placIQ RBSBBa_K193604 Version 1 (Component)GFP behind a constitutive promoter (placIQ) on pSB4A5
BBa_K1154006BBa_K1154006 Version 1 (Component)Mating pheromone-induced IGPD and constitutive LDH expression in yeast
BBa_K2087000BBa_K2087000 Version 1 (Component)CRISPR/Cas9 sgRNA Targeting Locus 618 on the DD96 Human Gene
BBa_K165100BBa_K165100 Version 1 (Component)Gli1 bs + LexA bs + mCYC + LexA repressor (mCherryx2 tagged) on pRS304*
BBa_K1363200BBa_K1363200 Version 1 (Component)Anti-LPS factor(LALF) regulated by lacI
BBa_K165101BBa_K165101 Version 1 (Component)Zif268-HIV bs + LexA bs + mCYC + Zif268-HIV repressor (mCherryx2 tagged) on pRS304*
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.
BBa_K563004BBa_K563004 Version 1 (Component)pTEF1, promoter of Translational elongation factor EF-1 alpha
BBa_K2092004BBa_K2092004 Version 1 (Component)alcR (incl RBS), ethanol-activated transcription factor from A. nidulans
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_K1172914BBa_K1172914 Version 1 (Component)Part 2 of the Biosafety-System TetOR alive (TetO GFP)