BBa_K2013016 1 BBa_K2013016 catechol 2，3dioxygenase 2016-10-11T11:00:00Z 2016-10-15T08:15:41Z This sequence is from Escherichia coli Strain MG1655 This part contains the sequence of coding catechol 2，3dioxygenase which can catalyze catechol into 2-hydroxymuconate semialdehyde.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:KOD Primer-F:5′-GAATTCGCGGCCGCTTCTAGAGTACTAGAGTCACACAGGAGGGTACTAGATG-3′ Primer-R:5′- CGCTACTAGTATTATTACCACGCGGTGTGATACAG-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 PstI restriction endonuclease. The second cutting procedure was performed with KpnI and NcoI 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 that encodes catechol 2，3dioxygenase false Demin Xu annotation2496051 1 dioxygenase range2496051 1 20 1171 annotation2496050 1 RBS range2496050 1 1 13 BBa_K2013016_sequence 1 tcacacaggagggtactagatggcgaaccgtatgctggtgaacgaaaccgcgtggttcggtcgtggtgcggtgggtgcgctgaccgacgaggttaagcgtcgtggctaccagaaagcgctgatcgtgaccgataagaccctggttcaatgcggtgtggttgcgaaggtgaccgacaaaatggatgcggcgggtctggcgtgggcgatctacgacggcgtggttccgaacccgaccattaccgtggttaaagaaggcctgggtgttttccagaacagcggtgcggactatctgatcgcgattggtggcggtagcccgcaagatacctgcaaggcgattggtatcattagcaacaacccggaatttgcggacgtgcgtagcctggagggcctgagcccgaccaacaaaccgagcgttccgatcctggcgattccgaccaccgcgggtaccgcggcggaagtgaccatcaactacgttattaccgatgaggaaaagcgtcgtaaattcgtgtgcgttgacccgcacgatatcccgcaggtggcgtttattgacgcggatatgatggacggtatgccgccggcgctgaaggcggcgaccggcgttgatgcgctgacccacgcgatcgaaggttatattacccgtggcgcgtgggcgctggcggacgcgctgcacatcaaggcgattgagatcattgcgggtgcgctgcgtggtagcgtggcgggtgacaaagatgcgggcgaggaaatggcgctgggccaatacgttgcgggcatgggtttcagcaacgtgggcctgggtctggttcacggtatggcgcacccgctgggtgcgttttataacaccccgcacggtgtggcgaacgcgatcctgctgccgcacgttatgcgttacaacgcggacttcaccggcgaaaagtatcgtgatattgcgcgtgtgatgggtgtgaaagttgaaggcatgagcctggaggaagcgcgtaacgcggcggtggaggcggtttttgcgctgaaccgtgacgttggtatcccgccgcacctgcgtgacgtgggcgttcgtaaagaggatattccggcgctggcgcaggcggcgctggacgatgtgtgcaccggcggtaacccgcgtgaggcgaccctggaagatatcgttgagctgtatcacaccgcgtggtaataa 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