Composite

Part:BBa_K3796209:Design

Designed by: Yang Feiyue   Group: iGEM21_CAU_China   (2021-09-24)


PgsiB-LacIq-terminator-Patp2-LacIq-terminator-Ptac-ndoA-terminator


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1492
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1492
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1492
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1492
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1492
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We added functional rbs sequences for Corynebacterium glutamicum between each promoter and coding region to ensure translation.

We added a short sequence of 8~11bp between each rbs and the following coding region to ensure higher translational efficiency.


Source

P-gsiB is initially from Bacillus subtilis and P-atp2 is initially from Corynebacterium glutamicum. In our work, these two promoters and following rbs are chemically synthesized.

LacIq and tac promoter are amplified from the plasmid pXMJ19.

The toxin gene ndoA is amplified from the genome of Bacillus subtilis subsp. subtilis str. 168.

T7 terminator is synthesized and rrnB terminators are amplified from the plasmid pXMJ19.

References