Device

Part:BBa_K2233002:Design

Designed by: Kyosuke Kita   Group: iGEM17_Kobe   (2017-10-21)


lacZ for monitoring amtB gene expression in B.subtilis chromosome


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 5434
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 592


Design Notes

Restriction site for EcoRI was removed by base substitution using IDT service. This part was sequenced, and there was a synonymous point mutation in CDS of lacZ.

Source

lacZ and cat genes (with promoter+RBS) were amplified from commercialized plasmids, pMutin2 and pSweet respectively. Sequence for recombination (Recombination site B) was synthesized by IDT. Recombination site A was amplified by PCR from B.subtilis strain 168 chromosome. These DNA fragments were assembled together by Gibson assembly method.

References