BBa_K165093BBa_K165093 Version 1 (Component)Zif268-HIV bs + TEF2 + Gli1 repressor (CFPx2 tagged) on pRS304*
BBa_K165098BBa_K165098 Version 1 (Component)Zif268-HIV bs + ADH1 + Gli1 repressor (CFPx2 tagged) on pRS304*
BBa_K165096BBa_K165096 Version 1 (Component)Zif268-HIV bs + MET25+ Gli1 repressor (CFPx2 tagged) on pRS304*
pSBBs0KBBa_K823026 Version 1 (Component)pSB<sub>Bs</sub>0K-P<sub>spac</sub> (replicative Bacillus subtilis expression vector; IPTG inducible
BBa_J06964BBa_J06964 Version 1 (Component)434 cI/lambda cI857 (E07700) RS flip-flop with EYFP
BBa_K177035BBa_K177035 Version 1 (Component)cI repressor from E. coli phage lambda (+LVA) under control of RBS.3 (medium)
BBa_K299300BBa_K299300 Version 1 (Component)J23100[promoter].K299107[TxbaIG].J61100[RBS] No b-brick scars between
BBa_K748004BBa_K748004 Version 1 (Component)P3+RBS+GFP+T. P3 can be activated by phosphorylated AgrA.
pSBBs1CBBa_K823021 Version 1 (Component)pSB<sub>Bs</sub>1C-<i>lacZ</i> (lacZ reporter vector for <i>B. subtilis</i>)
BBa_J06961BBa_J06961 Version 1 (Component)Lambda cI/LacI ts (mut 241) RS flip-flop with EYFP
BBa_J06962BBa_J06962 Version 1 (Component)Lambda cI/LacI ts (mut 265) RS flip-flop with EYFP
BBa_J06965BBa_J06965 Version 1 (Component)434 cI/lambda cI857 (Lambda II) RS flip-flop with EYFP
pSBBs2EBBa_K823027 Version 1 (Component)pSB<sub>Bs</sub>2E: Empty backbone for integration into Bacillus subtilis lacA locus
BBa_K901018BBa_K901018 Version 1 (Component)PTA with a sRBS
BBa_J119104BBa_J119104 Version 1 (Component)pT7-RBS-GFP-pLac+tRNA CCACC+pLac+tRNA AGGAC+pLac+tRNA CUACU
BBa_J119107BBa_J119107 Version 1 (Component)pT7-RBS-GFP-pLac+tRNA CCACC+pLac+tRNA AGGAC+pLac+tRNA CUACC
BBa_K1974033BBa_K1974033 Version 1 (Component)T7 Promoter+RBS+Hv1a+GS linker+snowdrop-lectin+linker+6X His-Tag
BBa_K1053209BBa_K1053209 Version 1 (Component)Pconst.- RBS-YF1/FixJ-DT- Pfixk2- taRNA- DT- Pconst.- HHR*- GFP- DT
pSBBs4SBBa_K823022 Version 1 (Component)pSB<sub>Bs</sub>4S: Empty backbone for integration into <i/>Bacillus subtilis thrC</i> locus
pSBBs1CBBa_K823023 Version 1 (Component)pSB<sub>Bs</sub>1C: Empty backbone for integration into <i>Bacillus subtilis</i> <i>amyE</i> locus
BBa_K165100BBa_K165100 Version 1 (Component)Gli1 bs + LexA bs + mCYC + LexA repressor (mCherryx2 tagged) on pRS304*
BBa_K165101BBa_K165101 Version 1 (Component)Zif268-HIV bs + LexA bs + mCYC + Zif268-HIV repressor (mCherryx2 tagged) on pRS304*
BBa_K180005BBa_K180005 Version 1 (Component)GoL - Primary plasmid (part 1)/RPS - Paper primary plasmid (part 1) [LuxR generator]
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.