Composite

Part:BBa_K4883007

Designed by: Yuqi Fu   Group: iGEM23_Hangzhou-BioX   (2023-06-29)
Revision as of 01:56, 6 October 2023 by Yuqi Fu (Talk | contribs)


RIB7-PTV-ADE4

This is a riboflavin synthase RIB7, connected with a PTV 2A self-cleaving peptide, followed by an IMP synthase ADE4.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 240
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 172



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. RIB7 (https://parts.igem.org/Part:BBa_K4883002 BBa_K4883002]) catalyzes the second step of the riboflavin (vitamin B2) biosynthesis pathway in S. cerevisiae. Co-overexpression of RIB7 and ADE4 could lead to overproduction of vitamin B2 in S. cerevisiae.

For the two genes to be expressed separately and normally, the genes are divided by a 2A self-cleaving peptide, PTV, which cleaves a longer peptide into two shorter peptides. PTV has been proven to have the second-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.  

[edit]
Categories
Parameters
None