Composite

Part:BBa_K3711057

Designed by: Jiacheng Shi   Group: iGEM21_HUST-China   (2021-10-03)


Panb1-α factor-crtB-AOX1 Terminator


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 124
    Illegal XhoI site found at 649
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1381
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1375


Description

This is a composite part for extracellular expression of phytoene synthase. crtB is expressed and participates in the production from two GGPP molecules to octahydrolycopene. Panb1 is a constitutive promoter in yeast, which is expressed under anaerobic conditions, while under aerobic conditions, Panb1, as a repression target of ROX1, is inhibited. When Panb1 initiates the expression, the signal peptide, α-factor, is used to express crtB outside of the cell.

Usage and Biology

crtB is derived from Erwinia and encodes octahydrolycopene synthase (PSY), which is involved in the synthesis of carotenoids. The early steps of carotenoid biosynthesis pathway include the synthesis of Geranylgeranyl pyrophosphate (GGPP), the condensation of two molecules of GGPP to octahydrolycopene, and desaturation of octahydrolycopene into plant fluorene, β-carotene, protolycopene and lycopene. crtB encodes octahydrolycopene synthase, which is responsible for the condensation of two molecules of GGPP into octahydrolycopene.
Carotenoids are widely found in nature. More than 630 different natural carotenoids have been identified. They are de novo synthesized from isoprene-like precursors, only in photosynthetic organisms and some microorganisms. The synthesis of carotenoids is encoded by plasmids or chromosome genes. The genes that encode carotenoid biosynthesis are clustered in a 12.4kb fragment. Genetic studies have shown that the expression of these genes requires CAMP.

Molecular cloning

Fig1. Plasmid construction and colony PCR results of Panb1-α factor-4CL-AOX1 Terminator, Panb1-α factor-crtI-AOX1 Terminator, Panb1-α factor-crtB-AOX1 Terminator and Panb1-α factor-crtE-AOX1 Terminator transformed E.coli
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Categories
Parameters
None