Part:BBa_K3425008:Design

pSB3K04: pOdd4 Loop Vector based on pSB3K3

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2723
  Illegal PstI site found at 12
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2723
  Illegal NheI site found at 1378
  Illegal PstI site found at 12
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2723
  Illegal XhoI site found at 171
  Illegal XhoI site found at 1014
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2723
  Illegal PstI site found at 12
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2723
  Illegal PstI site found at 12
  Illegal AgeI site found at 1464
  Illegal AgeI site found at 1787
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal SapI site found at 2729
  Illegal SapI.rc site found at 5
  

Design Notes

pOdd4 Loop Vector based on pSB3K3 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 pSB3K3 sequence with GIBSON_3K#1_Fw and GIBSON_3K#1_Rv_4A#1_Fw primers
• Part of the pSB3K3 sequence with GIBSON_3K#2_Fw_4A#2_Rv_4K#1_Fw and GIBSON_3K#2_Rv primers

The primers used for the Gibson assembly can be found in Team Uppsala 2020's Parts page.

Source

Gibson assembly from parts of pSB3K3 and BBa_J04454.