Biobrick compatibility

The Biobrick standard, or Biobrick RFC[10], is a popular standard because of its association with iGem. Though oftentimes cumbersome and time consuming to use, an extremely large amount of standard parts use this assembly. iGem’s website gives a good overview of this method.

However, I have purposely altered the standard in a few ways in order to make it better suited for compatibility with other methods.

Above is RFC[10], below is pKG. The prefixes of both are the same except for 1 modification, which is removal of the 1bp gap between NotI and XbaI because it is deemed unnecessary. The suffix differs more, as NotI is removed and an SbfI site is made by the addition of 2 nucleotides. This is done so both the prefix and suffix have an 8bp cutter as the outer restriction enzyme site. A homing endonuclease site is not additionally added because I-SceI is often used in selection schemes and other homing endonucleases are inefficient. Having 2 8bp cutters allows for DNAs assembled using alternative methods to be more open to additional biobrick cloning steps, as NotI and SbfI are far rarer than EcoRI and PstI. If required, the entire cassette can be replaced with a normal Biobrick cassette by cutting with EcoRI and PstI.