Overview
This year, BASIS-China contributed a total of 11 basic and 12 composite parts. It covers 7 kinds of terpene synthases producing pleasant aromas (α-pinene, β-pinene, limonene, farnesene, bisabolene, santalene and santalol). In parallel, we constructed a suicide switch for a cyanobacterial chassis capable of synthesizing terpenes, aiming to use cyanobacterial chassis to produce these delightful molecules in an environmentally friendly, safe and cheap way.
Composite Parts ///
We successfully built 8 composite parts for terpene production in Synechocystis sp. PCC 6803. Our composite parts are based on the promoter BBa_K3930029 and terminator BBa_B0015. This year, we will be competing for the Gold medal through our work on the conposite parts.
| Registry Code | Part Name | Short Description | Status | Length(bp) |
|---|---|---|---|---|
| BBa_K4880016 | Ptrc-theo-αPS | α-pinene synthase, expression induced by IPTG and theophylline | Successful colony PCR in Syn. sp. PCC 6803 | 2045 |
| BBa_K4880017 | Ptrc-theo-βPS | β-pinene synthase, expression induced by IPTG and theophylline | Syn. PCC 6803 GC-MS Correct Result | 2042 |
| BBa_K4880018 | Ptrc-theo-MsLIMS | Limonene synthase, expression induced by IPTG and theophylline | Successful colony PCR in Syn. sp. PCC 6803 | 1968 |
| BBa_K4880004 | Ptrc-theo-PaFS | Farnesene synthase, expression induced by IPTG and theophylline | Syn. PCC 6803 GC-MS Correct Result and BL21(DE3) correct protein expression | 1904 |
| BBa_K4880005 | Ptrc-theo-SaSS | Santalene synthase, expression induced by IPTG and theophylline | BL21(DE3) correct protein expression | 1871 |
| BBa_K4880006 | Ptrc-theo-AgBS | Bisabolene synthase, expression induced by IPTG and theophylline | Successful colony PCR in Syn. sp. PCC 6803 | 2615 |
| BBa_K4880007 | Ptrc-theo-PtPS | Pinene synthase, expression induced by IPTG and theophylline | Successful colony PCR in Syn. sp. PCC 6803 | 1272 |
| BBa_K4880015 | Ptrc-theo-SaSS-CYP | Santalene and santalol synthases, expression induced by IPTG and theophylline | Successful colony PCR in Syn. sp. PCC 6803 | 1356 |
Basic Parts ///
To construct the plasmids that allow Synechocystis sp. PCC 6803 to produce terpenes, iGEM BASIS-China 2023 used 12 basic parts, 5 of which are new ones that we have submitted.
| Registry Code | Part Name | Short Description | Status | Length(bp) |
|---|---|---|---|---|
| BBa_K4880000 | PaFS | Farnesene synthase | Syn. PCC 6803 GC-MS Correct Result and BL21(DE3) correct protein expression | 1743 |
| BBa_K4880001 | SaSS | Santalene synthase | BL21(DE3) correct protein expression | 1710 |
| BBa_K4880002 | AgBS | Bisabolene synthase | Successful colony PCR in Syn. sp. PCC 6803 | 2454 |
| BBa_K4880003 | PtPS | Pinene synthase | Successful colony PCR in Syn. sp. PCC 6803 | 1845 |
| BBa_K4880014 | CYP736A167 | Santalol synthase | Successful colony PCR in Syn. sp. PCC 6803 | 1503 |
| BBa_K517002 | αPS | α-pinene synthase | Successful colony PCR in Syn. sp. PCC 6803 | 1884 |
| BBa_K517003 | βPS | β-pinene synthase | Have positive GC-MS result in Syn. PCC 6803 | 1881 |
| BBa_J435283 | MsLIMS | Limonene synthase | Successful colony PCR in Syn. sp. PCC 6803 | 1800 |
Biocontainment System ///
To build our biocontainment system in Synechocystis sp. PCC 6803, 6 other basic parts that we have submitted.
| Registry Code | Part Name | Short Description | Status | Length(bp) |
|---|---|---|---|---|
| BBa_K4880020 | PpsbA2-sepA1-PisiA6803-sepT1 | PempK-like type II TA system where the toxin is controled by PisiA6803 | Did not get desired results when cultivating in liquid BG-11 medium lacking Fe3+ | 1737 |
| BBa_K4880021 | PpsbA2-sepA2-PisiA6803-sepT2 | VapC-like type II TA system where the toxin is controled by PisiA6803 | Did not get desired results when cultivating in liquid BG-11 medium lacking Fe3+ | 1821 |
| BBa_K4880022 | PpsbA2-sepA1-PisiA7942-sepT1 | PempK-like type II TA system where the toxin is controled by PisiA7942 | Circuit works | 1560 |
| BBa_K4880023 | PpsbA2-sepA2-PisiA7942-sepT2 | VapC-like type II TA system where the toxin is controled by PisiA7942 | Circuit works | 1664 |
| Registry Code | Part Name | Short Description | Status | Length(bp) |
|---|---|---|---|---|
| BBa_K4880008 | sepA1 | Antitoxin | Works | 216 |
| BBa_K4880009 | sepT1 | Toxin | Works | 351 |
| BBa_K4880010 | sepA2 | Antitoxin | Works | 447 |
| BBa_K4880011 | sepT2 | Toxin | Works | 384 |
| BBa_K4880012 | PisiA6803 | Fe3+ repressed promoter in Syn. sp. PCC 6803 | Biocontainment system using this promoter did not show desired result | 447 |
| BBa_K4880013 | PisiA7942 | Fe3+ repressed promoter in Syn. sp. PCC 7942 | Biocontainment system using this promoter did not show desired result | 270 |