TABLE S1. Insertions in known and putative essential genes(a) bnumber Gene Gene length Insertion site(b) Orientation(c) product b0058 yabO 660 454 S Ribosomal large subunit pseudouridine synthase A b0122 yacC 471 435 S orf, hypothetical protein b0167 glnD 2673 2325 S protein PII; uridylyltransferase acts on regulator of glnA b0215 dnaQ 732 486 AS DNA polymerase III, epsilon subunit b0245 b0245 342 59 S orf, hypothetical protein b0270 yagG 1383 104 S putative permease b0285 yagS 957 787 S orf, hypothetical protein b0320 yahF 1548 1273 S putative oxidoreductase subunit b0429 cyoD 330 171 S cytochrome o ubiquinol oxidase subunit IV b0469 apt 552 421 AS adenine phosphoribosyltransferase b0521 arcC 894 163 S putative carbamate kinase b0547 ybcN 456 158 AS orf, hypothetical protein b0592 fepB 957 181 AS ferric enterobactin (enterochelin) binding protein; periplasmic component b0636 ybeA 468 152 S orf, hypothetical protein b0652 gltL 726 461 S ATP-binding protein of glutamate/aspartate transport system b0659 ybeY 468 440 AS orf, hypothetical protein b0708 phrB 1419 823 AS deoxyribodipyrimidine photolyase (photoreactivation) b0733 cydA 1572 568 S cytochrome d terminal oxidase, polypeptide subunit I b0736 ybgC 405 378 AS orf, hypothetical protein b0798 ybiA 483 401 S orf, hypothetical protein b0810 glnP 660 560 S glutamine high-affinity transport system; membrane component b0846 b0846 537 349 S putative DEOR-type transcriptional regulator b0850 ybjC 288 215 AS orf, hypothetical protein b0865 ybjP 516 230 AS putative lipoprotein b0866 b0866 324 170 AS orf, hypothetical protein b0867 b0867 831 458 AS probable N-acetylmuramoyl-L-alanine amidase b0868 b0868 1050 732 S orf, hypothetical protein b0897 ycaC 627 245 S orf, hypothetical protein b0924 mukB 4461 4399 AS kinesin-like cell division protein involved in chromosome partitioning b0963 mgsA 468 293 S methylglyoxal synthase b0972 hyaA 1119 834 S hydrogenase-1 small subunit b0974 hyaC 708 460 AS probable Ni/Fe-hydrogenase 1 b-type cytochrome subunit b0975 hyaD 588 405 AS processing of HyaA and HyaB proteins b0987 ymcD 324 105 S orf, hypothetical protein b0995 torR 693 492 S response transcriptional regulator for torA (sensor TorS) b0998 torD 600 577 S part of trimethylamine-N-oxide oxidoreductase b0999 yccD 306 198 AS orf, hypothetical protein b1010 b1010 387 344 S hypothetical protein b1011 b1011 735 499 AS hypothetical isochorismatase family protein b1051 msyB 378 255 AS acidic protein suppresses mutants lacking function of protein export b1123 potD 1047 632 AS spermidine/putrescine periplasmic transport protein b1124 potC 795 672 S spermidine/putrescine transport system permease b1128 ycfD 1131 741 S orf, hypothetical protein b1144 ymfJ 285 221 AS hypothetical protein in lambdoid prophage e14 region b1175 minD 813 152 S cell division inhibitor, a membrane ATPase, activates minC b1189 dadA 1299 998 S D-amino acid dehydrogenase subunit b1204 pth 585 551 S peptidyl-tRNA hydrolase b1205 ychH 279 113 S orf, hypothetical protein b1213 b1213 393 97 AS orf, hypothetical protein b1237 hns 414 323 S DNA-binding protein HLP-II (HU, BH2, HD, NS); pleiotropic regulator b1246 oppD 1014 731 AS homolog of Salmonella ATP-binding protein of oligopeptide ABC transport system b1253 yciA 399 167 AS orf, hypothetical protein b1270 btuR 591 371 S cob(I)alamin adenolsyltransferase b1281 pyrF 738 237 AS orotidine-5'-phosphate decarboxylase b1306 pspC 360 260 AS phage shock protein: activates phage shock-protein expression b1308 pspE 315 101 AS phage shock protein b1322 ycjF 1062 653 S orf, hypothetical protein b1390 ydbP 747 327 AS phenylacetic acid degradation protein b1421 trg 1641 509 AS methyl-accepting chemotaxis protein III, ribose sensor receptor b1427 rimL 540 386 AS ribosomal-protein-serine acetyltransferase b1438 b1438 438 193 S orf, hypothetical protein b1441 b1441 1014 920 S putative ABC transporter ATP-binding protein b1442 b1442 942 50 AS putative ABC