Part:BBa_K1895998
SmtA metallothionein and LVA tag.
This is part BBa_K519010, SmtA which is a metallothionein that can bind to heavy metal ions. This variant has an LVA degradation tag before the stop codon which reduces its half-life in the cell.
Usage and Biology
As part of work on our arabinose controlled ‘variable resistor’. We synthesised two variants of our construct, one with and one without an ssRA degradation tag. The aim of this was to allow us to see if we can gain finer control over the resistance by increasing the rate of protein degradation. Both variants of the construct include a polyhistidine-tag to allow for protein purification from cultures.
This part is a component of the larger resistor part, consisting of the metallothionein SmtA (BBa_K519010) and a protein degradation tag. We have used an ssRA protein degradation tag. This causes the tagged protein to be degraded by the proteases ClpXP or ClpAP. There are a number of tag sequences, which are variants of AANDENYALAA, with the last three amino acids varying. The last three amino acids determine the half-life of the protein. LVA is a fast protein degradation tag. We chose to use this to ensure that the resistance is reduced quickly after removal of arabinose.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 93
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 93
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 93
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 93
Illegal AgeI site found at 121 - 1000COMPATIBLE WITH RFC[1000]
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