BBa_K2013010 1 BBa_K2013010 DCDDH 2016-10-11T11:00:00Z 2016-10-13T12:22:17Z This sequence is from a bacterium called Ideonella sakaiensis 201-F6 This part codes 1,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate dehydrogenase(DCDDH).DCDDH is a important enzyme for our project,which can hydrolyze 1,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate to protocatechuate. 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′-GAATTCGCGGCCGCTTCTAGAGTACTAGAGTCACACAAAAGGGTACTAGATG-3′ Primer-R:5′- CGCTACTAGTATTATTACGCGGTGCCTTCCTCAAC-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 1,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate dehydrogenase false Binglin Liu annotation2495651 1 RBS range2495651 1 1 13 annotation2496209 1 DCDDH range2496209 1 20 1003 BBa_K2013010_sequence 1 tcacacaaaagggtactagatgaacgcgaccccggcggcggttgctccgccgcgtctggcgctggcggtgggtgacccgaacggtatcggtccagaaatagctttaaaggcattagccgctcttccagcagccgcgagagggcgtatcacactgtacggtccggcggcggttctggaacgtaccgcgagccagctgggtcaagaggcgctgctgcgtgaacagccggtggttgacgcgggtgcgctgccggcgcaagcggcacgccctgggcgcattgatccggcggccggggcatccgcagtagcgtctgcctcagcggctattgaggcctgccgtcgcggcgaagctgatgccgtggtggcttgtccgcatcacgaaacagccatccaccaggcgggtattgcattctcgggttacccatcattagttgcgcgtgtgtgcgggcaacctgaagacagtgtattccttatgctggttgggggcggattacgtattgtgcacgcgactttacacgagtctgttgcacacgccctggggcgtttacagccagccttgatcgccgccgccgcacgtgcgggggcacgcgcttgcgctcgcttgggagtagcggacccccgtatcggcgtgtttggaatcaacccacacgcgagcgagggcggtttgtttggaccggaagatgcacaacatgtagaaccagctgtgcaggcccttcgtgccgaagggctgcgcgttgatgggccattaggcgcggacctgctgctggcgcagcgtcgtcacgacctgtacgtggcgatgctgcacgatcaaggccacatcccggttaaactgctggcgccgaacgcggcgagcgcgctgagcattggtgcgcaagtgctgctgagcagcgttggtcacggtagcgcgatggacatcgcgggtcgtggtattgccgatccgggtgcggtgctgcgtaccattgcgctgctgagcggtgcgggtgttgaggaaggcaccgcgtaataa 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