BBa_M36888 1 BBa_M36888 Sequence for sigma 32 transcription factor 2012-12-05T12:00:00Z 2015-05-08T01:14:06Z The particular part described here is specific to the genome of W3110 strain E. coli. The rpoH gene codes for sigma 32 (alternatively sigma H, RpoH) transcription factor, a component of RNA polymerase sigma 32 holoenzyme. This 32-kD stress factor belongs to the sigma-70 factor family and is involved in intracellular temperature regulation, initiating bacterial transcription of various chaperone proteins and proteases in response to sudden increases in ambient temperature. The gene is currently classified as essential, due to the fact that rpoH deletion mutants fail to grow at standard incubation temperatures. This part may be employed in heat shock-to-PoPs sensors by coupling expression of the rpoH gene to action of sigma 32-inducible promoters such as the htpG heat-shock promoter (part#: BBa_J45405). Transcriptional activation by rpoH transcripts is possible both within and without the sensor genetic circuit, as sigma factors can diffuse around the nucleus. false false _848_ 0 15030 9 Not in stock false Designs incorporating this part into a feedback amplification loop must beware of likely rpoH-deletion mutants. This is due to overproduction of sigma 32 under leaky promoters such as htpG (BBa_J45405). Less deletion mutants in E. coli are noted when plasmids containing rpoH are cloned at higher temperature. Consider two facts before using this part. First, rpoH transcript activity is reduced under excess heat shock protein production. As such, feedback loops using rpoH should employ weaker bicistronic ribosome binding sites so as to optimize heat-shock response. Second, sigma 32 is turned over rapidly, with a half-life under 1 minute. false Shamik Mascharak, Justo Caballero and Sibel Sayiner annotation2213640 1 STOP range2213640 1 853 861 annotation2213636 1 START range2213636 1 1 3 BBa_M36604 1 BBa_M36604 Heat-->PoPS (with postive feedback loop) 2012-12-05T12:00:00Z 2015-05-08T01:14:05Z htpG Heat Shock Promoter and Sequence for Sigma-32 Transcription Factor are from the E. coli genome. The bicistronic RBS is from the BIOFAB database. Find more information at their associated part page. This composite part is a sensor that focuses on creating a positive feedback loop to increase the PoPS signal for a desired actuator further downstream. In this design, Sigma-32, a sigma factor turned on when E. coli experiences heat shock, is the transcription factor for the htpG Heat Shock Promoter (BBa_J45504). Once active, the inducible promoter allows for, with the assistance of a medium strength bicistronic RBS (BBa_M36405), the transcription of rpoH (BBa_M36888), the sequence for sigma-32. Such a design creates a positive feedback loop, creating more sigma-32 from a small initial heat exposure. This is solely done in order to generate a larger PoPS signal, which would allow for a greater production of one's desired downstream production. Device input is heat exposure. Device output is a PoPS signal. false false _848_ 0 15029 9 Not in stock true It should be noted that E. coli has a built-in negative feedback to sigma-32, such that an abundance of sigma-32 turns off natural sigma-32 production. In regards to our promoter, this behavior leads to a sinusoidal relationship and eventually reaches a steady-state. Moreover, we found that creating a positive feedback loop in our construct increased the likelihood of mutations (making the DNA incredibly hard to synthesize) because the leakiness of the promoter caused a sigma-32 presence even when the cell wasn't under any stress. However, when we were first designing this composite, we wanted to produce significant sigma-32 in order to guarantee high expression of our selected actuator. false Sibel Sayiner, Justo Caballero, and Shamik Mascharak component2213639 1 BBa_M36888 component2213637 1 BBa_J45504 component2213638 1 BBa_M36405 annotation2213639 1 BBa_M36888 range2213639 