Part:BBa_K3726063:Design
CDS_SigB* (11801)
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 328
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 328
Illegal NheI site found at 955 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 328
Illegal XhoI site found at 364
Illegal XhoI site found at 601 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 328
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 328
- 1000INCOMPATIBLE 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 site 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.