Parts

This year，we have constructed 5 pairs quorum sensing system, 2 signal converter which can detect one kind of signal molecular and then transfer it into another. We also constructed 20 different receptor-promoter pairs, which can be used to measure the orthogonality of different quorum sensing promoters and receptors.

	Name	Type	Description	Designer	Length
W	BBa_K2315011	Measurement	LasR-pLas-GFP	FANG LUO	1913
W	BBa_K2315034	Measurement	LasR-pLas-GFP HSL inducible fluorescent actuator	FANG LUO, FANG BA	1953
	BBa_K2315001	Generator	promoter+RBS+lasR（with histag）+terminator	MingZhe Chen	994
	BBa_K2315002	Coding	LuxI with histag	FANG BA	597
	BBa_K2315003	Generator	pcons+LuxI (with histag)	FANG BA	795
	BBa_K2315004	Coding	LuxI	FANG BA	579
	BBa_K2315005	Generator	pcons+LuxI	FANG BA	640
	BBa_K2315006	Generator	RpaR-RHOPA-pLas-GFP	FANG BA	1917
	BBa_K2315007	Generator	RpaR-RHOPA-pRpa-GFP	FANG BA	1898
	BBa_K2315008	Generator	RpaR-RHOPA-pRhl-GFP	FANG BA	1893
	BBa_K2315009	Generator	RpaR-RHOPA-pLux-GFP	FANG BA	1897
	BBa_K2315010	Generator	RpaR-RHOPA-pTra-GFP	FANG BA	1896
	BBa_K2315012	Generator	pCon-LasR-pRhl-GFP	FANG BA	1881
	BBa_K2315013	Generator	pCon-RhlR-pRpa-GFP	FANG BA	1884
	BBa_K2315014	Generator	pCon-RhlR-pRhl-GFP	Fang Luo, FANG BA	1887
	BBa_K2315015	Generator	pCon-RhlR-pLux-GFP	FANG BA	1891
	BBa_K2315016	Generator	pCon-RhlR-pTra-GFP	FANG BA	1890
	BBa_K2315017	Generator	pCon-RhlR-pLas(Old)-GFP	FANG BA	1985
	BBa_K2315018	Generator	pCon-TraR-pRpa-GFP	FANG BA	1863
	BBa_K2315019	Generator	pCon-TraR-pLux-GFP	FANG BA	1870
	BBa_K2315020	Generator	pCon-TraR-pTra-GFP	FANG BA	1869
	BBa_K2315021	Generator	pCon-TraR-pLas(Old)-GFP	FANG BA	1964
	BBa_K2315022	Generator	pCon-LuxR-pRpa-GFP	FANG BA	1919
	BBa_K2315023	Generator	pCon-LuxR-pRhl-GFP	FANG BA	1914
	BBa_K2315024	Generator	pCon-LuxR-pLux-GFP	FANG BA	1900
	BBa_K2315025	Generator	pCon-RpaR-Sphingobium sp. EP60837-pLas-GFP	FANG BA	1932
	BBa_K2315026	Generator	pCon-RpaR-Sphingobium sp. EP60837-pRpa-GFP	FANG BA	1913
	BBa_K2315027	Generator	pCon-RpaR-Sphingobium sp. EP60837-pRhl-GFP	FANG BA	1908
