BBa_K2111000 1 BBa_K2111000 RI7_FLAG_OXTR 2016-10-18T11:00:00Z 2016-10-19T05:11:50Z The human oxytocin receptor sequence was found in the NCBI database, codon optimized for saccharomyce cerevisiae using http://www.jcat.de/ (with additional options to avoid cleavage sites EcoR1, Pst1, Spe1, Xba1. The sequence for the rat RI7 OR was found on the 2012 TU Delft iGEM team page and in Rhadhika 2007 (http://www.nature.com/nchembio/journal/v3/n6/full/nchembio882.html). This part is a chimeric receptor with a FLAG tag. It is composed of the ligand biding domain of the human oxytocin receptor, flanked by the N and C termini of the rat RI7 olfactory receptor. It was designed using the DIY receptor design guide created by the 2012 TU Delft iGEM team (http://2012.igem.org/Team:TU-Delft/receptordesign). This part is intended to express in the membrane of a yeast cell and - upon oxytocin ligand binding - activate the mating pathway and the inducible FUS1 promotor. This allows us to leverage a reporter protein under control of FUS1 to detect the presence of oxytocin. false false _2579_ 31485 31485 9 false We added a FLAG tag to facilitate SDS-PAGE. We added the yeast kozak sequence tacaaa to improve translation and codon optimized for saccharomyce cerevisiae. false Isaac Bean annotation2524246 1 RI7 C-terminus range2524246 1 1018 1116 annotation2523800 1 RI7 N-terminus range2523800 1 34 213 annotation2523657 1 FLAG tag range2523657 1 10 33 annotation2524445 1 Stop range2524445 1 1117 1122 annotation2522995 1 Kozak range2522995 1 1 6 annotation2524194 1 Start range2524194 1 7 9 annotation2524242 1 Human Oxytocin Receptor Binding Site range2524242 1 214 1017 BBa_K2111001 1 BBa_K2111001 PGK1_OXTR 2016-10-18T11:00:00Z 2016-10-19T06:46:31Z We used BBa_K1652000 from the 2015 University of Ottawa iGEM team. This is a composite of the PGK1 promoter (BBa_K1652000) and RI7_FLAG_OXTR (BBa_K2111000). The PGK1 (pPGK1Gx) is a S. cerevisiae optimized constitutive promoter to facilitate the expression of RI7_FLAG_OXTR. false false _2579_ 31485 31485 9 It's complicated false We used this promoter to test constitutive expression of our receptor. false Isaac Bean component2523738 1 BBa_K2111000 component2523736 1 BBa_K1652000 annotation2523738 1 BBa_K2111000 range2523738 1 881 2002 annotation2523736 1 BBa_K1652000 range2523736 1 1 872 BBa_K1652000 1 BBa_K1652000 pPGK1Gx 2015-09-07T11:00:00Z 2015-09-08T12:39:58Z S. cerevisiae PGK1 promoter with two gal4 binding sites close to the transcription start site (TSS) that cause the promoter to be repressed in the presence of Gal4 binding protein. false false _2069_ 18008 18008 9 false TSS in PGK1 is not well-defined, and there exists a region at the end of the promoter, about 100 bp long, that is very AT-rich. The promoter also contains two elements that resemble TATA boxes. false Martin Hanzel annotation2447044 1 Gal4 site range2447044 1 740 756 annotation2447037 1 pPGK1 range2447037 1 1 872 annotation2447051 1 Gal4 site range2447051 1 758 774 annotation2447052 1 TATA? range2447052 1 724 729 annotation2447053 1 TATA? range2447053 1 864 870 BBa_K2111001_sequence 1 