261 essential genes from Bacillus subtilis, codon optimized for Escherichia coli in MoClo-compatible high-copy ampicillin resistance backbones. Shipped in Escherichia coli Top10 in 3 96 well plates.

Discovery of genes essential for life is the first step in creating a truly synthetic organism. The Bacillus subtilis community has done an amazing job in curating a list of essential genes, and while the first published list goes back to 2003, the Bacillus subtilis community has continuously updated this list with results from new experiments. In 2016, a paper was published using a CRISPR-based method for knocking down essential genes in Bacillus subtilis. All the sgRNAs are available through the Bacillus Genetic Stock Center, which would allow for knockdown of native-sequence essential genes. We took the list of essential genes from that 2016 paper and synthesized 261 of the 289 genes through Twist Biosciences.

All the essential genes have been recorded for expression in Escherichia coli and have been made MoClo compatible. There are many possible applications for these genes: you could try to recreate essential gene pathways in-vitro, or purify the proteins for functional studies, or create a synthetic MoClo-compatible Bacillus subtilis genome using these genes and the sgRNAs available from the Bacillus Genetic Stock Center.

Recoding was done using scripts that stochastically assigned codons to amino acids. The codon table used was the FreqB table from "Design Parameters to Control Synthetic Gene Expression in Escherichia coli", which was used because it was shown to express proteins well and was recommended to us by JCVI. All genes where inserted into the BbsI:BbsI cloning sites of pOpen_build, which is individually available through our website.

**Documentation**

261 of the 289 essential genes from 2016 essential gene list are included in this distribution. The essential genes included in the distribution at attached in essential_included.csv, while the essential genes not included are in essential_not_included.csv. The spreadsheet BBF_to_uniprot.csv contains links from our genes to outside resources like uniprot, as well as detailed descriptions. The spreadsheet freegenes-distribution-bacillus-subtilis-essential-genes.csv includes a plate map for each plate that gets shipped in this distribution, with information on where exactly to find any gene. In the bsub_essential.zip file are genbank files for every gene. All genes are present on the plate in

*E. coli*glycerol stocks.

This product is made available under the unilateral OpenMTA.

261 unique parts

Plate Layout

**Instructions for Use:**

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

PlateSet: Bsubtilis-essentials-0 Bsubtilis-essentials-1 Bsubtilis-essentials-2

#### Genes

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

dnaN | DNA polymerase III subunit beta | NP_387883.1 |

gyrB | DNA gyrase subunit B | NP_387887.1 |

gyrA | DNA gyrase subunit A | NP_387888.1 |

guaB | Inosine-5'-monophosphate dehydrogenase | NP_387890.1 |

serS | serine--tRNA ligase | NP_387894.1 |

dnaX | DNA polymerase III subunit gamma/tau | NP_387900.2 |

holB | DNA polymerase III subunit delta | NP_387912.1 |

metS | methionine--tRNA ligase | NP_387919.1 |

ipk | 4-diphosphocytidyl-2C-methyl-D-erythritolkinase | NP_387927.1 |

glmU | bifunctional N-acetylglucosamine-1-phosphateuridyltransferase/glucosamine-1-phosphateacetyltransferase | NP_387931.1 |

prs | ribose-phosphate pyrophosphokinase | NP_387932.1 |

pth | peptidyl-tRNA hydrolase | NP_387934.1 |

divIC | cell division protein DivIC | NP_387943.1 |

tilS | tRNA(Ile)-lysidine synthase | NP_387948.1 |

hprT | hypoxanthine-guanine phosphoribosyltransferase | NP_387949.1 |

sul | dihydropteroate synthase | NP_387958.1 |

folB | dihydroneopterin aldolase | NP_387959.1 |

folK | 2-amino-4-hydroxy-6-hydroxymethyldihydropteridine pyrophosphokinase | NP_387960.1 |

