Difference between revisions of "Part:BBa M50055:Design"

Revision as of 08:18, 12 December 2016

Atrazine biosensor

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 760

Design Notes

We added an eGFP sequence taken from BBa_K598009 downstream of our atrazine riboswitch (BBa_M50048) so that the riboswitch would control translation of the eGFP. When we ordered the construct, DNA 2.0 also added an IPTG-inducible promoter to the beginning of our sequence. Typically, an RBS follows the promoter that is added to a plasmid. However, in this situation an RBS would be redundant and interfere with our riboswitch function; if there is an RBS before the UTR and aptamer, a ribosome could bind there and disrupt the shape of the riboswitch. For this reason, it was necessary to request a custom plasmid design that did not include the standard RBS.

Source

Riboswitch components: BBa_J89000

Atrazine aptamer: In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against Atrazine, IJMS International Journal of Molecular Sciences

eGFP sequence: BBa_K598009

References

https://parts.igem.org/Part:BBa_K598009

https://parts.igem.org/Part:BBa_J89000

Williams, R., Crihfield, C., Gattu, S., Holland, L., & Sooter, L. (2014). In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against Atrazine. IJMS International Journal of Molecular Sciences, 15(8), 14332-14347. doi:10.3390/ijms150814332

@@ Line 8: / Line 8: @@
 ===Design Notes===
-We added an eGFP sequence taken from BBa_K598009 downstream of our atrazine riboswitch (BBa_M50048) so that the riboswitch would control translation of the eGFP.
+We added an eGFP sequence taken from BBa_K598009 downstream of our atrazine riboswitch (BBa_M50048) so that the riboswitch would control translation of the eGFP. When we ordered the construct, DNA 2.0 also added an IPTG-inducible promoter to the beginning of our sequence. Typically, an RBS follows the promoter that is added to a plasmid. However, in this situation an RBS would be redundant and interfere with our riboswitch function; if there is an RBS before the UTR and aptamer, a ribosome could bind there and disrupt the shape of the riboswitch. For this reason, it was necessary to request a custom plasmid design that did not include the standard RBS.
 ===Source===