QIAprep Spin Miniprep Kit (Qiagen). Custom primers were obtained from
Integrated DNA Technologies (Table S3,
Information
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,
Information
verified by DNA sequencing at the ICMB Core Facility at the University
of Texas at Austin.