Team:OUC-China/Parts

• PROJECT
  • Description
  • Design
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  • Proof of Concept
• WET LAB
  • Results
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  • Measurement
• DRY LAB
  • Hardware
  • Model
• PARTS
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• HUMAN PRACTICE
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Basic Parts

This year, OUC-China designed 30 parts, 14 of them are basic parts and the rest of them are composite parts. Our composite parts include 7 basic circuits, 3 improved circuits, and 2 circuits that can test the function of the CRISPRi system. Most of them were tested by us to verify their function. We hope our parts will make it easier and more convenient to detect antibiotics in our environment, especially in the water environment. You can learn more about our project to see their specific function on our design page.

This year, we designed 14 basic parts. Once the basic parts were put together they can well form composite circuits to detect the antibiotics or other modules. It is worth noting that this year we choose a three-way junction dimeric Broccoli(3WJdB) aptamer as our reporter gene, whose transcription produced substantially more fluorescence than sfGFP.

Name	Type	Description	Length
BBa_K3777000	Reporter	3WJdB	155
BBa_K3777001	Coding	ctcs	519
BBa_K3777002	Coding	mphR	585
BBa_K3777003	Regulatory	inpnc-inap	13
BBa_K3777004	RNA	sgRNA for bs2	96
BBa_K3777005	Regulatory	BS2	23
BBa_K3777006	Regulatory	P(T7)-tetO	41
BBa_K3777007	Regulatory	P(T7)-ctco	48
BBa_K3777008	Regulatory	P(T7)-mphO	56
BBa_K3777009	Coding	tetM	2022
BBa_K3777011	Coding	mphA	906
BBa_K3777012	Regulatory	Pctcs	69
BBa_K3777027	Coding	dCas9	4107

Composite Parts

A series of composite parts have been designed this year, 11 of them are basic circuits and the rest of them are improved circuits, whose limit detection and background leakage are lower.

Basic circuits

This year, we constructed 11 basic circuits, which were used to test the antibiotics. The overuse of the antibitic is resulting in the contamination of freshwater. We began with TetR,ctcs,mphR, well-characterized model aTFs that bind the opreator sequences in the absence of their cognate ligands, and ligand-mediated de-repression of an aTF would result in a fluorescence signal.

Name	Type	Description	Length
BBa_K3777013	Composite	tetR-T7(tetO)-3WJdB	1057
BBa_K3777014	Composite	tetR-T7(teto)-tetM-3WJdB	3034
BBa_K3777015	Composite	tetR-T7(tetO)-sfGFP	1620
BBa_K3777016	Composite	tetR-T7(tetO)-tetM-sfGFP	3597
BBa_K3777017	Composite	ctcs-T7(ctcO)-3WJdB	959
BBa_K3777018	Composite	ctcs-tetM-T7(ctcO)-3WJdB	2936
BBa_K3777019	Composite	ctcs-T7(ctcO)-sfGFP	1522
BBa_K3777020	Composite	ctcs-T7(ctcO)-tetM-sfGFP	3499
BBa_K3777021	Composite	mphR-ermc-T7(mphO)-3WJdB	1810
BBa_K3777022	Composite	mphR-ermc-T7(mphO)-sfGFP	2373
BBa_K3777023	Composite	T7-3WJdB	318

Improved circuits

A shortcoming of our basic circuits is that it is limited by the natural sensitivity of the aTF. The typical concentrations of antibiotic pollution found in environmental water samples are always lower than the limit of its detection. Therefore, we designed A kleptamer RNA (KB2) sequence which can disrupt the folding of 3WJdB and lead to the loss of fluorescence. We hope our improved circuits can have lower background leakage.

Name	Type	Description	Length
BBa_K3777024	Composite	KB2-tetR-T7(tetO)-3WJdB-Ptet-sgRNA	1613
BBa_K3777025	Composite	KB2-ctcs-T7(ctcO)-3WJdB-Pctcs-sgRNA	1530
BBa_K3777026	Composite	KB2-mphR-ermc-T7(mphO)-3WJdB-PmphR-sgRNA	2437
BBa_K3777028	Composite	luxR-Plux-dCas9	5349
BBa_K3777029	Composite	luxR-Plux-dCas9-mphA	6461