actgtaattgcttttagttgtgtatttttagtgtgcaagtttctgtaaatcgattaatttttttttctttcctctttttattaaccttaatttttattttagattcctgacttcaactcaagacgcacagatattataacatctgcataataggcatttgcaagaattactcgtgagtaaggaaagagtgaggaactatcgcatacctgcatttaaagatgccgatttgggcgcgaatcctttattttggcttcaccctcatactattatcagggccagaaaaaggaagtgtttccctccttcttgaattgatgttaccctcataaagcacgtggcctcttatcgagaaagaaattaccgtcgctcgtgatttgtttgcaaaaagaacaaaactgaaaaaacccagacacgctcgacttcctgtcttcctattgattgcagcttccaatttcgtcacacaacaaggtcctagcgacggctcacaggttttgtaacaagcaatcgaaggttctggaatggcgggaaagggtttagtaccacatgctatgatgcccactgtgatctccagagcaaagttcgttcgatcgtactgttactctctctctttcaaacagaattgtccgaatcgtgtgacaacaacagcctgttctcacacactcttttcttctaaccaagggggtggtttagtttagtagaacctcgtgaaacttacatttacatatatataaacttgcataaacgggcgacagccctccgtcgggcgacagccctccgtaattcgtagtttttcaagttcttagatgctttctttttctcttttttacagatcatcaaggaagtaattatctactttttacaacaaatataaaacatactagagtacaaaatggactataaagatgacgatgataaggaacgtcgtaaccattccggtagagtgtctgagttcgtgttgttaggttttcccgcgccggccccccttcgtgtgttgctttttttcctgtctctactggcatatgtacttgtattaacggaaaacatgttgattattatagccatcagaaaccatcctactttacataagccgatgtataggacgacaagacaaaaacacagtcgtctatttttctttatgaagcatttaagtatagccgacttagtcgtcgctgtgtttcaggttctgccccagttactgtgggatattacattccgtttttacggccctgacttgctgtgtagattagtgaaatatctacaggtcgtaggtatgtttgcttcaacctatcttctattactaatgagtcttgatcgttgcttagctatatgccagccattgcgttcccttcgtaggaggacggatcgtcttgctgtccttgcgacgtggctaggctgtcttgtcgccagtgcaccgcaagtacatatcttcagtttacgtgaagtcgcggacggtgtctttgactgttgggcggtattcatccagccttggggtccaaaggcatacattacgtggattactctggccgtctatattgtgcctgtaatagttttggcagcatgttatggtctaatcagctttaagatctggcagaacctacgtctaaagactgctgccgctgcggctgctgaggctccggagggagcagctgccggggacggcggaagggtagccttggcacgtgtcagcagtgtaaaactaattagcaaagcaaaaataagaactgtgaagatgacattcattatcgttcttgccttcatcgtgtgttggacgcccttcttttttgtgcaaatgtggagtgtctgggatgcaaacgcaccaaaggaagcctcagcctttattattgtgatgttattggcgtcactaaacagttgttgcaatccctggatatacatgctgttctattgcttacgtaaccaagacgtcaaacgtgcgttaagaaggacgttacatctggcacaagatcaggaggcgaatactaataagggctctaagatcggctaataa BBa_K2111000_sequence 1 tacaaaatggactataaagatgacgatgataaggaacgtcgtaaccattccggtagagtgtctgagttcgtgttgttaggttttcccgcgccggccccccttcgtgtgttgctttttttcctgtctctactggcatatgtacttgtattaacggaaaacatgttgattattatagccatcagaaaccatcctactttacataagccgatgtataggacgacaagacaaaaacacagtcgtctatttttctttatgaagcatttaagtatagccgacttagtcgtcgctgtgtttcaggttctgccccagttactgtgggatattacattccgtttttacggccctgacttgctgtgtagattagtgaaatatctacaggtcgtaggtatgtttgcttcaacctatcttctattactaatgagtcttgatcgttgcttagctatatgccagccattgcgttcccttcgtaggaggacggatcgtcttgctgtccttgcgacgtggctaggctgtcttgtcgccagtgcaccgcaagtacatatcttcagtttacgtgaagtcgcggacggtgtctttgactgttgggcggtattcatccagccttggggtccaaaggcatacattacgtggattactctggccgtctatattgtgcctgtaatagttttggcagcatgttatggtctaatcagctttaagatctggcagaacctacgtctaaagactgctgccgctgcggctgctgaggctccggagggagcagctgccggggacggcggaagggtagccttggcacgtgtcagcagtgtaaaactaattagcaaagcaaaaataagaactgtgaagatgacattcattatcgttcttgccttcatcgtgtgttggacgcccttcttttttgtgcaaatgtggagtgtctgggatgcaaacgcaccaaaggaagcctcagcctttattattgtgatgttattggcgtcactaaacagttgttgcaatccctggatatacatgctgttctattgcttacgtaaccaagacgtcaaacgtgcgttaagaaggacgttacatctggcacaagatcaggaggcgaatactaataagggctctaagatcggctaataa BBa_K1652000_sequence 1 actgtaattgcttttagttgtgtatttttagtgtgcaagtttctgtaaatcgattaatttttttttctttcctctttttattaaccttaatttttattttagattcctgacttcaactcaagacgcacagatattataacatctgcataataggcatttgcaagaattactcgtgagtaaggaaagagtgaggaactatcgcatacctgcatttaaagatgccgatttgggcgcgaatcctttattttggcttcaccctcatactattatcagggccagaaaaaggaagtgtttccctccttcttgaattgatgttaccctcataaagcacgtggcctcttatcgagaaagaaattaccgtcgctcgtgatttgtttgcaaaaagaacaaaactgaaaaaacccagacacgctcgacttcctgtcttcctattgattgcagcttccaatttcgtcacacaacaaggtcctagcgacggctcacaggttttgtaacaagcaatcgaaggttctggaatggcgggaaagggtttagtaccacatgctatgatgcccactgtgatctccagagcaaagttcgttcgatcgtactgttactctctctctttcaaacagaattgtccgaatcgtgtgacaacaacagcctgttctcacacactcttttcttctaaccaagggggtggtttagtttagtagaacctcgtgaaacttacatttacatatatataaacttgcataaacgggcgacagccctccgtcgggcgacagccctccgtaattcgtagtttttcaagttcttagatgctttctttttctcttttttacagatcatcaaggaagtaattatctactttttacaacaaatataaaaca 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