Part collection

This year, we designed, built and characterized a set of parts. They were consisted of different parts collection and played different roles in different section of our parts. All the new parts had been submitted to The iGEM Parts Registry following the BioBricks assembly standard. On this page, we gave a brief description of our parts and we divided them into several sections according to the modules they worked in our project. For more detailed information about our parts, we highly recommend you to visit the corresponding parts page on The iGEM Parts Registry or turn to the Design and Engineering page of our wiki.

Basic Parts

ZJU-CHina 2023 Parts(Basic)
Module	Type	Designer	Code	Name	Length (bp)	Function and Description
Plant immunity	Regulatory	Qinyao Yan	BBa_K4628000	CaMV 35S promoter	346	In our experiment, it serves as a strong promoter for the expression of most proteins.
	Terminator		BBa_K4628001	CaMV poly(A) signal	177	In our experiment, it serves as a transcriptional terminator for the expression of most proteins.
	Coding		BBa_K4628002	CP(from TMV)	477	CP is the capsid protein of TMV, which is packaged on the outside of the viral nucleic acid to form protection.
			BBa_K4628003	Enhancer	351	A nanoantibody derived from alpaca that specifically recognizes GFP.
			BBa_K4628004	LaG16	393	A nanoantibody derived from alpaca that specifically recognizes GFP.
			BBa_K4628005	LaM2	381	A nanoantibody derived from alpaca that specifically recognizes mCherry.
			BBa_K4628006	LaM4	390	A nanoantibody derived from alpaca that specifically recognizes mCherry.
			BBa_K4628007	Pikm-1	3684	A kind of plant NLR receptor protein that can perform specific recognition and immune activation function.
			BBa_K4628008	Pikm-1*(Enhancer)	3813	Transforming the ID sequence of the original Pikm-1 to an Enhancer nanoantibody targeting GFP.
			BBa_K4628009	Pikm-2	3458	A kind of plant NLR helper protein that can dimerize with Pikm-1 to assist in its immune activation function.
TLS module	Reporter	Wenqing Ding	BBa_K4628010	eGFP	720	Enhanced Green Fluorescent Protein (eGFP) is a genetically engineered variant of the Green Fluorescent Protein (GFP)
	Reporter		BBa_K4628011	GUS	1815	It encodes β-glucosidase, a hydrolyzing enzyme that uses β-glucosinolates as substrates and whose product is readily detectable by the naked eye.
	RNA		BBa_K4628012	TLS(Ara.Ala.full)	73	The tRNA sequence of Arabidopsis thaliana Alanine.
			BBa_K4628013	TLS(Ara.Glu.full)	72	The tRNA sequence of Arabidopsis thaliana Glutamicacid.
			BBa_K4628014	TLS(Ara.Met.full)	75	The tRNA sequence of Arabidopsis thaliana methionine.
			BBa_K4628015	TLS(Nico.Met.full)	73	The tRNA sequence of Nicotiana benthamiana methionine.
			BBa_K4628016	TLS(Ara.Met.ΔΔt)	47	The modified tRNA sequence of Arabidopsis thaliana methionine, which deletes the two loop.
			BBa_K4628017	TLS(Ara.Met.ΔΔt.muti)	1441	Expecting to enhance mRNA binding to RNA motility-associated proteins by increasing the copy number of the TLS sequence, we replaced some of the repeats of SL24 with the RNA sequence of TLS (Ara.Mer.ΔΔt).
RdRP module	Regulatory	Linghao Xia	BBa_K4628018	subPromoter CP	211	Because part of the coding sequence of capsid protein constitutes the sequence of subPromoter CP, we mutant the start codon in the subpromoter(5,712–5,714,from ATG to AGA) in case of expressing a nonsense peptide.
	Regulatory		BBa_K4628019	subPromoter MP	135	This Part is derived from the subgenomic movement protein promoter in the TMV genome, which mediates the transcription of MP.
	Coding		BBa_K4628020	TMV movement protein	807	This Part is derived from the sequence encoding the 30 KDa protein in the TMV genome, which has movement protein activity and helps specific RNAs move between plant cells.
	Coding		BBa_K4628021	TMV RNA dependent RNA polymerase	4851	This Part is derived from the sequences encoding 126K and 183K proteins in the TMV genome, which has RNA-dependent RNA polymerase activity and can help the target RNA sequence replication and transcription.
Biosafety module	Regulatory	Junjie Li, Zhijian Yan	BBa_K4628022	50Spro	420	It’s a constitutive promoter in Agrobacterium tumefaciens str.C58 genome, encoding the synthetic 50S ribosomal protein L28[i]. The 420bp upstream sequence of this gene was selected as this promoter.
			BBa_K4628023	Pvbp2	526	The Pvbp2 promoter is an inducible promoter for VirD2-Binding Protein originated from Agrobacterium tumefaciens str.C58
			BBa_K4628024	Ptac/cuo	243	The Ptac promoter is a strong promoter in Agrobacterium strains. It can control the expression of the virE2 coding sequence. Studies have shown that virE2 expression can mediate T-DNA transfer, it requires IPTG induction and has a certain leakage[i]. Another study tested good controllability by designing a synthetic Ptac/CuO promoter by replacing the lac operator sequence in the Ptac promoter with the cumate operator sequence[ii].
	Coding		BBa_K4628025	Agrobacterium tumefaciens optimized miniSOG	321	This part derivers from BBa_K3111032. We replace codons in the original mRNA sequence with codons that are used frequently in Agrobacterium tumefaciens to ensure that the codons in the newly designed mRNA sequence are more compatible with the codon usage bias of our chassis, avoiding the emergence of rare codons.

