BBa_K812130BBa_K812130 Version 1 (Component)Citrine reporter with a Kozak sequence for expression in Xenopus
BBa_K2092004BBa_K2092004 Version 1 (Component)alcR (incl RBS), ethanol-activated transcription factor from A. nidulans
pCMV-ECFP-BBa_I763023 Version 1 (Component)LacI coding device with ECFP as a reporter regulated by pCMV
BBa_K1942000BBa_K1942000 Version 1 (Component)A shRNA corresponding DNA sequence for KRAS which could silence the gene
BBa_K1179015BBa_K1179015 Version 1 (Component)A U6_gRNA(Cr9) plasmid which produces Cr9 targeting guide RNA's for Cas9
BBa_J58011BBa_J58011 Version 1 (Component)Promoter which is activated by cI and CRP, using a transcription logic function type AND
BBa_K300096BBa_K300096 Version 1 (Component)Double phasin and intein separed by a flexible protein domain linker
BBa_K079016BBa_K079016 Version 1 (Component)RecA promoter with GFP reporter protein on a medium copy number plasmid
BBa_J22121BBa_J22121 Version 1 (Component)Lac Y gene under the rec A(SOS) promoter in plasmid pSB2K3
BBa_K371054BBa_K371054 Version 1 (Component)MPF(meta-prefix)+[GFP+10*GS+A] fusion protein+MSF(meta-suffix))
BBa_K779120BBa_K779120 Version 1 (Component)RNA Reporter top strand with quencher (RQ) and tag fluorophore (Alexa 488) MammoBlock
BBa_K1412088BBa_K1412088 Version 1 (Component)A combination of theophylline aptamer and taRNA that can response theophylline to regulate circuit
BBa_K831011BBa_K831011 Version 1 (Component)istR (inhibitor of SOS-induced toxicity by RNA) is small ncRNA of Escherichia coli K12
BBa_K831012BBa_K831012 Version 1 (Component)istR (inhibitor of SOS-induced toxicity by RNA) is small ncRNA of Escherichia coli K12
BBa_K1778002BBa_K1778002 Version 1 (Component)TRE-CYC1TATA is a recombinant promoter, which is constructed in order to make the Tet-on system func
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.