Biomedical Engineering Reference
In-Depth Information
Fig.
1
The mechanism of GoldenBraid. (
a
) TUs (e.g., 1, 2) assembled in complementary level
α
plasmids can
be used as entry vectors for a level
binary assembly, provided that they share a common BsmBI sticky end
(
encircled
C). Similarly, constructs assembled using paired level
Ω
Ω
plasmids can be used as entry vectors for
a subsequent level
can alternate
indefi nitely creating increasingly complex structures, as depicted by the
arrows
closing the double loop.
Encircled
K and S represent kanamycin resistance gene and spectinomycin resistance gene, respectively.
(
b
) Standard parts fl anked by fi xed BsaI cleavage sites (
boxed
) are multipartite assembled using level
α
assembly, as they share a BsaI sticky end (
squared
3). Level
α
and level
Ω
α
plasmids. Upon assembly, the newly assembled TU(Promoter:CodingSequence:Terminator) remains fl anked by
BsmBI cleavable sites (not depicted here)
