PUC19

pUC19 is a plasmid clonong vector created by Messing and co-workers in the University of California. p in the name stands for plasmid and UC represents the University in which it was created. It is circular and has 2686 base pairs^[1] and contains one amp^R gene, lac Z gene of E. coli and lac I gene. The MCS (multiple chemical sensitivity) region is splitted into the lac Z gene (codons 6-7 of lac Z are replaced by MCS), where various restriction sites for many restriction endonucleases are present. pUC18 is almost similar to pUC19, but possess different restriction sites.

The ori site or replicon, rep is derived from pMB1 vector. pUC vector is small but has a high copy number. The high copy number of pUC plasmids is a result of the lack of the rop gene and a single point mutation in rep of pMB1. The lac Z gene codes for β-galactosidase. The lac Z fragment, whose synthesis can be induced by IPTG, is capable of intra-allelic complementation with a defective form of β-galactosidase enzyme encoded by host chromosome (mutation lacZDM15). In the presence of IPTG in growth medium, bacteria synthesise both fragments of the enzyme and form blue colonies on media with X-Gal, by hydrolysing X-Gal. Insertion of DNA into the MCS located within the lac Z gene inactivates the N-terminal fragment of beta-galactosidase and abolishes intra-allelic complementation. Thus bacteria carrying recombinant plasmids in the MCS cannot hydrolyse X-Gal, giving rise to white colonies.

The sequence on pUC19 is^[2]

pUC19 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat 421 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct 481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 1021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 1081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 1141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 1201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 1261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 1321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 1381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 1441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 1501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 1561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 1621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 1681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 1741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 1801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 1861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 1921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 1981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 2041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 2101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 2161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 2221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 2281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 2341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 2401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 2461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 2521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 2581 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc 2641 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc

References

^[1] pUC18, pUC19: description & restriction map

^[2] Sequence of pUC19