MoClo system |
based on Golden Gate cloning |
90–100% for 10 fra gments |
50 kb (68 fragments) |
MASTER Ligation |
using MspJI, which specifically recognizes methylated 4-bp sites and generates a 4-bp arbitrary overhang |
NA |
29 kb |
DNA Assembler (any-gene-any-plasmid (AGAP)) |
based on yeast in vivo homologous recombination mechanism |
100% for 2 fragments |
50 kb |
Reiterative Recombination |
endonuclease-induced homologous recombination in conjunction with recyclable markers |
99% for less than 10 kb |
less than 10 kb, could be larger |
site-specific recombination-based tandem assembly (SSRTA) |
based on φBT1 integration system |
|
|
RecET-mediated linear-plus-linear homologous recombination (LLHR) |
linear-linear homologous recombination |
|
10–52 kb |
phage wBT1 integrase-mediated site-specific recombination |
requiring both homologous and site-specific recombinations. The homologous recombinations were used for targeted integration of the mutated attB and attP into Streptomyces chromosome, while the wBT1 integrase-mediated site-specific recombination was employed to excise targeted region of interest from the chromosome |
90% for 23 kb, 80% for 157 kb |
157 kb |
transformation-associated recombination (TAR) |
in vivo homologous recombination of Saccharomyces cerevisiae
|
|
21.3 kb and 67 kb |