Team:Chalmers-Gothenburg/Parts
About our parts
All basic parts were ordered with overhangs allowing for Gibson assembly of the composite BioBricks. All genes have been codon optimized for yeast and the restriction sites that are incompatible with the iGEM BioBrick standard have been removed. Once assembled, the composite BioBricks were used for integration into the genome of Saccharomyces cerevisiae. The use of dual promoters and two genes per plasmid reduces the amount of transformations needed for integration of the entire pathway, utilizing the EasyClone system as explained on the project design page.
In addition, constructs with His- and FLAG-tags were assembled to test whether the proteins were able to be expressed. The difference is the rather small addition of 24 base pairs at the end of each gene. The expression was tested through western blots, this is also described on the project design page.
Basic Parts
| Biobrick identifier | Description | Type | Verified function |
|---|---|---|---|
| BBa_K3054000 | pCCW12 promoter | Promoter | |
| BBa_K3054001 | pPGK1/pTEF1 dual promoter | Promoter | Partly (pTEF1) |
| BBa_K3054002 | pTDH3/pCIT2 dual promoter | Promoter | |
| BBa_K3054003 | pTEF1/pCCW12 dual promoter | Promoter | |
| BBa_K3054004 | pTEF1/pCIT2 dual promoter | Promoter | |
| BBa_K3054005 | pcbA5 coding region | Coding | |
| BBa_K3054006 | bphA1 coding region | Coding | |
| BBa_K3054007 | bphA2 coding region | Coding | |
| BBa_K3054008 | bphA3 coding region | Coding | |
| BBa_K3054009 | bphA4 coding region | Coding | |
| BBa_K3054010 | bphB coding region | Coding | |
| BBa_K3054011 | bphC coding region | Coding | |
| BBa_K3054012 | bphD coding region | Coding | |
| BBa_K3054013 | bphK coding region | Coding | |
| BBa_K3054014 | pcbA5 coding region + FLAG-tag | Coding | |
| BBa_K3054015 | bphA1 coding region + His-tag | Coding | Yes |
| BBa_K3054016 | bphA2 coding region + FLAG-tag | Coding | |
| BBa_K3054017 | bphA3 coding region + FLAG-tag | Coding | |
| BBa_K3054018 | bphA4 coding region + His-tag | Coding | |
| BBa_K3054019 | bphB coding region + FLAG-tag | Coding | |
| BBa_K3054020 | bphC coding region + His-tag | Coding | |
| BBa_K3054021 | bphD coding region + His-tag | Coding | |
| BBa_K3054022 | bphK coding region + FLAG-tag | Coding |
Composite Parts
| Biobrick identifier | Description | Type | Verified function |
|---|---|---|---|
| BBa_K3054023 | pcbA5 expressed by pCCW12 | Composite | |
| BBa_K3054024 | bphA1 expressed by pTEF1/ bphA2 expressed by pPGK1 | Composite | |
| BBa_K3054025 | bphA3 expressed by pCIT2/ bphA4 expressed by pTDH3 | Composite | |
| BBa_K3054026 | bphB expressed by pCCW12/ bphC expressed by pTEF1 | Composite | |
| BBa_K3054027 | bphD expressed by pTEF1/ bphK expressed by pCIT2 | Composite |
Further Reading
Check out the rest of our lab-related work by clicking on any of the images below! Go through the protocols used for the different types of Experiments performed, go through all of the 4 months of lab work documented in Notebook, or go straight for the actual Results of all our hard work. Under Contribution we have documented how we contributed to iGEM by further characterizing already existing BioBricks, while Safety will take you to the details of how we made sure to keep ourselves and others safe in the lab.
