KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER | ||
|---|---|---|---|---|
| Antibodies | ||||
| Monoclonal Anti-polyhistidine antibody produced in mouse | Sigma-Aldrich | Cat#H1029; RRID: AB_260015 | ||
| Goat Anti-Mouse IgG (H+L)-HRP Conjugate | Bio-Rad | Cat#1721011; RRID: AB_11125936 | ||
| Bacterial and Virus Strains | ||||
| E. coli BL21 Star™ (DE3) | ThermoFisher | Cat#C601003 | ||
| E. coli BL21 (DE3) | New England Biolabs | Cat#C2527I | ||
| E. coli DH5α | ThermoFisher | Cat#18265017 | ||
| E. coli C321.ΔA.759 | Martin et al., 2018 | N/A | ||
| E. coli 759.T7.int.lacUV5 | This paper | N/A | ||
| E. coli 759.T7.int.PtacI | This paper | N/A | ||
| E. coli 759.T7.int.Lpp5 | This paper | N/A | ||
| E. coli 759.T7.asl.lacUV5 | This paper | N/A | ||
| E. coli 759.T7.asl.PtacI | This paper | N/A | ||
| E. coli 759.T7.asl.Lpp5 | This paper | N/A | ||
| E. coli 759.T7.ΔkanR | This paper | N/A | ||
| E. coli 759.T7.ΔompT | This paper | N/A | ||
| E. coli 759.T7.K172L | This paper | N/A | ||
| E. coli 759.T7.K172G | This paper | N/A | ||
| E. coli 759.T7.D | This paper | N/A | ||
| E. coli 759.T7.D.ΔAbR | This paper | N/A | ||
| E. coli 759.T7.Opt | This paper | N/A | ||
| Chemicals, Peptides, and Recombinant Proteins | ||||
| T7 RNA polymerase | Prepared in-house | N/A | ||
| pAcF aminoacyl-tRNA synthetase | Prepared in-house | N/A | ||
| Potassium glutamate | Sigma-Aldrich | Cat#G1501 | ||
| Magnesium glutamate | Sigma-Aldrich | Cat#49605 | ||
| Ammonium glutamate | MP Biomedicals | Cat#02180595.1 | ||
| HEPES Buffer | Sigma-Aldrich | Cat#H3375 | ||
| PEG-8000 | Sigma-Aldrich | Cat#202452-500G | ||
| Dithiothreitol | Sigma-Aldrich | Cat#43816 | ||
| Putrescine | Sigma-Aldrich | Cat#P5780 | ||
| Spermidine | Sigma-Aldrich | Cat#S0266 | ||
| Glycerol | Sigma-Aldrich | Cat#G5516 | ||
| Folinic acid | Sigma-Aldrich | Cat#47612 | ||
| tRNA from E. coli MRE600 | Roche | Cat#10109541001 | ||
| Phosphoenol pyruvate | Roche | Cat#10108294 | ||
| NAD | Sigma-Aldrich | Cat#N8535 | ||
| CoA | Sigma-Aldrich | Cat#C3144 | ||
| Oxalic acid | Sigma-Aldrich | Cat#P0963 | ||
| Nuclease-free water | Ambion | Cat#AM9937 | ||
| ATP | Sigma-Aldrich | Cat#A2383 | ||
| GTP | Sigma-Aldrich | Cat#G8877 | ||
| UTP | Sigma-Aldrich | Cat#U6625 | ||
| CTP | Sigma-Aldrich | Cat#C1506 | ||
| L-Valine | Sigma-Aldrich | Cat#V0500 | ||
| L-Tryptophan | Sigma-Aldrich | Cat#T0254 | ||
| L-Phenylalanine | Sigma-Aldrich | Cat#P2126 | ||
| L-Isoleucine | Sigma-Aldrich | Cat#I2752 | ||
