Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Grant Robinson
Date created: 2011-09-26 11:00:00
Date modified: 2015-05-08 01:12:31
LacI-miRFF4 Repressor MammoBlock
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K511206_sequence (Version 1) |
Description
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.Notes
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.Source
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.| Sequence Annotation | Location | Component / Role(s) |
| LacI Repressor | 1,1080 | CDS feature/protein |