Coding

Part:BBa_K3697000

Designed by: Christopher Neimeth, Lauren Ramlan   Group: iGEM20_Stanford   (2020-10-22)
Revision as of 00:20, 24 October 2020 by Lramlan (Talk | contribs)


mCherry BSU

This is the coding sequence for producing a codon-optimized mCherry in B. subtilis. mCherry is a derivative of RFP. mCherry_BSU was created by codon-optimizing an E. coli mCherry for expression B. subtilis. When paired with a strong RBS and constitutive promotor (BBa_K3697010), mCherry_BSU can be quantified using a plate reader, and transformed B. subtilis colonies can be moderately red to the naked eye.

mCherry_BSU has an excitation peak at 585 nm and a peak emission at 615 nm. Color is readily visible in E. coli and B. subtilis in under 12 hours of incubation.

Fluorescence levels in B. subtilis and E. coli can be quantified using a fluorescent plate reader. Under high enough expression from a strong constitutive promotor such as pVeg (BBa_K143012), red colonies may be visible to the naked eye. E. coli expressing mCherry_BSU are red under natural light. E. coli liquid cultures may appear red to the naked eye, while B. subtilis liquid cultures likely will not appear red.

This protein can be used as a reporter in B. subtilis due to it's high visibility.

This part does not have a degradation tag.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4
    Illegal AgeI site found at 715
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None