Composite Parts

ZJU-CHina 2023 Parts(Composite)
Module	Type	Designer	Code	Name	Length (bp)	Function and Description
Plant immunity	Measurement	Qinyao Yan	BBa_K4628026	CP-GFP	1194	Insert the EGFP sequence after the capsid protein sequence in the TMV genome to construct a fusion protein.
	Generator		BBa_K4628027	Pikm-1 expression circuit	4166	Combined with CaMV 35S promoter, Pikm-1, CaMV poly(A) signal
			BBa_K4628028	Pikm-1(Enhancer) expression circuit	4295	Combined with CaMV 35S promoter, Pikm-1(Enhancer), CaMV poly(A) signal
			BBa_K4628029	Pikm-2 expression circuit	3644	Combined with CaMV 35S promoter, Pikm-2, CaMV poly(A) signal
			BBa_K4628030	eGFP expression circuit	1325	Combined with CaMV 35S promoter, eGFP, CaMV poly(A) signal
TLS module	Measurement	Wenqing Ding	BBa_K4628031	eGFP-TLS(Ara.Met.full)	795	Addition of the Arabidopsis thaliana methionine tRNA sequence to the 3'UTR region of the eGFP mRNA allowed the eGFP mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by fluorescent expression of eGFP.
			BBa_K4628032	eGFP-TLS(Ara.Met.ΔΔt)	767	Addition of the Arabidopsis thaliana methionine tRNA sequence with two loops deleted in the 3'UTR region of the eGFP mRNA allowed the eGFP mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by fluorescent expression of eGFP.
			BBa_K4628033	eGFP-TLS(Nico.Met.full)	793	Addition of the Nicotiana benthamiana methionine tRNA sequence to the 3'UTR region of the eGFP mRNA allowed the eGFP mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by fluorescent expression of eGFP.
			BBa_K4628034	GUS-TLS(Ara.Ala.full)	1888	Addition of the Arabidopsis thaliana Alanine tRNA sequence to the 3'UTR region of the GUS mRNA allowed the GUS mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by GUS staining reaction.
			BBa_K4628035	GUS-TLS(Ara.Glu.full)	1887	Addition of the Arabidopsis thaliana Glutamicacid tRNA sequence to the 3'UTR region of the GUS mRNA allowed the GUS mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by GUS staining reaction.
			BBa_K4628036	GUS-TLS(Ara.Met.full)	1890	Addition of the Arabidopsis thaliana methionine tRNA sequence to the 3'UTR region of the GUS mRNA allowed the GUS mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by GUS staining reaction.
			BBa_K4628037	GUS-TLS(Ara.Met.ΔΔt)	1862	Addition of the Arabidopsis thaliana methionine tRNA sequence with two loops deleted to the 3'UTR region of the GUS mRNA allowed the GUS mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by GUS staining reaction.
			BBa_K4628038	RuBy-TLS(Ara.Met.full)	4026	Splicing the Ruby part(BBa_K3900028) and the TLS(Ara.Met.full) part together.It allowed the Ruby mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by the RuBy-catalyzed production of betacyanin.
			BBa_K4628039	RuBy-TLS(Ara.Met.ΔΔt)	3998	Splicing the Ruby part(BBa_K3900028) and the TLS(Ara.Met.ΔΔt) part together.It allowed the RuBy mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by the RuBy-catalyzed production of betacyanin.
