BBa_K1355001 1 BBa_K1355001 Regulation and transport of mercury ions 2014-10-05T11:00:00Z 2015-07-23T04:05:30Z This construction is based on sequences present in the O26-CRL plasmid found in Escherichia coli O26. We added a reverse tryptophan operon terminator to ensure transcription termination of the merR messenger RNA. However we didn???t add transcription terminator after the merP gene, aiming connect this biobrick with others, enabling various functions related to mercury We call this part an ???Essential biobrick??? designed to be the key piece of any genetic construction related to mercury, targeting both a biosensor as a bioaccumulative. This biobrick has bidirectional promoter, the regulator MerR with a terminator reverse from the tryptophan operon and a forward MerP and MerT transporters. false false _1731_ 4206 15499 9 In stock true If is desirable to accumulate mercury, assemble this biobrick with a good RBS, a coding sequence of a peptide having affinity to metal and a terminator! Its function will be regulated by presence of mercury through MerR regulator. The bacteria will carry the mercury to cytoplasm until it is linked to the peptide having affinity to the metal, making a inactive mercury. Is not it amazing? This is the strategy that we use in our project! The so called ???Essential Biobrick??? was connected to biobrick for bioremediation, bioaccumulation and bio-detection! false Luna Barroco de Lacerda annotation2415127 1 Bidirecional promoter range2415127 1 486 514 annotation2415155 1 MerP range2415155 1 914 1189 annotation2415752 1 RBS range2415752 1 537 543 annotation2415126 1 MerR range2415126 1 44 480 annotation2415749 1 RBS range2415749 1 490 496 annotation2415134 1 Bidirecional promoter range2415134 1 510 538 annotation2397153 1 Trp Terminator range2397153 1 1 43 annotation2415143 1 RBS range2415143 1 901 907 annotation2415141 1 MerT range2415141 1 551 900 BBa_K1355003 1 BBa_K1355003 Mercury ions accumulator device 2014-10-05T11:00:00Z 2015-05-08T01:10:02Z BBa_K1355001; BBa_K346004 The Bioaccumulative biobrick is composed of our ???essential biobrick??? (BBa_K1355001) + a Metallothionein (BBa_K346004) available in the database of the iGEM since 2010.In the presence of mercury, The MerR protein will bind to it and MerT, MerP and Metal Binding Protein (MBP) proteins are synthesized and bioaccumulation will start! The MerP and MerT protein are responsible for the transport of mercury from the periplasm to the cytoplasm, leading him to MBP! The MBP binds to the mercury in the cytoplasm, rendering it inactive. false false _1731_ 0 15499 9 In stock true For this construct, we followed these steps: Extraction of plasmid DNA and DNA quantification of bacteria transformed with the Essential Biobrick (BBa_K1355001) and the bacteria transformed with the MBP Biobrick (BBa_K346004); Verifying the electrophoretic profile of the extracted plasmid DNA; Restriction enzyme digestion for Essential Biobrick with SpeI and PstI and for MBP Biobrick with PstI and XbaI; Checking the electrophoretic profile of digested samples; Purification of the Fragment of interest from the gel; Ligation the gel purification of MBP gene and digested Essential Biobrick; Transformation of ligation in DH5-alpha; Plasmid DNA extraction from bacteria transformed; Check the electrophoretic profile to see results of samples linked (no fragments); Digestion of Essential Biobrick + MBP Biobrick with EcoRI and PstI, aiming to analyze the fragment size to be isolated; Checking the electrophoretic profile of the digested sample to obtain results showing that the isolated fragment is the junction of Essential Biobrick +MBP Biobrick ; false Luna Barroco de Lacerda, Maria Clara Astolfi component2415267 1 BBa_K1355001 component2415272 1 BBa_K346004 annotation2415267 1 BBa_K1355001 range2415267 1 1 1189 annotation2415272 1 BBa_K346004 