BBa_K2013008 1 BBa_K2013008 TPA 1,2-dioxygenase 2016-10-11T11:00:00Z 2016-10-13T12:07:36Z This sequence is from a bacterium called Ideonella sakaiensis 201-F6 This part contains one of sequences coding (terephthalic acid )TPA 1,2-dioxygenase.This is the first. TPA is incorporated through the TPA transporter (TPATP) and catabolized by TPA 1,2-dioxygenase (TPADO).The result is that TPA is catabolized into 1,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate.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′-GAATTCGCGGCCGCTTCTAGAGTACTAGAGTCACACATGAGGGTACTAGATG-3′ Primer-R:5′- CAGCACATGGGCGTTTAATAATACTAGTAGCG-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 EcoRV restriction endonuclease. The second cutting procedure was performed with PstI 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 relates to encode TPA 1,2-dioxygenase. false Binglin Liu annotation2496206 1 ISF6_0227 range2496206 1 20 1267 annotation2495649 1 RBS range2495649 1 1 13 BBa_K2013008_sequence 1 tcacacatgagggtactagatgcaggacaccagcaccctgcaaccggtgcgttggagcggtccgggtctgacccgtatcccgtacggtgtttatcgtgatgcgcagctggcggcggaggaacaggcgcgtattttccaaggcgaaacctggaactacctgtgcctggaggcggaactgccggaggcgggtagctatcgtaccaccttcgtgggcgaaaccccggtggttgtggttcgtgacgcggatggtgaggtttacgcgtttgaaaaccgttgcgcgcaccgtggtgcgctgatcgcgctggagaagagcggccgtgcggaaaacttccagtgcgtgtaccacgcgtggagctataaccgtcaaggtgacctgaccggcgtggcgttcgagaagggtgttaaaggccgtggtggtatgccgccgggtttttgcaaggaagcgcacggtccgcgtaaactgcgtgtggcgaacttttgcggtctgctgttcggtagctttggcgatgaagttccgggtatcgaggaatatctgggcgaggaaatctgcgagcgtattgaacgtgttctgcacaagccggttgaggtgatcggtcgtttcacccaggcgctgccgaacaactggaaactgtacgtggaaaacgttcgtgacagctatcacgcgagcctgctgcacctgttctttaccacctttgagctgaaccgtctgagccaaaaaggtggcgtgatcgttgacgaaagcggtggcaaccacgtgagctacagcatggttgatgcgcaggcggagaaggacgcgagctaccgtgatcaaggtctgcgtagcgacgatgcgaactatcgtctgaaagacccgagcctgctggcgggcttccaggaatatggtgatggcgtgaccctgcaaatcctgagcgtgttcccgggttttgttctgcaacaaattcagaactgcctggcggtgcgtcaagttctgccgcgtggtgtggagcgtaccgaactgaactggacctacctgggctatgcggacgataccccggagcagcgtcaagttcgtctgaagcagagcaacctgatcggtccggcgggtttcattagcatggaagacggtgcggtgggtggctttgttcaacgtggtattgcgggtgcgggtgcgctggaggcggtggttgaaatgggtggcgatggcaccgcgagcagcgagggtcgtgcgaccgaagcgagcgtgcgtggtttttggaaagcgtaccgtcagcacatgggcgtttaataa 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