Part:BBa_K4047039
pGEM-SBPase (plasmid for deletion of SBPase)
This plasmid is for the deletion of glpX, encoding a bifunctional enzyme for fructose-1,6-bisphosphatase (FBPase) and sedoheptulose-1,7-bisphosphatase (SBPase) in Synechococcus elongatus PCC 7942. This plasmid was successfully integrated into transforming cells of this strain, detailed on the experience page, but was not able to achieve complete segregation (transformation of all genome copies) due to the essential nature of SBPase functionality for the cell. Successful integration was verified with PCR.
This plasmid inserts kanamycin resistance (BBa_K4047019) into the target gene, preventing functional enzyme synthesis and providing kanamycin resistance as a selective marker. This plasmid is built from the plasmid backbone BBa_K4047018, which also contains gentamicin resistance (BBa_K4047016).
Although this part lacks standard compatibility due to the enzymes available to our team, the plasmid does successfully integrate into the S. elongatus genome for the deletion of SBPase. We strongly recommend that future iGEM teams optimize this plasmid for compatibility and future work in engineering of the Calvin cycle.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1998
Illegal PstI site found at 2591 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1998
Illegal PstI site found at 2591 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1489
Illegal BglII site found at 4478
Illegal BamHI site found at 2345
Illegal XhoI site found at 2433
Illegal XhoI site found at 3675
Illegal XhoI site found at 4474 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1998
Illegal PstI site found at 2591 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1998
Illegal PstI site found at 2591
Illegal AgeI site found at 1857
Illegal AgeI site found at 4356 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2430
Illegal BsaI.rc site found at 864
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