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.