Team:Shanghaitech/Library


Collaborations:Library

This year, we have constructed several sensors and reporters as our inputs and executors to provide users more options when using the MagicBlock system. These blocks include lacI-lasI, which generates signals upon induction; pcons+GFP, which constitutively generates signals; lasR-GFP and lasR-mRFP, which present different fluorescence and lasR-amilCP, which gives bacteria resistance to chloramphenicol. Meanwhile, we also collected parts that can be reformed for our MagicBlock from other teams. For example, we have collected promoters that can sense nitrogen from UCAS, an auto-inducer quorum-sensing generator from SiCAU-China, a toolbox of recombinase from Fudan-China, cellulase and pectinase from LanZhou iGEM team, and Bt toxin protein from FAFU-China. In addition to part collection, we also reconstructed some MagicBlocks from previous characterized parts so that they can be integrated into our project. Thus our library was and will be continuously improved by users to increase the versatility.

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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

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