BBa_K1969013 1 BBa_K1969013 Nano-lantern(cAMP-1.6) regulated by GAL1 promoter 2016-10-09T11:00:00Z 2016-10-12T06:41:07Z The expression of Nano-lantern(cAMP-1.6) may affect the sensing resolution of intracellular cAMP level, so creating expression cassettes with different expression level is critical for the project. This cassette is one of median expression level. The sequence of Nano-lantern(cAMP-1.6) is cloned downstream the yeast GAL1 promoter and ended with an ADH1 terminator to generate a median expression cassette in yeast Saccharomyces cerevisiae. false false _2436_ 29963 29963 9 false Use a inducible promoter GAL1 to mediate the expression of reporter protein Nano-lantern(cAMP-1.6). false Yijie Lai component2495447 1 BBa_K1969000 component2495449 1 BBa_K1969011 component2495451 1 BBa_K1486025 component2495441 1 BBa_K1969004 component2495443 1 BBa_K1969010 annotation2495447 1 BBa_K1969000 range2495447 1 453 2555 annotation2495443 1 BBa_K1969010 range2495443 1 443 452 annotation2495441 1 BBa_K1969004 range2495441 1 1 442 annotation2495451 1 BBa_K1486025 range2495451 1 2576 2763 annotation2495449 1 BBa_K1969011 range2495449 1 2556 2575 BBa_K1969010 1 BBa_K1969010 Linker sequence between GAL1 promoter and Nano-lantern(cAMP-1.6) 2016-10-09T11:00:00Z 2016-10-12T06:38:15Z It's designed artificially. This is the linker sequence between GAL1 promoter and Nano-lantern(cAMP-1.6), it is left after the two functional part are ligated using recombinase. false false _2436_ 29963 29963 9 false As we originally don't have the 4 standard restriction enzymes require for the biobrick assembly, we use homologous recombination to link two functional parts together. false Yijie Lai annotation2489593 1 Linker sequence between GAL1 promoter and Nano-lantern(cAMP-1.6) range2489593 1 1 10 BBa_K1969004 1 BBa_K1969004 GAL1 promoter 2016-10-04T11:00:00Z 2016-10-08T08:45:50Z Saccharomyces cerevisiae genomic sequence. Transcription terminator for alcohol dehydrogenase 1. false false _2436_ 29963 29963 9 false We chose to PCR amplify it out of Saccharomyces cerevisiae genomic DNA. false Yijie Lai annotation2487974 1 GAL1 promoter range2487974 1 1 442 BBa_K1969011 1 BBa_K1969011 Linker sequence between ADH1 terminator and two coding sequences downstream GAL1 promoter 2016-10-09T11:00:00Z 2016-10-12T06:33:57Z It is designed artificially. This is the linker sequence between ADH1 terminator and Nano-lantern(cAMP-1.6) (downstream GAL1 promoter). false false _2436_ 29963 29963 9 false It is designed for homologous recombination with the backbone plasmid used to insert in the HIS3 site in yeast. false Yijie Lai annotation2495427 1 Linker sequence between ADH1 terminator and Nano-lantern(cAMP-1.6) (downstream GAL1 promoter) range2495427 1 1 20 BBa_K1969000 1 BBa_K1969000 Nano-lantern(cAMP-1.6) 2016-10-03T11:00:00Z 2016-10-05T10:23:22Z It is a engineered sequence created by professor Nagai and we obtained it from addgene (Plasmid #61245). Nano-lantern(cAMP-1.6) is a fusion protein designed by professor Nagai's group, It is a detector for changes in intracellular cAMP level. The reporter mainly contains three parts: the enhanced yellow fluorescent protein, Venus△C10, the mutated Renilla luciferase, RLuc8△N3, and the cAMP-recognition motif of human EPAC1 with a Q270E mutation(Saito, Chang et al. 2012). The yellow flourescent part is used to increase the emitted photon number of RLuc8△N3 by a Forster resonance energy transfer (FRET) mechanism, in which the excited energy of the luminescent substrate, coelenterazine, bound to RLuc is efficiently transferred intermolecularly to the acceptor Venus△C10. The cAMP-recognition motif of human EPAC1 is flanked by two split parts of RLuc8△N3 based on the complementation of split luciferase (CSL) mechanism, thus the conformational change in cAMP-recognition motif reconstitutes the catalytic activity of the split-Luc in a target-molecule-dependent fashion. Upon binding with cAMP, the fusion protein, Nano-lantern(cAMP-1.6), showed the signal increase of about 130%. We aimed to use the reporter to detect the cAMP surge upon stimulation by ligands of G-protein coupled receptors, so cloned Nano-lantern(cAMP-1.6) downstream three kinds of promoters and navigated its expression, localization and function in yeast Saccharomyces cerevisiae. false false _2436_ 29963 29963 9 false As it contains a YFP, we don't need to add any tag to monitor its cellular localization. false Yijie Lai annotation2487757 1 ATG range2487757 1 1 3 annotation2487756 1 Nano-lantern(cAMP-1.6) range2487756 1 1 2103 annotation2487758 1 TAA range2487758 1 2101 2103 BBa_K1486025 1 BBa_K1486025 ADH1 Terminator 2014-09-25T11:00:00Z 2015-05-08T01:10:42Z To edit To edit false false _1866_ 0 21042 9 Not in stock false To edit false EPFL iGem team 2014 annotation2418776 1 Adh1 terminator range2418776 1 1 188 BBa_K1969011_sequence 1 ggcgcgccacttctaaataa BBa_K1486025_sequence 1 gcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagtatgaggtcgctcttattgaccacacctctaccgg BBa_K1969004_sequence 1 cggattagaagccgccgagcgggcgacagccctccgacggaagactctcctccgtgcgtcctcgtcttcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggag BBa_K1969010_sequence 1 atgtcgtacg BBa_K1969000_sequence 1 atgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaa BBa_K1969013_sequence 1 cggattagaagccgccgagcgggcgacagccctccgacggaagactctcctccgtgcgtcctcgtcttcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggagatgtcgtacgatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaaggcgcgccacttctaaataagcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagtatgaggtcgctcttattgaccacacctctaccgg igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z