Part:BBa_J100282:Design

rClone Red Version 2 with RBS: Device for GGA Cloning and Testing RBS elements and Riboswitches

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 612
  Illegal AgeI site found at 724
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

When designing oligonucleotides for use with rClone Red V2, make sure they result in 5' overhang sticky ends that are CGAC (left) and GCGG (right). Also make sure the oligonucleotides do not contain binding sites for BsaI. Finally, make sure the RBS element ends immediately before the GCGG right sticky end. This will ensure a spacing of 6 bases between the RBS and the ATG start codon of RFP. Below is an example.