	BBa_K2315028	Generator	pCon-RpaR-Sphingobium sp. EP60837-pLux-GFP	FANG BA	1912
	BBa_K2315029	Generator	pCon-RpaR-Sphingobium sp. EP60837-pTra-GFP	FANG BA	1911
	BBa_K2315030	Generator	pCon-RpaR-Sphingobium sp. EP60837-pRpa-Mrfp1	FANG BA	2006
	BBa_K2315031	Generator	RhlI+lac	FANG BA	2282
	BBa_K2315032	Generator	pcon-lasR-plas-GFP	FANG BA	2004
	BBa_K2315033	Generator	pcon-lasI	FANG BA	825
	BBa_K2315035	Generator	pcon-rhlI	FANG BA	825
	BBa_K2315036	Generator	GFP-ter	FANG BA	879
	BBa_K2315037	Generator	lasR-ter	FANG BA	931
	BBa_K2315038	Generator	Pcons+B0034+lacI-ter	FANG BA	1341
	BBa_K2315039	Generator	lasI-lac	FANG BA	2282
	BBa_K2315040	Generator	lacI-ter	FANG BA	1288
	BBa_K2315041	Generator	pcon-traI	FANG BA	853
	BBa_K2315042	Generator	pcon-rpaI	FANG BA	869
	BBa_K2315043	Generator	plas-GFP-ter	FANG BA	1073
	BBa_K2315044	Generator	pcon-rpaR	FANG BA	946
	BBa_K2315045	Generator	pcon+luxI-2	FANG BA	777
	BBa_K2315046	Signalling	Rpa-Las signal molecule converter	FANG BA	1858
	BBa_K2315047	Generator	GFP+TER 去AAV	FANG BA	857
	BBa_K2315048	Generator	RhlR	FANG BA	937
	BBa_K2315049	Generator	J04500+TraI	FANG BA	883
	BBa_K2315050	Generator	J04500+RpaI	FANG BA	2386
	BBa_K2315051	Generator	TraI+Lac	FANG BA	2371
	BBa_K2315052	Generator	RpaI+Lac	FANG BA	2386
	BBa_K2315053	Generator	TraR+RFP	FANG BA	1836
	BBa_K2315054	Generator	RpaR+RFP	FANG BA	1880
	BBa_K2315055	Generator	TraR+terminator	FANG BA	860
	BBa_K2315056	Generator	RpaR+terminator	FANG BA	902
	BBa_K2315057	Generator	LasI lac诱导	FANG BA	2282
	BBa_K2315058	Generator	RhlI lac诱导	FANG BA	2315
	BBa_K2315059	Generator	pCon-TraI	FANG BA	853
	BBa_K2315060	Generator	pCon-RpaI	FANG BA	869
	BBa_K2315061	Generator	LasR-pLas-amilCP	FANG BA	1862
	BBa_K2315062	Generator	glnAp2+lasI	FANG BA	1069
	BBa_K2315063	Generator	asrCp+lasI	FANG BA	1197
	BBa_K2315064	Generator	glnHp2+lasI	FANG BA	1057
	BBa_K2315100	Coding	lasI with 6*his tag and double TAA	MingZhe Chen	627
	BBa_K2315101	Regulatory	pLas+b0034 without bar	FANG BA	169
	BBa_K2315102	Regulatory	Pcons+RBS (J23100+B0034) without bar	FANG BA	47
	BBa_K2315103	Coding	LasI with histag	FANG BA	627

