Part:BBa_K906010:Design
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In the design of this part, the RBS was taken from Synechocystis PCC 6803 so provide the following coding region ample translation. This RBS was also shown to work in E. coli by the 2010 USU iGEM team . The restriction sites were added to allow for modularity in the composite part K906103. Spacers between the restriction sites were added to prevent a decrease in cutting efficiency near the end of the linearized DNA (A phenomena noted on New England Biolab’s page: http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp#.UGzuRvl25go). Two restriction sites were added to allow for easy ligation of parts. Instead of requiring restriction sites with identical overhangs, the user can synthesize complementary primers with both required restriction sites. The user can then cut the primer with the same endonucleases as the DNA and, using the primer dimer as a “linker,” the DNA can be easily ligated together. In this way, the order of the genes may be easily interchanged so that future teams may be able to characterize the change in production caused by reordering the genes.
Source
From Synechocystis sp. PCC6803 sequence for PsbA2 promoter
http://www.ncbi.nlm.nih.gov/gene/951890