RED20 is a so-called "fail-safe genetic code" introduced by Calles and colleagues in 2019 [1]. RED20 encodes each of the 20 canonical amino acids with exactly one sense codon and terminates translation with the canonical three stop codons. RED20 therefore leaves 41 of the 64 triplet codons unassociated with any translation machinery, meaning they are "null codons". Recoding an organism in RED20 requires recoding all of its translated genes into RED20 as well as deleting its tRNA genes associated with RED20 null codons. The Free Genes E. coli RED20 essential gene distribution represents RED20 recoded versions of 245 of the 299 translated genes in the Keio collection, which represents the set of genes essential for life in E. coli as determined by single gene deletion viability [2].**What can it be used for**

The E. coli RED20 gene set can be used to research the engineering of RED20 encoded organisms and genetic systems. The E. coli RED20 gene set can also be used in E. coli based expression systems incorporating non-canonical amino acids. Because every gene in this gene set is encoded using only 20 sense codons and one of three stop codons, users of this gene set can reprogram any of the other 41 null codons using orthogonal translation machinery.**Where can I find more information**

- [1] Calles, J., Justice, I., Brinkley, D., Garcia, A. & Endy, D. Fail-safe genetic codes designed to intrinsically contain engineered organisms. Nucleic Acids Res. (2019). doi:10.1093/nar/gkz745
- [2] Baba, T. et al. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol. Syst. Biol. 2, 2006.0008 (2006).

This product is made available under the unilateral OpenMTA. Read Terms of Service at the bottom of the page for details.

245 unique parts

**Instructions for Use**

Culture at 37C overnight, with shaking at around 300rpm. The selection marker for all parts is ampicillin.

PlateSet: ecolired20-1 ecolired20-2 ecolired20-3

#### Genes

Gene | Name | NCBI ID |
---|---|---|

minD | inhibitor of FtsZ ring polymerization;chromosome-membrane tethering protein; membrane ATPase ofthe MinCDEE system | NP_415693.1 |

pth | peptidyl-tRNA hydrolase | NP_415722.1 |

prs | phosphoribosylpyrophosphate synthase | NP_415725.1 |

ispE | 4-diphosphocytidyl-2-C-methylerythritol kinase | NP_415726.1 |

lolB | lipoprotein localization factor | NP_415727.1 |

hemA | glutamyl tRNA reductase | NP_415728.1 |

prfA | peptide chain release factor RF-1 | NP_415729.1 |

kdsA | 3-deoxy-D-manno-octulosonate 8-phosphatesynthase | NP_415733.1 |

topA | NP_415790.1 | |

fabI | NP_415804.1 | |

dicA | Qin prophage; putative regulator for DicB | NP_416088.1 |

ydfB | Qin prophage; uncharacterized protein | NP_416090.2 |

tyrS | tyrosyl-tRNA synthetase | NP_416154.1 |

ydiL | putative HTH domain DNA-binding protein | NP_416204.2 |

pheS | NP_416229.1 | |

rplT | 50S ribosomal subunit protein L20 | NP_416231.1 |

infC | translation initiation factor IF-3 | NP_416233.1 |

nadE | NP_416254.1 | |

tsaB | NP_416321.1 | |

aspS | aspartyl-tRNA synthetase | NP_416380.1 |

argS | arginyl-tRNA synthetase | NP_416390.1 |

pgsA | phosphatidylglycerophosphate synthetase | NP_416422.1 |

yefM | antitoxin of the YoeB-YefM toxin-antitoxinsystem | NP_416521.2 |

yejM | essential inner membrane DUF3413domain-containing protein; lipid A production and membranepermeability factor | NP_416693.1 |

gyrA | NP_416734.1 | |

folC | bifunctional folylpolyglutamate synthase/dihydrofolate synthase | NP_416818.1 |

accD | NP_416819.1 | |

fabB | 3-oxoacyl-[acyl-carrier-protein] synthase I | NP_416826.1 |

gltX | glutamyl-tRNA synthetase | NP_416899.1 |

ligA | NP_416906.1 | |

hisS | histidyl tRNA synthetase | NP_417009.1 |

ispG | NP_417010.1 | |

suhB | inositol monophosphatase | NP_417028.1 |

tadA | tRNA-specific adenosine deaminase | NP_417054.2 |

era | NP_417061.1 | |

lepB | NP_417063.1 | |

rpoE | RNA polymerase sigma E factor | NP_417068.1 |

pssA | phosphatidylserine synthase;CDP-diacylglycerol-serine O-phosphatidyltransferase | NP_417080.4 |

