Difference between revisions of "Part:BBa K3332035"

Revision as of 10:24, 27 October 2020

E0020-ssrAtag（mf-lon）

Engineered cyan fluorescent protein tagged with ssrAtag(mf-lon), which can be recognised and degraded by mf-lon .We use it to model the feedback of mf-lon and check the function of mf-lon.

ssAtag（mf-lon）is tag pdt#1，which is recognized by mf-Lon hydrolases. It is attached to the 3’ end of the genes of ECFP. If there is mf-lon hydrolases, ECFP will be degraded.

Usage and Biology

Fig.1 Circuit

To construct this part, we moved pLtetO-1 (BBa_K3332034) ，E0020-ssrAtag（mf-lon）(BBa_K3332035) ，Terminator(BBa_B0034)，pTrc-2 derivative promoter (BBa_K3332040) and RBS(BBa_B0034) and mf-lon(BBa_K2333011) into the expression vector pUC57 by standard assembly. Then the ligation mixture was transformed into E. coli BL21(DE3) to characterize mf-Lon hydrolase.

Reference:

[1] Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ. 'Deadman' and 'Passcode' microbial kill switches for bacterial containment. Nat Chem Biol. 2016;12(2):82-86. doi:10.1038/nchembio.1979

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
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COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 Fig.1 Circuit
-To construct this part, we moved pLtetO-1 (BBa_K3332034) ，E0020-ssrAtag（mf-lon）(BBa_K3332035) ，Terminator(BBa_B0034)，pTrc-2 derivative promoter (BBa_K3332040) and RBS(BBa_B0034) and mf-lon(BBa_K2333011) into the expression vector pUC57 by standard assembly. Then the ligation mixture was transformed into E. coli BL21 to characterize mf-Lon hydrolase.
+To construct this part, we moved pLtetO-1 (BBa_K3332034) ，E0020-ssrAtag（mf-lon）(BBa_K3332035) ，Terminator(BBa_B0034)，pTrc-2 derivative promoter (BBa_K3332040) and RBS(BBa_B0034) and mf-lon(BBa_K2333011) into the expression vector pUC57 by standard assembly. Then the ligation mixture was transformed into E. coli BL21(DE3) to characterize mf-Lon hydrolase.
 ===Reference:===