Device

Part:BBa_J100272:Design

Designed by: Malcolm Campbell   Group: Campbell M Lab   (2016-06-21)


rClone Red Version 2: 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 1416
    Illegal AgeI site found at 1528
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 849
    Illegal BsaI.rc site found at 42


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.


Source

/

References