bamD | BamABCDE complex OM biogenesis lipoprotein | NP_417086.1 |

rplS | 50S ribosomal subunit protein L19 | NP_417097.1 |

trmD | NP_417098.1 | |

rpsP | 30S ribosomal subunit protein S16 | NP_417100.1 |

ffh | NP_417101.1 | |

grpE | heat shock protein | NP_417104.1 |

csrA | pleiotropic regulatory protein for carbon sourcemetabolism | NP_417176.1 |

alaS | alanyl-tRNA synthetase | NP_417177.1 |

ispF | NP_417226.1 | |

ispD | 4-diphosphocytidyl-2C-methyl-D-erythritolsynthase | NP_417227.1 |

ftsB | cell division protein | NP_417228.1 |

eno | enolase | NP_417259.1 |

pyrG | CTP synthetase | NP_417260.1 |

mazE | antitoxin of the ChpA-ChpR toxin-antitoxinsystem | NP_417263.1 |

lgt | phosphatidylglycerol-prolipoproteindiacylglyceryl transferase | NP_417305.1 |

prfB | peptide chain release factor RF-2 | NP_417367.1 |

fbaA | NP_417400.1 | |

pgk | phosphoglycerate kinase | NP_417401.1 |

metK | S-adenosylmethionine synthetase | NP_417417.1 |

yqgF | putative Holliday junction resolvase | NP_417424.1 |

plsC | 1-acyl-sn-glycerol-3-phosphate acyltransferase | NP_417490.1 |

mqsA | antitoxin for MqsR toxin; transcriptionalrepressor | NP_417493.1 |

parE | NP_417502.1 | |

ribB | NP_417513.1 | |

cca | NP_417528.1 | |

tsaD | NP_417536.1 | |

rpsU | 30S ribosomal subunit protein S21 | NP_417537.1 |

dnaG | DNA primase | NP_417538.1 |

rsmI | NP_417615.1 | |

yhbV | U32 peptidase family protein | NP_417628.2 |

ftsH | NP_417645.1 | |

rpmA | 50S ribosomal subunit protein L27 | NP_417652.1 |

rplU | 50S ribosomal subunit protein L21 | NP_417653.1 |

ispB | octaprenyl diphosphate synthase | NP_417654.1 |

murA | UDP-N-acetylglucosamine1-carboxyvinyltransferase | NP_417656.1 |

lptC | periplasmic membrane-anchored LPS-bindingprotein; LPS export protein | NP_417666.1 |

lptA | lipopolysaccharide export ABC transporterperiplasmic binding protein; Lipid A binding protein; LPSexport and assembly protein | NP_417667.1 |

rplM | 50S ribosomal subunit protein L13 | NP_417698.1 |

degS | NP_417702.1 | |

mreD | cell wall structural complex MreBCDtransmembrane component MreD | NP_417715.1 |

mreC | cell wall structural complex MreBCDtransmembrane component MreC | NP_417716.1 |

