Difference between revisions of "Part:BBa K2020052"
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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.
 
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.
  
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Compared to incorporation of tyrosine with [[Part:BBa_K2020050|wild type Mj Y-Synthetase]] in response to an amber codon:
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*Incorporation Efficiency: % (DMNBS incorporation value)
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*Incorporation Fidelity: %(descrimination againt tyrosine)
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===Full Set of DMNBS synthtetases===
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**[[Part:BBa_K2020052|DMNBS-RS Clone 1]]
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**[[Part:BBa_K2020053|DMNBS-RS Clone 2]]
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**[[Part:BBa_K2020054|DMNBS-RS Clone 3]]
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**[[Part:BBa_K2020055|DMNBS-RS Clone 4]]
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**[[Part:BBa_K2020056|DMNBS-RS Clone 5]]
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**[[Part:BBa_K2020057|DMNBS-RS Clone 6]]
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**[[Part:BBa_K2020058|DMNBS-RS Clone 7]]
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**[[Part:BBa_K2020059|DMNBS-RS Clone 8]]
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**[[Part:BBa_K2020060|DMNBS-RS Clone 9]]
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**[[Part:BBa_K2020061|DMNBS-RS Clone 10]]
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===Usage and Biology===
 
===Usage and Biology===
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==== incorporation of ncAA====
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====Assembly in a synthetase plasmid for incorporation of ncAA====
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[[File:T--Aachen--DMNBSRS.png|200px|thumb|left|alt text]]
 
[[File:T--Aachen--DMNBSRS.png|200px|thumb|left|alt text]]
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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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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 12:42, 17 October 2016


DMNBS-Synthetase for use in E.coli

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.

Compared to incorporation of tyrosine with wild type Mj Y-Synthetase in response to an amber codon:

  • Incorporation Efficiency: % (DMNBS incorporation value)
  • Incorporation Fidelity: %(descrimination againt tyrosine)

Full Set of DMNBS synthtetases

Usage and Biology

incorporation of ncAA

Assembly in a synthetase plasmid for incorporation of ncAA

alt text

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 flourescent reporter plasmid pFRY for the purpose of determining fidelity and efficiacy of synthetases for ncAA.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 84
    Illegal SapI.rc site found at 877