1 508 1368 annotation2213638 1 BBa_M36405 range2213638 1 414 501 annotation2213637 1 BBa_J45504 range2213637 1 1 405 BBa_J45504 1 BBa_J45504 htpG Heat Shock Promoter 2006-06-20T11:00:00Z 2015-08-31T04:08:49Z Chris Voigt Lab at UCSF This promoter is active at higher temperatures. false false _84_ 0 642 84 Not in stock false None. false Stephen Payne BBa_M36405 1 BBa_M36405 Bicistronic RBS (medium) pFAB1646 2012-12-05T12:00:00Z 2015-05-08T01:14:04Z More information can be found at BIOFAB.org. This is a bicistronic RBS, which consists of a constitutive RBS, and a leader sequence that includes the second RBS. It uses bicistronic junction architecture to tightly control translation. It contains two ribosome binding sites: one controls the gene of interest, and the another synthesizes a short leader protein. The latter bicistronic's translation prevents the former RBS from forming a secondary structure with the gene of interest. This ensures a more consistent expression level of your gene of interest. false false _848_ 0 15032 9 Not in stock false This RBS is of medium strength. false Justo A. Caballero, Shamik Mascharak and Sibel Sayiner BBa_M36604_sequence 1 cactgaagtgatcctcgccaccaaccccacggttgaaggtgaagctaccgctaactacattgccgagctttgcgcgcaatatgacgtggaagccagccgaatcgctcatggcgttccggttggcggcgagctggaaatggtcgacggcaccacgttgtcacactcccttgccgggcgtcataagattcgtttttaagcaaacgagagcaggatcacctgctctcgcttgaaattattctcccttgtccccatctctcccacatcctgtttttaaccttaaaatggcattattgaggtagacctacatgaaaggacaagaaactcgtggttttcagtcagaagtgaaacagcttctgcacctgatgatccattctctctattccaataaagaaatcttcctgcgtgtactagaggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcaggggatattttctgatgtactagatgactgacaaaatgcaaagtttagctttagccccagttggcaacctggattcctacatccgggcagctaacgcgtggccgatgttgtcggctgacgaggagcgggcgctggctgaaaagctgcattaccatggcgatctggaagcagctaaaacgctgatcctgtctcacctgcggtttgttgttcatattgctcgtaattatgcgggctatggcctgccacaggcggatttgattcaggaaggtaacatcggcctgatgaaagcagtgcgccgtttcaacccggaagtgggtgtgcgcctggtctccttcgccgttcactggatcaaagcagagatccacgaatacgttctgcgtaactggcgtatcgtcaaagttgcgaccaccaaagcgcagcgcaaactgttcttcaacctgcgtaaaaccaagcagcgtctgggctggtttaaccaggatgaagtcgaaatggtggcccgtgaactgggcgtaaccagcaaagacgtacgtgagatggaatcacgtatggcggcacaggacatgacctttgacctgtcttccgacgacgattccgacagccagccgatggctccggtgctctatctgcaggataaatcatctaactttgccgacggcattgaagatgataactgggaagagcaggcggcaaaccgtctgaccgacgcgatgcagggtctggacgaacgcagccaggacatcatccgtgcgcgctggctggacgaagacaacaagtccacgttgcaggaactggctgaccgttacggcgtttccgctgagcgtgtacgccagctggaaaagaacgcgatgaaaaaattgcgtgctgccattgaagcgtaatagtga BBa_J45504_sequence 1 cactgaagtgatcctcgccaccaaccccacggttgaaggtgaagctaccgctaactacattgccgagctttgcgcgcaatatgacgtggaagccagccgaatcgctcatggcgttccggttggcggcgagctggaaatggtcgacggcaccacgttgtcacactcccttgccgggcgtcataagattcgtttttaagcaaacgagagcaggatcacctgctctcgcttgaaattattctcccttgtccccatctctcccacatcctgtttttaaccttaaaatggcattattgaggtagacctacatgaaaggacaagaaactcgtggttttcagtcagaagtgaaacagcttctgcacctgatgatccattctctctattccaataaagaaatcttcctgcgtg BBa_M36405_sequence 1 gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcaggggatattttctgatg BBa_M36888_sequence 1 atgactgacaaaatgcaaagtttagctttagccccagttggcaacctggattcctacatccgggcagctaacgcgtggccgatgttgtcggctgacgaggagcgggcgctggctgaaaagctgcattaccatggcgatctggaagcagctaaaacgctgatcctgtctcacctgcggtttgttgttcatattgctcgtaattatgcgggctatggcctgccacaggcggatttgattcaggaaggtaacatcggcctgatgaaagcagtgcgccgtttcaacccggaagtgggtgtgcgcctggtctccttcgccgttcactggatcaaagcagagatccacgaatacgttctgcgtaactggcgtatcgtcaaagttgcgaccaccaaagcgcagcgcaaactgttcttcaacctgcgtaaaaccaagcagcgtctgggctggtttaaccaggatgaagtcgaaatggtggcccgtgaactgggcgtaaccagcaaagacgtacgtgagatggaatcacgtatggcggcacaggacatgacctttgacctgtcttccgacgacgattccgacagccagccgatggctccggtgctctatctgcaggataaatcatctaactttgccgacggcattgaagatgataactgggaagagcaggcggcaaaccgtctgaccgacgcgatgcagggtctggacgaacgcagccaggacatcatccgtgcgcgctggctggacgaagacaacaagtccacgttgcaggaactggctgaccgttacggcgtttccgctgagcgtgtacgccagctggaaaagaacgcgatgaaaaaattgcgtgctgccattgaagcgtaatagtga 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