Richard Lab:Bio-Brick Formats
Back to Protocols
Introduction
In general the Richard Lab uses the standard Bio-Brick format. However occasionally this format is incompatible with our goals (e.g. making protein fusions) or is unnecessarilly cumbersome (e.g. PCR overlap extension). To have a review on why (and how) we use these sites see the our New Standard Assembly page.
Formats
Biobrick Standard
This standard uses the typical biobrick assembly enzymes [EcoRI (GAATTC), XbaI (TCTAGA), SpeI (ACTAGT), and PstI (CTGCAG)] as well as NotI sites (GCGGCCGC) flanking the part. There is a slightly different prefix for protein coding parts to put the start codon in a better spot in relation to the RBS. During assembly the XbaI site of the "rear part" and the SpeI site of the "front" part eliminate eachother and form a "scar" which forever binds the two parts together.
- Prefix
- GAATTCgcggccgctTCTAGAg
- The EcoRI and XbaI sites are in CAPS while the NotI site is in bold.
- Protein Coding Prefix
- GAATTCgcggccgctTCTAG
- using this prefix the first base of the start codon (i.e. the "A" in "ATG") will complete the XbaI site.
- Suffix
- tACTAGTagcggccgCTGCAG
Short Biobrick
Protein Fusions
Back to Protocols