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<div class="col-lg-8 col-lg-offset-2"> | <div class="col-lg-8 col-lg-offset-2"> | ||
<h2>Improve.</h2> | <h2>Improve.</h2> | ||
− | <p>The Golden Gate modular cloning system we worked with provides many advantages over other assembling standards. However, the wildly used GFP | + | <p>The Golden Gate modular cloning system we worked with provides many advantages over other assembling standards. However, the wildly used GFP <a hre="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 a CDS: AATG at the 5’ end and GCTT at the 3’ end . </p> |
− | + | <p>The new, RFC[1000] compatible part was cloned into the pSB1C3 shipping vector and sequenced for verification. </p> | |
Revision as of 08:38, 25 September 2017
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 a CDS: AATG at the 5’ end and GCTT at the 3’ end .
The new, RFC[1000] compatible part was cloned into the pSB1C3 shipping vector and sequenced for verification.