| L-Leucine | Sigma-Aldrich | Cat#L8000 | ||
| L-Cysteine | Sigma-Aldrich | Cat#C7352 | ||
| L-Methionine | Sigma-Aldrich | Cat#M9625 | ||
| DL-Alanine | Sigma-Aldrich | Cat#A7627 | ||
| L-Arginine | Sigma-Aldrich | Cat#A8094 | ||
| L-Asparagine | Sigma-Aldrich | Cat#A0884 | ||
| L-Aspartic Acid | Sigma-Aldrich | Cat#A9256 | ||
| L-Glutamic acid | Sigma-Aldrich | Cat#G1501 | ||
| L-Glycine | Sigma-Aldrich | Cat#G7126 | ||
| L-Glutamine | Sigma-Aldrich | Cat#G3126 | ||
| L-Histidine | Sigma-Aldrich | Cat#H8000 | ||
| L-Lysine | Sigma-Aldrich | Cat#L5501 | ||
| L-Proline | Sigma-Aldrich | Cat#P0380 | ||
| L-Serine | Sigma-Aldrich | Cat#S4500 | ||
| L-Threonine | Sigma-Aldrich | Cat#T8625 | ||
| L-Tyrosine | Sigma-Aldrich | Cat#T3754 | ||
| Phusion™ polymerase | New England Biolabs | Cat#M0530L | ||
| SYBR® Safe | ThermoFisher | Cat#S33102 | ||
| Quick-Load® 1 kb DNA ladder | New England Biolabs | Cat#N0468S | ||
| Quick-Load® 100 bp DNA ladder | New England Biolabs | Cat#N0467S | ||
| T5 Exonuclease | New England Biolabs | Cat#M0363S | ||
| Taq Ligase | New England Biolabs | Cat#M0208L | ||
| dNTP Solution Mix | New England Biolabs | Cat#N0447L | ||
| Phusion™ HF Buffer | New England Biolabs | Cat#B0518S | ||
| Kanamycin | Sigma-Aldrich | Cat#K4000-25G | ||
| Carbenicillin | IBI Scientific | Cat#IB02025 | ||
| Chloramphenicol | Sigma-Aldrich | Cat#C0378-25G | ||
| IPTG | Sigma-Aldrich | Cat#I6758-10G | ||
| Tris-acetate | Sigma-Aldrich | Cat#T1258-250G | ||
| Tris | Sigma-Aldrich | Cat#T1503-500G | ||
| 14C Glycine | Perkin Elmer | Cat#NEC276E050UC | ||
| Scintillation fluid | MP Biomedicals | Cat#0188245304 | ||
| Bacto™ Tryptone | ThermoFisher | Cat#211705 | ||
| Bacto™ Yeast Extract | ThermoFisher | Cat#211929 | ||
| Sodium Chloride | Sigma-Aldrich | Cat#S9888 | ||
| Bacto™ Agar | VWR | Cat#90000-762 | ||
| Potassium phosphate monobasic | Sigma-Aldrich | Cat#P0662-500G | ||
| Potassium phosphate dibasic | Sigma-Aldrich | Cat#P3786-500G | ||
| Magnesium acetate | Sigma-Aldrich | Cat#M5661-250G | ||
| Potassium acetate | Sigma-Aldrich | Cat#P1190-1KG | ||
| L-p-acetylphenylalanine | Chem-Impex International Inc. | Cat#24756 | ||
| 4x NuPAGE LDS Sample Buffer | ThermoFisher | Cat#NP0007 | ||
| SeeBlue™ Plus2 Pre-stained Protein Standard | ThermoFisher | Cat#LC5925 | ||
| Methanol | Sigma-Aldrich | Cat#34860-1L-R | ||
| 2x Colony PCR Master mix | ThermoFisher | Cat#K0172 | ||
| Cutsmart® buffer | New England Biolabs | Cat#B7204S | ||
| USER™ Enzyme | New England Biolabs | Cat#M5505S | ||
