BBa_K2005091

BBa_K2005091 Version 1

Component

Source: http://parts.igem.org/Part:BBa_K2005091

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 08:04:15

CAN1, UV-maximized

Details

• Types: DnaRegion
• Roles: CDS; Coding
• Sequences: BBa_K2005091_sequence (Version 1)

Description

Coding sequence for CAN1 gene from yeast (Saccharomyces cerevisiae) with its sequence modified by our algorithm to maximize 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 introduce 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 make replacements for ones with higher mutation risks. 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 increase the predicted number of UV-mutation sites by 63%. This sequence also has 50 bp homology arms for genomic integration, and SNPs at CRISPR target sites.

Source

Modified from S. cerevisiae genomic sequences

Other Properties

• igem#status: Planning
• synbiohub#ownedBy: user/james
• synbiohub#ownedBy: user/myers
• synbiohub#topLevel: BBa_K2005091/1

Member of these Collections

• iGEM Parts Registry