Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jarrod Shilts
Date created: 2016-10-17 11:00:00
Date modified: 2016-10-18 08:01:20
CAN1, UV-resistant
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K2005090_sequence (Version 1) |
Description
Coding sequence for CAN1 gene from yeast (Saccharomyces cerevisiae) with its sequence modified by our algorithm to minimize its susceptibility to mutation. This sequence has sufficient flanking homology regions to allow for integration into yeast, and features base modifications to accommodate CRISPR homology-directed repair. CAN1 codes for an arginine permease that also confers susceptibility to the drug canavanine.We designed K2005090 as part of an in vivo selection system, where mutant cells could be positively enriched using canavanine to measure rates of mutation with our optimized genes.
Notes
To generate this artificial DNA sequence, we employed our custom-made algorithm for modifying gene sequences to eliminate mutagenic sites. These sites were identified based on prior research on DNA irradiation, where dipyrimidine motifs as particularly vulnerable. At these motifs, synonymous codon substitutions were made to eliminate mutation-prone nucleotide sequences and replace them with ones with a lower mutation risk. In addition, our algorithm used heuristics to factor in the effect different codons have on gene expression to ensure that the gene not only produced the same protein but produced it at similar levels. Overall we could lower the predicted number of UV-mutation sites by 39%.This sequence also has 50 bp homology arms for genomic integration, and SNPs at CRISPR target sites.