Plasmid

Part:BBa_K314204:Design

Designed by: Laura Austin   Group: iGEM10_Washington   (2010-10-22)

p4T6S::T7


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 332
    Illegal EcoRI site found at 2186
    Illegal EcoRI site found at 12859
    Illegal EcoRI site found at 13560
    Illegal EcoRI site found at 20163
    Illegal EcoRI site found at 20665
    Illegal EcoRI site found at 24358
    Illegal XbaI site found at 41958
    Illegal XbaI site found at 44779
    Illegal SpeI site found at 25862
    Illegal SpeI site found at 48309
    Illegal PstI site found at 1718
    Illegal PstI site found at 4778
    Illegal PstI site found at 6473
    Illegal PstI site found at 8721
    Illegal PstI site found at 8808
    Illegal PstI site found at 14226
    Illegal PstI site found at 16056
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 332
    Illegal EcoRI site found at 2186
    Illegal EcoRI site found at 12859
    Illegal EcoRI site found at 13560
    Illegal EcoRI site found at 20163
    Illegal EcoRI site found at 20665
    Illegal EcoRI site found at 24358
    Illegal NheI site found at 14694
    Illegal SpeI site found at 25862
    Illegal SpeI site found at 48309
    Illegal PstI site found at 1718
    Illegal PstI site found at 4778
    Illegal PstI site found at 6473
    Illegal PstI site found at 8721
    Illegal PstI site found at 8808
    Illegal PstI site found at 14226
    Illegal PstI site found at 16056
    Illegal NotI site found at 1
    Illegal NotI site found at 847
    Illegal NotI site found at 9090
    Illegal NotI site found at 10537
    Illegal NotI site found at 10700
    Illegal NotI site found at 15064
    Illegal NotI site found at 22265
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 332
    Illegal EcoRI site found at 2186
    Illegal EcoRI site found at 12859
    Illegal EcoRI site found at 13560
    Illegal EcoRI site found at 20163
    Illegal EcoRI site found at 20665
    Illegal EcoRI site found at 24358
    Illegal BglII site found at 13847
    Illegal BglII site found at 25843
    Illegal BglII site found at 28119
    Illegal BglII site found at 28221
    Illegal BglII site found at 36179
    Illegal BglII site found at 36368
    Illegal BglII site found at 37902
    Illegal BamHI site found at 353
    Illegal BamHI site found at 4845
    Illegal BamHI site found at 28473
    Illegal BamHI site found at 28851
    Illegal BamHI site found at 41336
    Illegal BamHI site found at 41952
    Illegal XhoI site found at 5951
    Illegal XhoI site found at 6482
    Illegal XhoI site found at 12346
    Illegal XhoI site found at 12739
    Illegal XhoI site found at 18729
    Illegal XhoI site found at 26791
    Illegal XhoI site found at 27404
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 332
    Illegal EcoRI site found at 2186
    Illegal EcoRI site found at 12859
    Illegal EcoRI site found at 13560
    Illegal EcoRI site found at 20163
    Illegal EcoRI site found at 20665
    Illegal EcoRI site found at 24358
    Illegal XbaI site found at 41958
    Illegal XbaI site found at 44779
    Illegal SpeI site found at 25862
    Illegal SpeI site found at 48309
    Illegal PstI site found at 1718
    Illegal PstI site found at 4778
    Illegal PstI site found at 6473
    Illegal PstI site found at 8721
    Illegal PstI site found at 8808
    Illegal PstI site found at 14226
    Illegal PstI site found at 16056
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 332
    Illegal EcoRI site found at 2186
    Illegal EcoRI site found at 12859
    Illegal EcoRI site found at 13560
    Illegal EcoRI site found at 20163
    Illegal EcoRI site found at 20665
    Illegal EcoRI site found at 24358
    Illegal XbaI site found at 41958
    Illegal XbaI site found at 44779
    Illegal SpeI site found at 25862
    Illegal SpeI site found at 48309
    Illegal PstI site found at 1718
    Illegal PstI site found at 4778
    Illegal PstI site found at 6473
    Illegal PstI site found at 8721
    Illegal PstI site found at 8808
    Illegal PstI site found at 14226
    Illegal PstI site found at 16056
    Illegal NgoMIV site found at 445
    Illegal NgoMIV site found at 651
    Illegal NgoMIV site found at 716
    Illegal NgoMIV site found at 937
    Illegal NgoMIV site found at 1528
    Illegal NgoMIV site found at 1671
    Illegal NgoMIV site found at 1748
    Illegal AgeI site found at 1327
    Illegal AgeI site found at 3893
    Illegal AgeI site found at 10078
    Illegal AgeI site found at 15232
    Illegal AgeI site found at 23042
    Illegal AgeI site found at 25214
    Illegal AgeI site found at 25911
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 8802
    Illegal BsaI site found at 10084
    Illegal BsaI site found at 11635
    Illegal BsaI site found at 11681
    Illegal BsaI site found at 12017
    Illegal BsaI site found at 18711
    Illegal BsaI site found at 25528
    Illegal BsaI.rc site found at 3890
    Illegal BsaI.rc site found at 6326
    Illegal BsaI.rc site found at 7457
    Illegal BsaI.rc site found at 25664
    Illegal BsaI.rc site found at 26422
    Illegal BsaI.rc site found at 27692
    Illegal BsaI.rc site found at 27893
    Illegal SapI site found at 20816
    Illegal SapI site found at 46183
    Illegal SapI site found at 47393
    Illegal SapI.rc site found at 2736
    Illegal SapI.rc site found at 5107
    Illegal SapI.rc site found at 35683
    Illegal SapI.rc site found at 38173
    Illegal SapI.rc site found at 38419
    Illegal SapI.rc site found at 41177


Design Notes

Note that this plasmid is for the purpose of making a strain that expresses the Type VI Secretion System - since the secretion system is complex and requires many genes which must be expressed together, BioBricking these genes individually does not make sense. Thus we have this part in a fosmid that can be expressed in the same strain as traditional BioBrick plasmids, such that proteins targeted for secretion can be expressed as BioBricks and then secreted by the secretion system. This is in line with iGEM, as the secretion system is present to essentially generate a background strain, then the user defines the proteins to be secreted by choosing the BioBricks and regulating them accordingly, allowing for a system that can be engineered to have a variety of different behaviors.

Source

Original source was from P. aeruginosa PA01 genome library--fosmid O24--from Mike Jacobs at University of Washington Department of Immunology.

References