Our BioBrick Collection

P_luxR: AHL-responsive promoter from Vibrio fischeri, positioned upstream of luxR; drives luxR transcription upon AHL, enabling positive feedback and signal amplification.

luxR: Quorum-sensing transcriptional regulator from V. fischeri; binds AHL to form an active complex that activates the pLuxI promoter and drives downstream gene expression.

P_luxI: LuxR-AHL-dependent promoter containing the lux box; activates transcription of downstream genes upon AHL induction (native promoter upstream of luxI).

phlA: Cell-lysis effector from Serratia liquefaciens; expression disrupts the bacterial membrane and causes lysis, serving as a kill-switch output when placed under P_LuxI control.

pUC18: High-copy pUC-family cloning backbone with pMB1/ColE1 origin, ampicillin resistance (bla), and lacZα MCS for blue–white screening.

pGL3-Basic: Firefly luciferase gene serving as a reporter, was originally on the pGL3-Basic vector. This vector provides a basis for the quantitative analysis of factors that poteneially regulate gene expression. These factors may be cis-acting elements, such as promoters and enhancers, or trans-acting factors, such as various DNA-binding proteins. In our project, P_fnrS promoters were inserted respectively into pGL3-Basic vector to determine the startup efficiency of promoter by detecting the expression under anaerobic conditions of firefly luciferase gene with the fluorescene intensity measurement.

Firefly luciferase: Luciferase from Photinus pyralis. By oxidation of substrate luciferin light is produced. Luciferase thus acts as a reporter protein when connected to other proteins or promoters.

AmpR(BsaI-free) (reverse): Ampicillin Resistance Gene, which is serving as a screening method to verify the success of the transformation.

AmpR Promoter: The AmpR promoter is responsible for initiating the transcription of the ampicillin resistance gene. The expression of the beta-lactamase enzyme, encoded by the AmpR gene, allows bacteria to survive in environments containing ampicillin by hydrolyzing the beta-lactam ring of the antibiotic, thereby inactivating it.

GmR: Gentamicin Resistance (GmR) gene confers resistance to the antibiotic gentamicin. In iGEM projects, it serves as a reliable selectable marker.

P_fnrS: The promoter was obtained from the Escherichia coli MG1655 genome and can be activated under anaerobic/hypoxic conditions to initiate the transcription of downstream genes.

yedQ: This part sequence originated from Escherichia coli encodes YedQ protein. YedQ is diguanylate cyclase.

pBBR1MCS-5: A low-copy-number, broad-host-range plasmid backbone with a gentamicin resistance marker. It is commonly used for stable gene expression in various bacterial hosts.

pUCP20: A high-copy-number plasmid backbone with an ampicillin resistance marker. It is often used in Pseudomonas and other Gram-negative bacteria for high-level gene expression.

Geneknock Cm R-thyA Knockout Long Homology Arm Targeting Fragment contains the chloramphenicol resistance gene (Cm R) derived from plasmid pKD3, flanked by two homologous fragments corresponding to the upstream and downstream regions of the bacterial thyA gene. This part is optimized for Lambda Red homologous recombination-mediated thyA gene knockout, building upon the principles of Lambda Red recombineering systems. The linear fragment is designed to be PCR-amplified and introduced into target bacterial strains (e.g., MC4100, BL21(DE3), DH5α) containing the pKD46 plasmid, which provides the necessary recombinases. Through homologous recombination, the fragment replaces the endogenous thyA gene with the chloramphenicol resistance cassette, enabling selection of thyA-deficient recombinant strains.

Geneknock Cm R-dapA Knockout Long Homology Arm Targeting Fragment Geneknock Cm R-dapA Knockout Long Homology Arm Targeting Fragment contains the chloramphenicol resistance gene (Cm R) derived from plasmid pKD3, flanked by two homologous fragments corresponding to the upstream and downstream regions of the bacterial dapA gene. This part is optimized for Lambda Red homologous recombination-mediated dapA gene knockout, building upon the principles of Lambda Red recombineering systems. The linear fragment is designed to be PCR-amplified and introduced into target bacterial strains (e.g., MC4100, BL21(DE3), DH5α) containing the pKD46 plasmid, which provides the necessary recombinases. Through homologous recombination, the fragment replaces the endogenous dapA gene with the chloramphenicol resistance cassette, enabling selection of dapA-deficient recombinant strains.

pKD3: A widely utilized suicide plasmid in bacterial genetics, characterized by its temperature-sensitive replication origin that enables plasmid curing at non-permissive temperatures, and it harbors the chloramphenicol resistance gene (cat) as a selectable marker, frequently employed for homologous recombination-mediated gene editing in Escherichia coli and related bacterial species due to its stable maintenance under selective pressure and efficient elimination post-recombination events.

pKD46: A commonly employed helper plasmid in bacterial genetic engineering, featuring a temperature-sensitive replication origin that facilitates its removal from host cells by shifting to non-permissive temperatures. It carries the araBp-controlled red operon (exo, bet, gam) derived from bacteriophage λ, which enables efficient homologous recombination in Escherichia coli and related species by mediating the uptake and processing of linear DNA substrates. Additionally, pKD46 encodes an ampicillin resistance gene (bla) as a selectable marker, ensuring stable maintenance under selective pressure while allowing for straightforward elimination post-recombination through temperature-mediated curing.