Part:BBa_K2797013:Design

High copy BioBrick assembly plasmid pSB1C3 with RFP internal standard

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 989
  Illegal suffix found in sequence at 1
  Illegal EcoRI site found at 3052
  Illegal XbaI site found at 3067
  Illegal SpeI site found at 1905
  Illegal PstI site found at 1919
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 989
  Illegal EcoRI site found at 3052
  Illegal NheI site found at 1017
  Illegal NheI site found at 1040
  Illegal SpeI site found at 2
  Illegal SpeI site found at 1905
  Illegal PstI site found at 16
  Illegal PstI site found at 1919
  Illegal NotI site found at 9
  Illegal NotI site found at 995
  Illegal NotI site found at 1912
  Illegal NotI site found at 3058
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 989
  Illegal EcoRI site found at 3052
  Illegal XhoI site found at 2036
  Illegal XhoI site found at 2928
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 989
  Illegal suffix found in sequence at 2
  Illegal EcoRI site found at 3052
  Illegal XbaI site found at 3067
  Illegal SpeI site found at 1905
  Illegal PstI site found at 1919
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 989
  Illegal EcoRI site found at 3052
  Illegal XbaI site found at 1004
  Illegal XbaI site found at 3067
  Illegal SpeI site found at 2
  Illegal SpeI site found at 1905
  Illegal PstI site found at 16
  Illegal PstI site found at 1919
  Illegal AgeI site found at 1627
  Illegal AgeI site found at 1739
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

pSB1C3 was linearized via 2-step PCR in a non-coding region a reasonable distance away from the MCS. Gibson ends were designed using NEBuilder and the RFP BioBrick was synthesized by IDT. Using Gibson Assembly, the RFP BioBrick was cloned into the pSB1C3 vector.

Source

The iGEM 2018 InterLab Study