Parts

1. Overview

1.1 Basic Parts

We expressed several enzymes related to phosphate metabolism from different microorganisms and conducted codon optimization in S. oneidensis: BBa_K5034205, BBa_K5034206, BBa_K5034209, BBa_K5034210, BBa_K5034212.
We improved PPK1 from original part BBa_K3022002, and the result is part BBa_K5034213. We conducted codon optimization based on S. oneidensis and successfully expressed PPK1 in S. oneidensis.

1.2 Composite Parts

We designed three composite parts using RBS with different strength(BBa_B0031, BBa_B0032 and BBa_B0034) and PPK1 gene(BBa_K5034213): BBa_K5034214, BBa_K5034215, BBa_K5034216, to control the expression of PPK1. With increased RBS strength, there is an increase in phosphorus accumulation ability and a decline in electricity-generating capacity.
We designed a composite part which combines PPK2(BBa_K5034205) and NADK(BBa_K5034206) gene together, with the help of dry lab modelling: BBa_K5034217. We found a notable enhancement in the polyphosphate, ATP and electroproduction capabilities.

1.3 Plasmid Parts

We designed pBBR1MCS-terminator plasmid for efficient gene expreesion in S. oneidensis: BBa_K5034201.
For each coding part(including composite parts), we inserted them into plasmid pBBR1MCS-terminator to construct expression vectors: BBa_K5034219, BBa_K5034220, BBa_K5034221, BBa_K5034222, BBa_K5034223, BBa_K5034225, BBa_K5034227, BBa_K5034229, BBa_K5034231.

2. Basic Parts

Registry code	Part name	Description
BBa_K5034205	PPK2	Reversibly convert Poly P and Pi, and favors the decomposition reaction.
BBa_K5034206	NADK	Convert Poly P to NADP
BBa_K5034209	PPN1	Convert Poly P into smaller polyphosphate fragments, but not completely into Pi.
BBa_K5034210	PPX	Thoroughly convert Poly P to Pi.
BBa_K5034212	PAP	Reversibly convert Poly P and AMP into ADP.
BBa_K5034213	PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P.

3. Composite Parts

Registry code	Part name	Description
BBa_K5034214	B0031-PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P. This part is activated by a weaker RBS(BBa_B0031).
BBa_K5034215	B0032-PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P. This part is activated by a medium RBS(BBa_B0032).
BBa_K5034216	B0034-PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P. This part is activated by an efficient RBS(BBa_B0034).
BBa_K5034217	PPK2-NADK	Reversibly convert Poly P and Pi. Combining PPK2 gene(BBa_K5034205) and NADK gene(BBa_K5034206) together to show effects.

4. Plasmid Parts

Registry code	Part name	Description
BBa_K5034201	pBBR1MCS-terminator	Plasmid backbone.
BBa_K5034219	pBBR1MCS-terminator-PAP	Reversibly convert Poly P and AMP into ADP. PAP gene(BBa_K5034212) which is from Acinetobacter johnsonii and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201).
BBa_K5034220	pBBR1MCS-terminator-B0031-PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P. This part is activated by a weaker RBS. PPK1 gene(BBa_K5034213) which is from Citrobacter freundii and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201) with the BBa-B0031 RBS(BBa_B0031).
BBa_K5034221	pBBR1MCS-terminator-B0032-PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P. This part is activated by a medium RBS. PPK1 gene(BBa_K5034213) which is from Citrobacter freundii and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201) with the BBa-B0032 RBS(BBa_B0032).
BBa_K5034222	pBBR1MCS-terminator-B0034-PPK1	Reversibly convert Poly P and Pi but process favors the generation of Poly P. This part is activated by an efficient RBS. PPK1 gene(BBa_K5034213) which is from Citrobacter freundii and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201) with the BBa-B0034 RBS(BBa_B0034).
BBa_K5034223	pBBR1MCS-terminator-B0034-PPK2-B0034-NADK	Reversibly convert Poly P and Pi with PPK2 and convert Poly P and NAD with NADP. Combining PPK2(BBa_K5034205) and NADK(BBa_K5034206) together to show effects. PPK2 gene and NADK gene are both linked with BBa-B0034 RBS(BBa_B0034) and together expressed in the pBBR1MCS-terminator plasmid(BBa_K5034201).
BBa_K5034225	pBBR1MCS-terminator-NADK	Convert Poly P to NADP. NADK gene(BBa_K5034206) which is from Mycobacterium tuberculosis H37Rv and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201).
BBa_K5034227	pBBR1MCS-terminator-PPK2	Reversibly convert Poly P and Pi, and favors the decomposition reaction. PPK2 gene(BBa_K5034205) which is from Pseudomonas aeruginosa and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201).
BBa_K5034229	pBBR1MCS-terminator-PPX	Thoroughly convert Poly P to Pi. PPX gene(BBa_K5034210) which is from Yeast and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201).
BBa_K5034231	pBBR1MCS-terminator-PPN1	Convert Poly P into smaller polyphosphate fragments, but not completely into Pi. PPN1 gene(BBa_K5034209) which is from Saccharomyces cerevisiae and we performed codon optimization on is expressed in pBBR1MCS-terminator plasmid(BBa_K5034201).