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 annotation2489538 1 protein linker range2489538 1 1172 1215 annotation2489537 1 TetX range2489537 1 1 1171 annotation2489539 1 GFP range2489539 1 1216 1935 BBa_K2150013_sequence 1 atgaaaaaaaccatgcgtatcgacaccgacaaacagatgaacctgctgtctgacaaaaacgttgcgatcatcggtggtggtccggttggtctgaccatggcgaaactgcttcagcagaacggtatcgacgtttctgtttacgaacgtgacaacgaccgtgaagcgcgtatcttcggtggtaccctggacctgcacaaaggttctggtcaggaagcgatgaaaaaagcgggtctgcttcagacctactacgacctggcgctgccgatgggtgttaacatcgcggacaaaaaaggtaacatcctgtctaccaaaaacgttaaaccggaaaaccgtttcgacaacccggaaatcaaccgtaacgacctgcgtgcgatcctgctgaactctctggaaaacgacaccgttatctgggaccgtaaactggttatgctggaaccgggtaaaaaaaaatggaccctgaccttcgaaaacaaaccgtctgaaaccgcggacctggttatcctggcgaacggtggtatgtctaaagttcgtaaattcgttaccgacaccgaagttgaagaaaccggtaccttcaacatccaggcggacatccaccagccggaaatcaactgcccgggtttcttccagctgtgcaacggtaaccgtctgatggcgtctcaccagggtaacctgctgttcgcgaacccgaacaacaacggtgcgctgcacttcggtatctctttcaaaaccccggacgaatggaaaaaccagacccaggttgacttccagaaccgtaactctgttgttgacttcctgctgaaagagttctctgactgggacgaacgttacaaagaactgatccacaccaccctgtctttcgttggtctggcgacccgtatcttcccgctggaaaaaccgtggaaatctaaacgtccgctgccgatcaccatgatcggtgacgcggcgcacctgatgccgccgttcgcgggtcagggtgttaactctggtctggttgacgcgctgatcctgtctgacaacctggcggacggtaaattcaactctatcgaagaagcggttaaaaactacgaacagcagatgttcatgtacggtaaagaagcgcaggaagaatctacccagaacgaaatcgaaatgttcaaaccggacttcaccttccagcagctgctgaacgttggtggaggaggttctggaggcggtggaagtggtggcggaggtagcatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa 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