Overview
We designed 30 parts to accomplish our goal this year. Among them, 23 parts are basic parts, and the rest are composite parts. We shared characterization of some important parts. All parts that we used had been submitted to the iGEM Registry. The basic parts contain the enzyme components we have characterized, promoters and signal peptides, and SZ1, SZ2 to construct fusion protein. Composite parts are closely related to the expression of PET hydrolases. If you want to get more information about our parts, we strongly recommend you visit the specific part pages.
1.Basic parts
| Name | Remark | Type | Length |
|---|---|---|---|
| BBa_K5249000 | template plasmid | Plasmid | 8894bp |
| BBa_K5249001 | -alpha-factor secretion signal | Signalling | 267bp |
| BBa_K5249002 | IsPETasePA | Coding | 824bp |
| BBa_K5249003 | FAST-PETase-212/277 | Coding | 893bp |
| BBa_K5249004 | MHETase | Coding | 1841bp |
| BBa_K5249005 | AOX1 terminator | Terminator | 247bp |
| BBa_K5249007 | GAP | Regulatory | 477bp |
| BBa_K5249008 | AOX1 | Regulatory | 944bp |
| BBa_K5249009 | MF-alpha-1 | Signalling | 267bp |
| BBa_K5249010 | SZ1 | Device | 144bp |
| BBa_K5249011 | SZ2 | Device | 150bp |
| BBa_K5249012 | PETase | Coding | 795bp |
| BBa_K5249013 | RGG | Coding | 501bp |
| BBa_K5249014 | mCherry | Coding | 705bp |
| BBa_K5249022 | 92FAST-PETase-212/277 | Coding | 893bp |
| BBa_K5249023 | 169FAST-PETase-212/277 | Coding | 893bp |
| BBa_K5249024 | 190FAST-PETase-212/277 | Coding | 893bp |
| BBa_K5249025 | 212FAST-PETase-212/277 | Coding | 893bp |
| BBa_K5249026 | 223FAST-PETase-212/277 | Coding | 893bp |
| BBa_K5249027 | 29IsPETasePA | Coding | 824bp |
| BBa_K5249028 | 59IsPETasePA | Coding | 824bp |
| BBa_K5249029 | 122IsPETasePA | Coding | 824bp |
| BBa_K5249030 | 183IsPETasePA | Coding | 824bp |
| BBa_K5249032 | GAP-MF | Regulatory | 744bp |
| BBa_K5249033 | AOX1-MF | Regulatory | 1211bp |
In this project, we use some basic components such as interacting peptides SZ1 and SZ2, promoters AOXm, GAP, AOX1, signal peptides MF, α-signal peptides, and enzymes which contains PETase, IsPETasePA and it’s single site mutants with S29A, T59S, T122P, N183A mutant position sites. Among them, THR-122-PRO is our BEST NEW BASIC PART.
2.Composite parts
Moreover, the multi-enzyme complex can better improve some specific properties of our enzyme, so, we create MHETase - IsPETase PA and MHETase- FAST-PETase-212/277 composite parts, which can further degrade PET into EG and TPA. We also construct a double copy of IsPETasePA to increase the expression of plastic degrading enzyme.
| Name | Remark | Type | Length |
|---|---|---|---|
| BBa_K5249019 | MHETase - IsPETasePA | Coding | 5774bp |
| BBa_K5249020 | MHETase- FAST-PETase-212/277 | Coding | 5844bp |
| BBa_K5249021 | IsPETase PA2c | Coding | 4557bp |