In addition, we also collect parts from other Team, and tried to transform them adapting to our system.

And here is our part collection list

name	Team	E-mail	Function Description	experience
glnAp2+riboJ+RBS	UCAS	ucasigem@163.com	A sigma54-dependent promoter together with its regulating region, responsing to the changes of nitrogen concentration in the environment. Originally regulate the transcription of glnA.	Part:BBa_K2287001
glnHp2	UCAS	ucasigem@163.com	A sigma54-dependent promoter together with its regulating region, responsing to the changes of nitrogen concentration in the environment. Originally regulate the transcription of glnH.	Part:BBa_K2287002
astCp	UCAS	ucasigem@163.com	A sigma54-dependent promoter together with its regulating region, responsing to the changes of nitrogen concentration in the environment. Originally regulate the transcription of astC.	Part:BBa_K2287003
celluse	Lanzhou	347684025@qq.com	celluse	Part:BBa_K2377002
pectinase	Lanzhou	347684025@qq.com	pectinase	Part:BBa_K2377003
φBT1 integrase	Fudan-China	igem@fudan.edu.cn	This ORF codes for the steptomyces phage φBT1 integrase, which catalyzes a site specific recombination between φBT1 attB and attP sequences.	Part:BBa_K2460001
φBT1 attB	Fudan-China	igem@fudan.edu.cn	the sequence which can be recoginized by the steptomyces phage φBT1 integrase	Part:BBa_K2460002
φBT1 attP	Fudan-China	igem@fudan.edu.cn	the sequence which can be recoginized by the steptomyces phage φBT1 integrase	Part:BBa_K2460003
φRv1 integrase	Fudan-China	igem@fudan.edu.cn	This ORF codes for the mycobacterium phage φRv1 integrase, which catalyzes a site specific recombination between φRv1 attB and attP sequences.	Part:BBa_K2460004
φRv1 attB	Fudan-China	igem@fudan.edu.cn	the sequence which can be recoginized by the steptomyces phage φRv1 integrase	Part:BBa_K2460005
φRv1 attP	Fudan-China	igem@fudan.edu.cn	the sequence which can be recoginized by the steptomyces phage φRv1 integrase	Part:BBa_K2460006
TG1 integrase	Fudan-China	igem@fudan.edu.cn	This ORF codes for the steptomyces phage TG1 integrase, which catalyzes a site specific recombination between TG1 attB and attP sequences.	Part:BBa_K2460007
TG1 attB	Fudan-China	igem@fudan.edu.cn	the sequence which can be recoginized by the steptomyces phage TG1 integrase	Part:BBa_K2460008
TG1 attP	Fudan-China	igem@fudan.edu.cn	the sequence which can be recoginized by the steptomyces phage TG1 integrase	Part:BBa_K2460009
4A5-R-IG(△LVA)	SiCAU-China	linjj23@126.com	It's a positive feedback system, which could appear strong green fluorescence by background expression in BL21(DE3) or other host.	Part:BBa_K2311002
4A5-R-IG(△LVA)-AiiA	SiCAU-China	linjj23@126.com	It's a positive feedback system, which could appear strong green fluorescence by background expression in BL21(DE3) or other host. But this system may need more time to show postive feedback effect than the positive feedback system without AiiA control	Part:BBa_K2311001
Cry4Aa4 + Extended FMDV	FAFU-China	jacarandasmile@163.com	Cry4Aa is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex by bioassay. The Cry protein is consisted of three functional domains. Domain I is a seven α-helices bundle. It can insert itself into a membrane by using its hydrophobic helices α4 and α5 to insert into the phospholipid bilayer. The pore formation occurs on its α3 helix. Domain II and domain III are two β-sheets which are involved in the receptor interactions. Domain II contains extremely variable loops, which are the binding site of the receptor. Domain III has the function of stabilizing the toxin. Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer ，Then the oligomer binds to a GPI-anchored APN or ALP. Secondly, the previous binding induces the oligomer insertion into the lipid rafts membrane. A formation of ion permeable pore is followed by the insertion which allows small molecules to pass through the membrane. The membrane potential inevitably changes greatly, causing the swelling of cell and finally breaking down. When the cell lysis reaches to a certain degree, the midgut necrosis and epithelial denaturation follow. Then, the alkaline hypertonic inclusions in midgut enters into hemocoel and the pH of haemolymph rises causing paralysis of larvae and finally death. To increase the express level in Chlamydomonas reintmrdtii, we synthesis Chlamydomonas reintmrdtii codon optimized Cyt1. Meanwhile, we added 2A peptide sequence at the end of 5’ According to the published paper and the result of Swiss-model, there is no effect to the toxicity of Cry or Cyt. Users can use infusion technology to link the express vector to express toxins	Part:BBa_K2074021
Cry10Aa4 + Extended FMDV	FAFU-China	jacarandasmile@163.com	Cry10Aa is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex and Aedes by bioassay. The Cry protein is consisted of three functional domains. Domain I is a seven α-helices bundle. It can insert itself into a membrane by using its hydrophobic helices α4 and α5 to insert into the phospholipid bilayer. The pore formation occurs on its α3 helix. Domain II and domain III are two β-sheets which are involved in the receptor interactions. Domain II contains extremely variable loops, which are the binding site of the receptor. Domain III has the function of stabilizing the toxin. Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer.	Part:BBa_K2074022
cyt1	FAFU-China	jacarandasmile@163.com	Cyt1 is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex and Aedes by bioassay. Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer，Then the oligomer binds to a GPI-anchored APN or ALP. Secondly, the previous binding induces the oligomer insertion into the lipid rafts membrane. A formation of ion permeable pore is followed by the insertion which allows small molecules to pass through the membrane. The membrane potential inevitably changes greatly, causing the swelling of cell and finally breaking down. When the cell lysis reaches to a certain degree, the midgut necrosis and epithelial denaturation follow. Then, the alkaline hypertonic inclusions in midgut enters into hemocoel and the pH of haemolymph rises causing paralysis of larvae and finally death. To increase the express level in Chlamydomonas reintmrdtii, we synthesis Chlamydomonas reintmrdtii codon optimized Cyt1. Meanwhile, we added 2A peptide sequence at the end of 5’ According to the published paper and the result of Swiss-model, there is no effect to the toxicity of Cry or Cyt. Users can use infusion technology to link the express vector to express toxins.	Part:BBa_K2074024
cyt2	FAFU-China	jacarandasmile@163.com	Cyt2 is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex and Aedes by bioassay.Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer	Part:BBa_K2074025

Team:Shanghaitech/Parts

Parts