pUC18, pUC19

pUC18 and pUC19 vectors are small, high copy number, E.coli plasmids, 2686 bp in length. They are identical except that they contain multiple cloning sites (MCS) arranged in opposite orientations. pUC18/19 plasmids contain: (1) the pMB1 replicon rep responsible for the replication of the plasmid (source plasmid pBR322). The high copy number of pUC plasmids is a result of the lack of the rop gene and a single point mutation in the replicon rep of pMB1; (2) the bla gene, coding for -lactamase, that confers resistance to ampicillin (source plasmid pBR322). It differs from that of pBR322 by two point mutations; (3) the region of E.coli lac operon containing a CAP protein binding site, promoter Plac, lac repressor binding site and the 5-terminal part of the lacZ gene encoding the N-terminal fragment of -galactosidase (source M13mp18/19). This fragment, whose synthesis can be induced by IPTG, is capable of intra-allelic () complementation with a defective form of -galactosidase encoded by the host (mutation (lacZ) M15 ). In the presence of IPTG, bacteria synthesize both fragments of the enzyme and form blue colonies on media with X-Gal. Insertion of DNA into the MCS located within the lacZ gene (codons 6-7 of lacZ are replaced by MCS) inactivates the N-terminal fragment of -galactosidase and abolishes -complementation. Bacteria carrying recombinant plasmids therefore give rise to white colonies. The map shows enzymes that cut pUC18/19 DNA once. Thermo Scientific enzymes are shown in orange. The coordinates refer to the position of the first nucleotide in each recognition sequence. The exact positions of the genetic elements are shown on the map (termination codons included). The bla gene nucleotides 2486-2418 (complementary strand) code for a signal peptide. The LacZ polypeptide corresponding to wt -galactosidase and essential for blue/white screening ends at nt position 236 (compl. strand); another 30 codons in the same reading frame are derived from pBR322. The indicated rep region is sufficient to promote replication. DNA replication initiates at position 866 (+/- 1) and proceeds in the direction indicated. Plasmids carrying the pMB1 and ColE1 replicons are incompatible, but they are fully compatible with those carrying the p15A replicon (pACYC177, pACYC184). pMB1-derived plasmids can be amplified using chloramphenicol.

GenBank/EMBL Accession Numbers

For pUC18 L09136; for pUC19 L09137.

Additional Information
CAP protein binding site 591-554 (compl. strand); mRNA (LacZ) starts at nt position 507 (compl. strand); lac repressor binding site 507-487 (compl. strand). Enzymes which cut pUC18 DNA once: HindIII* 399 AatII 2617 KpnI* 438 Acc65I* 438 LguI 683 AflIII 806 NdeI 183 BamHI* 429 NmeAIII 1822 BcgI 2215 PaeI* 405 BsaXI 659 PdmI 2294 BstAPI 179 PfoI 46 BveI* 413 PscI 806 CaiI 1217 PspFI 1110 Cfr9I* 434 PstI* 411 Cfr10I 1779 SacI* 444 Eam1105I 1694 SalI* 417 Ecl136II* 444 ScaI 2177 Eco24I* 444 SdaI* 410 Eco31I 1766 SmaI* 434 Eco88I* 434 SspI 2501 EcoO109I 2674 SspDI 235 EcoRI* 450 XapI* 450 EheI 235 XbaI* 423 GsuI 1784 XmiI* 417 HincII* 417 * MCS

Multiple Cloning Site of pUC18

M13/pUC sequencing primer, 17-mer (-20), (#S0100) 399 HindIII 5 - G TAA AAC GAC GGC CAG TGC CAA GCT TGC 3 - C ATT TTG CTG CCG GTC ACG GTT CGA ACG Val Val Ala Leu Ala Leu Ser Ala PaeI ATG TAC His PstI HincII Cfr9I Ecl136II SdaI SalI Eco88I Acc65I Eco24I EcoRI BveI XmiI XbaI BamHI SmaI KpnI SacI XapI 455 CCT GCA GGT CGA CTC TAG AGG ATC CCC GGG TAC CGA GCT CGA ATT CGT AAT CAT GGT CAT AGC TGT TTC CTG - 3 GGA CGT CCA GCT GAG ATC TCC TAG GGG CCC ATG GCT CGA GCT TAA GCA TTA GTA CCA GTA TCG ACA AAG GAC - 5 Arg Cys Thr Ser Glu Leu Pro Asp Gly Pro Val Ser Ser Ser Asn Thr Ile Met Thr Met M13/pUC reverce sequencing primer, 17-mer (-26), (#S0101)

