Difference between revisions of "Part:BBa K1065302:Design"
Line 8: Line 8:
 
This part has been created first by inserting the RBS sequence between pFixK2 and amilGFP (in Bba_K952003) through mutagenesis PCR  with a specific primer.  <br>Furthermore we decided to regulate transcription with an upstream promoter, pLac (BBa_R0010).
 
This part has been created first by inserting the RBS sequence between pFixK2 and amilGFP (in Bba_K952003) through mutagenesis PCR  with a specific primer.  <br>Furthermore we decided to regulate transcription with an upstream promoter, pLac (BBa_R0010).
  
 +
 +
Part  <partinfo>Bba_k952003</partinfo> was missing an RBS before the amilGFP CDS. We inserted the RBS by mutagenesis, added a pLAC promoter in front of the device and charachterized it.
 
===Source===
 
===Source===
  

Revision as of 16:20, 24 September 2013

Blue light sensing device without inverter for the production of amilGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 770
    Illegal NgoMIV site found at 842
    Illegal NgoMIV site found at 932
    Illegal NgoMIV site found at 950
    Illegal NgoMIV site found at 1462
    Illegal NgoMIV site found at 1755
    Illegal NgoMIV site found at 1849
    Illegal AgeI site found at 484
    Illegal AgeI site found at 1630
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1519
    Illegal BsaI.rc site found at 383


Design Notes

This part has been created first by inserting the RBS sequence between pFixK2 and amilGFP (in Bba_K952003) through mutagenesis PCR with a specific primer.
Furthermore we decided to regulate transcription with an upstream promoter, pLac (BBa_R0010).


Part BBa_K952003 was missing an RBS before the amilGFP CDS. We inserted the RBS by mutagenesis, added a pLAC promoter in front of the device and charachterized it.

Source

The part consist of:
Bba_K952003: the entire blue light sensing device with fluorescent output.
BBa_R0010: pLac promoter (LacI regulated) RBS sequence

References

1) Moglich A, Ayers RA and Moffat K. (2009) Design and Signaling Mechanism of Light-Regulated Histidine Kinases. J. Mol. Bio. 385, 5, 1433-1444. 2)Ohlendorf, R., Vidavski, R.R., Eldar, A., Moffat, K. & Möglich, A.(2012). From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression. J. Mol. Biol., 416: 534: 542