BBa_I746107 1 BBa_I746107 I746101 (AIP sensor infrastructure) + I746105 (Inducible P2 + GFP reporter) 2007-10-22T11:00:00Z 2015-08-31T04:08:04Z The original sequences for agrC and agrA were taken from the S. aureus Sequencing Project (results for strain NCTC8325) at Oklahoma University (http://www.genome.ou.edu/staph.html). The sequences were then codon-optimised for E. coli using GeneDesigner from DNA2.0 (http://www.dna20.com), and then synthesised by that company. Released HQ 2013 This part is the AIP sensor infrastructure ([http://partsregistry.org/wiki/index.php/Part:BBa_I746101 I746101]) from the <i>agr</i> system with an AIP-inducible promoter P2 and GFP reporter ([http://partsregistry.org/wiki/index.php/Part:BBa_I746105 I746105]) downstream. A constitutive/inducible promoter may be added upstream of this part, which will cause production of the components of the signal transduction pathway (AgrC and AgrA); when phosphorylated AgrA is present, the AIP-sensitive promoter (P2) is activated and GFP ([http://partsregistry.org/wiki/index.php/Part:BBa_E0840 E0840]) is output. The original part was made in order to transfer the S. aureus oligopeptide-based quorum sensing system into a BioBrick-compatible signalling mechanism. In the natural system, the signalling oligopeptide (termed AIP) is made from AgrD by the membrane-located enzyme AgrB. It is then detected by the membrane-located AgrC, which phosphorylates AgrA which then has DNA-binding activity and upregulates transcription of the promoters termed P2 and P3 in the agr locus. There are four known variants of AIP with different molecular structures and cross-inhibitory activity; this BioBrick generates group I AIP. The agr P2 operon is an autocatalytic sensory transduction system in Staphylococcus aureus. P2 promoter regulates the synthesis of AgrB (a transmembrane protein) and AgrD (precursor of AIP autoinducing peptide). AIP binds to AgrC, a membrane binding receptor and it phosphorylates AgrA. The phosphorylated AgrA increases the activity of agr P2 promoter about 50-fold. false false _116_ 0 2127 9 In stock false N/A true Lovelace Soirez component1952701 1 BBa_B0010 component1952703 1 BBa_B0012 component1952707 1 BBa_I746104 component1952713 1 BBa_B0010 component1952715 1 BBa_B0012 component1952709 1 BBa_B0030 component1952712 1 BBa_E0040 component1952700 1 BBa_I746100 annotation1952707 1 BBa_I746104 range1952707 1 2066 2161 annotation1952712 1 BBa_E0040 range1952712 1 2191 2910 annotation1952700 1 BBa_I746100 range1952700 1 1 1920 annotation1952709 1 BBa_B0030 range1952709 1 2170 2184 annotation1952701 1 BBa_B0010 range1952701 1 1929 2008 annotation1952713 1 BBa_B0010 range1952713 1 2919 2998 annotation1952715 1 BBa_B0012 range1952715 1 3007 3047 annotation1952703 1 BBa_B0012 range1952703 1 2017 2057 BBa_I746104 1 BBa_I746104 P2 promoter in agr operon from S. aureus 2007-08-30T11:00:00Z 2015-08-31T04:08:04Z The section of the sequence constituting the P2 promoter is subject to rather more debate than the sequences of the coding regions. [http://jb.asm.org/cgi/content/full/186/22/7549 Koenig, Robbin L., Ray, Jessica L., Maleki, Soheila J., Smeltzer, Mark S., Hurlburt, Barry K. Staphylococcus aureus AgrA Binding to the RNAIII-agr Regulatory Region J. Bacteriol. 2004 186: 7549-7555] shows the areas of the sequence to which phosphorylated AgrA binds while L Rao, R K Karls, and M J Betley: In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus. Bacteriol. 1995 May; 177(10): 2609???2614. shows the transcription start sites; we took the overall extent of the promoter region from that labelled in Morfeldt, E., K. Tegmark, and S. Arvidson. 1996. Transcriptional control of the agr-dependent virulence gene regulator, RNAIII, in Staphylococcus aureus. Mol. Microbiol. 