BBa_K541815 1 BBa_K541815 B.subtilis and E.coli Episomal Shuttle Vector with LALF and SacB signal peptide 2011-09-20T11:00:00Z 2015-05-08T01:12:38Z Horseshoe crab Lipopolysaccharide (LPS), or endotoxin, is the major mediator of septic shock, a serious complication of Gram-negative bacterial infections in humans. Molecules that bind LPS and neutralize its biological effects or enhance its clearance could have important clinical applications. Limulus anti-LPS factor (LALF) binds LPS tightly, and, in animal models, reduces mortality when administered before or after LPS challenge or bacterial infection. The wedge- shaped molecule has a striking charge distribution and amphipathicity that suggest how it can insert into membranes. The binding site for LPS probably involves an extended amphipathic loop, and it has been proposed that two mammalian LPS-binding proteins will have a similar loop. The amphipathic loop structure may be used in the design of molecules with therapeutic properties against septic shock. false false _708_ 0 9573 9 It's complicated false Optimiz<ed for B.subtilis false Mustafa Elitok component2136649 1 BBa_K090401 component2136658 1 BBa_K541515 annotation2136649 1 BBa_K090401 range2136649 1 1 5208 annotation2136658 1 BBa_K541515 range2136658 1 5217 5753 BBa_K090401 1 pBSEP1 Gram-positive Shuttle Vector for Episomal Expression 2008-10-27T12:00:00Z 2015-05-08T01:08:37Z This part was made via InFusion assembly from many separate pieces. The backbone comes from PAD123 (Bacillus Stock Center catalog number ECE166), and contains replication origins for B. subtilis and E. coli, as well as resistance markers for Ampicillin and Chloramphenicol. This episomal plasmid is capable of transformation into both Gram-negative and Gram-positive organisms. It replicates as an episomal plasmid and does not integrate into the genome. For selection in E. coli, use Ampicillin, 100 ??g per mL. For selection in B. subtilis (and other similar gram-positive organisms), use Chloramphenicol, 5 ??g per mL (not 35). false false _188_ 0 3501 9 Not in stock false We had to remove a PstI site inside the coding region of the beta-lactamase gene coding for Ampicillin resistance, and thus the backbone had to be PCRed from two parts, with a joining primer filling in cut site with a degenerate codon. false Daniel Goodman annotation1992774 1 puc9 Plasmid (E. coli rep origin) range1992774 1 399 649 annotation1992770 1 Biobrick Prefix range1992770 1 5187 5208 annotation1992769 1 Biobrick Suffix range1992769 1 1 21 annotation1992771 1 Beta-Lactamase range1992771 1 4120 4981 annotation1992772 1 Chloramphenicol Acetyl-Transferase range1992772 1 2176 2924 annotation1992773 1 Gram Positive Replication Protein range1992773 1 723 1742 BBa_K541515 1 BBa_K541515 Limulus anti-LPS factor (LALF) for Bacillus subtilis 2011-07-12T11:00:00Z 2015-05-08T01:12:38Z Horseshoe crabs (Limulus polyphemus and Tachypleus tridentatus) are ancient arachnids that possess a primitive circulatory system, the hemolymph, containing only one kind of cell, the hemocyte. Exposure of hemocytes to bacterial endotoxins [lipopolysaccharide (LPS)] results in the activation of an intracellular coagulation cascade (Iwanaga et al., 1986), a defense against microbial invasion Constitutive promoter veg(BBa_K143012) coupled to the strong Ribosome Binding Site spoVG(BBa_K143021) from B. subtilis. These parts have been taken from 2008_Imperial_Collage. Pveg is a constitutive promoter that constitutively expresses the P43 protein in B. subtilis. SpoVG is an endogenous ribosome binding site from B. subtilis. The sequence of the spoVG ribosome binding site is AAAGGUGGUGA which is complementary to the sequence UUUCCUCCACU from the 3' region of the 16s rRNA from B. subtilis. SacB is a signal peptide used in the Sec-SRP (secretory signal recognition particle) pathway by B. subtilis. Signal peptides are responsible for directing preproteins (secretory proteins with a signal peptide region attached) through an appropriate secretory pathway. In the case of the Sec-SRP signal peptide, they direct preproteins from the cytoplasm into the growth medium. Lipopolysaccharide (LPS), or endotoxin, is the major mediator of septic shock, a serious complication of Gram-negative bacterial