Part:BBa_K4579000

P_Tet* promoter + RBS

Introduction

The 2023 UT Austin iGEM Team’s Parts Collection includes a multitude of parts necessary for engineering bacteria to secrete microcins, a type of small antimicrobial peptide. Specifically, our team has designed parts that allow us to engineer a modular two-plasmid microcin secretion system that secretes putative novel microcins predicted by bioinformatics analysis. The first plasmid—the ‘microcin’ plasmid—contains the microcin and a signal peptide, while the second plasmid—the ‘secretion system’ plasmid—contains genes for two proteins of the E. coli microcin V type I secretion system (T1SS) machinery collectively referred to as cvaAB. Our parts can be easily assembled into transcriptional units to express any of our current novel microcins either constitutively or under inducible control.

Usage and Biology

P_name is an inducible promoter which functions as a Type 2 part in the BTK/YTK standard. This part undergoes BsaI digestion to produce a linear part with overhangs characteristic of a Type 2 part in the BTK/YTK standard. This part consists of the Pname promoter upstream of a ribosome binding site and hammerhead ribozyme (HHRz) sequence. This promoter can be bound by ProteinName (link to BBa for that regulator) which acts as a(n) activator/repressor that binds to/is removed from the promoter when bound by the inducer molecule [inducer], allowing for the selective induction of transcription in cells containing both P_name and the proteinGene gene.

Figure 1. BTK/YTK part types

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]