| anhydrotetracycline | Sigma-Aldrich | Cat#37919-100MG-R | ||
| rhamnose | Sigma-Aldrich | Cat#83650-10G | ||
| Trichloroacetic acid | Sigma-Aldrich | Cat#T6399-500G | ||
| Sodium Hydroxide | Sigma-Aldrich | Cat#221465-500G | ||
| 200-proof ethanol | Sigma-Aldrich | Cat#E7023-6X500ML | ||
| Gel drying solution | Bio-Rad | Cat#1610752 | ||
| LC-MS Grade methanol | Fisher Scientific | Cat#A456-4 | ||
| LC-MS Grade water | Fisher Scientific | Cat#W6-4 | ||
| LC-MS Grade acetonitrile | Fisher Scientific | Cat#A955-4 | ||
| LC-MS Grade formic acid | Fisher Scientific | Cat#A1171-AMP | ||
| NuPAGE MOPS SDS running buffer | ThermoFisher | Cat#NP0001 | ||
| Critical Commercial Assays | ||||
| HiSpeed® Plasmid Maxi Kit | Qiagen | Cat#12643 | ||
| Ni-NTA Agarose | Qiagen | Cat#30210 | ||
| MagStrep “type3” XT beads 5% suspension | IBA Life Sciences | Cat#2-4090-002; Cat#2-1003-100; Cat#2-1042-025 | ||
| E.Z.N.A.® Plasmid Mini Kit | Omega Biotek | Cat#D6943-01 | ||
| E.Z.N.A.® Gel Extraction Kit | Omega Biotek | Cat#D2500-01 | ||
| Immun-Blot® Opti-4CN™ Colorimetric Kit | Bio-Rad | Cat#1708235 | ||
| Oligonucleotides | ||||
| See Table S1 | ||||
| Recombinant DNA | ||||
| pKD4 | Datsenko and Wanner (2000). | N/A | ||
| pAR1219 | Davanloo et al., 1984. | N/A | ||
| pDPtacIAcRSTT1 | de Boer et al., 1983. | N/A | ||
| pDTT1-Lpp5-EF-Tu | Inouye and Inouye (1985). Gan et al., 2017. | N/A | ||
| pY71-sfGFP | Bundy and Swartz (2010). | N/A | ||
| pY71-sfGFP-T216amb | Bundy and Swartz (2010). | N/A | ||
| pY71-sfGFP-2amb | Hong et al., 2014b. | N/A | ||
| pY71-sfGFP-5amb | Hong et al., 2014b. | N/A | ||
| pY71-pAcFRS | Hong et al., 2014b. | N/A | ||
| pEVOL-pAcF | Young et al., 2010. | N/A | ||
| pY71-T7-tz-o-tRNA | Hong et al., 2014b. | N/A | ||
| pUC-T7-int.lacUV5 | This paper | N/A | ||
| pUC-T7-asl.lacUV5 | This paper | N/A | ||
| pUC-T7-asl.PtacI | This paper | N/A | ||
| pUC-T7-asl.Lpp5 | This paper | N/A | ||
| p15a-T7-asl.Lpp5 | This paper | N/A | ||
| p15a-T7-asl.PtacI | This paper | N/A | ||
| p15a-T7-int.Lpp5 | This paper | N/A | ||
| pMA7CR_2.0 | Ronda et al., 2016 | N/A | ||
| pMAZ-SK | Ronda et al., 2016 | N/A | ||
| pMAZ-His | This paper | N/A | ||
| pMAZ-Cure | This paper | N/A | ||
| pY71-FI-ELP20 | Martin et al., 2018 | N/A | ||
| pY71-FI-ELP30 | Martin et al., 2018 | N/A | ||
| pY71-FI-ELP40 | Martin et al., 2018 | N/A | ||
| pY71-FI-ELP20X | Martin et al., 2018 | N/A | ||
| pY71-FI-ELP30X | Martin et al., 2018 | N/A | ||
| pY71-FI-ELP40X | Martin et al., 2018 | N/A | ||