BBa_K2066010
BBa_K2066010 Version 1 (Component)
Tet Monomer C w/ 64 bp spacer for ICA

BBa_K137088
BBa_K137088 Version 1 (Component)
optimized (TA) repeat constitutive promoter with 19 bp between -10 and -35 elements

BBa_K137087
BBa_K137087 Version 1 (Component)
optimized (TA) repeat constitutive promoter with 17 bp between -10 and -35 elements

BBa_K137090
BBa_K137090 Version 1 (Component)
optimized (A) repeat constitutive promoter with 17 bp between -10 and -35 elements

BBa_K137091
BBa_K137091 Version 1 (Component)
optimized (A) repeat constitutive promoter with 18 bp between -10 and -35 elements

BBa_K137089
BBa_K137089 Version 1 (Component)
optimized (TA) repeat constitutive promoter with 21 bp between -10 and -35 elements

BBa_K137086
BBa_K137086 Version 1 (Component)
optimized (TA) repeat constitutive promoter with 15 bp between -10 and -35 elements

BBa_K137085
BBa_K137085 Version 1 (Component)
optimized (TA) repeat constitutive promoter with 13 bp between -10 and -35 elements

BBa_K855006
BBa_K855006 Version 1 (Component)
pvdQ gene with a silent mutation at 1494 bp to remove the internal PstI site

BBa_K855005
BBa_K855005 Version 1 (Component)
pvdQ gene with a silent mutation at 1491 bp to remove the internal PstI site

BBa_K1051801
BBa_K1051801 Version 1 (Component)
Targeted to HUBI gene ATG downstream position of the 12 bp sgRNA, cooperate to dCas 9 protein, inhib

BBa_K1199044
BBa_K1199044 Version 1 (Component)
HheCW249P 2,3-DCP(2,3-dichloropropanol)->Glycerol

Intein_assisted_Bisection_Mapping
Intein_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.

SEGA
SEGA_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.