BBa_K2005002

BBa_K2005002 Version 1

Component

Source: http://parts.igem.org/Part:BBa_K2005002

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 08:18:30

LacZ-alpha, UV-maximized

Details

• Types: DnaRegion
• Roles: Reporter; engineered_region
• Sequences: BBa_K2005002_sequence (Version 1)

Description

LacZ alpha fragment with its coding DNA redesigned to maximize its susceptibility to UV-induced DNA mutation. 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 radiological damage, which identified dipyrimidine motifs as particularly vulnerable. 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%

Assembly

This sequence was synthesized and ligated into pSB1C3. We confirmed that this construct was synthesized and integrated correctly by sequencing.

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 radiological damage, which identified dipyrimidine motifs as particularly vulnerable. 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%

Source

Synthetic variant of Bba_I732006

Other Properties

• igem#status: Planning
• synbiohub#ownedBy: user/james
• synbiohub#ownedBy: user/myers
• synbiohub#topLevel: BBa_K2005002/1

Member of these Collections

• iGEM Parts Registry