BBa_K290001
BBa_K290001 Version 1 (Component)
constitutive RhlR with bicistronic LuxI - GFP controlled by pRhl

BBa_R4030
BBa_R4030 Version 1 (Component)
PoPS/RiPS Generator composed of the Tet promoter and a strong RBS (R0040.E0030)

BBa_J58015
BBa_J58015 Version 1 (Component)
Mutated promoter activated by OmpR-P with the reporter GFP

pCMV-ECFP-
BBa_I763023 Version 1 (Component)
LacI coding device with ECFP as a reporter regulated by pCMV

BBa_K165100
BBa_K165100 Version 1 (Component)
Gli1 bs + LexA bs + mCYC + LexA repressor (mCherryx2 tagged) on pRS304*

BBa_K300096
BBa_K300096 Version 1 (Component)
Double phasin and intein separed by a flexible protein domain linker

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.

BBa_K2144011
BBa_K2144011 Version 1 (Component)
Coding sequence for Nuclease with His6 and LPXTG tag regulated by T7-promoter

BBa_K165101
BBa_K165101 Version 1 (Component)
Zif268-HIV bs + LexA bs + mCYC + Zif268-HIV repressor (mCherryx2 tagged) on pRS304*

BBa_K2123117
BBa_K2123117 Version 1 (Component)
Novel RFP device regulated by mercury: MerR (regulatory protein) + Stationary phase with mer operato

BBa_K079021
BBa_K079021 Version 1 (Component)
LacI repressor and GFP reporter proteins under the control of the J23118 costitutive promoter and La

BBa_K831011
BBa_K831011 Version 1 (Component)
istR (inhibitor of SOS-induced toxicity by RNA) is small ncRNA of Escherichia coli K12

BBa_K831012
BBa_K831012 Version 1 (Component)
istR (inhibitor of SOS-induced toxicity by RNA) is small ncRNA of Escherichia coli K12

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.