Difference between revisions of "Part:BBa K1332009"

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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)<br>
 
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)<br>
 
[[File:prefix.png|500px|]]<br>
 
[[File:prefix.png|500px|]]<br>
 +
<b>Figure 1</b><br>
 
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)<br>
 
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)<br>
 
[[File:suffix.png|500px|]]<br>
 
[[File:suffix.png|500px|]]<br>
 +
<b>Figure 2</b><br>
 
A circular mRNA consists of RBS, the protein coding sequence and 56bp fragments of the mRNA circularization device. (figure 3)<br>
 
A circular mRNA consists of RBS, the protein coding sequence and 56bp fragments of the mRNA circularization device. (figure 3)<br>
 
[[File:cyc-seq.png|500px|]]<br>
 
[[File:cyc-seq.png|500px|]]<br>
 +
<b>Figure 3. Red"GT" is removed.</b><br>
 
Adjust the length of a circular mRNA to multiples of 3 to keep a pattern of the reading frame.
 
Adjust the length of a circular mRNA to multiples of 3 to keep a pattern of the reading frame.
  

Revision as of 17:58, 10 October 2014

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)
Prefix.png
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)
Suffix.png
Figure 2
A circular mRNA consists of RBS, the protein coding sequence and 56bp fragments of the mRNA circularization device. (figure 3)
Cyc-seq.png
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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]