Difference between revisions of "Part:BBa K2323006"

Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2323006 short</partinfo>
 
<partinfo>BBa_K2323006 short</partinfo>
 +
This GFP (E0040) is tagged with a protein degradation tag (pdt) called LAA. This causes the protein to degrade in E.coli with a first-order rate of 0.0327 per min, from the ClpP machinery of E.coli. It is under the control of a pTet promoter (aTc inducible) and the RBS is B0034. The terminator is B0015.
  
 
+
<!-- -->
pTet promoter, RBS B0034, GFP E0040, LAA degradation tag, Terminator B0015.  
+
This biobrick is part of a collection of degradation tags, characterized on the page [https://parts.igem.org/Part:BBa_K2323003 BBa_K2323003]. See also [https://parts.igem.org/Part:BBa_K2323005 BBa_K2323005], [https://parts.igem.org/Part:BBa_K2323007 BBa_K2323007], [https://parts.igem.org/Part:BBa_K2323008 BBa_K2323008].
This GFP is tagged by an LAA degradation tag to be recognized by ClpP protease from E.coli
+
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 13: Line 13:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2323006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2323006 SequenceAndFeatures</partinfo>
 +
===Plasmid composition===
 +
[[File:BBa_K2323006.png| frame | 400px | center | The collection is cloned from [https://parts.igem.org/Part:BBa_I13522 BBa_I13522] with overhang PCR using 5'-phosphated primers that contain the tag as an overhang. Primers are indicated with the map ([https://benchling.com/s/9bY7WB1X read-only benchling map]). Cloning was confirmed with sequencing. ]]
  
  

Revision as of 13:40, 30 October 2017


GFP-LAA This GFP (E0040) is tagged with a protein degradation tag (pdt) called LAA. This causes the protein to degrade in E.coli with a first-order rate of 0.0327 per min, from the ClpP machinery of E.coli. It is under the control of a pTet promoter (aTc inducible) and the RBS is B0034. The terminator is B0015.

This biobrick is part of a collection of degradation tags, characterized on the page BBa_K2323003. See also BBa_K2323005, BBa_K2323007, BBa_K2323008.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 724

Plasmid composition

The collection is cloned from BBa_I13522 with overhang PCR using 5'-phosphated primers that contain the tag as an overhang. Primers are indicated with the map (read-only benchling map). Cloning was confirmed with sequencing.