CMV promoter - MLS - GFP - hGH polyA tail
In characterization of Mitochondrial Leader Sequence(MLS) (BBa_K1119001), the CDS of MLS was assembled in frame with that of GFP reporter using Freiburg’s RFC25 format(BBa_K648013). The translation unit was driven by CMV promoter (BBa_K1119006) and terminated by hGH polyA signal (BBa_K404108). This construct was used for characterization of MLS BioBrick(BBa_K1119001) .
This MLS-GFP generator (BBa_K1119009) was transfected into HEK293FT cells. Mitochondria were stained after transfection and co-localization was determined by area of signal that overlapped.
To provide a positive control, CDS of EGFP from pEGFP-N1 (Clontech) was inserted downstream and in frame with the CDS of the MLS in the commercial plasmid pCMV/myc/mito, (Invitrogen, Carlsbard, CA). A negative control was made by GFP generator that does not contains the CDS of MLS (BBa_K1119008).
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 614
- 23COMPATIBLE WITH RFC
- 25Illegal NgoMIV site found at 628
Illegal AgeI site found at 1462
- 1000Illegal BsaI site found at 1865
Illegal BsaI.rc site found at 1367