Difference between revisions of "Part:BBa K2020051"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | ====Incorporation of DMNBS==== | ||
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| + | ====Assembly in a synthetase plasmid for incorporation of ncAA==== | ||
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| + | [[File:T--Aachen--Mj YRS CUA.jpg|200px|thumb|left|pACYC derived plasmid with Mj tyrosyl synthetase and cognate tRNA]] | ||
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| + | Most synthetases are used with low copy plasmids (e.g. pACYC). Assemble the tRNA and the synthetase into a low copy plasmid, each one with an own promoter and one terminator for both. (See picture). If your application is not for incorporation into a protein but for use with a second plasmid, make shure to use replicons from different incompatibility groups, eg. ColE1 and p15A and different selection markers. A second plasmid could be the [[Part:BBa_K2020040|flourescent reporter plasmid pFRY]] for the purpose of determining fidelity and efficiacy of synthetases for ncAA. | ||
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Revision as of 09:15, 15 October 2016
wild type tyrosyl synthetase for use in E.coli with amber anticodon and Y32G
This is a DMNBS-synthetase to be used as a orthogonal synthetase in E.coli. This part can be used together with the cognate tRNA BBa_K2020042 to incorporate DMNBS in response to an amber stop codon.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 84
Illegal SapI.rc site found at 877