Plasmid

Part:BBa_J100315

Designed by: Dylan Maghini   Group: Campbell M Lab   (2017-05-07)


J100288 with RFP replacing LuxAB

In their 2014 paper "Negative selection and stringency modulation in phage-assisted continuous evolution," Carlson et al., designed part J100265 (pJC173b) to be used as an accessory plasmid in phage-assisted continuous evolution (PACE). This part comes from a collection of 8 plasmids numbers J100264 - J100271. PACE employs M13 phage and E. coli in a chemostat. J100288 was based on J100265 and contains a deletion of 210 bp, encoding 70 amino acids, from gene III. The mutant gene III encodes a protein III that acts as a dominant negative for M13 phage reproduction. J100315 is based on J100288, but with an insertion of RFP in place of LuxAB through the use of PCR and GGA.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1018
    Illegal PstI site found at 1812
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1018
    Illegal PstI site found at 1812
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2481
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1018
    Illegal PstI site found at 1812
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1018
    Illegal PstI site found at 1812
    Illegal AgeI site found at 3458
    Illegal AgeI site found at 3570
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3804
    Illegal SapI site found at 2648


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