Biobrick standard

This standard defines the required sequence properties for a Biobrick(tm) standard biological part. It does not define any functional characteristics of the parts, nor does it motivate any aspect of these standards. All sequences defined herein are specified in the 5' to 3' direction.

1. A Biobrick compatible standard biological part consists of a DNA fragment potentially conveying informational or functional properties to a composite structure assembled from multiple parts. The current assembly process requires certain sequence properties

for the part and the surrounding DNA.

1. Allowed sequences within Biobrick parts include any DNA sequence which does not contain the following subsequences:

  EcoRI site:   GAATTC
  XbaI site:    TCTAGA
  SpeI site:    ACTAGT
  PstI site:    CTGCAG
  NotI site:    GCGGCCGC

1. Biobrick Suffix

• Each Biobrick part must contain precisely this sequence immediately following the 3' end of the part:

  T ACTAGT A GCGGCCG CTGCAG

1. Biobrick Prefix:

To allow the construction of ribosomal binding sequences 5' of coding regions, the prefix used for coding regions is distinguished from the sequence for non-coding Biobrick parts.

• • For non-coding Biobrick parts (the default) the Biobrick part must contain precisely the following sequence immediately 5' of the part:

     GAATTC GCGGCCGC T TCTAGA G

• • For Biobrick parts coding for proteins, the Biobrick part must contain precisely the following sequence immediately 5' of the ATG start of the coding region:

     GAATTC GCGGCCGC T TCTAG

• • Parts containing start codons other than ATG must be modified to use ATG as the start codon. We highly recommend, but do not require, that all coding regions terminate with in-frame TAATAA stop codons, replacing other stop codons (TGA, TAG).

1. Plasmid context

Biobrick parts must be supplied in plasmids compatible with registry and assembly procedures. The pSBxxxx series of plasmids comply with these requirements. Other plasmids may be compliant with the following constraints:

• • Antibiotic resistance: All plasmids must carry at least one of the following antibiotic resistance markers:

     Ampicillin
     Chloramphenicol
     Kanamycin
     Tetracycline