BBa_K1795000 1 BBa_K1795000 E. Coli Codon Optimized dCAS9, solo 2015-09-16T11:00:00Z 2015-09-17T09:39:49Z The basic template for the sgRNA sequence was taken from the supplementary information of Qi LS, Larson MH, Gilbert LA, et al. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell. 2013;152(5):1173-1183. doi:10.1016/j.cell.2013.02.022. This sgRNA sequence was designed to target the Promoter Bba_R0010 with the N20 sequence atgttgtgtggaattgtgag. The part is under the control of the promoter Bba_J23100. If this gRNA is expressed in a cell with a form of the dCas9 protein, the protein will be targeted to the N20 sequence and repress gene expression. Expression of this part is repressed in the presence of LacI and CAP protein. false false _2220_ 27446 21426 9 false Ensure the promoter targeted by this part is not used within this part. false Andrew D Halleran, Joseph L Maniaci 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_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 annotation1961222 1 BBa_R0010 range1961222 1 1 200 annotation1961226 1 LacI binding site range1961226 1 166 200 annotation1961221 1 end of LacI coding region (inactive) range1961221 1 1 88 annotation1961223 1 CAP binding site range1961223 1 89 126 annotation1961224 1 -35 range1961224 1 137 142 annotation1961227 1 start range1961227 1 173 173 annotation1961225 1 -10 range1961225 1 161 166 BBa_B0015 1 BBa_B0015 double terminator (B0010-B0012) 2003-07-16T11:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 Double terminator consisting of BBa_B0010 and BBa_B0012 false true _1_ 0 24 7 In stock false true Reshma Shetty component1916610 1 BBa_B0010 component1916612 1 BBa_B0012 annotation1916612 1 BBa_B0012 range1916612 1 89 129 annotation1916610 1 BBa_B0010 range1916610 1 1 80 BBa_K1795001 1 BBa_K1795001 E. Coli Codon Optimized fdCAS9 under R0010 2015-09-16T11:00:00Z 2015-09-17T09:39:24Z Thanks to team iGEM13_SJTU-BioX-Shanghai for original, non-codon optimized dCas9 sequence. This sequence codes for dCas9 and is codon optimized for E. Coli. It is under the promoter R0010 and has RBS B0034 and DT B0015. false false _2220_ 27446 27446 9 false false Andrew D Halleran, Joseph L Maniaci component2467996 1 BBa_B0034 component2467997 1 BBa_K1795000 component2467988 1 BBa_R0010 component2468004 1 BBa_B0015 annotation2467988 1 BBa_R0010 range2467988 1 1 200 annotation2467997 1 BBa_K1795000 range2467997 1 227 4339 annotation2467996 1 BBa_B0034 range2467996 1 209 220 annotation2468004 1 BBa_B0015 range2468004 1 4348 4476 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_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_R0010_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacaca BBa_B0034_sequence 1 aaagaggagaaa BBa_K1795001_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagaaagaggagaaatactagatggacaaaaaatacagtatcggcctggcgatcggcaccaattcagttggctgggcagttattactgatgaatacaaggtcccctccaaaaagtttaaagtcctgggcaacaccgatcgtcacagcatcaaaaaaaatctgatcggagccttgttattcgacagtggtgagacggcagaagccacccgtttgaagcgtactgcccgccgtcgctacacacgtcgtaagaaccgtatctgttacttgcaggaaattttctcaaatgaaatggcgaaagtggacgacagcttcttccaccgccttgaagagtcattcttagttgaggaagacaaaaaacacgaacgtcatcctatctttggcaatatcgtcgatgaagtcgcgtatcacgaaaagtacccgaccatttatcacttgcgtaaaaaactggtcgactcgaccgacaaagcagacctgcgtctgatttatttagcattggcccatatgattaaatttcgcggccacttcttaatcgaaggcgacttgaatccggacaactcggatgtggataaattgttcattcagctggtccagacgtataatcagctttttgaggaaaacccgatcaacgcatcgggcgttgatgccaaggcgattttatccgcacgcctgtcgaaaagccgccgtctggaaaatctcatcgcacaactgccgggcgaaaagaaaaacggtctgttcgggaacttgattgcactgtccctgggcctgaccccaaactttaaaagcaatttcgacttggccgaagatgcgaagctgcagctgagtaaagacacctatgatgacgacttagataatttgttggcgcagattggtgatcaatacgccgacctctttctcgcagcgaaaaatctgagtgatgcgatcttattatctgatatcctgcgcgtcaacaccgaaattaccaaagcaccgctgtccgccagtatgatcaagcgctacgacgagcatcatcaagatctgacccttctcaaagcgttagtgcgccagcagctgccggaaaaatacaaagagatcttcttcgaccagagtaaaaacggatatgcaggttacattgatggcggcgcatcacaggaagaattttataaatttattaaaccaattttagagaaaatggatggtactgaagaactccttgtcaaactgaaccgcgaagatcttctgcgcaaacaacgcacgtttgacaacggttccattccgcatcaaattcatttgggagaactccatgctattctgcgtcgtcaggaagatttctatccgtttttgaaggataatcgtgagaagattgagaaaatcctgacatttcgtatcccttactatgtcggtcctctggcgcgcggcaacagccgctttgcatggatgactcgtaaaagcgaggaaacgattaccccgtggaatttcgaagaagtagtggacaaaggagcgtcggcgcagagttttattgaacgcatgaccaatttcgataaaaatctgccgaacgaaaaagtactgccaaaacactctctgctgtatgaatacttcaccgtgtacaatgaactcactaaggtcaaatatgtaacagaaggtatgcgtaaacccgcgtttttaagcggggaacaaaaaaaggccatcgtagaccttctcttcaaaacgaaccgcaaggtaaccgtgaaacagttaaaagaggactatttcaaaaaaatcgaatgttttgacagcgtggagatctcaggtgtggaagatcgcttcaacgcttcactcggaacttaccatgatctgttaaagatcattaaagacaaggattttcttgataacgaagagaacgaagacattttagaggatatcgtcctgacactgactttgtttgaagatcgcgaaatgattgaagaacgcctgaaaacctatgcgcatctgtttgatgacaaagtcatgaaacagctgaaacgtcgtcgctatactggctggggccgtctgagtcgcaaactcattaacggtattcgtgacaaacaaagtggtaaaacgattctggattttcttaaaagcgacggatttgcgaatcgtaacttcatgcaattaatccacgacgattcgctgacatttaaagaggacattcaaaaagctcaggtgagtggccagggtgacagcttgcatgagcatattgcgaacttagctggtagccctgccattaaaaaaggcattttgcagactgtaaaagtggtggacgaactggttaaggtgatgggccgccacaaaccagaaaatatcgttattgaaatggcacgcgaaaaccagactacgcaaaaaggtcagaaaaactctcgtgaacgtatgaaacgcatcgaagaaggaatcaaagaactgggtagccagatcctgaaagaacacccggtggaaaacacgcaactgcagaacgaaaaactgtatctctattatttacaaaatggtcgcgacatgtatgtggatcaagaattggatattaatcgcttgtccgattatgacgtcgatgcgattgttccgcagtccttcctgaaagatgacagcattgataacaaagttttaactcgctcggacaaaaaccgcgggaaatctgataacgtcccgagcgaggaggtagtgaaaaagatgaaaaactattggcgccagctgctgaacgcgaaactcatcacccagcgtaaatttgataaccttacaaaagcagaacgtggtggtttgtccgagctggataaggcgggcttcatcaaacgccagctcgtagagacgcgccaaatcactaaacatgtggcgcaaatcctggatagccgtatgaatacgaagtacgatgagaatgacaaactgattcgcgaagtcaaagtgatcaccctgaaatctaagttggtgagcgattttcgcaaagactttcagttttataaggttcgcgagattaacaactatcatcatgcgcacgatgcctacttaaatgccgtggtgggcaccgctctcatcaaaaaatacccaaaattggagtcagagtttgtttacggtgattataaagtttatgatgttcgtaaaatgatcgcaaaaagcgagcaggagatcggaaaagcaacagccaagtattttttttatagcaatattatgaattttttcaaaacagaaatcaccttagcgaatggcgaaatccgtaagcgccctcttattgaaacaaatggggaaaccggcgaaattgtctgggataagggccgcgattttgccactgtgcgtaaagtgctgtctatgccacaggttaatatcgtgaaaaaaaccgaggtacagaccggtgggtttagtaaagaaagcattctgcccaagcgcaacagcgataaactgattgcgcgcaaaaaagattgggatccgaaaaaatatggtggcttcgattcgcctactgttgcgtattcggtcctcgtagtggctaaggtagaaaaaggtaaatctaaaaaattaaaaagcgtaaaagaattactgggaattacgattatggaacgtagctcgtttgagaaaaacccgattgacttcctggaagcgaaaggatataaagaagttaaaaaagatctgatcattaagctgccgaaatactcgctctttgaactcgaaaacggtcgtaaacgcatgctggcatcagcaggcgagctgcagaaaggcaacgagcttgcgttgccctccaaatacgtcaatttcttgtatctggctagtcattacgaaaaattgaaaggatccccggaagacaacgaacaaaaacaactgttcgtggaacagcacaaacattatcttgacgagatcattgaacagatctctgaattttccaaacgtgttattctggccgatgccaacctggacaaggttctttctgcgtacaacaaacatcgcgacaaaccaatccgcgagcaagctgaaaatattattcatttatttacactgacaaacctgggtgcgccggcggcgtttaaatatttcgatacaaccatcgatcgcaagcgttacacctctaccaaagaagtgctggatgccaccctgattcatcaaagcattacgggtctgtacgaaacgcgcattgatctgagtcagctgggcggggactaagtcgactactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_B0012_sequence 1 tcacactggctcaccttcgggtgggcctttctgcgtttata BBa_K1795000_sequence 1 atggacaaaaaatacagtatcggcctggcgatcggcaccaattcagttggctgggcagttattactgatgaatacaaggtcccctccaaaaagtttaaagtcctgggcaacaccgatcgtcacagcatcaaaaaaaatctgatcggagccttgttattcgacagtggtgagacggcagaagccacccgtttgaagcgtactgcccgccgtcgctacacacgtcgtaagaaccgtatctgttacttgcaggaaattttctcaaatgaaatggcgaaagtggacgacagcttcttccaccgccttgaagagtcattcttagttgaggaagacaaaaaacacgaacgtcatcctatctttggcaatatcgtcgatgaagtcgcgtatcacgaaaagtacccgaccatttatcacttgcgtaaaaaactggtcgactcgaccgacaaagcagacctgcgtctgatttatttagcattggcccatatgattaaatttcgcggccacttcttaatcgaaggcgacttgaatccggacaactcggatgtggataaattgttcattcagctggtccagacgtataatcagctttttgaggaaaacccgatcaacgcatcgggcgttgatgccaaggcgattttatccgcacgcctgtcgaaaagccgccgtctggaaaatctcatcgcacaactgccgggcgaaaagaaaaacggtctgttcgggaacttgattgcactgtccctgggcctgaccccaaactttaaaagcaatttcgacttggccgaagatgcgaagctgcagctgagtaaagacacctatgatgacgacttagataatttgttggcgcagattggtgatcaatacgccgacctctttctcgcagcgaaaaatctgagtgatgcgatcttattatctgatatcctgcgcgtcaacaccgaaattaccaaagcaccgctgtccgccagtatgatcaagcgctacgacgagcatcatcaagatctgacccttctcaaagcgttagtgcgccagcagctgccggaaaaatacaaagagatcttcttcgaccagagtaaaaacggatatgcaggttacattgatggcggcgcatcacaggaagaattttataaatttattaaaccaattttagagaaaatggatggtactgaagaactccttgtcaaactgaaccgcgaagatcttctgcgcaaacaacgcacgtttgacaacggttccattccgcatcaaattcatttgggagaactccatgctattctgcgtcgtcaggaagatttctatccgtttttgaaggataatcgtgagaagattgagaaaatcctgacatttcgtatcccttactatgtcggtcctctggcgcgcggcaacagccgctttgcatggatgactcgtaaaagcgaggaaacgattaccccgtggaatttcgaagaagtagtggacaaaggagcgtcggcgcagagttttattgaacgcatgaccaatttcgataaaaatctgccgaacgaaaaagtactgccaaaacactctctgctgtatgaatacttcaccgtgtacaatgaactcactaaggtcaaatatgtaacagaaggtatgcgtaaacccgcgtttttaagcggggaacaaaaaaaggccatcgtagaccttctcttcaaaacgaaccgcaaggtaaccgtgaaacagttaaaagaggactatttcaaaaaaatcgaatgttttgacagcgtggagatctcaggtgtggaagatcgcttcaacgcttcactcggaacttaccatgatctgttaaagatcattaaagacaaggattttcttgataacgaagagaacgaagacattttagaggatatcgtcctgacactgactttgtttgaagatcgcgaaatgattgaagaacgcctgaaaacctatgcgcatctgtttgatgacaaagtcatgaaacagctgaaacgtcgtcgctatactggctggggccgtctgagtcgcaaactcattaacggtattcgtgacaaacaaagtggtaaaacgattctggattttcttaaaagcgacggatttgcgaatcgtaacttcatgcaattaatccacgacgattcgctgacatttaaagaggacattcaaaaagctcaggtgagtggccagggtgacagcttgcatgagcatattgcgaacttagctggtagccctgccattaaaaaaggcattttgcagactgtaaaagtggtggacgaactggttaaggtgatgggccgccacaaaccagaaaatatcgttattgaaatggcacgcgaaaaccagactacgcaaaaaggtcagaaaaactctcgtgaacgtatgaaacgcatcgaagaaggaatcaaagaactgggtagccagatcctgaaagaacacccggtggaaaacacgcaactgcagaacgaaaaactgtatctctattatttacaaaatggtcgcgacatgtatgtggatcaagaattggatattaatcgcttgtccgattatgacgtcgatgcgattgttccgcagtccttcctgaaagatgacagcattgataacaaagttttaactcgctcggacaaaaaccgcgggaaatctgataacgtcccgagcgaggaggtagtgaaaaagatgaaaaactattggcgccagctgctgaacgcgaaactcatcacccagcgtaaatttgataaccttacaaaagcagaacgtggtggtttgtccgagctggataaggcgggcttcatcaaacgccagctcgtagagacgcgccaaatcactaaacatgtggcgcaaatcctggatagccgtatgaatacgaagtacgatgagaatgacaaactgattcgcgaagtcaaagtgatcaccctgaaatctaagttggtgagcgattttcgcaaagactttcagttttataaggttcgcgagattaacaactatcatcatgcgcacgatgcctacttaaatgccgtggtgggcaccgctctcatcaaaaaatacccaaaattggagtcagagtttgtttacggtgattataaagtttatgatgttcgtaaaatgatcgcaaaaagcgagcaggagatcggaaaagcaacagccaagtattttttttatagcaatattatgaattttttcaaaacagaaatcaccttagcgaatggcgaaatccgtaagcgccctcttattgaaacaaatggggaaaccggcgaaattgtctgggataagggccgcgattttgccactgtgcgtaaagtgctgtctatgccacaggttaatatcgtgaaaaaaaccgaggtacagaccggtgggtttagtaaagaaagcattctgcccaagcgcaacagcgataaactgattgcgcgcaaaaaagattgggatccgaaaaaatatggtggcttcgattcgcctactgttgcgtattcggtcctcgtagtggctaaggtagaaaaaggtaaatctaaaaaattaaaaagcgtaaaagaattactgggaattacgattatggaacgtagctcgtttgagaaaaacccgattgacttcctggaagcgaaaggatataaagaagttaaaaaagatctgatcattaagctgccgaaatactcgctctttgaactcgaaaacggtcgtaaacgcatgctggcatcagcaggcgagctgcagaaaggcaacgagcttgcgttgccctccaaatacgtcaatttcttgtatctggctagtcattacgaaaaattgaaaggatccccggaagacaacgaacaaaaacaactgttcgtggaacagcacaaacattatcttgacgagatcattgaacagatctctgaattttccaaacgtgttattctggccgatgccaacctggacaaggttctttctgcgtacaacaaacatcgcgacaaaccaatccgcgagcaagctgaaaatattattcatttatttacactgacaaacctgggtgcgccggcggcgtttaaatatttcgatacaaccatcgatcgcaagcgttacacctctaccaaagaagtgctggatgccaccctgattcatcaaagcattacgggtctgtacgaaacgcgcattgatctgagtcagctgggcggggactaagtcgac BBa_B0015_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata 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