BBa_K1969003 1 BBa_K1969003 Full length S. cerevisiae ADH1 promoter 2016-10-04T11:00:00Z 2016-10-08T08:10:33Z a a false false _2436_ 29963 29963 9 false a false Yijie Lai annotation2486020 1 ADH1 promoter range2486020 1 1 705 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 annotation2487758 1 TAA range2487758 1 2101 2103 annotation2487756 1 Nano-lantern(cAMP-1.6) range2487756 1 1 2103 annotation2487757 1 ATG range2487757 1 1 3 BBa_K1969007 1 BBa_K1969007 Nano-lantern(cAMP-1.6) regulated by ADH1 promoter 2016-10-07T11:00:00Z 2016-10-12T06:37:08Z Use a constitutive promoter ADH1 to mediate the expression of reporter protein Nano-lantern(cAMP-1.6). The sequence of Nano-lantern(cAMP-1.6) is cloned downstream the yeast ADH1 promoter and ended with an ADH1 terminator to generate a expression cassette in yeast Saccharomyces cerevisiae. false false _2436_ 29963 29963 9 false 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 high expression level. false Yijie Lai component2495439 1 BBa_K1486025 component2495437 1 BBa_K1969009 component2495429 1 BBa_K1969003 component2495431 1 BBa_K1969008 component2495435 1 BBa_K1969000 annotation2495437 1 BBa_K1969009 range2495437 1 2839 2867 annotation2495429 1 BBa_K1969003 range2495429 1 1 705 annotation2495431 1 BBa_K1969008 range2495431 1 706 735 annotation2495439 1 BBa_K1486025 range2495439 1 2868 3055 annotation2495435 1 BBa_K1969000 range2495435 1 736 2838 BBa_K1969009 1 BBa_K1969009 The linker sequence between ADH1 terminator and different coding sequences 2016-10-08T11:00:00Z 2016-10-12T06:28:04Z It's artificially designed . This is the linker sequence between ADH1 terminator and Nano-lantern(cAMP-1.6) to introduce the AvrII and AscI unique cut sites. false false _2436_ 29963 29963 9 false We need AvrII and AscI unique cut sites to assemble the parts together. false Yijie Lai annotation2489076 1 Linker between ADH1 terminator and Nano-lantern(cAMP-1.6) range2489076 1 1 29 BBa_K1969008 1 BBa_K1969008 The linker sequence between ADH1 promoter and Nano-lantern(cAMP-1.6) 2016-10-08T11:00:00Z 2016-10-12T09:32:39Z It's artificially designed. This DNA sequence is used to link the ADH1 promoter and Nano-lantern(cAMP-1.6) together and introduce the restriction site for ClaI. false false _2436_ 29963 29963 9 false We add a sequence downstream the ADH1 promoter to give a space between the 5' cap and cds in the transcript, and meanwhile introduce the restriction site for ClaI. false Yijie Lai annotation2489055 1 Linker between ADH1 promoter and Nano-lantern(cAMP-1.6) range2489055 1 1 30 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_K1486025_sequence 1 gcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagtatgaggtcgctcttattgaccacacctctaccgg BBa_K1969008_sequence 1 atctcatatacaatgtctatcccaatcgat BBa_K1969000_sequence 1 atgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaa BBa_K1969009_sequence 1 cctaggtgaggcgcgccacttctaaataa BBa_K1969003_sequence 1 atccttttgttgtttccgggtgtacaatatggacttcctcttttctggcaaccaaacccatacatcgggattcctataataccttcgttggtctccctaacatgtaggtggcggaggggagatatacaatagaacagataccagacaagacataatgggctaaacaagactacaccaattacactgcctcattgatggtggtacataacgaactaatactgtagccctagacttgatagccatcatcatatcgaagtttcactaccctttttccatttgccatctattgaagtaataataggcgcatgcaacttcttttctttttttttcttttctctctcccccgttgttgtctcaccatatccgcaatgacaaaaaaatgatggaagacactaaaggaaaaaattaacgacaaagacagcaccaacagatgtcgttgttccagagctgatgaggggtatctcgaagcacacgaaactttttccttccttcattcacgcacactactctctaatgagcaacggtatacggccttccttccagttacttgaatttgaaataaaaaaaagtttgctgtcttgctatcaagtataaatagacctgcaattattaatcttttgtttcctcgtcattgttctcgttccctttcttccttgtttctttttctgcacaatatttcaagctataccaagcatacaatcaact BBa_K1969007_sequence 1 atccttttgttgtttccgggtgtacaatatggacttcctcttttctggcaaccaaacccatacatcgggattcctataataccttcgttggtctccctaacatgtaggtggcggaggggagatatacaatagaacagataccagacaagacataatgggctaaacaagactacaccaattacactgcctcattgatggtggtacataacgaactaatactgtagccctagacttgatagccatcatcatatcgaagtttcactaccctttttccatttgccatctattgaagtaataataggcgcatgcaacttcttttctttttttttcttttctctctcccccgttgttgtctcaccatatccgcaatgacaaaaaaatgatggaagacactaaaggaaaaaattaacgacaaagacagcaccaacagatgtcgttgttccagagctgatgaggggtatctcgaagcacacgaaactttttccttccttcattcacgcacactactctctaatgagcaacggtatacggccttccttccagttacttgaatttgaaataaaaaaaagtttgctgtcttgctatcaagtataaatagacctgcaattattaatcttttgtttcctcgtcattgttctcgttccctttcttccttgtttctttttctgcacaatatttcaagctataccaagcatacaatcaactatctcatatacaatgtctatcccaatcgatatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaacctaggtgaggcgcgccacttctaaataagcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagtatgaggtcgctcttattgaccacacctctaccgg 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