Composite

Part:BBa_K5091012

Designed by: Angelina Oblak   Group: iGEM24_ULethbridge   (2024-10-01)


pmsC-pmsB Expression Construct

This composite part is designed to co-express the pmsC and pmsB genes from Pseudomonas fluorescens, which work together to biosynthesize salicylic acid. The construct includes the following components:

araC-pBAD Promoter (BBa_I0500): This promoter is inducible by arabinose and allows for controlled expression of both pmsC and pmsB. The arabinose-inducible system is useful for regulating gene expression levels, preventing unnecessary metabolic burden on the host before induction.

RBS (BBa_B0030): A strong ribosome binding site used for efficient translation of the first gene, pmsC, ensuring proper initiation of protein synthesis.

pmsC Gene (BBa_K5091008): This gene encodes isochorismate synthase, the first enzyme in the pathway that converts chorismate to isochorismate, a precursor to salicylic acid. It plays a critical role in salicylic acid biosynthesis.

RBS (BBa_B0034): A standard ribosome binding site used for the translation of the second gene, pmsB, ensuring efficient translation initiation for this downstream gene.

pmsB Gene (BBa_K5091009): This gene encodes isochorismate pyruvate lyase, the second enzyme in the pathway. It converts isochorismate into salicylic acid, completing the biosynthetic process.

Terminator (BBa_B0012): A reliable double terminator that ensures proper termination of transcription, increasing mRNA stability and preventing read-through, which is essential for efficient gene expression.

This construct is ideal for projects aiming to produce salicylic acid in microbial systems, particularly for applications in plant defense and biocontrol strategies. The arabinose-inducible promoter allows for tight regulation of salicylic acid production, preventing potential toxicity or metabolic stress in the host until induction.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1351
    Illegal PstI site found at 1732
    Illegal PstI site found at 2110
    Illegal PstI site found at 2529
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal PstI site found at 1351
    Illegal PstI site found at 1732
    Illegal PstI site found at 2110
    Illegal PstI site found at 2529
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 1893
    Illegal BamHI site found at 2091
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1351
    Illegal PstI site found at 1732
    Illegal PstI site found at 2110
    Illegal PstI site found at 2529
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1351
    Illegal PstI site found at 1732
    Illegal PstI site found at 2110
    Illegal PstI site found at 2529
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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Categories
Parameters
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