lysS | lysine--tRNA ligase | NP_387963.1 |

ispD | 2-D-methyl-D-erythritol 4-phosphatecytidylyltransferase | NP_387971.1 |

ispF | 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase | NP_387972.1 |

gltX | glutamate--tRNA ligase | NP_387973.1 |

cysS | cysteine--tRNA ligase | NP_387975.1 |

secE | preprotein translocase subunit SecE | NP_387981.1 |

rplJ | 50S ribosomal protein L10 | NP_387985.2 |

rplL | 50S ribosomal protein L7/L12 | NP_387986.1 |

rpsL | 30S ribosomal protein S12 | NP_387991.2 |

rpsG | 30S ribosomal protein S7 | NP_387992.2 |

fusA | elongation factor G | NP_387993.2 |

tufA | elongation factor Tu | NP_387994.1 |

rpsJ | 30S ribosomal protein S10 | NP_387996.1 |

rplC | 50S ribosomal protein L3 | NP_387997.1 |

rplD | 50S ribosomal protein L4 | NP_387998.1 |

rplB | 50S ribosomal protein L2 | NP_388000.2 |

rpsS | 30S ribosomal protein S19 | NP_388001.1 |

rpsC | 30S ribosomal protein S3 | NP_388003.2 |

rplP | 50S ribosomal protein L16 | NP_388004.1 |

rpsQ | 30S ribosomal protein S17 | NP_388006.1 |

rplN | 50S ribosomal protein L14 | NP_388007.1 |

rplX | 50S ribosomal protein L24 | NP_388008.1 |

rplE | 50S ribosomal protein L5 | NP_388009.1 |

rpsN | 30S ribosomal protein S14 | NP_388010.1 |

rpsH | 30S ribosomal protein S8 | NP_388011.2 |

rplF | 50S ribosomal protein L6 | NP_388012.1 |

rplR | 50S ribosomal protein L18 | NP_388013.2 |

rpsE | 30S ribosomal protein S5 | NP_388014.1 |

rpmD | 50S ribosomal protein L30 | NP_388015.1 |

secY | protein translocase subunit SecY | NP_388017.1 |

adk | adenylate kinase | NP_388018.1 |

infA | translation initiation factor IF-1 | NP_388020.1 |

rpsM | 30S ribosomal protein S13 | NP_388022.2 |

rpsK | 30S ribosomal protein S11 | NP_388023.1 |

rpoA | DNA-directed RNA polymerase subunit alpha | NP_388024.1 |

rplQ | 50S ribosomal protein L17 | NP_388025.1 |

rplM | 50S ribosomal protein L13 | NP_388030.2 |

rpsI | 30S ribosomal protein S9 | NP_388031.1 |

glmM | phosphoglucosamine mutase | NP_388058.1 |

glmS | glutamine--fructose-6-phosphateaminotransferase | NP_388059.1 |

ycgG | hypothetical protein | NP_388192.2 |

nadE | NH(3)-dependent NAD(+) synthetase | NP_388195.1 |

aroK | shikimate kinase | NP_388197.2 |

ddl | D-alanine--D-alanine ligase | NP_388337.1 |

murF | UDP-N-acetylmuramoyl-tripeptide--D-alanyl-D-alanine ligase | NP_388338.1 |

acpS | holo-[acyl-carrier-protein] synthase | NP_388343.1 |

alrA | alanine racemase 1 | NP_388345.1 |

yddT | hypothetical protein | NP_388391.1 |

ydiC | hypothetical protein | NP_388473.1 |

gcp | tRNA threonylcarbamoyladenosine biosynthesisprotein Gcp | NP_388475.1 |

groES | chaperonin | NP_388483.2 |

groEL | chaperonin | NP_388484.1 |

ydiP | BsuMI modification methylase subunit YdiP | NP_388488.1 |

ligA | DNA ligase | NP_388544.1 |

gatC | Glutamyl-tRNA(Gln) amidotransferase subunit C | NP_388549.1 |

gatA | Glutamyl-tRNA(Gln) amidotransferase subunit A | NP_388550.1 |

gatB | aspartyl/glutamyl-tRNA amidotransferase subunitB | NP_388551.2 |

yerQ | diacylglycerol kinase | NP_388554.1 |

yezG | hypothetical protein | YP_003097692.1 |

cspR | Putative tRNA (cytidine(34)-2'-O)-methyltransferase | NP_388774.2 |

yhdL | anti-sigma-M factor YhdL | NP_388832.1 |

plsC | 1-acyl-sn-glycerol-3-phosphate acyltransferase | NP_388835.1 |

prsA | foldase protein PrsA | NP_388876.1 |

hemE | uroporphyrinogen decarboxylase | NP_388893.1 |

hemH | ferrochelatase | NP_388894.1 |

hemY | protoporphyrinogen oxidase | NP_388895.1 |

yisB | hypothetical protein | NP_388947.1 |

fabF | 3-oxoacyl-[acyl-carrier-protein] synthase 2 | NP_389016.1 |

trpS | tryptophan--tRNA ligase | NP_389024.1 |

ppnK | inorganic polyphosphate/ATP-NAD kinase 1 | NP_389043.1 |

hemD | hypothetical protein | NP_389105.1 |

yjzJ | hypothetical protein | YP_003097711.1 |

xkdB | phage-like element PBSX protein XkdB | NP_389134.2 |

xkdC | phage-like element PBSX protein XkdC | NP_389135.2 |

mgtE | magnesium transporter MgtE | NP_389213.1 |

patA | N-acetyl-LL-diaminopimelate aminotransferase | NP_389283.2 |

dapH | 2,3,4,5-tetrahydropyridine-2,6-dicarboxylate N-acetyltransferase | NP_389301.2 |