transporter permease protein b1443 b1443 795 248 S putative ABC transporter permease protein b1448 b1448 519 345 S hypothetical acetyltransferase b1452 b1452 1062 46 S putative receptor b1463 b1463 846 802 S N-hydroxyarylamine O-acetyltransferase b1477 yddM 363 286 AS orf, hypothetical protein b1477 yddM 363 302 S orf, hypothetical protein b1523 b1523 360 225 S orf, hypothetical protein b1531 marA 390 248 S multiple antibiotic resistance; transcriptional activator of defense systems b1534 ydeF 1188 426 AS putative transport protein b1556 b1556 291 172 S lysis protein S homolog from lambdoid prophage Qin b1563 relE 288 190 AS orf, hypothetical protein b1565 b1565 306 134 S orf, hypothetical protein b1578 b1578 657 426 AS orf, hypothetical protein b1582 b1582 327 76 S orf, hypothetical protein b1586 b1586 348 145 S orf, hypothetical protein b1589 b1589 618 258 AS putative dimethyl sulfoxide reductase, Fe-S subunit b1609 rstB 1302 684 AS sensor histidine protein kinase (RstA regulator) b1610 tus 930 836 AS DNA-binding protein; inhibition of replication at Ter sites b1618 uidR 591 203 AS repressor for uid operon b1654 ydhD 348 227 S orf, hypothetical protein b1671 b1671 720 520 AS putative oxidoreductase, Fe-S subunit b1684 ydiC 369 146 S orf, hypothetical protein b1686 b1686 411 145 AS orf, hypothetical protein b1695 b1695 1206 169 AS putative oxidoreductase b1698 ydiR 939 393 S putative flavoprotein b1700 ydiT 294 244 AS orf, hypothetical protein b1704 aroH 1047 215 AS 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase b1722 b1722 759 217 S orf, hypothetical protein b1738 celA 321 238 S PEP-dependent phosphotransferase enzyme IV for cellobiose, arbutin, and salicin b1741 b1741 888 599 AS putative excinuclease subunit b1742 b1742 639 472 AS orf, hypothetical protein b1745 b1745 1344 503 AS succinylarginine dihydrolase b1747 b1747 1035 315 S arginine succinyltransferase b1756 b1756 654 151 AS putative ATP-binding component of a transport system b1764 selD 1044 193 S selenophosphate synthase, H(2)Se added to acrylyl-tRNA b1791 yeaN 1182 456 S putative amino acid/amine transport protein b1846 yebE 660 417 AS orf, hypothetical protein b1848 yebG 291 119 S orf, hypothetical protein b1863 ruvC 522 86 S Holliday junction nuclease; resolution of structures; repair b1864 yebC 741 201 AS orf, hypothetical protein b1865 ntpA 453 241 S dATP pyrophosphohydrolase b1871 yecP 972 275 S putative enzyme b1884 cheR 861 229 AS response regulator for chemotaxis; protein glutamate methyltransferase b1887 cheW 504 25 S positive regulator of CheA protein activity b1890 motA 888 546 AS proton conductor component of motor; no effect on switching b1900 araG 1515 835 AS ATP-binding component of high-affinity L-arabinose transport system b1901 araF 990 512 AS L-arabinose-binding periplasmic protein b1929 yedE 1206 119 S putative transport system permease protein b1976 b1976 837 631 AS orf, hypothetical protein b2009 sbmC 474 365 S SbmC protein b2042 galF 894 520 AS homolog of Salmonella UTP--glucose-1-P uridyltransferase, probably a UDP-gal transferase b2073 b2073 660 308 AS orf, hypothetical protein b2080 b2080 372 121 S orf, hypothetical protein b2091 gatD 1041 479 S galactitol-1-phosphate dehydrogenase b2114 metG 2034 1637 S methionine tRNA synthetase b2150 mglB 999 322 AS galactose-binding transport protein; receptor for galactose taxis b2179 yejE 1026 874 S putative transport system permease protein b2231 gyrA 2628 496 S DNA gyrase, subunit A, type II topoisomerase b2244 yfaD 900 450 AS orf, hypothetical protein b2246 b2246 1329 343 S putative transport protein b2267 elaA 462 249 S orf, hypothetical protein b2313 cvpA 489 334 S membrane protein required for colicin V production b2387 b2387 327 162 AS putative PTS system enzyme IIB component b2423 cysW 450 271 S sulfate transport system permease W protein b2450 b2450 840 14 S orf, hypothetical protein b2456 cchB 288 167 S detox protein b2479 gcvR 639 128 S