BBa_K2013005 1 BBa_K2013005 RBS and terephthalic acid(TPA) transporter 2016-10-11T11:00:00Z 2016-10-12T11:39:51Z This sequence is from a bacterium called Ideonella sakaiensis 201-F6 The sequence in this part is one of the sequences encoding TPA transporter. Because of it,TPA will be transported from extracellular to intracellular.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′- CGCTACTAGTATTATTAATCTTCTTGAAACGCGTCCT-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 SpeI restriction endonuclease. The second cutting procedure was performed with NcoI and PstI 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 TPA transporter. false Binglin Liu annotation2495645 1 RBS range2495645 1 1 13 annotation2496128 1 ISF6_0077 range2496128 1 20 1528 BBa_K2013005_sequence 1 tcacacaaaagggtactagatggacctgctgaacaacctggcgctgggcttcggtgtggcgtttaccctgcaaaacctggcgtacgcgttcctgggctgcattttgggaactctgatcggtgtcctgcctggccttgggccagtagcaactatcgctatgttgcttcccagcatctacgcgctggacgcgaccccggcgctgattatgctggcgggtatctactatggcgcgcaatatggtggcagcaccaccgcgatcctgattaacgttccgggcgagagcagcagcgtggttaccgccattgatggctaccagatggctcgccagggccgcgctgggccagcactggctgcggctgggttaggtagcttctttgcgggctgcgtgggcaccattgttattgcggcgttcgcgcccccgttgaccgaactggcctttaagtttggaccggcggagtatttttcgttaatggtactggggttgattggtgctgtcgtactggcctccggcagccttctgaaagcaattagcatgattatccttgggcttcttttggcgcagatcaatacagatgtaattagcggaacggctcgctatagttttgatatcccagaattaacagacggaatcggctttgttaccatcgcaatgggtgttttcggatttggagaaatcattgcaaatcttggtcagccagccgagaatcgcgaagttttcaccaaagacgtgaagggactgtggccgacccgtgaagactttaagctggcttggccagcagtagtacgtgggacggcattgggaagtttattaggtgtgttaccgggcgggggcgcattactttccagtttcgcgtcgtacacgatggagaaaaaaatggcaggcgataagggtcgttttggcaagggtgatatccgtggcgttgcgggtccggaaagcgcgaacaacgcgggtgcgcagaccagcttcattccgatgctgaccctgggtatcccgccgaacgcggtgatggcgctgatggttggcgcgatgaccattaaagatatccagccgggtccgcaagtgatgaccagcaacccgaccctgttttggggcctgattgcgagcatgtgggtgggtaacctgatgctggttatcctgaacctgccgatgatcggtatttggatcaagctgctgaccgtgccgtaccgttacctgttcccggcgattaccctgttttgctgcatcggcgtttacaccctgaacaacaacaacttcgacgtgtatatgaccgcggcgttcgcggttattggttacatcttttataagctgagctgcgaggcggccccactgttattgggttttattcttggtcccatgatggaggagaaccttcgtcgtgccttgttgctgagccgtggcgattggagcacctttgtgacccgtccgctgagcgcgggtctgctgctggcggcggttgcgatgattgtgatcgttatgctgccgagcatcaagaaaaagcgtgaggacgcgtttcaagaagattaataa 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