dapL | N-acetyldiaminopimelate deacetylase | NP_389302.1 |

rnjA | ribonuclease J 1 | NP_389336.1 |

pdhA | pyruvate dehydrogenase E1 component subunitalpha | NP_389341.1 |

ylaN | hypothetical protein | NP_389367.1 |

ftsW | lipid II flippase FtsW | NP_389368.1 |

coaD | phosphopantetheine adenylyltransferase | NP_389385.1 |

ftsL | cell division protein FtsL | NP_389398.1 |

pbpB | penicillin-binding protein 2B | NP_389399.2 |

murE | UDP-N-acetylmuramoyl-L-alanyl-D-glutamate--2,6-diaminopimelate ligase | NP_389401.1 |

mraY | phospho-n-acetylmuramoyl-pentapeptide-transferase | NP_389402.2 |

murD | UDP-N-acetylmuramoylalanine--D-glutamate ligase | NP_389403.1 |

murG | UDP-N-acetylglucosamine--N-acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N-acetylglucosamine transferase | NP_389405.2 |

murB | UDP-N-acetylenolpyruvoylglucosamine reductase | NP_389406.1 |

divIB | cell division protein DivIB | NP_389407.1 |

ftsA | cell division protein FtsA | NP_389411.2 |

ftsZ | cell division protein FtsZ | NP_389412.2 |

ileS | isoleucine--tRNA ligase | NP_389426.2 |

coaBC | coenzyme A biosynthesis bifunctional proteinCoaBC | NP_389452.1 |

priA | Primosomal protein N' | NP_389453.1 |

yloU | hypothetical protein | NP_389465.1 |

plsX | phosphate acyltransferase | NP_389471.1 |

fabD | malonyl CoA-acyl carrier protein transacylase | NP_389472.1 |

fabG | 3-oxoacyl-[acyl-carrier-protein] reductase FabG | NP_389473.1 |

acpP | acyl carrier protein | NP_389474.1 |

rnc | ribonuclease 3 | NP_389475.1 |

ffh | signal recognition particle protein | NP_389480.1 |

rpsP | 30S ribosomal protein S16 | NP_389481.1 |

trmD | tRNA (guanine-N(1)-)-methyltransferase | NP_389485.1 |

rplS | 50S ribosomal protein L19 | NP_389486.1 |

rbgA | ribosome biogenesis GTPase A | NP_389487.1 |

rpsB | 30S ribosomal protein S2 | NP_389531.1 |

tsf | elongation factor Ts | NP_389532.1 |

pyrH | uridylate kinase | NP_389533.2 |

frr | ribosome-recycling factor | NP_389534.1 |

uppS | undecaprenyl pyrophosphate synthase | NP_389535.1 |

cdsA | phosphatidate cytidylyltransferase | NP_389536.1 |

dxr | 1-deoxy-D-xylulose 5-phosphate reductoisomerase | NP_389537.2 |

proS | proline--tRNA ligase | NP_389539.1 |

nusA | transcription termination/antiterminationprotein NusA | NP_389542.1 |

infB | translation initiation factor IF-2 | NP_389545.1 |

ribC | riboflavin biosynthesis protein RibC | NP_389549.1 |

rpsO | 30S ribosomal protein S15 | NP_389550.1 |

dapG | aspartokinase 1 | NP_389558.2 |

pgsA | CDP-diacylglycerol--glycerol-3-phosphate3-phosphatidyltransferase | NP_389574.2 |

ymdA | ribonuclease Y | NP_389578.1 |

nrdI | ribonucleotide reductase stimulatory proteinNrdI | NP_389619.1 |

nrdF | ribonucleoside-diphosphate reductase subunitbeta | NP_389621.1 |

yneF | hypothetical protein | NP_389674.1 |

yneS | glycerol-3-phosphate acyltransferase | NP_389689.1 |

odhB | dihydrolipoyllysine-residue succinyltransferasecomponent of 2-oxoglutarate dehydrogenase complex | NP_389818.2 |