mreB | NP_417717.2 | |

accB | NP_417721.1 | |

accC | NP_417722.1 | |

tsaC | NP_417741.1 | |

def | peptide deformylase | NP_417745.1 |

fmt | NP_417746.1 | |

rplQ | 50S ribosomal subunit protein L17 | NP_417753.1 |

rpoA | NP_417754.1 | |

rpsD | 30S ribosomal subunit protein S4 | NP_417755.1 |

rpsK | 30S ribosomal subunit protein S11 | NP_417756.1 |

secY | preprotein translocase membrane subunit | NP_417759.1 |

rplO | 50S ribosomal subunit protein L15 | NP_417760.1 |

rpsE | 30S ribosomal subunit protein S5 | NP_417762.1 |

rplR | 50S ribosomal subunit protein L18 | NP_417763.1 |

rpsH | 30S ribosomal subunit protein S8 | NP_417765.1 |

rpsN | 30S ribosomal subunit protein S14 | NP_417766.1 |

rplE | 50S ribosomal subunit protein L5 | NP_417767.1 |

rplX | 50S ribosomal subunit protein L24 | NP_417768.1 |

rplN | 50S ribosomal subunit protein L14 | NP_417769.1 |

rpmC | 50S ribosomal subunit protein L29 | NP_417771.1 |

rplP | 50S ribosomal subunit protein L16 | NP_417772.1 |

rpsC | 30S ribosomal subunit protein S3 | NP_417773.1 |

rplV | 50S ribosomal subunit protein L22 | NP_417774.1 |

rplB | 50S ribosomal subunit protein L2 | NP_417776.1 |

rplW | 50S ribosomal subunit protein L23 | NP_417777.1 |

rplD | 50S ribosomal subunit protein L4 | NP_417778.1 |

rpsL | 30S ribosomal subunit protein S12 | NP_417801.1 |

trpS | tryptophanyl-tRNA synthetase | NP_417843.1 |

yrfF | NP_417857.1 | |

asd | NP_417891.1 | |

ftsX | putative ABC transporter permease | NP_417919.1 |

ftsE | cell division ATP-binding protein | NP_417920.1 |

yhhQ | DUF165 family inner membrane protein | NP_417928.1 |

glyQ | NP_418017.1 | |

gpsA | NP_418065.1 | |

waaU | lipopolysaccharide core biosynthesis | NP_418080.1 |

waaA | NP_418090.1 | |

coaD | pantetheine-phosphate adenylyltransferase | NP_418091.1 |

rpmB | 50S ribosomal subunit protein L28 | NP_418094.1 |

dfp | NP_418096.4 | |

dut | deoxyuridinetriphosphatase | NP_418097.1 |

gmk | guanylate kinase | NP_418105.1 |

gyrB | YP_026241.1 | |

dnaN | NP_418156.1 | |

dnaA | NP_418157.1 | |

rpmH | 50S ribosomal subunit protein L34 | NP_418158.1 |

rnpA | protein C5 component of RNase P | NP_418159.1 |

tnaB | tryptophan transporter of low affinity | NP_418165.1 |

glmS | L-glutamine:D-fructose-6-phosphateaminotransferase | NP_418185.1 |

glmU | fused N-acetyl glucosamine-1-phosphateuridyltransferase/glucosamine-1-phosphate acetyltransferase | NP_418186.1 |

rho | transcription termination factor | NP_418230.1 |

wzyE | putative ECA polysaccharide chain elongationprotein | NP_418241.1 |

hemD | uroporphyrinogen III synthase | NP_418248.1 |

hemC | hydroxymethylbilane synthase | YP_026260.1 |

ubiB | NP_418279.1 | |

hemG | NP_418292.1 | |

yihA | cell division GTP-binding protein | NP_418301.3 |

ftsN | essential cell division protein | NP_418368.1 |

murI | glutamate racemase | NP_418402.2 |

murB | NP_418403.1 | |

birA | NP_418404.1 | |

coaA | pantothenate kinase | NP_418405.1 |

secE | preprotein translocase membrane subunit | NP_418408.1 |

nusG | transcription termination factor | NP_418409.1 |

rplJ | 50S ribosomal subunit protein L10 | NP_418412.1 |

rplL | 50S ribosomal subunit protein L7/L12 | NP_418413.1 |

rpoB | NP_418414.1 | |

ubiA | p-hydroxybenzoate octaprenyltransferase | NP_418464.1 |

plsB | glycerol-3-phosphate O-acyltransferase | NP_418465.4 |

lexA | transcriptional repressor of SOS regulon | NP_418467.1 |

dnaB | replicative DNA helicase | NP_418476.1 |

ssb | single-stranded DNA-binding protein | NP_418483.1 |

alsK | D-allose kinase | NP_418508.1 |

groS | NP_418566.1 | |

orn | oligoribonuclease | NP_418586.4 |

tsaE | NP_418589.1 | |

rpsR | 30S ribosomal subunit protein S18 | NP_418623.1 |

chpS | antitoxin of the ChpBS toxin-antitoxin system | NP_418645.2 |

ppa | inorganic pyrophosphatase | NP_418647.1 |

valS | valyl-tRNA synthetase | NP_418679.1 |

lptF | lipopolysaccharide export ABC permease | NP_418682.1 |

lptG | lipopolysaccharide export ABC permease | NP_418683.4 |

dnaC | DNA biosynthesis protein | NP_418781.1 |

ribF | bifunctional riboflavin kinase/FAD synthetase | NP_414566.1 |

ileS | isoleucyl-tRNA synthetase | NP_414567.1 |

lspA | NP_414568.1 | |

ispH | NP_414570.1 | |

folA | dihydrofolate reductase | NP_414590.1 |

lptD | NP_414596.1 | |

ftsL | membrane bound cell division leucine zipperseptum protein | NP_414625.1 |

ftsI | transpeptidase involved in septal peptidoglycansynthesis; penicillin-binding protein 3 | NP_414626.1 |

murF | UDP-N-acetylmuramoyl-tripeptide:D-alanyl-D-alanine ligase | NP_414628.1 |

mraY | phospho-N-acetylmuramoyl-pentapeptidetransferase | NP_414629.1 |

murD | UDP-N-acetylmuramoyl-L-alanine:D-glutamateligase | NP_414630.1 |

ftsW | putative lipid II flippase; integral membraneprotein; FtsZ ring stabilizer | NP_414631.1 |