Multiple Cloning Site of pUC19

M13/pUC sequencing primer, 17-mer (-20), (#S0100) Ecl136II Cfr9I HincII PstI EcoRI Eco24I Acc65I Eco88I SalI BveI 396 XapI SacI KpnI SmaI BamHI XbaI XmiI SdaI PaeI HindIII 452 5 - G TAA AAC GAC GGC CAG TGA ATT CGA GCT CGG TAC CCG GGG ATC CTC TAG AGT CGA CCT GCA GGC ATG CAA GCT TGG CGT AAT CAT GGT CAT AGC TGT TTC CTG - 3 3 - C ATT TTG CTG CCG GTC ACT TAA GCT CGA GCC ATG GGC CCC TAG GAG ATC TCA GCT GGA CGT CCG TAC GTT CGA ACC CGA TTA GTA CCA GTA TCG ACA AAG GAC - 5 Val Val Ala Leu Ala Leu Ser Ala His Arg Cys Thr Ser Glu Leu Pro Asp Gly Pro Val Ser Ser Ser Asn Thr Ile Met Thr Met M13/pUC reverce sequencing primer, 17-mer (-26), (#S0101)

Reference 1. Yanisch-Perron, C., et al., Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors, Gene, 33, 103-119, 1985.

pUC18 DNA Restriction Sites

Enzyme DrdI AatII Acc65I AflIII Alw21I Alw26I Alw44I BamHI BauI BccI BceAI BcgI BcnI BmrI BfmI BfuI BglI Bme1390I BpuEI BsaWI BsaXI BseDI BseGI BseLI BseMI BseMII BseNI BseSI BseXI Bsh1236I Bsh1285I BshNI BspLI BspPI BstAPI BsuRI BtsI BveI CaiI EaeI Cfr9I Cfr10I Cfr13I CseI Csp6I DraI Eam1104I Eam1105I EciI Ecl136II Eco24I Eco31I Eco47I Eco57I Eco88I EcoO109I EcoP15I EcoRI EcoRII EheI Esp3I FokI FspBI GsuI HaeII Number of recognition sites 1 91 2617 438 806 177 51 177 429 979 1735 387 2215 47 364 411 1015 245 47 912 1012 659 354 77 47 1753 171 365 177 41 2 276 235 235 429 179 287 593 413 1217 388 434 1779 286 108 168 1563 290 1694 878 444 444 1766 1837 1333 434 2674 1214 450 354 235 51 77 424 1784 235 2 908 3 4 5 6 7 8 9 10 11 12 Enzyme Hin1I Hin1II HincII HindIII HinfI HphI Hpy8I Hpy99I Hpy188I HpyAV HpyF3I KpnI LguI Lsp1109I LweI MaeIII MbiI MboII MmeI MspA1I MvaI NdeI NmeAIII NmuCI NsbI PaeI PagI PdmI PfeI PfoI PscI Psp1406I PspFI PstI PsuI PvuI PvuII RsaI RseI SacI SalI ScaI SchI SdaI SduI SmaI SmoI FauI SspI SspDI TaaI TaiI TaqI TasI TatI TauI TscAI TseI TspDTI TspGWI VspI XapI XbaI XceI XmiI Number of recognition sites 1 235 38 417 399 420 12 177 372 31 270 171 438 683 41 78 57 496 291 996 112 354 183 1822 57 256 405 1526 2294 641 46 806 1924 1110 411 429 276 306 168 1947 444 417 2177 420 410 177 434 912 114 2501 235 19 374 418 451 167 150 392 41 639 2149 576 450 423 37 417 2 2235 406 641 21 417 385 156 1036 1081 254 153 348 737 685 1180 306 545 368 1919 2534 781 2297 1447 2066 628 439 2106 1458 2178 2465 1544 1556 2324 2341 3 2617 461 706 1550 1120 907 918 1304 1490 327 208 368 2538 1456 628 833 1956 2639 4 807 781 1777 1608 1701 996 1346 1656 630 229 1163 1547 1146 954 2167 5 1527 1177 2173 2366 1964 1349 1817 2196 711 894 1226 2302 1391 967 6 2018 1694 2399 1483 2054 2622 729 1946 1342 2380 2332 7 2028 8 2106 9 2142 10 2535 11 2640 12

