Difference between revisions of "Part:BBa K551001"
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===Functional Parameters===
 
===Functional Parameters===
 
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<partinfo>BBa_K551001 parameters</partinfo>
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We performed a thermal sensitivity experiment in liquid medium in which we measured the plasmid loss frequency as a function of time of culture at 42°C.  We compared the viability (CFU/ml: colony forming units per ml of culture) of MC1061/pIN grown at 30°C and 42°C and plated on LB and LB Amp medium. If the plasmid is lost, then the bacteria won't grow on the LB Amp medium plates.

Revision as of 16:20, 21 September 2011

plasmid of insertion and deletion

The pINDEL plasmid is composed of 2 functional units:

(i) the IN function which is composed of the gam, exo and bet genes coding for the lambda Red recombinase system (ii) the DEL function which is based on the flp gene encoding the FLP site-specific recombinase and expressed at 42°C

Another feature of pINDEL is that its replication origin is not functional at 42°C due to the thermosensitivity of the RepA101Ts protein. This protein initiates replication at the ori site in permissive conditions (30°C) and is inactive at 42°C.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 396
    Illegal EcoRI site found at 3708
    Illegal EcoRI site found at 5217
    Illegal EcoRI site found at 6322
    Illegal XbaI site found at 423
    Illegal SpeI site found at 6004
    Illegal SpeI site found at 8956
    Illegal PstI site found at 435
    Illegal PstI site found at 4712
    Illegal PstI site found at 4959
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 396
    Illegal EcoRI site found at 3708
    Illegal EcoRI site found at 5217
    Illegal EcoRI site found at 6322
    Illegal SpeI site found at 6004
    Illegal SpeI site found at 8956
    Illegal PstI site found at 435
    Illegal PstI site found at 4712
    Illegal PstI site found at 4959
    Illegal NotI site found at 9275
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 396
    Illegal EcoRI site found at 3708
    Illegal EcoRI site found at 5217
    Illegal EcoRI site found at 6322
    Illegal BglII site found at 632
    Illegal BglII site found at 1728
    Illegal BamHI site found at 417
    Illegal BamHI site found at 3647
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 396
    Illegal EcoRI site found at 3708
    Illegal EcoRI site found at 5217
    Illegal EcoRI site found at 6322
    Illegal XbaI site found at 423
    Illegal SpeI site found at 6004
    Illegal SpeI site found at 8956
    Illegal PstI site found at 435
    Illegal PstI site found at 4712
    Illegal PstI site found at 4959
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 396
    Illegal EcoRI site found at 3708
    Illegal EcoRI site found at 5217
    Illegal EcoRI site found at 6322
    Illegal XbaI site found at 423
    Illegal SpeI site found at 6004
    Illegal SpeI site found at 8956
    Illegal PstI site found at 435
    Illegal PstI site found at 4712
    Illegal PstI site found at 4959
    Illegal AgeI site found at 3482
    Illegal AgeI site found at 5139
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 837
    Illegal BsaI.rc site found at 9958
    Illegal SapI site found at 989
    Illegal SapI site found at 1787
    Illegal SapI site found at 3464


Functional Parameters

resistance100 g/ml

We performed a thermal sensitivity experiment in liquid medium in which we measured the plasmid loss frequency as a function of time of culture at 42°C. We compared the viability (CFU/ml: colony forming units per ml of culture) of MC1061/pIN grown at 30°C and 42°C and plated on LB and LB Amp medium. If the plasmid is lost, then the bacteria won't grow on the LB Amp medium plates.