Part:BBa_K1332009
mRNA circularization device (3 side) (endless translation)
This part consists of the 5´ side of the intron in td gene of T4 phage and a double terminator. The protein coding sequence that is inserted between this device and mRNA circularization device (5’ side) can be circularized. If you circularized the protein coding sequence (It’s a stop codon have been removed.), you can get a circular mRNA that is translated semi-permanently.
How to use
You need modifying the prefix and suffix of a protein coding sequence.
There is a stop codon “TAG” in a restriction site in the prefix, so insert “AG” just before the stop codon to shift a reading frame. (figure 1)
Figure 1
There is a translation termination codon in the protein coding frame, so delete a part of the sequence to remove the translation termination codon. (figure 2)
Figure 2
A circular mRNA consists of RBS, the protein coding sequence and 56bp fragments of the mRNA circularization device. (figure 3)
Figure 3. Red"GT" is removed.
Adjust the length of a circular mRNA to multiples of 3 to keep a pattern of the reading frame.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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