Part:BBa_K1598008

J23101 promoter + RBS + Human FDXR + RBS + Human FDX1 + RBS + Human CYP11A1 + rrnb double terminator

Also called Cytochrome P450scc, Cholesterol Side Chain Cleavage Enzyme (P450scc) is a mitochondrial enzyme that converts cholesterol into pregnenolone, which is a neurosteroid as well as the precursor to a variety of steroid hormones and neurotransmitters. The gene that encodes it is CYP11A1. Furthermore, the catalysis involves 3 monooxygenase reactions, which requires two electron transport proteins: ferrodoxin (FDX1) and ferredoxin reductase (FDXR). Our biobrick consists of the iGEM prefix followed by the J23101 constitutive promoter, RBS, FDXR gene, RBS, FDX1 GENE, RBS, CYP11A1 gene, rrnb double terminator, and the iGEM suffix. This will be ligated onto a PSB1C3 backbone.

Emerging preclinical and clinical evidence suggests that pregnenolone may be a promising novel therapeutic candidate in schizophrenia. It has also been shown to reduce the side effects of benzodiazepines. Furthermore, cholesterol side chain cleavage is the 1st step in the biosynthesis of steroids such as testosterone, progesterone, cortisol, estrogen, etc. Therefore, this biobrick has a plethora of applications, and is in a way several biobricks in 1.

Usage and Biology

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 3275
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 565
  Illegal XhoI site found at 1457
  Illegal XhoI site found at 3710
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1653
  Illegal AgeI site found at 1090
  
• 1000
  COMPATIBLE WITH RFC[1000]