BBa_K2020001 1 pelB+subE subtilisin E gene with leader sequence pelB 2016-10-03T11:00:00Z 2016-10-04T07:47:20Z The sequence of the signal peptide pelB was obtained through the Registry (BBa_J32015: http://parts.igem.org/Part:BBa_J32015). Together with the subtilisin E gene (BBa_K2020000: http://parts.igem.org/Part:BBa_K2020000) it was ordered from IDT. This composite part consists of the leader sequence pelB (BBa_J32015) and the subtilisin E gene (BBa_K2020000). It can be used to express subtilisin E in Escherichia coli and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins. false false _2487_ 30763 30763 9 false The sequence of the subtilisin E gene from Bacillus subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. Moreover, the leader sequence pelB begins with a start odon and was placed in front of the subtilisin E gene to generate a functional coding sequence that can be expressed in E. coli. false Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert component2485824 1 BBa_K2020000 component2485821 1 BBa_J32015 annotation2485824 1 BBa_K2020000 range2485824 1 67 1125 annotation2485821 1 BBa_J32015 range2485821 1 1 66 BBa_K2020002 1 ExpSubE expression system for subtilisin E in E. coli 2016-10-04T11:00:00Z 2016-10-05T10:39:31Z The sequences of the BioBrick parts were obtained through the Registry. The construct was ordered at IDT. add links This composite part consists of the promoter [http://parts.igem.org/Part:BBa_R0010 BBa_R0010], the RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034], the newly created BioBrick part [http://parts.igem.org/Part:BBa_K2020001 BBa_K2020001] and the terminator [http://parts.igem.org/Part:BBa_B0010 BBa_B0010]. BioBrick BBa_K2020001 is a composite part itself and includes the secretion tag pelB ([http://parts.igem.org/Part:BBa_J32015 BBaJ32015]) and a subtilisin E gene optimized for E. coli codon usage ([http://parts.igem.org/Part:BBa_K2020000 BBa_K2020000]). Once introduced into Escherichia coli, this BioBrick is able to produce subtilisin E and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins. false false _2487_ 30763 30763 9 false The sequence of the subtilisin E gene from Bacillus subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. false Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert component2486152 1 BBa_K2020001 component2486138 1 BBa_R0010 component2486153 1 BBa_B0010 component2486146 1 BBa_B0034 annotation2486153 1 BBa_B0010 range2486153 1 1360 1439 annotation2486138 1 BBa_R0010 range2486138 1 1 200 annotation2486146 1 BBa_B0034 range2486146 1 209 220 annotation2486152 1 BBa_K2020001 range2486152 1 227 1351 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_R0010 1 LacI promoter (lacI regulated) 2003-01-31T12:00:00Z 2015-05-08T01:14:14Z The Plac insert was PCR'd from the MG1655 strain of E.coli K12. Released HQ 2013 Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) <p> The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG. false true _1_ 0 24 7 In stock false <P> <P><P> LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and this regulator. This part is incompatible with environments containing lactose or lactose analogs. true annotation1961223 1 CAP binding site range1961223 1 89 126 annotation1961225 1 -10 range1961225 1 161 166 annotation1961224 1 -35 range1961224 1 137 142 annotation1961222 1 BBa_R0010 range1961222 1 1 200 annotation1961221 1 end of LacI coding region (inactive) range1961221 1 1 88 annotation1961227 1 start range1961227 1 173 173 annotation1961226 1 LacI binding site range1961226 1 166 200 BBa_K2020000 1 subE subtilisin E gene 2016-10-03T11:00:00Z 2016-10-04T07:43:31Z The sequence wasoriginally obtained from a wildtype Bacillus subtilis and was ordered from IDT. This gene can be used to express subtilisin E in Escherichia coli. Subtilisin E is an alkaline serine protease which non-specifically deigests proteins. To use this part a suitable leader sequence has to be placed in front as the sequence of this BioBrick does not contain a start codon. false false _2487_ 30763 30763 9 false The sequence from B. subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. false Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert annotation2485808 1 propeptide range2485808 1 1 230 annotation2485809 1 subtilisin E range2485809 1 231 1059 BBa_J32015 1 PelB PelB leader sequence; directs protein to E. coli periplasmic membrane 2006-08-24T11:00:00Z 2015-08-31T04:08:46Z Novagen Released HQ 2013 The pelB leader sequence is a sequence of amino acids which when attached to a protein, directs the protein to the periplasmic membrane of E. coli, where the sequence is removed by pelB peptidase. It is used to direct coat protein-antigen fusions to the cell surface. false true _50_ 0 495 50 In stock true . true Austen Heinz annotation1898182 1 PelB range1898182 1 1 66 BBa_B0034 1 BBa_B0034 RBS (Elowitz 1999) -- defines RBS efficiency 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 RBS based on Elowitz repressilator. false true _1_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. <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 for this part: <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. annotation23325 1 conserved range23325 1 5 8 BBa_B0010_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc BBa_R0010_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacaca BBa_K2020001_sequence 1 atgaaatacctgctgccgaccgctgctgctggtctgctgctcctcgctgcccagccggcgatggccgcgggcaaaagcagcaccgaaaaaaaatatattgtgggctttaaacagaccatgagcgcgatgagcagcgcgaaaaaaaaagatgtgattagcgaaaaaggcggcaaagtgcagaaacagtttaaatatgtgaacgcggcggcggcgaccctggatgaaaaagcggtgaaagaactgaaaaaagatccgagcgtggcgtatgtggaagaagatcatatcgcgcatgaatatgcccagagcgtgccgtatggcattagccagattaaagccccggcgctgcatagccagggctataccggcagcaacgtgaaagtggcggtgattgatagcggcattgatagcagccatccggatctgaacgtgcgcggcggcgcgagctttgtgccgagcgaaaccaacccgtatcaggatggcagcagccatggcacccatgtggcgggcaccattgcggcgctgaacaacagcattggcgtgctgggcgtgagcccgagcgcgagcctgtatgcggtgaaagtgctggatagcaccggcagcggccagtatagctggattattaacggcattgaatgggcgattagcaacaacatggatgtgattaacatgagcctgggcggcccgaccggcagcaccgcgctgaaaaccgtggtggataaagcggtgagcagcggcattgtggtggcggcggcggcgggcaacgaaggcagcagcggcagcaccagcaccgtgggctatccggcgaaatatccgtcaacgattgctgttggcgccgtaaattcaagcaatcagcgtgcgtcattctcatccgcaggtagcgaactggatgtgatggcgccgggcgtgagcattcagagcaccctgccgggcggcacctatggcgcgtataacggcaccagcatggcgaccccgcatgtggcgggcgcggcggcgctgattctgagcaaacatccgacctggaccaacgcgcaggtgcgcgatcgcctggaaagcaccgcgacctatctgggcaacagcttttattatggcaaaggcctgattaacgtgcaggcggcggcgcagtaa BBa_B0034_sequence 1 aaagaggagaaa BBa_K2020002_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagaaagaggagaaatactagatgaaatacctgctgccgaccgctgctgctggtctgctgctcctcgctgcccagccggcgatggccgcgggcaaaagcagcaccgaaaaaaaatatattgtgggctttaaacagaccatgagcgcgatgagcagcgcgaaaaaaaaagatgtgattagcgaaaaaggcggcaaagtgcagaaacagtttaaatatgtgaacgcggcggcggcgaccctggatgaaaaagcggtgaaagaactgaaaaaagatccgagcgtggcgtatgtggaagaagatcatatcgcgcatgaatatgcccagagcgtgccgtatggcattagccagattaaagccccggcgctgcatagccagggctataccggcagcaacgtgaaagtggcggtgattgatagcggcattgatagcagccatccggatctgaacgtgcgcggcggcgcgagctttgtgccgagcgaaaccaacccgtatcaggatggcagcagccatggcacccatgtggcgggcaccattgcggcgctgaacaacagcattggcgtgctgggcgtgagcccgagcgcgagcctgtatgcggtgaaagtgctggatagcaccggcagcggccagtatagctggattattaacggcattgaatgggcgattagcaacaacatggatgtgattaacatgagcctgggcggcccgaccggcagcaccgcgctgaaaaccgtggtggataaagcggtgagcagcggcattgtggtggcggcggcggcgggcaacgaaggcagcagcggcagcaccagcaccgtgggctatccggcgaaatatccgtcaacgattgctgttggcgccgtaaattcaagcaatcagcgtgcgtcattctcatccgcaggtagcgaactggatgtgatggcgccgggcgtgagcattcagagcaccctgccgggcggcacctatggcgcgtataacggcaccagcatggcgaccccgcatgtggcgggcgcggcggcgctgattctgagcaaacatccgacctggaccaacgcgcaggtgcgcgatcgcctggaaagcaccgcgacctatctgggcaacagcttttattatggcaaaggcctgattaacgtgcaggcggcggcgcagtaatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc BBa_J32015_sequence 1 atgaaatacctgctgccgaccgctgctgctggtctgctgctcctcgctgcccagccggcgatggcc BBa_K2020000_sequence 1 gcgggcaaaagcagcaccgaaaaaaaatatattgtgggctttaaacagaccatgagcgcgatgagcagcgcgaaaaaaaaagatgtgattagcgaaaaaggcggcaaagtgcagaaacagtttaaatatgtgaacgcggcggcggcgaccctggatgaaaaagcggtgaaagaactgaaaaaagatccgagcgtggcgtatgtggaagaagatcatatcgcgcatgaatatgcccagagcgtgccgtatggcattagccagattaaagccccggcgctgcatagccagggctataccggcagcaacgtgaaagtggcggtgattgatagcggcattgatagcagccatccggatctgaacgtgcgcggcggcgcgagctttgtgccgagcgaaaccaacccgtatcaggatggcagcagccatggcacccatgtggcgggcaccattgcggcgctgaacaacagcattggcgtgctgggcgtgagcccgagcgcgagcctgtatgcggtgaaagtgctggatagcaccggcagcggccagtatagctggattattaacggcattgaatgggcgattagcaacaacatggatgtgattaacatgagcctgggcggcccgaccggcagcaccgcgctgaaaaccgtggtggataaagcggtgagcagcggcattgtggtggcggcggcggcgggcaacgaaggcagcagcggcagcaccagcaccgtgggctatccggcgaaatatccgtcaacgattgctgttggcgccgtaaattcaagcaatcagcgtgcgtcattctcatccgcaggtagcgaactggatgtgatggcgccgggcgtgagcattcagagcaccctgccgggcggcacctatggcgcgtataacggcaccagcatggcgaccccgcatgtggcgggcgcggcggcgctgattctgagcaaacatccgacctggaccaacgcgcaggtgcgcgatcgcctggaaagcaccgcgacctatctgggcaacagcttttattatggcaaaggcctgattaacgtgcaggcggcggcgcagtaa 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