Table 1
Strains and plasmids
Cell line
Manufacturer
Genotype
Efficiency™ DH10B™
competent cells
Thermo fisher Scientific
F
À
mcrA
Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1
araD139
Δ(ara, leu)7697 galU galK
λ
À
rpsL nupG/pMON14272/pMON7124
DH10B
ΔcobB ΔpyrF
Spinck et al. 2020 [24]
[DH10B]
pyrF cobB
BL21 (DE3)
Novagen
fhuA2 [lon] ompT gal (λ DE3) [dcm]
ΔhsdS λ DE3
¼ λ sBamHIo
ΔEcoRI-B
int::(lacI::PlacUV5::T7 gene1) i21
Δnin5
pPylT-URA3
Spinck et al. 2020 [24]
scURA3 cloned in pPylT
pPylT-URA3K93TAG-PylS
Spinck et al. 2020 [24]
PylS cloned cistronically in pPylT-URA3K93TAG
pPylT-URA3K93TAG-AcKRS3
Spinck et al. 2020 [24]
pPylT-URA3K93TAG-AcKRS3
pBK-His
6 -CobB
Spinck et al. 2018 [25]
pBK-His
6 -CobB
pFluc
Spinck et al. 2018 [25]
FlucK529TAG-His
6 cloned in pCDF-PylT
pBK-AcKRS3opt
Spinck et al. 2018 [25]
Encodes AcKRS3 with mutations for improved PylT binding
pBK-PylS
Neumann et al. 2008 [26]
Encodes
M. barkeri pyrrolysyl-tRNA synthetase
A Directed Evolution System for Lysine Deacetylases
321