21:1227???1237. (not available online). Released HQ 2013 The agr P2 operon is an autocatalytic sensory transduction system in Staphylococcus aureus. P2 promoter regulates the synthesis of AgrB (a transmembrane protein) and AgrD (precursor of AIP autoinducing peptide). AIP binds to AgrC, a membrane binding receptor and it phosphorylates AgrA. The phosphorylated AgrA increases the expression level from P2 promoter. false false _116_ 0 2121 9 In stock false Codon usage has been optimised for E. coli (which also seems to give good results for B. subtilis true Zhizhen Zhao BBa_B0012 1 BBa_B0012 TE from coliphageT7 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>. Released HQ 2013 Transcription terminator for the <i>E.coli</i> RNA polymerase. false false _1_ 0 24 7 In stock false <P> <P>Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG).<P> Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator. false Reshma Shetty annotation1686 1 T7 TE range1686 1 8 27 annotation7020 1 BBa_B0012 range7020 1 1 41 annotation1687 1 stop range1687 1 34 34 annotation1690 1 polya range1690 1 28 41 BBa_E0040 1 GFP green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212 2004-09-29T11:00:00Z 2016-01-26T02:09:38Z Released HQ 2013 GFP (mut3b) [note that this part does not have a barcode] false true _11_1_ 4206 61 7 In stock false true jcbraff annotation1934520 1 GFP protein range1934520 1 1 720 BBa_I746100 1 BBa_I746100 AIP sensor infrastructure (agrC + agrA from S. aureus with RBSes) 2007-10-19T11:00:00Z 2015-08-31T04:08:04Z The original sequences for agrC and agrA were taken from the ''S. aureus'' Sequencing Project (results for strain NCTC8325) at Oklahoma University (http://www.genome.ou.edu/staph.html). The sequences were then codon-optimised for ''E. coli'' using GeneDesigner from DNA2.0 (http://www.dna20.com), and then synthesised by that company. This part was made in order to transfer the S. aureus oligopeptide-based quorum sensing system into a BioBrick-compatible signalling mechanism. In the natural system, the signalling oligopeptide (termed AIP) is made from AgrD by the membrane-located enzyme AgrB. It is then detected by the membrane-located AgrC, which phosphorylates AgrA which then has DNA-binding activity and upregulates transcription of the promoters termed P2 and P3 in the agr locus. There are four known variants of AIP with different molecular structures and cross-inhibitory activity; this BioBrick generates group I AIP. This part contains the coding regions for group I agrC and agrA (in that order) from ''S. aureus'' strain NCTC8325, prefixed by the B0034 RBS and separated by a BamHI site. false false _116_ 0 1851 9 Not in stock false N/A false David Wyatt annotation1951374 1 B0034-sequence RBS range1951374 1 1 12 annotation1951378 1 agrA coding sequence range1951378 1 1291 1920 annotation1951375 1 B0034-sequence RBS range1951375 1 1273 1284 annotation1951376 1 BamHI site range1951376 1 1267 1272 annotation1951377 1 agrC coding sequence range1951377 1 19 1266 BBa_B0030 1 BBa_B0030 RBS.1 (strong) -- modified from R. Weiss 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>. false true _44_46_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix (&quot;orig&quot; in figure 4-14 of Ron Weiss thesis). <p>No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info <a href="mailto:(bchow@media.mit.edu)">Brian Chow</a> true Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003. annotation7025 1 BBa_B0030 range7025 1 1 15 annotation1702 1 RBS range1702 1 8 12 annotation1701 1 RBS-1\Strong range1701 1 1 15 BBa_B0010 1 BBa_B0010 T1 from E. coli rrnB 2003-11-19T12:00:00Z 2015-08-31T04:07:20Z Transcriptional terminator consisting of a 64 bp stem-loop. false false _1_ 0 24 7 In stock false true Randy Rettberg annotation7018 1 BBa_B0010 range7018 1 1 80 annotation4184 1 stem_loop range4184 1 12 55 BBa_B0010_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc BBa_I746104_sequence 1 attaaatacaaattacatttaacagttaagtatttatttcctacagttaggcaatataatgataaaagattgtactaaatcgtataatgacagtga BBa_I746107_sequence 1 