BBa_K2013013 1 BBa_K2013013 PCA 3.4-dioxygenase 2016-10-11T11:00:00Z 2016-10-13T12:29:54Z This sequence is from a bacterium called Ideonella sakaiensis 201-F6 This part contains one of sequences that code Protocatechuate(PCA)3.4-dioxygenase.when PCA3.4-dioxygenase works, the resultant Protocatechuate will be ring-cleaved.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′- CGCTACTAGTATTATTAACGCGCATCACCCGCCG-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 PCA3.4-dioxygenase false Binglin Liu annotation2495675 1 RBS range2495675 1 1 13 annotation2495676 1 ISF6_0627 range2495676 1 20 790 BBa_K2013013_sequence 1 tcacacaggagggtactagatgggccgtgcgccggaaccgggtgatgaatttctgatgcgtgacaccaccagccatccgccggcgtacgcgccggcgtataaaaccagcgtgctgcgcagcccgcgtaacgcgctgatcagcctgcaacagagcctgagcgaggtgaccggtccggtgttccgtgcggaggaactgggtccgctggataacgacctgatcctgaactgcgcgcacccggatcaaccgcatggtctgccgattggtgaacgtatcgtggttcacggttacgttcgcgaccagttcggtcgtccggttcgtaatgcgctggttgaagtgtggcaggcgaacgcgagcggtcgttatcgtcacaagaaagaccagtatatcggtgcgctggacccgaacttcggcggttgcggtcgtaccctgaccgatgcggacggtcattacgtttaccgtaccattcgtccgggcccgtacccgtggcgtaaccgtatcaacgattggcgtccggcgcatattcactacgcgatcagcggtgacgcgtgggcgcaacgtctgattacccagatgtacttcgagggcgatccgctgatcgcgagctgcccgattctgcaaaccgttccgagcgaagagcagattcgtggcctggttgcgctgcaagatccgggtgcgtttgtgcagctggacagccgtgcgtaccgtttcgacatcgttctgcgtggtcagcgtgcgacctggtttgagaaccgcgttcaaccgctgccggcgggtgatgcgcgttaataa 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