range2415272 1 1198 1676 BBa_K346004 1 BBa_K346004 RBS(B0034)_MBP(lead metal binding peptide egineered from PbrR)+Terminator(B0015) 2010-10-13T11:00:00Z 2016-01-13T10:29:19Z yes Released HQ 2013 This part is combined with a rbs, a mbp which codes for the metal binding peptide and a terminator. false false _457_ 4206 6045 9 In stock true to be given false Junyi Jiao annotation2100340 1 mbp(lead binding peptide) range2100340 1 19 345 annotation2100341 1 BBa_B0015 range2100341 1 351 479 annotation2100339 1 conserved range2100339 1 5 8 annotation2100338 1 BBa_B0034 range2100338 1 1 12 BBa_K346004_sequence 1 aaagaggagaaatactagatgcaggattgcggtgaagtcaatatgctcttggatgagcacatccgtcaggtcgaatctcggatcggagctttgctcgaactgaagcaccatttggtggaactgcgcgaagcctgttctggtgccaggcccgcccaatcgtgcgggattctgcacggactgtcggactcttccggacaggattgcggtgaagtcaatatgctcttggatgagcacatccgtcaggtcgaatctcggatcggagctttgctcgaactgaagcaccatttggtggaactgcgcgaagcctgttctggtgccaggcccgcccaatcgtgcgggtaatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_K1355003_sequence 1 aaagaaagttaaaatgccgccagcggaactggcggctgtgggactaaggcatagctgaccttgccaggcctgcttcgccctgtagtgacgcgatcaacgggcaggaaacattcccctttcgtgcatggcaggcgcacacgagttcagacagcacggtttccatgcgcgccaagtcggccatcttctcgcgcacgtccttgagcttgtgttcggccaggctgctggcctcctcgcagtgggtgccatcgtcgagccgcaacagctcggcaatctcgtccagactgaaccccagccgctgtgccgatttcacgaatttcacccgaaccacgtccgcctccccatagcggcggatgctgccgtaaggcttgtccggttcccgcaacaggcccttgcgctgatagaagcggattgtctccacgttgaccccggccgccttggcaaaaacgccaatggtcaggttttccaaattattttccatatcgcttgactccgtacatgagtacggaagtaaggttacgctatccaatccaaattcaaaagggccaacgtatgtctgaaccacaaaacgggcgcggtgcgctcttcgccggcgggctggccgccattcttgcatcgacctgctgcctggggccgctagtactggtcgccctgggcttctccggtgcttggatcggcaacctgacggtgctggaaccctatcgaccgttgttcatcggcgcggcgctagtggcgctgttcttcgcctggaagcggatttaccggcccgtgcaggcatgcaagccaggtgaggtctgcgcgattccgcaggtgcgcgccacctacaagctgattttctggatcgtggccgtgctggtcctggtcgcgcttggatttccctatgtcgttccatttttctattaaccaggagttcatcatgaagaaactgtttgcctcccttgccctcgccgccgctgttgccccggtgtgggccgctacccagaccgtcacgctagcggttcccggcatgacttgcgccgcctgcccgatcacagtcaagaaagcgctctccaaggtcgaaggcgtgagcaaggtcgatgtgggcttcgagaagcgcgaggccgtcgtcacttttgacgacaccaaggccagcgtacagaagctgaccaaggccaccgcagacgccggctatccgtccagcgtcaagcagtgatactagagaaagaggagaaatactagatgcaggattgcggtgaagtcaatatgctcttggatgagcacatccgtcaggtcgaatctcggatcggagctttgctcgaactgaagcaccatttggtggaactgcgcgaagcctgttctggtgccaggcccgcccaatcgtgcgggattctgcacggactgtcggactcttccggacaggattgcggtgaagtcaatatgctcttggatgagcacatccgtcaggtcgaatctcggatcggagctttgctcgaactgaagcaccatttggtggaactgcgcgaagcctgttctggtgccaggcccgcccaatcgtgcgggtaatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_K1355001_sequence 1 aaagaaagttaaaatgccgccagcggaactggcggctgtgggactaaggcatagctgaccttgccaggcctgcttcgccctgtagtgacgcgatcaacgggcaggaaacattcccctttcgtgcatggcaggcgcacacgagttcagacagcacggtttccatgcgcgccaagtcggccatcttctcgcgcacgtccttgagcttgtgttcggccaggctgctggcctcctcgcagtgggtgccatcgtcgagccgcaacagctcggcaatctcgtccagactgaaccccagccgctgtgccgatttcacgaatttcacccgaaccacgtccgcctccccatagcggcggatgctgccgtaaggcttgtccggttcccgcaacaggcccttgcgctgatagaagcggattgtctccacgttgaccccggccgccttggcaaaaacgccaatggtcaggttttccaaattattttccatatcgcttgactccgtacatgagtacggaagtaaggttacgctatccaatccaaattcaaaagggccaacgtatgtctgaaccacaaaacgggcgcggtgcgctcttcgccggcgggctggccgccattcttgcatcgacctgctgcctggggccgctagtactggtcgccctgggcttctccggtgcttggatcggcaacctgacggtgctggaaccctatcgaccgttgttcatcggcgcggcgctagtggcgctgttcttcgcctggaagcggatttaccggcccgtgcaggcatgcaagccaggtgaggtctgcgcgattccgcaggtgcgcgccacctacaagctgattttctggatcgtggccgtgctggtcctggtcgcgcttggatttccctatgtcgttccatttttctattaaccaggagttcatcatgaagaaactgtttgcctcccttgccctcgccgccgctgttgccccggtgtgggccgctacccagaccgtcacgctagcggttcccggcatgacttgcgccgcctgcccgatcacagtcaagaaagcgctctccaaggtcgaaggcgtgagcaaggtcgatgtgggcttcgagaagcgcgaggccgtcgtcacttttgacgacaccaaggccagcgtacagaagctgaccaaggccaccgcagacgccggctatccgtccagcgtcaagcagtga 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