Part:BBa_K1520024

Pcons2-rbs-lacI-Ter-Plac-rbs-luxI-Ter-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-rbs-luxR-Ter

Plasmid in positive feedback verification: Pcons2-rbs-lacI-Ter-Plac-rbs-luxI-Ter-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-rbs-luxR-Ter It is a positive feedback plasmid using luxI and luxR as key to feedback. Introducing IPTG will produce rfp, luxI and luxR, and luxI together with luxR will work back to enhance production of rfp.

The blue line in the picture shows the fluorescence intensity of the bacteria with this plasmid, which serves as a control in our experiment. The yellow line and purple line show the fluorescence intensity of the bacteria with different positive feedback plasmids.The yellow line is the one with this part, from the picture, we can conclude that this part can achieve the best expression efficiency,which is much better than the control (purple) and the luxI(blue).

From left to right, Marker(1Kb),

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter,

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-Ter,

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-Ter,

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-rbs-luxR-Ter.

The substances of electrophoresis are made from PCR of plasmid with VF2 and VR as their primers.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 2380
  Illegal NheI site found at 2403
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1208
  Illegal BglII site found at 2222
  Illegal BglII site found at 4818
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3358