Part:BBa_K2170060
Improved Killswitch (CMV-GMK_TK90-polyA) in RFC[10]
Since the Killswitchs of Slovenia 2012 BBa_K782063 and Freiburg 2007 BBa_K404113 both contain a forbidden PstI site we wanted to improve these parts, because safety mechanisms are of extremely high importance. This is why we created the RFC[10] compatible BioBrick BBa_K2170150 (pCMV-GMK_TK90), and this part (pCMV-GMK_TK90-polyA), that contains an additional poly adenylation site for improved mRNA stability. These improved parts are characterized with respect to mRNA expression level and functionality in cell culture.
Quantitative analysis by mRNA measuring of Poly-A-sequence addition showed 4.6-fold higher mRNA stability in comparison to the non-Poly-A-construct. [http://2016.igem.org/Team:LMU-TUM_Munich/Safety More information]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 576
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1241
Illegal NgoMIV site found at 2146 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2669
Illegal SapI.rc site found at 633
None |