Plasmids (Table S2, Supporting Information) were isolated using the
QIAprep Spin Miniprep Kit (Qiagen). Custom primers were obtained from
Integrated DNA Technologies (Table S3, Supporting
Information). PCR reagents were purchased from Takara, New
England Biolabs, or Stratagene, and PCR products were isolated using
a QIAquick PCR Purification Kit (Qiagen). DNA fragments were isolated
from 0.7% agarose gels using the QIAquick Gel Extraction Kit (Qiagen).
All other DNA-modifying enzymes were purchased from New England Biolabs
and used according to the manufacturer’s protocols. When necessary,
purity and yield of extracted DNA were monitored using a NanoDrop
instrument (GE Healthcare). To create pQlink plasmids used for the
coexpression of multiple genes, reagents purchased from Invitrogen
were used for ligation-independent cloning (LIC) and as previously
described.41 (link) All plasmids constructed in
this study were transformed into a chemically competent E.
coli XL-1 Blue storage strain before transformation into
the strain used in experiments (Table S2, Supporting
Information). Plasmid promoter and gene-insert sequences were
verified by DNA sequencing at the ICMB Core Facility at the University
of Texas at Austin.