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 component1916612 1 BBa_B0012 component1916610 1 BBa_B0010 annotation1916612 1 BBa_B0012 range1916612 1 89 129 annotation1916610 1 BBa_B0010 range1916610 1 1 80 BBa_K2150031 1 pT7 T7 promoter without RBS 2016-10-11T11:00:00Z 2016-10-12T09:06:12Z PCR amplified from BBa_K525998 This part is the T7 promoter without RBS. It has been reported that T7 promoter doesn't work with E.coli RNA polymerase, but only with T7 RNA polymerase(T7RNAP). However, we observed leakage of it in the absence of T7RNAP. false false _2620_ 31283 31283 9 false We deleted the RBS in BBa_K525998, so that RBS with different efficiency can be used. false Jianyi Huang 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 annotation1690 1 polya range1690 1 28 41 annotation7020 1 BBa_B0012 range7020 1 1 41 annotation1686 1 T7 TE range1686 1 8 27 annotation1687 1 stop range1687 1 34 34 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_K2150013 1 tetX-GFP A fusion protein combining tetX(BBa_K2150101) with GFP(BBa_E0040) 2016-10-08T11:00:00Z 2016-10-19T01:06:12Z The source of tetX is the same as (xxx), the protein linker is provided by our advisor Li Hua, and the GFP is from BBa_E0040. We combine this three parts using overlap extension polymerase chain reaction (OE-PCR). This protein(noted as tetX-GFP) combines a tetracycline-degrading enzyme (tetX monooxygenase, ) with a green fluorescent protein (GFPmt3, BBa_E0040). TetX-GFP has activities of both the tetX and the GFP. As a degradation enzyme, it can degrade tetracycline(tc) and its analogs as we describe (here), and also give the tetracycline resistance to E.coli. As a green fluorescent protein, it inherits the normal function of BBa_E0040. However, the expression quantity of this protein may be smaller than BBa_E0040 under the same promoter or it may take longer time for tetX-GFP to mature, for we notice that when it is expressed under the same promoter as BBa_E0040, the fluorescent intensity(FI) of tetX-GFP is weaker than BBa_E0040 at the same growth stage of E.coli, and it takes longer for E.coli expressing tetX-GFP to reach the same FI as E.coli expressing BBa_E0040. We use tetX-GFP as a monitor of the quantity of tetX expressed in E.coli. TetX-GFP can also act as a reporter of whether our system is working (meaning whether our machine is degrading tetracycline). When tetX-GFP is expressed under a tetracycline-regulated promoter such as BBa_R0040, it will become a visible sensor of tetracycline in the presence of tetR, with no need for additional tetracycline-resistance genes. false false _2620_ 29891 29891 9 false The protein linker is 3*GGGGS which provides flexibility to the fusion protein. When choosing the codon for this linker, we need to avoid repeated sequence which may cause problems when assemble these parts. false Yang Ye annotation2489537 1 TetX range2489537 1 1 1171 annotation2489538 1 protein linker range2489538 1 1172 1215 annotation2489539 1 GFP range2489539 1 1216 1935 BBa_K2150017 1 BBa_K2150017 pT7 TetX-GFP(fusion protein) 2016-10-11T11:00:00Z 2016-10-12T07:14:21Z pT7 promoter with RBS comes from BBa_K525998,DT from BBa_B0015. As for fusion protein tetX-GFP, tetX is the same as (xxx), the protein linker is provided by our advisor Li Hua, and the GFP is from BBa_E0040. We combine this three parts using overlap extension polymerase chain reaction (OE-PCR). pT7+RBS+TetX-GFP(fusion protein)+DT. TetX and GFP are combined with a 3*GGGGS linker to form a fusion protein. false false _2620_ 31283 31283 9 false