yomL | hypothetical protein | NP_390015.1 |

sunI | hypothetical protein | NP_390032.1 |

dfrA | dihydrofolate reductase | NP_390064.1 |

dnaD | DNA replication protein DnaD | NP_390116.1 |

panC | pantothenate synthetase | NP_390123.1 |

birA | bifunctional protein BirA | NP_390125.1 |

cca | CCA-adding enzyme | NP_390126.1 |

dapB | 4-hydroxy-tetrahydrodipicolinate reductase | NP_390130.1 |

aroE | 3-phosphoshikimate 1-carboxyvinyltransferase | NP_390141.1 |

aroF | chorismate synthase | NP_390152.3 |

hepT | heptaprenyl diphosphate synthase component 2 | NP_390155.1 |

ubiE | demethylmenaquinone methyltransferase | NP_390156.1 |

hepS | heptaprenyl diphosphate synthase component 1 | NP_390157.1 |

folE | GTP cyclohydrolase 1 | NP_390159.1 |

hbs | DNA-binding protein HU 1 | NP_390160.1 |

gpsA | glycerol-3-phosphate dehydrogenase | NP_390164.1 |

engA | GTPase Der | NP_390165.1 |

cmk | cytidylate kinase | NP_390170.1 |

resC | cytochrome c biogenesis protein ResC | NP_390194.2 |

scpA | segregation and condensation protein A | NP_390203.1 |

ypuF | hypothetical protein | NP_390204.1 |

yqjK | ribonuclease Z | NP_390265.1 |

dxs | 1-deoxy-D-xylulose-5-phosphate synthase | NP_390307.1 |

folD | bifunctional protein FolD | NP_390311.1 |

yqhY | hypothetical protein | NP_390313.2 |

accC | acetyl-CoA carboxylase biotin carboxylasesubunit | NP_390314.2 |

accB | acetyl-CoA carboxylase biotin carboxyl carrierprotein subunit | NP_390315.1 |

ispG | 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (flavodoxin) | NP_390386.1 |

