Plasmid_Backbone

Part:BBa_K3425016:Design

Designed by: N�ria Garriga Alonso   Group: iGEM20_UofUppsala   (2020-10-19)


pSB4A04: pOdd4 Loop Vector based on pSB4A5


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3404
    Illegal PstI site found at 12
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3404
    Illegal PstI site found at 12
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3404
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3404
    Illegal PstI site found at 12
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3404
    Illegal PstI site found at 12
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal SapI site found at 3410
    Illegal SapI.rc site found at 5


Design Notes

pOdd4 Loop Vector based on pSB4A5 for level 3 assemblies. A Gibson assembly experiment was designed, using three parts amplified via PCR:

  • The mRFP1 device (BBa_J04454) from pSB1K04 with VF2 and VR primers
  • Part of the pSB4A5 sequence with GIBSON_3K#1_Rv_4A#1_Fw and GIBSON_4A#1_Rv_4K#1_Rv primers
  • Part of the pSB4A5 sequence with GIBSON_4A#2_Fw_4K#2_Fw and GIBSON_3K#2_Fw_4A#2_Rv_4K#1_Fw primers
The primers used for the Gibson assembly can be found in Team Uppsala 2020's Parts page.


Source

Gibson assembly from parts of pSB4A5 and BBa_J04454.

References