Part:BBa_K4883009

RIB1-ERBV-1-RIB7-PTV-ADE4

This is a riboflavin synthase RIB1, followed by a PTV 2A self-cleaving peptide, another riboflavin synthase RIB7, an ERBV-1 2A self-cleaving peptide, and an IMP synthase ADE4.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1335
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 428
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 448
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1267
  

Usage and Biology

ADE4 (BBa_K4883000) catalyzes the first step of the purine nucleotide biosynthetic pathway in Saccharomyces cerevisiae, forming IMP. IMP is the precursor of GTP, and GTP is the precursor of vitamin B2. RIB1 (BBa_K4883001) and RIB7 (BBa_K4883002) catalyze the first and second steps of the riboflavin (vitamin B2) biosynthesis pathway in S. cerevisiae. Co-overexpression of ADE4, RIB1, and RIB7 could lead to overproduction of vitamin B2 in S. cerevisiae.

For the three genes to be expressed separately and normally, the genes are divided by two 2A self-cleaving peptides, ERBV-1 and PTV, which cleave a longer peptide into two shorter peptides. ERBV-1 and PTV have been proven to have the highest cleavage efficiency in S. cerevisiae (Souza-Moreira et al., 2018).

References

Souza-Moreira TM, Navarrete C, Chen X, Zanelli CF, Valentini SR, Furlan M, Nielsen J, Krivoruchko A. Screening of 2A peptides for polycistronic gene expression in yeast. FEMS Yeast Res. 2018 Aug 1;18(5). doi: 10.1093/femsyr/foy036. PMID: 29617770.