TY - JOUR
T1 - Construction of Small Genes with Repetitive Elements Using Oligo Extension and Golden Gate Assembly
AU - Nguyen, Michael T.A.
AU - Andersen, Ebbe Sloth
AU - Pothoulakis, Georgios
N1 - Publisher Copyright:
© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2025
Y1 - 2025
N2 - Gene synthesis efficiency has greatly improved in recent years but is limited when it comes to repetitive sequences and results in synthesis failure or delays by DNA synthesis vendors. Here, we describe a method for the assembly of small synthetic genes with repetitive elements: First, a gene of interest is split in silico into small synthons of up to 80 base pairs flanked by Golden Gate-compatible overhangs. Then synthons are made by oligo extension and finally assembled into a synthetic gene by Golden Gate assembly.
AB - Gene synthesis efficiency has greatly improved in recent years but is limited when it comes to repetitive sequences and results in synthesis failure or delays by DNA synthesis vendors. Here, we describe a method for the assembly of small synthetic genes with repetitive elements: First, a gene of interest is split in silico into small synthons of up to 80 base pairs flanked by Golden Gate-compatible overhangs. Then synthons are made by oligo extension and finally assembled into a synthetic gene by Golden Gate assembly.
KW - Gene synthesis
KW - Golden Gate cloning
KW - Molecular cloning
KW - Repetitive DNA sequences
KW - Synthetic biology
UR - http://www.scopus.com/inward/record.url?scp=85205606140&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-4220-7_12
DO - 10.1007/978-1-0716-4220-7_12
M3 - Journal article
C2 - 39363074
AN - SCOPUS:85205606140
SN - 1064-3745
VL - 2850
SP - 219
EP - 227
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -