Plasmid

Part:BBa_K734006:Design

Designed by: Ben Slater   Group: iGEM12_Austin_Texas   (2012-10-01)


pReporter.permaflipped control for ZombiE.coli


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2394
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
    Illegal NgoMIV site found at 5477
    Illegal NgoMIV site found at 5637
    Illegal AgeI site found at 568
    Illegal AgeI site found at 846
    Illegal AgeI site found at 958
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3474
    Illegal BsaI.rc site found at 70
    Illegal SapI.rc site found at 4556


Design Notes

In testing, induction with IPTG does not allow RFP expression, possibly due to extensive secondary structure on the RNA transcripts blocking the RBS.


Source

LasI: from pSB1A2-K081009

RFP: from pSB1AK3-K081014

double terminator: from pSB1AK3-B0015

backbone: pET21-derived

References