ispH | 4-hydroxy-3-methylbut-2-enyl diphosphatereductase | NP_390395.2 |

dnaG | DNA primase | NP_390400.2 |

glyS | glycine--tRNA ligase beta subunit | NP_390404.2 |

glyQ | glycine--tRNA ligase alpha subunit | NP_390405.1 |

era | GTPase Era | NP_390407.1 |

holA | hypothetical protein | NP_390434.1 |

nadD | nicotinate-nucleotide adenylyltransferase | NP_390442.1 |

yqeH | hypothetical protein | NP_390445.1 |

yqaE | XRE family transcriptional regulator | NP_390512.1 |

alaS | alanine--tRNA ligase | NP_390618.1 |

mnmA | tRNA-specific 2-thiouridylase MnmA | NP_390627.3 |

aspS | aspartate--tRNA ligase | NP_390633.1 |

nifS | cysteine desulfurase NifS | NP_390666.1 |

rpmA | 50S ribosomal protein L27 | NP_390672.1 |

rplU | 50S ribosomal protein L21 | NP_390674.1 |

mreD | rod shape-determining protein MreD | NP_390679.1 |

mreC | cell shape-determining protein MreC | NP_390680.2 |

mreB | rod shape-determining protein MreB | NP_390681.2 |

folC | folylpolyglutamate synthase | NP_390686.1 |

hemB | delta-aminolevulinic acid dehydratase | NP_390691.1 |

hemC | porphobilinogen deaminase | NP_390693.1 |

hemA | glutamyl-tRNA reductase | NP_390695.1 |

engB | GTP-binding protein EngB | NP_390697.1 |

racE | glutamate racemase | NP_390717.2 |

trxA | thioredoxin | NP_390728.1 |

pheT | phenylalanine--tRNA ligase beta subunit | NP_390741.1 |

pheS | phenylalanine--tRNA ligase alpha subunit | NP_390742.1 |

ysgA | tRNA/rRNA methyltransferase YsgA | NP_390743.1 |

rplT | 50S ribosomal protein L20 | NP_390763.1 |

infC | translation initiation factor IF-3 | NP_390765.1 |

dnaI | primosomal protein DnaI | NP_390776.1 |

dnaB | replication initiation and membrane attachmentprotein | NP_390777.1 |

coaE | dephospho-CoA kinase | NP_390784.1 |

accA | acetyl-CoA carboxylase carboxyltransferasesubunit alpha | NP_390798.1 |

accD | acetyl-CoA carboxylase carboxyltransferasesubunit beta | NP_390799.2 |

rpsD | 30S ribosomal protein S4 | NP_390844.1 |

tyrS | tyrosine--tRNA ligase 1 | NP_390845.1 |

murC | UDP-N-acetylmuramate--L-alanine ligase | NP_390857.1 |

asnB | asparagine synthetase 1 | NP_390932.1 |

metK | S-adenosylmethionine synthase | NP_390933.1 |

mntA | manganese-binding lipoprotein MntA | NP_390955.1 |

menC | o-succinylbenzoate synthase | NP_390956.1 |

menE | 2-succinylbenzoate-CoA ligase | NP_390957.1 |

menD | 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate synthase | NP_390960.1 |

mrpA | Na(+)/H(+) antiporter subunit A | NP_391038.2 |

mrpB | Na(+)/H(+) antiporter subunit B | NP_391039.1 |

mrpC | Na(+)/H(+) antiporter subunit C | NP_391040.2 |

mrpD | Na(+)/H(+) antiporter subunit D | NP_391041.3 |

mrpE | Na(+)/H(+) antiporter subunit E | YP_054591.2 |

mrpF | Na(+)/H(+) antiporter subunit F | NP_391043.2 |

mrpG | Na(+)/H(+) antiporter subunit G | NP_391044.1 |

yumC | ferredoxin--NADP reductase 2 | NP_391091.2 |

dapF | diaminopimelate epimerase | NP_391097.1 |

sufB | FeS cluster assembly protein SufB | NP_391146.1 |

iscU | NifU-like protein | NP_391147.1 |

sufS | cysteine desulfurase | NP_391148.1 |

sufD | FeS cluster assembly protein SufD | NP_391149.1 |

sufC | ABC transporter ATP-binding protein | NP_391150.1 |

rghRA | HTH-type transcriptional repressor RghR | NP_391246.1 |

eno | enolase | NP_391270.1 |

pgm | 2,3-bisphosphoglycerate-independent phosphoglycerate mutase | NP_391271.1 |

gapA | glyceraldehyde-3-phosphate dehydrogenase 1 | NP_391274.1 |

trxB | thioredoxin | NP_391359.1 |

prfB | peptide chain release factor 2 | NP_391409.1 |

secA | protein translocase subunit SecA | NP_391410.1 |

tagO | undecaprenyl-phosphate N-acetylglucosaminyl1-phosphate transferase | NP_391433.1 |

tuaB | teichuronic acid biosynthesis protein TuaB | NP_391440.1 |

mnaA | UDP-N-acetylglucosamine 2-epimerase | NP_391446.1 |

tagH | teichoic acids export ATP-binding protein TagH | NP_391451.1 |

tagG | teichoic acid translocation permease proteinTagG | NP_391452.1 |

tagF | CDP-glycerol:poly | NP_391453.1 |

tagD | glycerol-3-phosphate cytidylyltransferase | NP_391455.1 |

tagA | N-acetylmannosaminyltransferase | NP_391456.1 |

tagB | CDP-glycerol:glycerophosphateglycerophosphotransferase | NP_391457.1 |

mbl | MreB-like protein | NP_391522.2 |

murAA | UDP-N-acetylglucosamine1-carboxyvinyltransferase 1 | NP_391557.1 |

prfA | peptide chain release factor 1 | NP_391582.1 |

pyrG | CTP synthase | NP_391596.1 |

rodA | rod shape-determining protein RodA | NP_391691.1 |

menA | 1,4-dihydroxy-2-naphthoate octaprenyltransferase | NP_391728.1 |

yxlC | hypothetical protein | NP_391748.1 |

walK | sensor histidine kinase | NP_391920.1 |

walR | transcriptional regulator | NP_391921.1 |

dnaC | replicative DNA helicase | NP_391924.2 |

ppaC | manganese-dependent inorganic pyrophosphatase | NP_391935.1 |

rpsR | 30S ribosomal protein S18 | NP_391969.2 |

ssbA | single-stranded DNA-binding protein | NP_391970.1 |

rnpA | ribonuclease P protein component | NP_391985.1 |

#### Bionet

The bionet enables open peer-peer exchange of functional biomaterials and associated data. This product may also be available from bionet nodes that are more convenient to you. Here are other bionet nodes who may be willing to provide you this specific product.Name | Contact |
---|---|

Amjed Alsultan | amjed.talib {at} qu {dot} edu {dot} iq |

Jordan Gonzalez | jgonzalez {at} thecitizensciencelab {dot} org |

Dong Nguyen Tam | NA |