Expression level of tetX can be measured by the fluorescence intensity of GFP and the expression of tetracycline-degrading enzyme tetX can be amplified by T7 RNA polymerase. false Jianyi Huang component2495556 1 BBa_B0034 component2495554 1 BBa_K2150031 component2495567 1 BBa_B0015 component2495560 1 BBa_K2150013 annotation2495554 1 BBa_K2150031 range2495554 1 1 20 annotation2495560 1 BBa_K2150013 range2495560 1 47 1981 annotation2495556 1 BBa_B0034 range2495556 1 29 40 annotation2495567 1 BBa_B0015 range2495567 1 1990 2118 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 annotation4184 1 stem_loop range4184 1 12 55 annotation7018 1 BBa_B0010 range7018 1 1 80 BBa_B0010_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc BBa_B0034_sequence 1 aaagaggagaaa BBa_K2150031_sequence 1 taatacgactcactataggg BBa_K2150017_sequence 1 taatacgactcactatagggtactagagaaagaggagaaatactagatgaaaaaaaccatgcgtatcgacaccgacaaacagatgaacctgctgtctgacaaaaacgttgcgatcatcggtggtggtccggttggtctgaccatggcgaaactgcttcagcagaacggtatcgacgtttctgtttacgaacgtgacaacgaccgtgaagcgcgtatcttcggtggtaccctggacctgcacaaaggttctggtcaggaagcgatgaaaaaagcgggtctgcttcagacctactacgacctggcgctgccgatgggtgttaacatcgcggacaaaaaaggtaacatcctgtctaccaaaaacgttaaaccggaaaaccgtttcgacaacccggaaatcaaccgtaacgacctgcgtgcgatcctgctgaactctctggaaaacgacaccgttatctgggaccgtaaactggttatgctggaaccgggtaaaaaaaaatggaccctgaccttcgaaaacaaaccgtctgaaaccgcggacctggttatcctggcgaacggtggtatgtctaaagttcgtaaattcgttaccgacaccgaagttgaagaaaccggtaccttcaacatccaggcggacatccaccagccggaaatcaactgcccgggtttcttccagctgtgcaacggtaaccgtctgatggcgtctcaccagggtaacctgctgttcgcgaacccgaacaacaacggtgcgctgcacttcggtatctctttcaaaaccccggacgaatggaaaaaccagacccaggttgacttccagaaccgtaactctgttgttgacttcctgctgaaagagttctctgactgggacgaacgttacaaagaactgatccacaccaccctgtctttcgttggtctggcgacccgtatcttcccgctggaaaaaccgtggaaatctaaacgtccgctgccgatcaccatgatcggtgacgcggcgcacctgatgccgccgttcgcgggtcagggtgttaactctggtctggttgacgcgctgatcctgtctgacaacctggcggacggtaaattcaactctatcgaagaagcggttaaaaactacgaacagcagatgttcatgtacggtaaagaagcgcaggaagaatctacccagaacgaaatcgaaatgttcaaaccggacttcaccttccagcagctgctgaacgttggtggaggaggttctggaggcggtggaagtggtggcggaggtagcatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_B0012_sequence 1 tcacactggctcaccttcgggtgggcctttctgcgtttata BBa_K2150013_sequence 1 atgaaaaaaaccatgcgtatcgacaccgacaaacagatgaacctgctgtctgacaaaaacgttgcgatcatcggtggtggtccggttggtctgaccatggcgaaactgcttcagcagaacggtatcgacgtttctgtttacgaacgtgacaacgaccgtgaagcgcgtatcttcggtggtaccctggacctgcacaaaggttctggtcaggaagcgatgaaaaaagcgggtctgcttcagacctactacgacctggcgctgccgatgggtgttaacatcgcggacaaaaaaggtaacatcctgtctaccaaaaacgttaaaccggaaaaccgtttcgacaacccggaaatcaaccgtaacgacctgcgtgcgatcctgctgaactctctggaaaacgacaccgttatctgggaccgtaaactggttatgctggaaccgggtaaaaaaaaatggaccctgaccttcgaaaacaaaccgtctgaaaccgcggacctggttatcctggcgaacggtggtatgtctaaagttcgtaaattcgttaccgacaccgaagttgaagaaaccggtaccttcaacatccaggcggacatccaccagccggaaatcaactgcccgggtttcttccagctgtgcaacggtaaccgtctgatggcgtctcaccagggtaacctgctgttcgcgaacccgaacaacaacggtgcgctgcacttcggtatctctttcaaaaccccggacgaatggaaaaaccagacccaggttgacttccagaaccgtaactctgttgttgacttcctgctgaaagagttctctgactgggacgaacgttacaaagaactgatccacaccaccctgtctttcgttggtctggcgacccgtatcttcccgctggaaaaaccgtggaaatctaaacgtccgctgccgatcaccatgatcggtgacgcggcgcacctgatgccgccgttcgcgggtcagggtgttaactctggtctggttgacgcgctgatcctgtctgacaacctggcggacggtaaattcaactctatcgaagaagcggttaaaaactacgaacagcagatgttcatgtacggtaaagaagcgcaggaagaatctacccagaacgaaatcgaaatgttcaaaccggacttcaccttccagcagctgctgaacgttggtggaggaggttctggaggcggtggaagtggtggcggaggtagcatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa 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