Part:BBa_K2945012

Cut 4 Primer: Primer opposite the Prefix Binding Site

Cut 2 Primer: Primer opposite the Suffix Binding Site This primer is to be used when creating new BioBricks via Gibson assembly. Currently, creating a new BioBrick requires getting the sequence between the Prefix and Suffix elements. However, these elements can actually anneal to each other, making Biobricks assembly with Gibson using them as overhangs quite tricky. This also means that enzymatic digestion cannot be used to effectively create new BioBricks. This primers site opposite the VF2 primer binding site and points towards the plasmid. This allows it to be used to linearize pSB1C3 so that it has homology with BioBrick-like inserts that contain the VF2 site. This is designed for synthesized BioBricks from either IDT or Twist as these sequences can be added to the desired BioBrick and then can be assembled to pSB1C3 using Gibson Assembly.

This Primer is Compatible for Linearization by PCR with the Cut 3 Primer that is opposite the VF2 Binding Site (BBa_K2945011)

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 11
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 11
  Illegal NotI site found at 3
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 11
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 11
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 11
  
• 1000
  COMPATIBLE WITH RFC[1000]