Device

Part:BBa_K2170060

Designed by: Max Mustermann   Group: iGEM16_LMU-TUM_Munich   (2016-10-11)


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]

Muc16 Killswicht 001.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 576
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1241
    Illegal NgoMIV site found at 2146
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2669
    Illegal SapI.rc site found at 633


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Categories
Parameters
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