BBa_K1973008 1 BBa_K1973008 nahR-Psal-nasF-yhjH 2016-10-11T11:00:00Z 2016-10-12T03:34:58Z Molecular cloning. <i>nahR</i> (<i>Pseudomonas putida</i>, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the P<i>sal</i> promoter (see BBa_J61051) in the presence of salicylate. <i>yhjH</i> (see BBa_K1973034) is expressed from the P<i>sal</i> promoter in the presence of this molecule. <i>nasF</i> (see BBa_K1973001) is an attenuator of transcription elongation (terminator) from <i>Klebsiella pneumoniae</i> that decreases the basal expression from the P<i>sal</i> promoter. false false _2440_ 30017 30017 9 false We digested BBa_K1973034 as a vector (EcoRI and XbaI) and BBa_K1973005 as an insert (EcoRI and SpeI), and then we ligate them. false Alvaro Escobar Doncel component2493582 1 BBa_K1973005 component2493586 1 BBa_K1973034 annotation2493582 1 BBa_K1973005 range2493582 1 1 1151 annotation2493586 1 BBa_K1973034 range2493586 1 1160 1948 BBa_K1973033 1 BBa_K1973033 nahR-Psal (inducible promoter) 2016-10-08T11:00:00Z 2016-10-09T11:17:26Z It was obtained by PCR from a plasmid containing this sequence. The PCR primers added the prefix and suffix needed. nahR (Pseudomonas putida, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the Psal promoter (see BBa_J61051) in the presence of salicylate. false false _2440_ 30003 30003 9 false We had included the RBS BBa_B0034 in the PCR primers, as well as the scar it should have appeared in the cloning of the DNA piece. false Laura Claret Fern??ndez annotation2489302 1 nahR range2489302 1 1 900 annotation2489301 1 Psal range2489301 1 901 1039 BBa_K1973034 1 BBa_K1973034 yhjH 2016-10-08T11:00:00Z 2016-10-09T11:36:10Z It was obtained by PCR from the genome of E.coli DH5 alpha. yhjH encodes the YhjH protein, a c-diGMP (cyclic diguanilate monophosphate) phosphodiesterase. c-diGMP regulates both the formation and dispersion of the biofilm, depending on the quantity of it that is present inside the cell. High levels of c-diGMP will increase the biofilm formation, and low levels of it will induce its dispersion. To manipulate this, we will make use of a phosphodiesterase, that will destroy the c-diGMP, and a diguanilate cyclase, that will produce it, YhjH and PleD* respectively. false false _2440_ 30003 30003 9 false We have inlcuded the sequence of RBS BBa_B0034 by means of the PCR primers used for the amplification, as well as the scar it should have been in case of the clonation of this part. false Laura Claret Fern??ndez annotation2489356 1 scar range2489356 1 13 18 annotation2489355 1 BBa_B0034 range2489355 1 1 12 annotation2489357 1 yhjH range2489357 1 19 789 BBa_K1973005 1 BBa_K1973005 nahR-Psal-nasF 2016-10-10T11:00:00Z 2016-10-11T05:51:56Z Molecular cloning. <i>nahR</i> (<i>Pseudomonas putida</i>, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the P<i>sal</i> promoter (see BBa_J61051) in the presence of salicylate. <i>nasF</i> (see BBa_K1973001) is an attenuator of transcription elongation (terminator) from <i>Klebsiella pneumoniae</i> that decreases the basal expression from the P<i>sal</i> promoter. false