Regulatory
P-erm

Part:BBa_K3183000:Design

Designed by: Natasha Cooke   Group: iGEM19_Oxford   (2019-09-20)


Erythromycin Constitutive Promoter

The part was identified in the article by Lizier et al; the gene was codon optimised for L. reuteri and synthesized by IDT. The part was leter assembled by Gibson Assembly into composite parts such as: BBa_K3183103, BBa_K3183100 etc., which were finally assembled into the pTRKH3 vector (BBa_K3183050), again by Gibson Assembly


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]


Design Notes

This promoter was specifically selected as an ideal promoter for L. reuteri because it conformed to the RF10, RF12, RF21, RF23, RF25, and RF1000 standards.

Source

It is the erythromycin ribosomal methylase (ermB) promoter from the broad-host range plasmid pAMβ1 isolated from Enterococcus faecalis. We ordered the sequence as a gBlock from IDT.

References

Lizier, Michela, et al. “Comparison of Expression Vectors in Lactobacillus Reuteri  Strains.” FEMS Microbiology Letters, vol. 308, no. 1, Aug. 2010, pp. 8–15., doi:10.1111/j.1574-6968.2010.01978.x.