Coding

Part:BBa_K3332035

Designed by: Ruomeng Bi   Group: iGEM20_XMU-China   (2020-10-14)


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

caption

Fig 1. pLtetO_B0034_ecfp-ssrAtag(mf-lon)_B0015_pTrc-2 derivative_B0034_mf-lon_B0015

To construct this part, we moved pLtetO-1 (BBa_K3332034) ,E0020-ssrAtag(mf-lon)(BBa_K3332035) ,Terminator(BBa_B0015),pTrc-2 derivative promoter (BBa_K3332040), 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]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
abs
biology
colorCyan
directionForward
emissionCFP
emit476
excitation
excite439
kegg
lum
proteinECFP
swisspro