infections in humans. Molecules that bind LPS and neutralize its biological effects or enhance its clearance could have important clinical applications. Limulus anti-LPS factor (LALF) binds LPS tightly, and, in animal models, reduces mortality when administered before or after LPS challenge or bacterial infection. The wedge- shaped molecule has a striking charge distribution and amphipathicity that suggest how it can insert into membranes. The binding site for LPS probably involves an extended amphipathic loop, and it has been proposed that two mammalian LPS-binding proteins will have a similar loop. The amphipathic loop structure may be used in the design of molecules with therapeutic properties against septic shock. false false _708_ 0 4260 9 Not in stock true During our design, we have looked out that this anti-LPS factor gene should be transfered into gram positive bacteria (i.e. B.subtilis) because of absence of LPS in gram positive bacteria. false Cihan TA&#350;TAN annotation2123337 1 rbs range2123337 1 106 117 annotation2123340 1 LALF range2123340 1 226 537 annotation2123335 1 Sigma A-35 range2123335 1 63 68 annotation2123338 1 BBa_K143021 range2123338 1 106 117 annotation2123339 1 SacB range2123339 1 124 219 annotation2123336 1 Sigma A -10 range2123336 1 86 91 annotation2123334 1 BBa_K143053 range2123334 1 1 117 annotation2123333 1 BBa_K143012 range2123333 1 1 97 BBa_K541515_sequence 1 aattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgttactagagaaaggtggtgaatactagatgaacatcaaaaaatttgcaaaacaggcgacagttctgacatttacaacagcactgcttgcaggcggcgcaacacaagcatttgcaaaagaaacatactagatggatggaatttggacacaactgatttttaccctggtgaagaaccttgctacactttggcaatccggcgattttcagttccttgaccatgaatgccattaccgtattaaacctacgtttagacgcttgaagtggaagtataaaggaaaattttggtgcccgtcttggaccagcattactggacgtgcgacaaagtcatcacgctccggtgccgtcgaacattcggtccgaaattttgttgggcaagcgaagtcgtcaggcctgataacgcaaagacaagcggaacagtttattagtcaatacaattaataa BBa_K541815_sequence 1 tactagtagcggccgctgcagaagcttgagtaggacaaatccgccgagcttcgacgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagaacaaagaatacaagaaaatatttacaaaaaatcaatttaacaattccttaaaacatgcaggaattgacgatttaaacaatattagctttgaacaattcttatctcttttcaatagctataaattatttaataagtaagttaagggatgcataaactgcatcccttaacttgtttttcgtgtgcctattttttgtgaatcgctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgtctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggctggcttaactatgcggcatcagagcagattgtactgagagtgcaccatacaaaacatatttcaacacaatacaaatgggttagttaaaaaagcaggccttctaaaggtctgctttttttatttgattatgtaatttttaatgccaggatgccaataagccataacctcaaatgcaccatttgcaacctcgtcatcttcttcctcaatcttgaccagatcgccgtcctccgcatcaccaagattcagctctttatgtatctccttcaaaatgccaccgtatccaattaaccttcgagctgccaacgcatcatccaagtaaagcaccgtgttcagattgtcttcagtcaccttattaccgcgcacacaatccgtatccttaaccggatatttagagatttcgcgaacagctttttgctccatcattgcgttccgcacatcgttttcaatctgttcagcgtcaatcttagctttacctttcactcgacgaatatcgacaattggagtgtaatccaatttcatcgcctttttccaaaggctcgtccactccgcctgcttaatatagtttttcccaaaataatttttccttactggtatcaacacatgaaaatgaggatgatatgtatcttcttcatgatttttggtaatctctaaagctctgaaaaatccaagaaccgaagtttttacttttttgtactggaacagtttcctaaagccttccatcatcgcagaaatttgtggcttcagccgttctccctttacatttcgaatcgtcagcgtgagaaaaatccatccgcagccgtactgtctattggcttcctctacgatcaacttattgtgataagcaatttttaacgacctgcgccacgcacacatcggacataacctcactttacaaaaatgggcttgatacagttttaacttgcccgtctccgggtctctcttaaacgaaagatactctgcacaactaattagtttttcagcctttttgccatagtaaggtgccccaatcttactctctaacgcttcgtaatgctccgccatgaggttcgtccgtctctttttccccttccaatcccgctttttacctgttgcggttttatcttcgaggatgctataatcattttcagatgaataaatcaacaaaaaaactccttctgagctagttctctagcattctattattttgattcgacaccttaataatagcagaaggagtttttacctgtcaaagaaccatcaaacccttgatacacaaggctttgacctaattttgaaaaatgatgttgtttctatatagtatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaacacaaaagaccacattttttaatgtggtctttattcttcaactaaagcacccattagttcaacaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaatattgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaattcactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagacaattggaagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataaccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatataaattttaccctgcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactggttacaatagcgacggagagttaggttattgggataagttagagccactttatacaatttttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaagagttttatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttcccaaaacacctatacctgaaaatgctttttctctttctattattccatggacttcatttactgggtttaacttaaatatcaataataatagtaattaccttctacccattattacagcaggaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcattctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagataggcctaatgactggcttttataatatgagataatgccgactgtactttttacagtcggttttctaatgtcactaacctgccccgttagtcgccattcgccagctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcaatgctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgccgcaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaacacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgtcttcaagaattcgcggccgcttctagagtactagagaattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgttactagagaaaggtggtgaatactagatgaacatcaaaaaatttgcaaaacaggcgacagttctgacatttacaacagcactgcttgcaggcggcgcaacacaagcatttgcaaaagaaacatactagatggatggaatttggacacaactgatttttaccctggtgaagaaccttgctacactttggcaatccggcgattttcagttccttgaccatgaatgccattaccgtattaaacctacgtttagacgcttgaagtggaagtataaaggaaaattttggtgcccgtcttggaccagcattactggacgtgcgacaaagtcatcacgctccggtgccgtcgaacattcggtccgaaattttgttgggcaagcgaagtcgtcaggcctgataacgcaaagacaagcggaacagtttattagtcaatacaattaataa BBa_K090401_sequence 1 tactagtagcggccgctgcagaagcttgagtaggacaaatccgccgagcttcgacgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagaacaaagaatacaagaaaatatttacaaaaaatcaatttaacaattccttaaaacatgcaggaattgacgatttaaacaatattagctttgaacaattcttatctcttttcaatagctataaattatttaataagtaagttaagggatgcataaactgcatcccttaacttgtttttcgtgtgcctattttttgtgaatcgctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgtctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggctggcttaactatgcggcatcagagcagattgtactgagagtgcaccatacaaaacatatttcaacacaatacaaatgggttagttaaaaaagcaggccttctaaaggtctgctttttttatttgattatgtaatttttaatgccaggatgccaataagccataacctcaaatgcaccatttgcaacctcgtcatcttcttcctcaatcttgaccagatcgccgtcctccgcatcaccaagattcagctctttatgtatctccttcaaaatgccaccgtatccaattaaccttcgagctgccaacgcatcatccaagtaaagcaccgtgttcagattgtcttcagtcaccttattaccgcgcacacaatccgtatccttaaccggatatttagagatttcgcgaacagctttttgctccatcattgcgttccgcacatcgttttcaatctgttcagcgtcaatcttagctttacctttcactcgacgaatatcgacaattggagtgtaatccaatttcatcgcctttttccaaaggctcgtccactccgcctgcttaatatagtttttcccaaaataatttttccttactggtatcaacacatgaaaatgaggatgatatgtatcttcttcatgatttttggtaatctctaaagctctgaaaaatccaagaaccgaagtttttacttttttgtactggaacagtttcctaaagccttccatcatcgcagaaatttgtggcttcagccgttctccctttacatttcgaatcgtcagcgtgagaaaaatccatccgcagccgtactgtctattggcttcctctacgatcaacttattgtgataagcaatttttaacgacctgcgccacgcacacatcggacataacctcactttacaaaaatgggcttgatacagttttaacttgcccgtctccgggtctctcttaaacgaaagatactctgcacaactaattagtttttcagcctttttgccatagtaaggtgccccaatcttactctctaacgcttcgtaatgctccgccatgaggttcgtccgtctctttttccccttccaatcccgctttttacctgttgcggttttatcttcgaggatgctataatcattttcagatgaataaatcaacaaaaaaactccttctgagctagttctctagcattctattattttgattcgacaccttaataatagcagaaggagtttttacctgtcaaagaaccatcaaacccttgatacacaaggctttgacctaattttgaaaaatgatgttgtttctatatagtatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaacacaaaagaccacattttttaatgtggtctttattcttcaactaaagcacccattagttcaacaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaatattgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaattcactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagacaattggaagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataaccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatataaattttaccctgcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactggttacaatagcgacggagagttaggttattgggataagttagagccactttatacaatttttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaagagttttatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttcccaaaacacctatacctgaaaatgctttttctctttctattattccatggacttcatttactgggtttaacttaaatatcaataataatagtaattaccttctacccattattacagcaggaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcattctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagataggcctaatgactggcttttataatatgagataatgccgactgtactttttacagtcggttttctaatgtcactaacctgccccgttagtcgccattcgccagctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcaatgctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgccgcaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaacacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgtcttcaagaattcgcggccgcttctagag 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