Coding

Part:BBa_K3726064:Design

Designed by: Santiago Barrag�n Ariza   Group: iGEM21_MADRID_UCM   (2021-10-07)

CDS_SigB (7942)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 328
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 328
    Illegal NheI site found at 955
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 328
    Illegal BglII site found at 219
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 328
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 328
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

In the upstream region the bases taatc corresponds with 5bp a spacer sequence to improve ribosomal translation, while AATG overhang creates an start codon that forces translation initiation of the downstream coding sequence.

In the downstream region, the extra “aa” bases allow the creation of an additional stop codon (TAA) at the end of the upstream coding sequence, ending the translation of the desired CDS in accordance with the Marburg Collection design guidelines.

Source

Coding sequence of this sigma factor has been obtained from Synechococcus elongatus PCC 7942 genome through PCR.

References

D. Kaczmarzyk, J. Anfelt, A. Särnegrim and E. Hudson, "Overexpression of sigma factor SigB improves temperature and butanol tolerance of Synechocystis sp. PCC6803", 2021.