Composite

Part:BBa_K3711062

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


Panb1-α factor-ACC-AOX1 Terminator


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1902
    Illegal NheI site found at 2400
    Illegal NheI site found at 2442
  • 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 1360
    Illegal NgoMIV site found at 2120
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1022
    Illegal BsaI.rc site found at 2020
    Illegal BsaI.rc site found at 2324


Description

This is a composite part for extracellular expression of Acetyl CoA carboxylase. ACC is expressed and participates in the production from acetyl CoA to malonyl coenzyme. 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 ACC outside of the cell.

Usage and Biology

Acetyl-CoA carboxylase (ACC) is a biotin enzyme that can catalyze the reaction of "acetyl-CoA+ATP+HCO3→malonyl-CoA+ADP+Pi". It exists widely in nature. ACC is a rate-limiting enzyme for ab initio synthesis of fatty acids, which catalyzes acetyl-CoA to malonyl-CoA, which eventually forms C16 acyl-CoA. ACC can be divided into multi-subunit ACC and multi-functional ACC. Polysubunit ACC exists in plants and bacteria and consists of four subunits, namely, biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP) and two subunits of carboxyltransferase (CT), α-CT and β-CT. Multifunctional ACC mostly exists in eukaryotes. ACC has been used in the drug design of obesity, diabetes and plant herbicides, and is also a target gene for some crops.

Molecular cloning

Fig1. Plasmid construction and colony PCR results of Panb1-α factor-CUS-AOX1 Terminator, Panb1-α factor-ACC-AOX1 Terminator and Panb1-α factor-PepACS-AOX1 Terminator transformed E.coli
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Categories
Parameters
None