444 1766 1120 2363 1859 1292 82 1744 1071 2542 1813 82 1174 1159 433 321 648 1935 1081 391 1120 254 4 719 438 429 430 389 2092 645 1741 908 439 1582 684 1024

1120 2531 2366 2670 2146 434 1262 354 1451 1990 434 1679 822 1490 606 2366 327 107 1143 550 438 1373 646 2120 2087 1820 1486 2178 2274 2488 1852

2281 2684

2366

2414 1618 2429 2064 2075 2195

435 1940 434 2319 545 1860 840 1656 1209 630 652 2066 1647 550 1447 820

1185

1881

2232

1148 2156 1625 2489

1213 2386 1956

1216 2014

1422 2167

1750 2355

2116

435

545

833

954

967

1185

1881

2232

966 2147 1006 2196 1222 711 654 2215 836 1459 831

1285 1339 729 852 875 1544 849 1745 1148 1433 1647 1557 1283 1863 1213 1763 1741 2021 1741 1906 1216 2256 1782 2324 1821 2170 1422 2588 1993 2342 2088 2345 1750 2116

2583 2675

1837 2236

2059

2675

706 444 1174 124 54 1509 448 487 2177 732 593 254 1575 2466 635 405

1177 1120 1451 284 262 1925 906 504 850 702 327 1677 1870 806

1694 2281 2319 598 768 2298 2350 579 1005 1208 630 1980 2366 655 839 2618 1567 2089 1221 711 1309 1873 2211 1492 729 1622 2128 2440 1641 1148 1746 1213 2093 1216 2120 1422 2137

2059 2381 1330 545 2683 321 1301 680 1423 833 1679 1554 1050 954 1860 1889 967 2147

1750

2116

There are no restriction sites in pUC18 DNA for the following enzymes: AanI, AarI, AbsI, AdeI, AjiI, AjuI, AlfI, AloI, ApaI, BaeI, BbvCI, BclI, BcuI, BglII, BoxI, BpiI, BplI, Bpu10I, Bpu1102I, BseJI, BseRI, BsgI, BshTI, Bsp68I, Bsp119I, Bsp120I, Bsp1407I, BspOI, BspTI, Bst1107I, BstXI, Bsu15I, BtgI, BtgZI, Cfr42I, CpoI, CspCI, Eco32I, Eco47III, Eco52I, Eco72I, Eco81I, Eco91I, Eco105I, Eco130I, Eco147I, FalI, FaqI, FseI, FspAI, Kpn2I, KspAI, MlsI, MluI, Mph1103I, MreI, MssI, MunI, Mva1269I, NcoI, NheI, NotI, OliI, PacI, PasI, PauI, PdiI, Pfl23II, Ppu21I, Psp5II, PspXI, PsrI, PsyI, SanDI, SexAI, SfaAI, SfiI, SgfI, SgrAI, SgrDI, SgsI, SmiI, SrfI, Van91I, XagI, XcmI, XhoI, XmaJI.

Indicates restriction sites of Thermo Scientific enzymes that are sensitive (cleavage completely blocked or partially inhibited) to overlapping Dam/Dcm methylation. Complete cleavage is achieved with DNA substrates isolated from dam or dcm hosts.