Team:Uppsala Universitet/Parts
Parts Overview
The basic parts used in the project were codon optimised for Pichia pastoris and synthesised by IDT. Modifications of basic parts such as addition of ribosome standby sites was performed by inverse-mutagenesis PCR. Construction of composite parts was achieved by gibson assembly.
All used parts are presented in Table 1 and 2. Descriptions of all parts can be found in the Basic parts or Composite parts. The team chose F2A as the best basic part created in the project. The self-cleaving mechanism, the functionality of which we proved in P. pastoris, can be used in a multitude of future projects to coexpress proteins in one mRNA strand in a eukaryotic expression system.
As its best composite part, the Uppsala Team nominates Expression construct HRP-2A-AAO. In this part functional coexpression of horseradish peroxidase and aryl-alcohol oxidase was shown.
Table 1: Parts used for gold medal criteria
| # | Name | Type | Description |
|---|---|---|---|
| 1 | BBa_K3105111 | Basic part | RBS with ribosome standby site |
| 2 | BBa_K3105666 | Composite part | AsPink with ribosome standby site upstream of RBS |
Table 2: Parts used for main project
| # | Name | Type | Description |
|---|---|---|---|
| 1 | BBa_K3105668 | Basic part | Horseradish peroxidase |
| 2 | BBa_K3105667 | Basic part | Aryl-alcohol oxidase |
| 3 | BBa_K3105684 | Composite part | Expression construct HRP |
| 4 | BBa_K3105685 | Composite part | Expression construct AAO |
| 5 | BBa_K3105681 | Composite part | Expression construct LiP |
| 6 | BBa_K3105683 | Composite part | Expression construct Glox |
| 7 | BBa_K3105682 | Composite part | Expression construct MnP |
| 8 | BBa_K3105669 | Basic part | F2A |
| 9 | BBa_K3105670 | Basic part | eGFP |
| 10 | BBa_K3105671 | Composite part | HRP-2A-eGFP |
| 11 | BBa_K3105672 | Composite part | HRP-2A-AAO |
| 11 | BBa_K3105678 | Composite part | AAO-2A-eGFP |
| 12 | BBa_K3105677 | Composite part | Expression construct HRP-2A-eGFP |
| 13 | BBa_K3105680 | Composite part | Expression construct AAO-2A-eGFP |
| 14 | BBa_K3105679 | Composite part | Expression construct HRP-2A-AAO |
