. Author manuscript; available in PMC: 2017 Jun 1.

Published in final edited form as: Biochemistry. 2015 Oct 5;54(41):6392–6401. doi: 10.1021/acs.biochem.5b00942

Table 1.

Plasmids used in this study

Plasmid	Description	Source
pUC8	High-copy number plasmid containing the pBR322 origin of replication, the multiple cloning site of M13mp7, and conferring Amp^R. Gene expression is driven by an upstream lac promoter.	⁶⁵
pKK17	EcoRI_HindIII fragment of K. aerogenes ureDABCEFG ligated into similarly digested pKK223-3 containing the pBR322 origin of replication, an upstream tac promoter, downstream rrnB ribosomal terminator sequence, and conferring Amp^R	^{13, 66}
pKK17D*, -V37L, -Y42D, -E46A, -E46Q, -C48A, -H49A, -H54A, -I59Y, -D63A, -D63Q, -L65I, -L65W, -S85K, -K86A, -Y88V, -Y88F, -R89A, -W111Y, -T128E, -D142A, -R148M, -E153A, -E153Q, -R163A, -E165A, -D169A, -E176A, -E176Q, -T196K, -R211A	EcoRI-AgeI ureD mutant fragments from pMF001L* ligated into similarly digested pKK17	This study
pKKG	PstI-KpnI ureG_Str fragment ligated into similarly digested pKK17 resulting in replacement of UreG with one containing a C-terminal Strep-II tag (ureDABCEFG_Str)	¹¹
pKKD*G, -D63A, -D63Q. –S85K, -D142A, -E176A, -E176Q, R211A	EcoRI-AgeI ureD mutant fragments from pMF001L* ligated into similarly digested pKKG	This study
pEC002	pMal-c2X derived vector using the pBR322 origin of replication and conferring Amp^R for the cytosolic overproduction of maltose binding protein fused at the N-terminus of UreD. Gene expression is governed by an upstream tac promoter and a downstream rrnB ribosomal terminator sequence.	⁸
pMF001	EcoRI-HindIII ureD fragment from pEC002 ligated into similarly digested pUC8	This study
pMF001L	EcoRI-AgeI 5’UTR-ureD fragment from pKK17 ligated into similarly digested pMF001	This study