false _2440_ 0 30017 30017 9 It's complicated false We digested BBa_K1973001 as a vector (EcoRI and XbaI) and BBa_K1973033 as an insert (EcoRI and SpeI), and then we ligate them. false Alvaro Escobar Doncel component2493061 1 BBa_K1973001 component2493059 1 BBa_K1973033 annotation2493061 1 BBa_K1973001 range2493061 1 1048 1151 annotation2493059 1 BBa_K1973033 range2493059 1 1 1039 BBa_K1973001 1 BBa_K1973001 nasF 2016-10-08T11:00:00Z 2016-10-09T11:39:45Z It was obtained by PCR from a plasmid containing its sequence. nasF is an attenuator of transcription elongation (terminator) from Klebsiella pneumoniae that decreases the basal expression from the Psal promoter. The nasR protein disrupts the terminator structure and allows the expression again, providing an useful tool for tight ON/OFF expression systems. false false _2440_ 30003 30003 9 false No special considerations. false Laura Claret Fern??ndez annotation2489358 1 nasF transcription terminator range2489358 1 1 104 BBa_K1973034_sequence 1 aaagaggagaaatactagatgataaggcaggttatccagcgaataagcaaccctgaagcaagcatcgagagcttgcaggaacggcgtttttggttgcagtgtgagcgtgcttacacctggcagccgatctatcaaacatgcgggcggttaatggccgtggagctattaacggtggtcacgcatcccttgaacccttcgcaacgcctgccgccggatcgctattttactgaaatcaccgtcagccatcggatggaggttgtgaaagagcagattgatttgctggcgcaaaaagccgacttctttatagagcacggcctgctggcatcggtcaatattgatggccctacgctcatcgccctgcgtcagcaaccaaaaatcctgcgccagattgagcgtcttccctggctgcgtttcgaactggtggagcatatccgtctgccgaaagattcaacctttgcctcgatgtgtgaatttggcccgctgtggctggatgattttggtaccgggatggcaaatttctctgcgctaagtgaagtgcgttatgactacatcaaaatcgcgcgagaactgtttgtgatgctgcgtcagtcgccggaaggacgcacactcttttctcagcttttacatctaatgaatcgctattgtcgcggggtgattgtcgagggcgtagaaacgccggaagagtggcgtgatgttcagaactcgcccgcattcgccgcacaaggctggtttctttcacgcccggcaccgatagaaacgctgaatacggcggttctggcgctataataa BBa_K1973033_sequence 1 tcaatccgtaaacaggtcaaacatcagttgccgcaaccaaatattggctaggtccttgtggtacttcgcatgccagaacatgttgatggctatttcaggcaagacgactgggtgcggcaaggcgcttaggccgaagggctccacgcagcagtcggctaaacgtatcggcacagtggcgagcagatcggtgcgctggaggatgtggccaacggcggcgaagtgcggcacttccagacggatgtcgcgccggatgccgacccgtgtcatgtacgtgtccacctcgccgtggccggtgccagcggcgatgacacgcacgtggccgtaggaacagaagcgctccagagtcaggggttcgcgggtgactggatggtccttgcgacataggcacacgtagtgattctggagcagccggcgctgaaagaagccagtttgcagattgggaagcaggcccacggccaagtccacggttccgttctgcaaggcctgcatcaggctcatcgaactgtcgcgcaccgtactgatcacgcaattgggggcctggtgagccagcacatccatcagccgcggcatgaagtagatctcgccaatgtcggtcatggccagggtgaaggtacgctcgctggtcagcggatcgaagctttcatggtgctgtagggcgttgcgcagtgcgtgcatggccgaagtgacgggctcggccagatgcgcggcatagggtgtgggttccattccctgatgtgtgcgcacgaagagtgggtcctgtagcgaggtgcgcaggcgtttcagcgcattgctcacggcaggctgggtcaggcccaggttctccgcagtgatagagacgcgtctgtcgaccagcaactggttgaacaccaccagcaggtttaaatccaggtcacgcagttccatggggcctcgcttgggttattgctggtgcccggccgggcgcaatattcatgttgatgatttattatatatcgagtggtgtatttatcaatattgtttgctccgttatcgttattaacaagtcatcaataaagccatc BBa_K1973008_sequence 1 