murG | N-acetylglucosaminyl transferase | NP_414632.1 |

ftsQ | NP_414635.1 | |

ftsA | ATP-binding cell division FtsK recruitmentprotein | NP_414636.1 |

ftsZ | GTP-binding tubulin-like cell division protein | NP_414637.1 |

lpxC | UDP-3-O-acyl N-acetylglucosamine deacetylase | NP_414638.1 |

secM | regulator of secA translation | NP_414639.2 |

secA | NP_414640.1 | |

can | carbonic anhydrase | NP_414668.1 |

folK | 2-amino-4-hydroxy-6-hydroxymethyldihyropteridinepyrophosphokinase | NP_414684.1 |

hemL | NP_414696.1 | |

erpA | iron-sulfur cluster insertion protein | NP_414698.1 |

rpsB | 30S ribosomal subunit protein S2 | NP_414711.1 |

tsf | translation elongation factor EF-Ts | NP_414712.1 |

pyrH | uridylate kinase | NP_414713.1 |

frr | ribosome recycling factor | NP_414714.1 |

dxr | 1-deoxy-D-xylulose 5-phosphate reductoisomerase | NP_414715.1 |

ispU | undecaprenyl pyrophosphate synthase | NP_414716.1 |

cdsA | CDP-diglyceride synthase | NP_414717.2 |

rseP | NP_414718.1 | |

bamA | BamABCDE complex OM biogenesis outer membranepore-forming assembly factor | NP_414719.1 |

lpxD | NP_414721.1 | |

lpxA | UDP-N-acetylglucosamine acetyltransferase | NP_414723.1 |

lpxB | tetraacyldisaccharide-1-P synthase | NP_414724.1 |

accA | NP_414727.1 | |

tilS | NP_414730.1 | |

proS | prolyl-tRNA synthetase | NP_414736.1 |

yafN | antitoxin of the YafO-YafN toxin-antitoxinsystem | NP_414767.1 |

yagG | CP4-6 prophage; putative sugar transporter | NP_414804.1 |

hemB | NP_414903.4 | |

secD | SecYEG protein translocase auxillary subunit | NP_414942.1 |

secF | SecYEG protein translocase auxillary subunit | NP_414943.1 |

ribD | NP_414948.1 | |

ribE | riboflavin synthase beta chain | NP_414949.1 |

thiL | thiamine monophosphate kinase | NP_414951.1 |

dxs | NP_414954.1 | |

ispA | geranyltranstransferase | NP_414955.1 |

dnaX | NP_415003.1 | |

adk | adenylate kinase | NP_415007.1 |

hemH | ferrochelatase | NP_415008.1 |

lpxH | NP_415057.1 | |

cysS | cysteinyl-tRNA synthetase | NP_415059.1 |

folD | NP_415062.1 | |

entD | phosphopantetheinyltransferase component ofenterobactin synthase multienzyme complex | NP_415115.2 |

mrdB | cell wall shape-determining protein | NP_415167.1 |

mrdA | NP_415168.1 | |

nadD | NP_415172.1 | |

holA | NP_415173.1 | |

lptE | NP_415174.1 | |

lnt | apolipoprotein N-acyltransferase | NP_415190.1 |

glnS | glutamyl-tRNA synthetase | NP_415206.1 |

fldA | flavodoxin 1 | NP_415210.1 |

cydA | NP_415261.2 | |

infA | translation initiation factor IF-1 | NP_415404.1 |

serS | seryl-tRNA synthetase | NP_415413.1 |

msbA | lipid ABC transporter permease/ATPase | NP_415434.1 |

lpxK | NP_415435.1 | |

kdsB | 3-deoxy-manno-octulosonate cytidylyltransferase | NP_415438.1 |

mukF | NP_415442.1 | |

mukE | NP_415443.2 | |

asnS | asparaginyl tRNA synthetase | NP_415450.1 |

fabA | beta-hydroxydecanoyl thioester dehydrase | NP_415474.1 |

yceQ | uncharacterized protein | NP_415603.2 |

fabD | malonyl-CoA-[acyl-carrier-protein] transacylase | NP_415610.1 |

fabG | 3-oxoacyl-[acyl-carrier-protein] reductase | NP_415611.1 |

acpP | NP_415612.1 | |

tmk | thymidylate kinase | NP_415616.1 |

holB | NP_415617.1 | |

lolC | lipoprotein-releasing system transmembraneprotein | NP_415634.1 |

lolE | lipoprotein-releasing system transmembraneprotein | NP_415636.1 |

purB | adenylosuccinate lyase | NP_415649.1 |

minE | cell division topological specificity factor | NP_415692.1 |