BBa_K2013003 1 BBa_K2013003 RBS and MHETase 2016-10-11T11:00:00Z 2016-10-12T11:26:25Z This sequence is from a bacterium called Ideonella sakaiensis 201-F6 This part contains the coding sequence of MHETase which can hydrolyzes MHET to TPA and EG. MHETase is second enzymes on the downstream of PETase in the bacterium Ideonella sakaiensis 201-F6 that Japanese scientists found.MHET is degraded into two kinds of natural environment harmless substances: terephthalic acid and ethylene glycol. We carried out a codon optimization on this part.In addition，We carried out codon optimization on this part. Experimental Validation This part is validated through Four ways: amplification, PCR, Enzyme cutting and Sequence. Amplification Enzyme:Q5 Primer-F:5′- GAATTCGCGGCCGCTTCTAGAGTACTAGAGTCACACAAAAGGGTACTAGATG-3′ Primer-R:5′- CGCTACTAGTATTATTACGGCGGAGCCGCGCAC-3′ Results PCR Enzyme:Taq Primer-F:5′-CCACCTGACGTCTAAGAAAC-3′ Primer-R:5′-GTATTACCGCCTTTGAGTGA-3′ Results Double digestion After the assembly ,the plasmid was transferred into the Competent E. coli top10. After culturing overnight in LB,we minipreped the plasmid for double digestion .The first cutting procedure was performed with EcoRI and HindIII restriction endonuclease. The second cutting procedure was performed with PstI and BaHI restriction endonuclease. The plasmid was cutted in a 25μL system at 37 ℃ for 1 hours. The Electrophoresis was performed on a 1％ Agarose glu. Results false false _2480_ 32794 32794 9 false This is a Translational units part encoding MHETase. false Binglin Liu annotation2495606 1 RBS range2495606 1 1 13 annotation2495607 1 MHETase range2495607 1 20 1842 BBa_K2013003_sequence 1 tactagagtcacacaaaagggtactagatgcagaccaccgtgaccaccatgctgctggcgagcgttgcgctggcggcgtgcgcgggtggcggtagcaccccgctgccgctgccgcagcaacagccgccgcaacaggagccgccgccgccgccggtgccgctggcgagccgtgcggcgtgcgaagcgctgaaggatggtaacggtgacatggtgtggccgaacgcggcgaccgtggttgaggttgcggcgtggcgtgatgcggcgccggcgaccgcgagcgcggcggcgctgccggagcactgcgaagttagcggtgcgatcgcgaagcgtaccggtattgacggctacccgtatgaaatcaaatttcgtctgcgtatgccggcggagtggaacggtcgtttctttatggaaggcggtagcggtaccaacggcagcctgagcgcggcgaccggtagcatcggcggtggccaaattgcgagcgcgctgagccgtaactttgcgaccattgcgaccgatggtggccacgacaacgcggtgaacgataacccggacgcgctgggtaccgttgcgttcggcctggatccgcaggcgcgtctggacatgggttacaacagctatgatcaagtgacccaggcgggcaaggcggcggttgcgcgtttctacggtcgtgcggcggacaaaagctattttatcggttgcagcgagggtggccgtgaaggcatgatgctgagccaacgtttcccgagccactacgatggtatcgtggcgggtgcgccgggttatcagctgccgaaggcgggtattagcggcgcgtggaccacccaaagcctggcgccggcggcggtgggtctggacgcgcagggcgttccgctgattaacaaaagctttagcgacgcggatctgcacctgctgagccaagcgatcctgggtacctgcgatgcgctggacggtctggcggatggcattgttgacaactaccgtgcgtgccaggcggcgttcgatccggcgaccgcggcgaacccggcgaacggtcaagcgctgcaatgcgtgggtgcgaagaccgcggactgcctgagcccggtgcaagttaccgcgatcaaacgtgcgatggcgggtccggttaacagcgcgggcaccccgctgtacaaccgttgggcgtgggatgcgggtatgagcggcctgagcggtaccacctataaccaaggttggcgttcctggtggctgggcagctttaacagcagcgcgaacaacgcgcagcgtgtgagcggtttcagcgcgcgtagctggctggttgactttgcgaccccgccggagccgatgccgatgacccaggtggcggcgcgtatgatgaagttcgactttgatatcgacccgctgaaaatttgggcgaccagcggtcaattcacccagagcagcatggattggcacggtgcgaccagcaccgatctggcggcgtttcgtgaccgtggtggcaagatgatcctgtaccacggtatgagcgacgcggcgttcagcgcgctggataccgcggactactatgaacgtctgggtgcggcgatgccgggtgcggcgggtttcgcgcgtctgtttctggtgccgggtatgaaccattgcagcggtggtccgggcaccgatcgttttgacatgctgaccccgctggttgcgtgggttgagcgtggtgaagcgccggatcaaattagcgcgtggagcggcaccccgggttatttcggtgttgcggcgcgtacccgtccgctgtgcccgtacccgcagatcgcgcgttataaaggtagcggcgacattaacaccgaggcgaacttcgcgtgcgcggctccgccgtaataa 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