Team:Shanghaitech/Parts


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.

NameTypeDescriptionDesignerLength
  WBBa_K2315011MeasurementLasR-pLas-GFP FANG LUO1913
  WBBa_K2315034MeasurementLasR-pLas-GFP HSL inducible fluorescent actuator FANG LUO, FANG BA1953
  BBa_K2315001Generatorpromoter+RBS+lasR(with histag)+terminatorMingZhe Chen994
  BBa_K2315002CodingLuxI with histagFANG BA597
  BBa_K2315003Generatorpcons+LuxI (with histag)FANG BA795
  BBa_K2315004CodingLuxIFANG BA579
  BBa_K2315005Generatorpcons+LuxIFANG BA640
  BBa_K2315006GeneratorRpaR-RHOPA-pLas-GFPFANG BA1917
  BBa_K2315007GeneratorRpaR-RHOPA-pRpa-GFPFANG BA1898
  BBa_K2315008GeneratorRpaR-RHOPA-pRhl-GFP FANG BA1893
  BBa_K2315009GeneratorRpaR-RHOPA-pLux-GFP FANG BA1897
  BBa_K2315010GeneratorRpaR-RHOPA-pTra-GFP FANG BA1896
  BBa_K2315012GeneratorpCon-LasR-pRhl-GFP FANG BA1881
  BBa_K2315013GeneratorpCon-RhlR-pRpa-GFP  FANG BA1884
   BBa_K2315014GeneratorpCon-RhlR-pRhl-GFP  Fang Luo, FANG BA1887
  BBa_K2315015GeneratorpCon-RhlR-pLux-GFP  FANG BA1891
  BBa_K2315016GeneratorpCon-RhlR-pTra-GFP  FANG BA1890
  BBa_K2315017GeneratorpCon-RhlR-pLas(Old)-GFP  FANG BA1985
  BBa_K2315018GeneratorpCon-TraR-pRpa-GFP  FANG BA1863
  BBa_K2315019GeneratorpCon-TraR-pLux-GFP  FANG BA1870
  BBa_K2315020GeneratorpCon-TraR-pTra-GFP  FANG BA1869
  BBa_K2315021GeneratorpCon-TraR-pLas(Old)-GFP FANG BA1964
  BBa_K2315022GeneratorpCon-LuxR-pRpa-GFP  FANG BA1919
  BBa_K2315023GeneratorpCon-LuxR-pRhl-GFP FANG BA1914
  BBa_K2315024GeneratorpCon-LuxR-pLux-GFP  FANG BA1900
  BBa_K2315025GeneratorpCon-RpaR-Sphingobium sp. EP60837-pLas-GFP FANG BA1932
  BBa_K2315026GeneratorpCon-RpaR-Sphingobium sp. EP60837-pRpa-GFP FANG BA1913
  BBa_K2315027GeneratorpCon-RpaR-Sphingobium sp. EP60837-pRhl-GFP FANG BA1908
  BBa_K2315028GeneratorpCon-RpaR-Sphingobium sp. EP60837-pLux-GFP FANG BA1912
  BBa_K2315029GeneratorpCon-RpaR-Sphingobium sp. EP60837-pTra-GFP FANG BA1911
  BBa_K2315030GeneratorpCon-RpaR-Sphingobium sp. EP60837-pRpa-Mrfp1 FANG BA2006
  BBa_K2315031GeneratorRhlI+lac FANG BA2282
  BBa_K2315032Generatorpcon-lasR-plas-GFP FANG BA2004
  BBa_K2315033Generatorpcon-lasI FANG BA825
  BBa_K2315035Generatorpcon-rhlI FANG BA825
  BBa_K2315036GeneratorGFP-ter FANG BA879
  BBa_K2315037GeneratorlasR-ter FANG BA931
   BBa_K2315038GeneratorPcons+B0034+lacI-terFANG BA1341
  BBa_K2315039GeneratorlasI-lac FANG BA2282
  BBa_K2315040GeneratorlacI-ter FANG BA1288
  BBa_K2315041Generatorpcon-traI FANG BA853
  BBa_K2315042Generatorpcon-rpaI FANG BA869
  BBa_K2315043Generatorplas-GFP-ter FANG BA1073
  BBa_K2315044Generatorpcon-rpaR FANG BA946
  BBa_K2315045Generatorpcon+luxI-2 FANG BA777
   BBa_K2315046SignallingRpa-Las signal molecule converterFANG BA1858
  BBa_K2315047GeneratorGFP+TER 去AAV FANG BA857
  BBa_K2315048GeneratorRhlR FANG BA937
  BBa_K2315049GeneratorJ04500+TraI FANG BA883
  BBa_K2315050GeneratorJ04500+RpaI FANG BA2386
  BBa_K2315051GeneratorTraI+LacFANG BA2371
  BBa_K2315052GeneratorRpaI+LacFANG BA2386
  BBa_K2315053GeneratorTraR+RFP FANG BA1836
  BBa_K2315054GeneratorRpaR+RFP FANG BA1880
  BBa_K2315055GeneratorTraR+terminator FANG BA860
  BBa_K2315056GeneratorRpaR+terminator FANG BA902
   BBa_K2315057GeneratorLasI lac诱导FANG BA2282
  BBa_K2315058GeneratorRhlI lac诱导FANG BA2315
  BBa_K2315059GeneratorpCon-TraI FANG BA853
  BBa_K2315060GeneratorpCon-RpaI FANG BA869
  BBa_K2315061GeneratorLasR-pLas-amilCPFANG BA1862
  BBa_K2315062GeneratorglnAp2+lasIFANG BA1069
  BBa_K2315063GeneratorasrCp+lasIFANG BA1197
  BBa_K2315064GeneratorglnHp2+lasIFANG BA1057
  BBa_K2315100CodinglasI with 6*his tag and double TAAMingZhe Chen627
   BBa_K2315101RegulatorypLas+b0034 without barFANG BA169
   BBa_K2315102RegulatoryPcons+RBS (J23100+B0034) without barFANG BA47
  BBa_K2315103CodingLasI with histagFANG BA627

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

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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

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