BBa_S01839BBa_S01839 Version 1 (Component)--Specify Parts List--
BBa_K1773008BBa_K1773008 Version 1 (Component)pLux/cI Right promoter with strong RBS
BBa_S01902BBa_S01902 Version 1 (Component)--Specify Parts List--
BBa_K329007BBa_K329007 Version 1 (Component)Strong RBS (B0034) - LuxR (C0062)
BBa_K2128200BBa_K2128200 Version 1 (Component)MAHS with strong promoter/RBS
BBa_K1172914BBa_K1172914 Version 1 (Component)Part 2 of the Biosafety-System TetOR alive (TetO GFP)
BBa_K136011BBa_K136011 Version 1 (Component)Strong Constitutive Promoter - GFP Tripart
BBa_K909004BBa_K909004 Version 1 (Component)cI with strong ribosomal binding site
BBa_J70084BBa_J70084 Version 1 (Component)Adds 6 his suffix, using BioScaffold part J70030 (PpiI) in pSB1AT3
BBa_K1106009BBa_K1106009 Version 1 (Component)LuxR generator with a strong RBS
[OriTR]+[RBBa_K1439002 Version 1 (Component)This part contains a reporter gene BBa_J04450, combined with OriTR. Used to test plasmid mobility.
BBa_K2123206BBa_K2123206 Version 1 (Component)Strong RBS + Organomercurial Lyase (MerB) for E. coli
BBa_K1497197BBa_K1497197 Version 1 (Component)B0034-CHI - Chalcone Isomerase from Petunia with strong RBS
BBa_K779121BBa_K779121 Version 1 (Component)Short DNA reporter top strand (with RQ quencher) MammoBlock
BBa_K779116BBa_K779116 Version 1 (Component)Short RNA reporter bottom strand (with ROX fluorophore) MammoBlock
BBa_K779120BBa_K779120 Version 1 (Component)RNA Reporter top strand with quencher (RQ) and tag fluorophore (Alexa 488) MammoBlock
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.