A seamless and iterative DNA assembly method named PS-Brick and its assisted metabolic engineering for threonine and 1-propanol production

Table 1 The collections of endonuclease-assisted DNA assembly methods

Method	REs used in the method^b	RE Sub-type	Scar sequence	Iterative Assembly
BioBrick [19]	SpeI ∩ PstI ∩ XbaI ∩ EcoRI	IIP	8 bp	Yes
BglBrick [24]	EcoRI ∩ BglII ∩ BamHI ∩ XhoI	IIP	6 bp	Yes
ePathBrick [22]	SpeI ∩ XbaI ∩ NheI ∩ AvrII	IIP	6 bp	Yes
YaliBrick [28]	SpeI ∩ SalI ∩ XbaI ∩ AvrII	IIP	6 bp	Yes
Quick Gene Assembly [14]	SpeI ∩ EcoRI ∩ XbaI	IIP	6 bp	Yes
iBrick [25]	I-SceI ∩ PI-PspI	Homing endonucleases	21 bp	Yes
C-Brick [17]	FnCpf1 endonuclease	CRISPR–Cas nucleases	6 bp	Yes
CCTL [27]	FnCpf1 endonuclease	CRISPR–Cas nucleases	> 8 bp	Yes
PODAC [13]	BsaI	IIS	8 bp	Yes
Golden Gate [30]	BsaI	IIS	4 bp	No
GoldenBraid [12]	BsaI ∩ BsmBI	IIS	4 bp	Yes^c
MoClo [32]	BsaI ∩ BpiI	IIS	4 bp	Yes^c
Start-Stop Assembly [42]	SapI ∩ BsaI ∩ BbsI	IIS	Scarless	Yes
PS-Brick^a	(One of hundreds of IIP REs generating overhangs of two or more bases) ∩ (BmrI ∪ BciVI ∪ HphI ∪ MlyI)	IIP and IIS	Scarless	Yes

^aThis study
^b∩ Denotes all of the REs before and after this logical AND symbol are simultaneously used, and ∪ denotes only one among the REs before and after this logical OR symbol is used. For example, “SpeI ∩ PstI ∩ XbaI ∩ EcoRI” indicates that the four given REs are simultaneously used in BioBrick standard, and all of the four RE sites are required to be eliminate from the insertion parts. “BmrI ∪ BciVI ∪ HphI ∪ MlyI” indicates that any one of the four REs can be adopted in the PS-Brick, providing broad available options
^cElaborate plasmid libraries and complex protocols are required for iterative workflows of Golden Braid and MoClo systems

ISSN: 2731-3654