transcriptional regulation of gcv operon b2518 ndk 432 234 AS nucleoside diphosphate kinase b2539 hcaA2 519 173 AS small terminal subunit of phenylpropionate dioxygenase b2540 hcaC 321 181 S ferredoxin subunit of phenylpropionate dioxygenase b2567 rnc 681 562 S RNase III, ds RNA b2582 trxC 420 258 AS putative thioredoxin-like protein b2618 b2618 309 189 S orf, hypothetical protein b2717 hycI 471 106 AS protease involved in processing C-terminal end of the large subunit of hydrogenase 3 b2720 hycF 543 147 AS probable iron-sulfur protein of hydrogenase 3 (part of FHL complex) b2732 ygbA 354 205 AS orf, hypothetical protein b2737 b2737 1167 597 S orf, hypothetical protein b2748 b2748 312 26 S orf, hypothetical protein b2811 ygdK 444 230 AS orf, hypothetical protein b2828 lgt 876 552 S phosphatidylglycerol-prolipoprotein diacylglyceryl transferase; a major membrane phospholipid b2909 ygfB 585 216 S orf, hypothetical protein b2926 pgk 1164 880 S phosphoglycerate kinase b2935 tktA 1992 546 S transketolase 1 isozyme b2947 gshB 951 563 S glutathione synthetase b2948 yqgE 636 459 S orf, hypothetical protein b2957 ansB 1047 688 AS periplasmic L-asparaginase II b2990 hybG 249 121 S hydrogenase-2 operon protein: may effect maturation of large subunit of hydrogenase-2 b2993 hybD 495 63 S probable processing element for hydrogenase-2 b3002 yqhA 495 275 S orf, hypothetical protein b3033 yqiB 423 187 AS putative enzyme b3066 dnaG 1746 798 S DNA biosynthesis; DNA primase b3095 yqjA 663 206 AS orf, hypothetical protein b3119 tdcR 345 93 AS threonine dehydratase operon activator protein b3133 agaV 510 288 AS N-acetylgalactosamine-specific IIB component 2 (EIIB-AGA) b3164 pnp 2205 556 S polynucleotide phosphorylase; cytidylate kinase activity b3202 rpoN 1434 324 AS RNA polymerase, sigma(54 or 60) factor; nitrogen and fermentation regulation b3204 ptsN 492 284 AS phosphotransferase system enzyme IIA, regulates N metabolism b3219 yhcF 717 500 AS putative transcriptional regulator b3226 yhcK 792 385 S Nan regulator b3234 degQ 1368 833 AS serine endoprotease b3237 argR 471 118 AS repressor of arg regulon; cer-mediated site specific recombination b3306 rpsH 393 300 S 30S ribosomal subunit protein S8, and regulator b3344 yheM 360 70 AS orf, hypothetical protein b3386 rpe 678 386 S D-ribulose-5-phosphate 3-epimerase b3463 ftsE 669 284 S ATP-binding component of a membrane-associated complex involved in cell division b3500 gor 1353 1092 AS glutathione oxidoreductase b3560 glyQ 912 254 S glycine tRNA synthetase, alpha subunit b3607 cysE 822 108 AS serine acetyltransferase b3639 dfp 1293 1182 AS flavoprotein affecting synthesis of DNA and pantothenate metabolism b3706 thdF 1365 378 S GTPase involved in synthesis of hypermodified nucleoside, tRNA 5-methylaminomethyl-2-thiouridine; b3731 atpC 420 369 AS membrane-bound ATP synthase, F1 sector, epsilon-subunit b3736 atpF 471 159 S membrane-bound ATP synthase, F0 sector, subunit b b3778 rep 2022 508 AS rep helicase, a single-stranded DNA dependent ATPase b3816 corA 951 798 S Mg2+ transport, system I b3860 dsbA 627 232 S protein disulfide isomerase I, essential for cytochrome c synthesis and formate-dependent reduction b3902 rhaD 825 509 S rhamnulose-phosphate aldolase b3958 argC 1005 955 AS N-acetyl-gamma-glutamylphosphate reductase b3995 yjaE 477 306 S regulator of sigma D b4030 yjbA 411 179 S orf, hypothetical protein b4054 tyrB 1194 309 AS tyrosine aminotransferase, tyrosine repressible b4155 yjeA 1008 398 S putative lysyl-tRNA synthetase b4262 yjgQ 1086 1066 S orf, hypothetical protein b4302 sgcA 432 387 AS putative PTS system enzyme II A component b4373 rimI 447 402 S acyltransferase for 30S ribosomal subunit protein S18; acetylation of N-terminal alanine (a) Based on essential gene list of Gerdes et al. (22). (b) Last position in the gene from the translation start before the insertion. (c) Orientation of transposon relative to KanR transcription. AS, antisense; S, sense.