Difference between revisions of "Team:BOKU-Vienna/Improve"
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<h2>Improve.</h2><br> | <h2>Improve.</h2><br> | ||
<p>The Golden Gate modular cloning system we worked with provides many advantages over other assembling standards. However, the wildly used GFP <a href="http://parts.igem.org/Part:BBa_E0040" target="_blank">BBa_E0040</a> part is not compatible with this standard (RFC[1000]). Therefore, we decided to improve this part by silently mutating a BsaI restriction site to comply with RFC[1000]. Additionally we added the special RFC[1000] <a href="http://parts.igem.org/partsdb/scars.cgi" target="_blank">prefix and suffix </a> with BsaI restriction sites. After digestion they create so called <a href="https://2014.igem.org/Team:BostonU/MoClo" target="_blank">fusion sites</a>, sticky ends which are specific for coding sequences: AATG at the 5’ end and GCTT at the 3’ end . </p> | <p>The Golden Gate modular cloning system we worked with provides many advantages over other assembling standards. However, the wildly used GFP <a href="http://parts.igem.org/Part:BBa_E0040" target="_blank">BBa_E0040</a> part is not compatible with this standard (RFC[1000]). Therefore, we decided to improve this part by silently mutating a BsaI restriction site to comply with RFC[1000]. Additionally we added the special RFC[1000] <a href="http://parts.igem.org/partsdb/scars.cgi" target="_blank">prefix and suffix </a> with BsaI restriction sites. After digestion they create so called <a href="https://2014.igem.org/Team:BostonU/MoClo" target="_blank">fusion sites</a>, sticky ends which are specific for coding sequences: AATG at the 5’ end and GCTT at the 3’ end . </p> | ||
| − | <p>The new, RFC[1000] compatible part <a href="http://parts.igem.org/Part:BBa_K2294444" target="_blank">BBa_K2294444</a> was cloned into the pSB1C3 shipping vector | + | <p>The new, RFC[1000] compatible part <a href="http://parts.igem.org/Part:BBa_K2294444" target="_blank">BBa_K2294444</a> was cloned into the pSB1C3 shipping vector.</p> |
Revision as of 16:51, 1 October 2017
Improve
Improve.
The Golden Gate modular cloning system we worked with provides many advantages over other assembling standards. However, the wildly used GFP BBa_E0040 part is not compatible with this standard (RFC[1000]). Therefore, we decided to improve this part by silently mutating a BsaI restriction site to comply with RFC[1000]. Additionally we added the special RFC[1000] prefix and suffix with BsaI restriction sites. After digestion they create so called fusion sites, sticky ends which are specific for coding sequences: AATG at the 5’ end and GCTT at the 3’ end .
The new, RFC[1000] compatible part BBa_K2294444 was cloned into the pSB1C3 shipping vector.
