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 08:36:01
LacZ-alpha, oxidation-maximized
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K2005004_sequence (Version 1) |
Description
LacZ alpha fragment with its coding DNA redesigned to maximize its susceptibility to oxidative DNA damage. Compared with standard LacZ, this gene is therefore more likely to mutate into diverse forms.Sequence Design
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 poly-guanine sites by 27% and had an increase in overall guanines.
Assembly
This sequence was synthesized and ligated into pSB1C3. We confirmed that this construct was synthesized and integrated correctly by sequencing.