| E. coli Strains |
| DH5α |
F−endAI recAl hsdRl7 (rK− mK−) deoR supE44 thi-J gyrA96 relA
|
Gibco-BRL |
| BL21 Star (DE3) |
F−ompT hsdSB(rB−mB−) gal dcm rne131 (DE3) |
Life Technologies |
| ArcticExpress |
F−ompT hsdS(rB− mB−) dcm+ TetRgal endA Hte [cpn10 cpn60 GentR] |
Agilent Technologies |
| Plasmids |
| pACYCDuet-1 |
expression vector, T7 promoter, CamR
|
Novagen |
| pET-45b(+) |
expression vector, T7 promoter, AmpR
|
Novagen |
| pMIG-RadAi |
Pho RadA intein flanked by short extein sequences cloned into ClaI/SphI sites between MBP and GFP coding sequences in pACYCDuet-1 backbone |
Present study |
| pFL-RadAi |
full-length intein-containing radA, cloned into BamHI/XhoI sites of pET-45b(+) |
Present study |
| pRadA |
inteinless radA constructed based on pFL-Radi |
Present study |
| pFL-RadAi-AA |
full-length intein-containing radA, C153A, N324A splicing inactive intein mutant |
Present study |
| pM1 |
full-length intein-containing radA, R358A, E360A, R361A, R363A, E364A helix 1 extein multiple mutant |
Present study |
| pM2 |
R358A, R361A helix 1 extein multiple mutant |
Present study |
| pM3 |
E360A, R363A, E364A helix 1 extein multiple mutant |
Present study |
| pM4 |
R358A helix 1 extein single point mutant |
Present study |
| pM5 |
E360A helix 1 extein single point mutant |
Present study |
| pM6 |
R361A helix 1 extein single point mutant |
Present study |
| pM7 |
E364A helix 1 extein single point mutant |
Present study |
| pM8 |
R503A helix 1 single point extein mutant |
Present study |
| pM9 |
E354A extein single point catalytic mutant, impaired ATPase activity |
Present study |
| pM10 |
Q465A extein single point catalytic mutant, impaired ATPase activity |
Present study |
| pM11 |
E77A, K79A, E80A extein multiple mutant |
Present study |
| pM12 |
E57A, K58A, R60A, E61A extein multiple mutant |
Present study |