aaagaggagaaatactagatgattctgatgttcaccattccagcaattattagcggtattaagtacagcaagctggattatttcttcatcatcgtaatctctaccctgtccctgttcctgttcaaaatgttcgattccgcgagcctgattatcctgacttccttcatcatcatcatgtatttcgtaaaaatcaaatggtatagcatcctgctgatcatgacctcccagattatcctgtactgtgccaactacatgtatatcgttatctacgcgtacatcacgaaaatctccgattctatcttcgttatctttccgtctttcttcgtggtgtacgttacgatctctattctgttttcctatattatcaatcgcgttctgaaaaagatctctactccgtacctgattctgaacaaaggttttctgatcgtcatctccactatcctgctgctgaccttctctctgttcttcttttactctcagatcaactccgatgaggcaaaagtcatccgtcagtacagcttcatctttatcggtattactatcttcctgagcattctgactttcgttatcagccagttcctgctgaaagaaatgaagtacaagcgcaaccaagaagaaatcgagacttactacgagtacaccctgaagattgaagcgatcaacaacgaaatgcgtaaatttcgtcacgactacgttaacattctgaccaccctgagcgaatacatccgtgaagacgacatgcctggcctgcgtgattatttcaacaaaaacattgtgccgatgaaagataacctgcaaatgaacgctattaagctgaacggtatcgaaaacctgaaagtacgtgagatcaagggtctgattaccgctaaaatcctgcgtgcgcaggaaatgaacatcccaattagcatcgaaatcccagacgaagtttctagcattaacctgaacatgattgacctgtctcgtagcatcggcatcatcctggacaacgctatcgaagcttccactgaaatcgacgacccgattattcgcgttgcttttatcgagagcgaaaactccgtgacctttatcgtgatgaacaaatgcgcggacgatatcccgcgtattcacgaactgttccaggaatctttcagcaccaagggtgaaggtcgcggcctgggcctgtccaccctgaaagagatcgcggataacgccgacaacgttctgctggacactattatcgaaaacggtttcttcattcagaaagtcgaaatcatcaataactaataaggatccaaagaggagaaatactagatggaaatcgcactggctacggacaatccatatgaggttctggaacaggctaaaaacatgaacgacatcggctgctatttcctggatattcagctgagcaccgacatcaacggtatcaaactgggctctgaaatccgtaaacatgatccggttggtaacattattttcgtgacttctcattctgagctgacctatctgacgttcgtatacaaggttgcagccatggatttcatctttaaagacgatccggcagagctgcgtacgcgcatcatcgactgtctggaaactgcgcacacgcgcctgcaactgctgtccaaagataactccgtggaaaccattgaactgaaacgtggcagcaacagcgtttacgttcagtacgatgatatcatgttcttcgaaagctccactaagtcccaccgtctgatcgcgcatctggataatcgtcagatcgagttctatggcaacctgaaagagctgagccagctggacgatcgcttcttccgttgtcataactccttcgttgttaaccgtcacaacatcgaatccatcgactccaaagagcgtattgtatacttcaagaacaaagaacactgttatgcgtctgtacgcaatgtcaagaaaatctaataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagattaaatacaaattacatttaacagttaagtatttatttcctacagttaggcaatataatgataaaagattgtactaaatcgtataatgacagtgatactagagattaaagaggagaaatactagatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_B0030_sequence 1 attaaagaggagaaa BBa_E0040_sequence 1 atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa BBa_I746100_sequence 1 aaagaggagaaatactagatgattctgatgttcaccattccagcaattattagcggtattaagtacagcaagctggattatttcttcatcatcgtaatctctaccctgtccctgttcctgttcaaaatgttcgattccgcgagcctgattatcctgacttccttcatcatcatcatgtatttcgtaaaaatcaaatggtatagcatcctgctgatcatgacctcccagattatcctgtactgtgccaactacatgtatatcgttatctacgcgtacatcacgaaaatctccgattctatcttcgttatctttccgtctttcttcgtggtgtacgttacgatctctattctgttttcctatattatcaatcgcgttctgaaaaagatctctactccgtacctgattctgaacaaaggttttctgatcgtcatctccactatcctgctgctgaccttctctctgttcttcttttactctcagatcaactccgatgaggcaaaagtcatccgtcagtacagcttcatctttatcggtattactatcttcctgagcattctgactttcgttatcagccagttcctgctgaaagaaatgaagtacaagcgcaaccaagaagaaatcgagacttactacgagtacaccctgaagattgaagcgatcaacaacgaaatgcgtaaatttcgtcacgactacgttaacattctgaccaccctgagcgaatacatccgtgaagacgacatgcctggcctgcgtgattatttcaacaaaaacattgtgccgatgaaagataacctgcaaatgaacgctattaagctgaacggtatcgaaaacctgaaagtacgtgagatcaagggtctgattaccgctaaaatcctgcgtgcgcaggaaatgaacatcccaattagcatcgaaatcccagacgaagtttctagcattaacctgaacatgattgacctgtctcgtagcatcggcatcatcctggacaacgctatcgaagcttccactgaaatcgacgacccgattattcgcgttgcttttatcgagagcgaaaactccgtgacctttatcgtgatgaacaaatgcgcggacgatatcccgcgtattcacgaactgttccaggaatctttcagcaccaagggtgaaggtcgcggcctgggcctgtccaccctgaaagagatcgcggataacgccgacaacgttctgctggacactattatcgaaaacggtttcttcattcagaaagtcgaaatcatcaataactaataaggatccaaagaggagaaatactagatggaaatcgcactggctacggacaatccatatgaggttctggaacaggctaaaaacatgaacgacatcggctgctatttcctggatattcagctgagcaccgacatcaacggtatcaaactgggctctgaaatccgtaaacatgatccggttggtaacattattttcgtgacttctcattctgagctgacctatctgacgttcgtatacaaggttgcagccatggatttcatctttaaagacgatccggcagagctgcgtacgcgcatcatcgactgtctggaaactgcgcacacgcgcctgcaactgctgtccaaagataactccgtggaaaccattgaactgaaacgtggcagcaacagcgtttacgttcagtacgatgatatcatgttcttcgaaagctccactaagtcccaccgtctgatcgcgcatctggataatcgtcagatcgagttctatggcaacctgaaagagctgagccagctggacgatcgcttcttccgttgtcataactccttcgttgttaaccgtcacaacatcgaatccatcgactccaaagagcgtattgtatacttcaagaacaaagaacactgttatgcgtctgtacgcaatgtcaagaaaatctaataa BBa_B0012_sequence 1 tcacactggctcaccttcgggtgggcctttctgcgtttata 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