BBa_K1413001 1 sfGFP-DmpR P0 promoter-RBS B0032-sfGFP- Terminator B0015 - Pr promoter-DmpR 2014-09-29T11:00:00Z 2015-05-08T01:10:18Z This part is a composition of two parts from Peking iGEM 2013: BBa_K1031222 (sfGFP) & BBa_K1031211 (DmpR) Professor V.Shringler provided 2013 Peking iGEM Team with pVI401 composed of Pr-DmpR on vector pVI39. This part consist on a sensor of phenolic compounds based on a transcription factorof the Ntrc family , DmpR. Found in Pseudomonas sp. strain CF 600, DmpR regulates expression from the Po promoter, which drives transcription from one single large operon for phenol degradation (dmpKLMNOPQBCDEFGHI). With GFP attached to P0 promoter, it is then possible to evaluate presence of phenol by fluorescence analysis, if DmpR is expressed. Thus this part is basically composed of [P0 promoter + sfGFP + Pr promoter + DmpR] and allows sensing of phenolic compounds thanks to a constitutively expressed DmpR that will activate expression of GFP. We characterized this biosensor using phenol as effector. false false _1791_ 0 22607 9 It's complicated true We performed an assembly of the two parts by : -digesting BBa_K1031222 with EcorI and XbaI while digesting BBa_K1031211 by EcorI and SpeI. -ligating the two digestion products. BBa_K1031211 (Dmpr) had an extra Pst1 restriction site at position 226 (position 0 being EcoR1 restriction site). We thus performed several trials of site directed mutagenesis and finally managed to modify a single nucletide in the Pst1 site, with respect to the amino acids sequence. false Nandjafot MENDY annotation2392556 1 sfGFP range2392556 1 199 929 annotation2392555 1 RBS Junction range2392555 1 193 198 annotation2392553 1 RBS Junction range2392553 1 172 179 annotation2392558 1 SpeI/XbaI mixed site range2392558 1 1059 1066 annotation2392561 1 BBa_K1413001 range2392561 1 1 2847 annotation2392559 1 PR Promoter range2392559 1 1082 1155 annotation2392557 1 Terminator B0015 range2392557 1 930 1058 annotation2392552 1 P0 Promoter range2392552 1 1 171 annotation2392554 1 RBS B0032 range2392554 1 180 192 annotation2392560 1 DmpR range2392560 1 1156 2847 BBa_K1413001_sequence 1 taagcatttgctcaagcggccttgggcaattgatcaaatgcttaaaaagtctgcgcaagcgcggcttaatttcgctcgctccgatcattctaaaaattagaaacacattgaaaaacattaccttgaagtctgttttcagaccttggcacagccgttgcttgatgtcctgcgtactagagtcacacaggaaagactagaatgcgtaaaggcgaagagctgttcactggtgtcgtccctattctggtggaactggatggtgatgtcaacggtcataagttttccgtgcgtggcgagggtgaaggtgacgcaactaatggtaaactgacgctgaagttcatctgtactactggtaaactgccggtaccttggccgactctggtaacgacgctgacttatggtgttcagtgctttgctcgttatccggaccatatgaagcagcatgacttcttcaagtccgccatgccggaaggctatgtgcaggaacgcacgatttcctttaaggatgacggcacgtacaaaacgcgtgcggaagtgaaatttgaaggcgataccctggtaaaccgcattgagctgaaaggcattgactttaaagaagacggcaatatcctgggccataagctggaatacaattttaacagccacaatgtttacatcaccgccgataaacaaaaaaatggcattaaagcgaattttaaaattcgccacaacgtggaggatggcagcgtgcagctggctgatcactaccagcaaaacactccaatcggtgatggtcctgttctgctgccagacaatcactatctgagcacgcaaagcgttctgtctaaagatccgaacgagaaacgcgatcatatggttctgctggagttcgtaaccgcagcgggcatcacgcatggtatggatgaactgtacaaatgatgataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagaggatgaccccatctggaatcgccgcctgccttgcgctatagcggcgaccctgattttccccatctaaaaataaataggggcctcgcttacatgccgatcaagtacaagcctgaaatccagcactccgatttcaaggacctgaccaacctgatccacttccagagcatggaaggcaagatctggcttggcgaacagcgcatgctgttgcttcagttttcagcgatggccagctttcgccgggaaatggtcaataccctgggcatcgaacgcgccaagggcttgttcctgcgccatggttaccagtccggcctgaaggatgccgaactggccaggaagctgagaccgaatgccagcgaagtcggcatgttcctcgctgggccgcagatgcattcactcaagggtctggtcaaggtccgccccaccgagctcgatatcgacaaggaatacgggcgcttctatgccgagatggagtggatcgactccttcgaggtggaaatctgccagaccgacctggggcagatgcaagacccggtgtgctggactctgctcggctacgcctgcgcctattcctcggcgttcatgggccgggaaatcatcttcaaggaagtcagctgcggcggctgcggcggcgacaagtgccgggtcattggcaagccggccgaagagtgggacgacgttgccagcttcaaacagtatttcaagaacgaccccatcatcgaggaactctacgagttgcaatcgcaactgttgtcgctgcgtaccaacctcgacaaacaggaaggccagtactacggcatcggtcagaccccggcctaccagaccgtgcgcaatatgatggacaaggccgcacagggcaaagtctcggtgctgctgcttggcgagaccggggtcggcaaggaggtcatcgcgcgtagcgtgcacctgcgcagcaaacgcgccgccgagccctttgtcgcggtgaactgtgcggcgatcccgccggacctgatcgagtccgaattgttcggcgtggaaaaaggcgccttcaccggcgcctcccagtcacgcatgggccgcttcgagcgggccgacaagggcaccatcttccttgacgaggtgatcgaactcagcccgcgcgctcaggccagtctgctgcgcgtgctgcaagaaggcgagctggagcgagttggcgacaaccgcacgcgcaagatcgacgtaagggttatcgccgccacccacgaggacctggccgaagcggtcaaggccgggcgttttcgcgccgacctgtactaccgactgaacgttttcccggtggcgatcccggcgttgcgcgaacgccgcgaggacattccactgctggttgagcacttccttcagcgcttccaccaggagtacggcaagagaaccctcggcctttcagacaaagccctggaggcctgcctgcattacagttggccgggcaatatccgtgagctggagaacgtcatcgagcgcggcatcatcctcaccgatccgaacgaaagcatcagcgtgcaggcgctgttcctacgggcgccggaagagccgcagaccgccagcgagcgggtgtcgttggacggcgtgctgattcagccaggcaatggccagggcagttggatcagccagttgttgagcagcggcctgagcctcgacgagatcgaggaaagcctgatgcgcgaagccatgcaacaggccaaccaaaacgtctccggtgccgcgcgcttgctcggcctaagccgaccggcactggcctatcggctgaagaaaatcggcatcgaaggctag 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