BBa_K1969014 1 BBa_K1969014 Nano-lantern(cAMP-1.6) regulated by CUP1 promoter 2016-10-11T11:00:00Z 2016-10-12T09:09:11Z It's a combination of two basic parts. The sequence of Nano-lantern(cAMP-1.6) is cloned downstream the yeast CUP1 promoter without a terminator to generate a low expression cassette in yeast Saccharomyces cerevisiae. false false _2436_ 29963 29963 9 false As the CUP1 promoter is a promoter of low expression, and the insertion site at yeast genome has a terminator of its own, we don't cloned any terminator downstream the coding sequence. false Yijie Lai component2493896 1 BBa_K1969005 component2493898 1 BBa_K1969012 component2493902 1 BBa_K1969000 annotation2493902 1 BBa_K1969000 range2493902 1 304 2406 annotation2493896 1 BBa_K1969005 range2493896 1 1 235 annotation2493898 1 BBa_K1969012 range2493898 1 236 303 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_K1969005 1 BBa_K1969005 CUP1 promoter 2016-10-07T11:00:00Z 2016-10-08T09:05:47Z The genomic sequence from Saccharomyces cerevisiae. This is the full length of CUP1 promoter in yeast Saccharomyces cerevisiae. As the major copper-activated metallothionine in yeast, Cup1p binds and sequesters cuprous copper(I), Cu+, providing the principal method of removing this metal ion from the cell. CUP1 transcription is specifically induced by the copper-dependent transcription activator Cup2p (more commonly known as Ace1p) in response to high levels of copper ions and by Hsf1p in response to heat shock, glucose starvation and oxidation stress. In the presence of copper, Cup1p is also capable of antioxidant activity and thus contributes a significant, albeit minor, role to oxygen radical detoxification, especially in the absence of Cu,Zn-superoxide dismutase Sod1p. false false _2436_ 29963 29963 9 false As the promoter sequence is rather short, we may use recombinase to inset it upstream different coding sequence to control the expression level. false Yijie Lai annotation2487976 1 CUP1 promoter range2487976 1 1 235 BBa_K1969012 1 BBa_K1969012 Linker sequence between CUP1 promoter and Nano-lantern(cAMP-1.6) 2016-10-09T11:00:00Z 2016-10-12T06:40:08Z It's designed artificially. This is the linker sequence between CUP1 promoter and Nano-lantern(cAMP-1.6) to introduce ClaI, MseI sites. false false _2436_ 29963 29963 9 false It is used to introduce ClaI, MseI sites and left by homologous recombination. false Yijie Lai annotation2489595 1 Linker sequence between CUP1 promoter and Nano-lantern(cAMP-1.6) range2489595 1 1 68 BBa_K1969005_sequence 1 ctagttagaaaaagacatttttgctgtcagtcactgtcaagagattcttttgctggcatttcttctagaagcaaaaagagcgatgcgtcttttccgctgaaccgttccagcaaaaaagactaccaacgcaatatggattgtcagaatcatataaaagagaagcaaataactccttgtcttgtatcaattgcattataatatcttcttgttagtgcaatatcatatagaagtcatc BBa_K1969000_sequence 1 atgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaa BBa_K1969012_sequence 1 gaaatagatattaagaaaaacaaactgtacaatcaatcaatcaatcatcacataaaatgtctatcgat BBa_K1969014_sequence 1 ctagttagaaaaagacatttttgctgtcagtcactgtcaagagattcttttgctggcatttcttctagaagcaaaaagagcgatgcgtcttttccgctgaaccgttccagcaaaaaagactaccaacgcaatatggattgtcagaatcatataaaagagaagcaaataactccttgtcttgtatcaattgcattataatatcttcttgttagtgcaatatcatatagaagtcatcgaaatagatattaagaaaaacaaactgtacaatcaatcaatcaatcatcacataaaatgtctatcgatatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaa igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z