tcaatccgtaaacaggtcaaacatcagttgccgcaaccaaatattggctaggtccttgtggtacttcgcatgccagaacatgttgatggctatttcaggcaagacgactgggtgcggcaaggcgcttaggccgaagggctccacgcagcagtcggctaaacgtatcggcacagtggcgagcagatcggtgcgctggaggatgtggccaacggcggcgaagtgcggcacttccagacggatgtcgcgccggatgccgacccgtgtcatgtacgtgtccacctcgccgtggccggtgccagcggcgatgacacgcacgtggccgtaggaacagaagcgctccagagtcaggggttcgcgggtgactggatggtccttgcgacataggcacacgtagtgattctggagcagccggcgctgaaagaagccagtttgcagattgggaagcaggcccacggccaagtccacggttccgttctgcaaggcctgcatcaggctcatcgaactgtcgcgcaccgtactgatcacgcaattgggggcctggtgagccagcacatccatcagccgcggcatgaagtagatctcgccaatgtcggtcatggccagggtgaaggtacgctcgctggtcagcggatcgaagctttcatggtgctgtagggcgttgcgcagtgcgtgcatggccgaagtgacgggctcggccagatgcgcggcatagggtgtgggttccattccctgatgtgtgcgcacgaagagtgggtcctgtagcgaggtgcgcaggcgtttcagcgcattgctcacggcaggctgggtcaggcccaggttctccgcagtgatagagacgcgtctgtcgaccagcaactggttgaacaccaccagcaggtttaaatccaggtcacgcagttccatggggcctcgcttgggttattgctggtgcccggccgggcgcaatattcatgttgatgatttattatatatcgagtggtgtatttatcaatattgtttgctccgttatcgttattaacaagtcatcaataaagccatctactagaggagtgaataaaaggttttgggcagcgcgccaatggcggcgcgtatgtccagggataaaggcgtccagcggtgcgtaagcaccgccgggcgctttttttttgcgctactagagaaagaggagaaatactagatgataaggcaggttatccagcgaataagcaaccctgaagcaagcatcgagagcttgcaggaacggcgtttttggttgcagtgtgagcgtgcttacacctggcagccgatctatcaaacatgcgggcggttaatggccgtggagctattaacggtggtcacgcatcccttgaacccttcgcaacgcctgccgccggatcgctattttactgaaatcaccgtcagccatcggatggaggttgtgaaagagcagattgatttgctggcgcaaaaagccgacttctttatagagcacggcctgctggcatcggtcaatattgatggccctacgctcatcgccctgcgtcagcaaccaaaaatcctgcgccagattgagcgtcttccctggctgcgtttcgaactggtggagcatatccgtctgccgaaagattcaacctttgcctcgatgtgtgaatttggcccgctgtggctggatgattttggtaccgggatggcaaatttctctgcgctaagtgaagtgcgttatgactacatcaaaatcgcgcgagaactgtttgtgatgctgcgtcagtcgccggaaggacgcacactcttttctcagcttttacatctaatgaatcgctattgtcgcggggtgattgtcgagggcgtagaaacgccggaagagtggcgtgatgttcagaactcgcccgcattcgccgcacaaggctggtttctttcacgcccggcaccgatagaaacgctgaatacggcggttctggcgctataataa BBa_K1973001_sequence 1 gagtgaataaaaggttttgggcagcgcgccaatggcggcgcgtatgtccagggataaaggcgtccagcggtgcgtaagcaccgccgggcgctttttttttgcgc BBa_K1973005_sequence 1 tcaatccgtaaacaggtcaaacatcagttgccgcaaccaaatattggctaggtccttgtggtacttcgcatgccagaacatgttgatggctatttcaggcaagacgactgggtgcggcaaggcgcttaggccgaagggctccacgcagcagtcggctaaacgtatcggcacagtggcgagcagatcggtgcgctggaggatgtggccaacggcggcgaagtgcggcacttccagacggatgtcgcgccggatgccgacccgtgtcatgtacgtgtccacctcgccgtggccggtgccagcggcgatgacacgcacgtggccgtaggaacagaagcgctccagagtcaggggttcgcgggtgactggatggtccttgcgacataggcacacgtagtgattctggagcagccggcgctgaaagaagccagtttgcagattgggaagcaggcccacggccaagtccacggttccgttctgcaaggcctgcatcaggctcatcgaactgtcgcgcaccgtactgatcacgcaattgggggcctggtgagccagcacatccatcagccgcggcatgaagtagatctcgccaatgtcggtcatggccagggtgaaggtacgctcgctggtcagcggatcgaagctttcatggtgctgtagggcgttgcgcagtgcgtgcatggccgaagtgacgggctcggccagatgcgcggcatagggtgtgggttccattccctgatgtgtgcgcacgaagagtgggtcctgtagcgaggtgcgcaggcgtttcagcgcattgctcacggcaggctgggtcaggcccaggttctccgcagtgatagagacgcgtctgtcgaccagcaactggttgaacaccaccagcaggtttaaatccaggtcacgcagttccatggggcctcgcttgggttattgctggtgcccggccgggcgcaatattcatgttgatgatttattatatatcgagtggtgtatttatcaatattgtttgctccgttatcgttattaacaagtcatcaataaagccatctactagaggagtgaataaaaggttttgggcagcgcgccaatggcggcgcgtatgtccagggataaaggcgtccagcggtgcgtaagcaccgccgggcgctttttttttgcgc 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