BBa_K511206 1 BBa_K511206 LacI-miRFF4 Repressor MammoBlock 2011-09-26T11:00:00Z 2015-05-08T01:12:31Z This part is a concatenation of a mammalian codon-optimized version of the LacI repressor originally derived from Escherichia coli and the synthetic FF4 miRNA sequence. This part encodes a dual transcriptional/translational repression system including a protein that binds tightly to both the lactose operator and an intronic micro RNA (miRNA) that binds tightly to the FF4 sequence. LacI-miRFF4 is best used with an FF4-tagged protein that is transcribed by a promoter that contains a lactose operator, allowing for combined repression at a transcriptional and translational level. Because transcriptional repression from LacI and miRNA-mediated degradation of FF4-tagged mRNA are relieved once the respective elements have been degraded or diluted in a cell, the LacI-miRFF4 construct is particularly well-suited to circuits that require strong but transient repression, such as a fast toggle switch. false false _674_ 0 6719 9 It's complicated false Because the FF4 miRNA was designed to bind mRNA in a three-dimensional manner, the FF4 sequence has significant secondary structure. This makes it difficult to design primers that will capture the miRFF4 region. To work around this problem, we suggest using restriction cloning rather than PCR-based methods to isolate either the LacI-miRFF4 part or the miRFF4 region specifically. false Grant Robinson annotation2143922 1 LacI Repressor range2143922 1 1 1080 BBa_K511206_sequence 1 atgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagccaaaaaagaagagaaaggtctaataggtgctcgcttcggcagcacatatactatgttgaatgaggcttcagtactttacagaatcgttgcctgcacatcttggaaacacttgctgggattacttcttcaggttaacccaacagaaggctcgagtgctgttgacagtgagcgccgcttgaagtttttaattaaatagtgaagccacagatgtatttaattaaagacttcaagcggtgcctactgcctcggagaattcaaggggctactttaggagcaattatcttgtttactaaaactgaataccttgctatctctttgatacatttttacaaagctgaattaaaatggtataaattaaatcacttttttcaattg 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