			BBa_K4628040	RuBy-TLS(Nico.Met.full)	4024	Splicing the Ruby part(BBa_K3900028) and the TLS(Nico.Met.full) part together.It allowed the RuBy mRNA to acquire the ability to move long distances throughout the plant body, and this process can be reported by the RuBy-catalyzed production of betacyanin.
RdRP module	Device	Linghao Xia	BBa_K4628041	pGreenⅡ 0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP	9901	Agrobacterium transforming the plasmid was used to transect tobacco instantaneously, and GFP was transcribed and translated through subPromoter CP. At the same time, RNA-dependent RNA polymerase enables the transcribed RNA to self-replicate, thus improving the existence time of specific RNA and the expression amount of target protein.
			BBa_K4628042	pGreenⅡ 0800-TMV RNA dependent RNA polymerase-subPromoter MP-EGFP	9026	Agrobacterium transforming the plasmid was used to transect tobacco instantaneously, and GFP was transcribed and translated through subPromoter MP. At the same time, RNA-dependent RNA polymerase enables the transcribed RNA to self-replicate, thus improving the existence time of specific RNA and the expression amount of target protein
			BBa_K4628043	pGreenⅡ 0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP-TLS(Ara.Met.full)-UTR	9975	On the basis of pGreen Ⅱ0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP, We inserted a tRNA-like sequence (TLS (Ara.Met.full)) after TMV 3 'UTR.
			BBa_K4628044	pGreenⅡ 0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP-UTR-TLS(Ara.Met.full)	9975	On the basis of pGreen Ⅱ0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP, We inserted a tRNA-like sequence (TLS (Ara.Met.full)) after TMV 3 'UTR.
			BBa_K4628045	pGreenⅡ 0800-TMV RNA dependent RNA polymerase- subPromoter MP-EGFP- TLS(Ara.Met.full) -UTR	9100	On the basis of pGreen Ⅱ0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP, We inserted a tRNA-like sequence (TLS (Ara.Met.full)) before TMV 3 'UTR.
			BBa_K4628046	pGreenⅡ 0800-TMV RNA dependent RNA polymerase- subPromoter MP-EGFP-UTR -TLS(Ara.Met.full)	9100	On the basis of pGreen Ⅱ0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP, We inserted a tRNA-like sequence (TLS (Ara.Met.full)) after TMV 3 'UTR.
Biosafety module	Generator	Junjie Li, Zhijian Yan	BBa_K4628047	50Spro-KillerRed	1272	KillerRed (BBa_K1184000) is a red fluorescent protein that generates reactive oxygen species (ROS) under yellow-green light (540-585 nm)[i]. This component serves a crucial role in preventing engineered bacteria from inadvertently escaping into the environment .
Biosafety module	Generator	Junjie Li, Zhijian Yan	BBa_K4628048	50Spro-miniSOG	871	This phototoxic protein, derived from LOV protein, requires light induction and produces toxic substances under blue light excitation to kill bacteria[i]. It also emits green fluorescence for easy observation. To facilitate the use of this protein in Agrobacterium, its codon has been optimized.