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Team:Warwick/Parts
From 2014.igem.org
5' Promoter
This is derived from the 5i UTR of the HCV virus strain 1b isolate Con1. The secondary structure of this sequence acts as a binding site and initiates replication of the system by RdRp. It also includes the first 16 amino acids of the first gene of HCV as this has been shown to increase the efficacy of binding of the RdRp to the 5' promoter.
IRES
This acts as an initiation for eukaryotic ribosomes and begins translation of the following protein sequence. We compared two different IRESs; the classical EMCV IRES used in many papers and has been shown to be compatible with replicons
The NKRF IRES derived from the 3’UTR of the mammalian NF-kappaB repressing factor was also tested as during investigation as to the efficacy and strength of the EMCV IRES, NKRF derived IRES was shown to be 30-fold more efficient however never used in Huh7, that we chose to design our replicon for.
MS2 Box
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Neomycin Resistance
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Aptazyme
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siRNA
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MS2 Coat Protein
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P2A
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RdRp
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3' Promoter
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