Parts

Our collection of parts designed and characterized for this project, organized into functional sections.

Part Collection Overview

Our part collection is grouped into three main sections:

  • Section 1: Primary system components that produce and secrete the antimicrobial peptide nisin (NisA + maturation/export machinery).
  • Section 2: Control & regulatory components, including quorum-sensing input (ComDE→PcomE) and our L-Asp–linked containment (AalR→MazE/MazF).
  • Section 3: Validation & benchmark plasmids used to verify expression, secretion, and functional activity under induction/tri-plasmid setups.
Diagram of LactoLock Part Design

Section 1: Core Functional Parts

Core parts that synthesize, mature, and export nisin.

Part Number Description
BBa_253HSFRE pET-21a(+)-nisA (AmpR, T7). Structural gene for pre-NisA under T7 promoter; C-terminal 6xHis for purification. Substrate for NisB/C → NisT export → NisP leader cleavage to yield active nisin.
BBa_25LMDN1U pRSFDuet-1-nisB-nisP (KanR, T7). NisB dehydratase (Ser/Thr → Dha/Dhb) + NisP extracellular protease (leader cleavage); synchronous IPTG induction.
BBa_25AY9ZAD pACYCDuet-1-nisC-nisT (CmR, T7). NisC cyclase for (methyl)lanthionine ring formation; NisT ABC exporter for Sec-independent pre-NisA export; compatible ori/markers for tri-plasmid system.

Section 2: Control & Regulatory Parts

Quorum-sensing input for context-specific AMP expression and containment components for chassis control.

Part Number Description
BBa_25KZ8SGU pMG36e_P32_comDE + PcomE→nisA (ErmR). Constitutive ComD/ComE TCS (P32) senses S. mutans CSP; ComE activates cognate promoter driving pre-NisA only in presence of CSP (context-specific deployment).
BBa_25PQ3LAU AalR LysR-type TF (from A. baylyi) sensing L-aspartate; links L-Asp to downstream transcription from aspA operator (ATGC–N7–GCAT). Used to couple L-Asp to MazE expression in our containment switch.
BBa_259O18OF MazE antitoxin driven by aspA promoter under AalR control; neutralizes MazF RNase when L-Asp present (toothpaste context), tuning survival vs. kill.
BBa_25X2APSD MazF toxin (pET-21(+), T7, LacI). Endoribonuclease that halts translation; expressed with weak RBS to minimize basal toxicity; used as the lethal leg of our containment switch.
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