Coding

Part:BBa_K3726063:Design

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-07)
Revision as of 13:53, 7 October 2021 by Sbarraga (Talk | contribs)

CDS_SigB* (11801)


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 XhoI site found at 364
    Illegal XhoI site found at 601
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 328
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 328
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 88

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.

additionally, we have removed an internal BsmbI site in the base 862 in order to make this pat MoClo compatible with the marburg collection. However, the Sapl and BbsI sites should be removed in order to make this part RFC1000 compatible.

Source

Coding sequence of this sigma factor has been obtained from Synechococcus elongatus PCC 11801 genome through PCR. This sequence is codified in the gen DOP62_11145 of Synechococcus elongatus PCC 11801 genome.

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.