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GenBank/EMBL accession numbers J01749, K00005, L08654, M10282, M10283, M10286, M10356, M10784, M10785, M10786, M33694, V01119.
The plasmid pBR322 is one of the most commonly used E.coli cloning vectors. pBR322 is 4361 bp in length and contains: (1) the replicon rep responsible for the replication of plasmid (source - plasmid pMB1); (2) rop gene coding for the Rop protein, which promotes conversion of the unstable RNA I - RNA II complex to a stable complex and serves to decrease copy number (source - plasmid pMB1); (3) bla gene, coding for beta-lactamase that confers resistance to ampicillin (source - transposon Tn3); (4) tet gene, encoding tetracycline resistance protein (source - plasmid pSC101).
The circular sequence is numbered such that 1 is the first T of the unique EcoRI site GAATTC and the count increases first through the tet gene, the pMB1 material, and finally through the Tn3 region. The map shows enzymes that cut pBR322 DNA once. Enzymes produced by Fermentas are shown in blue. The coordinates refer to the position of first nucleotide in each recognition sequence.
The exact position of genetic elements is shown on the map (termination codons included). The bla gene nucleotides 4153-4085 (complementary strand) code for a signal peptide. The indicated rep region is sufficient to promote replication. DNA replication initiates at position 2533 (+/- 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.
References
Bolivar, F., Rodriguez, R.L., Greene, P.J., Betlach, M.C., Heyneker. H,L. and Boyer, H.W., Construction and characterization of new cloning vehicles. II. A multipurpose cloning system, Gene 2, 95-113, 1977.
Covarrubias, L., Cervantes, L., Covarrubias, A., Soberon, X., Vichido, I., Blanco, A., Kupersztoch-Portnoy, Y.M. and Bolivar, F., Construction and characterization of new cloning vehicles. V. Mobilization and coding properties of pBR322 and several deletion derivatives including pBR327 and pBR328, Gene 13, 25-35, 1981.
Peden, K.W., Revised sequence of the tetracycline-resistance gene of pBR322, Gene 22, 277-280, 1983.
Sutcliffe, J.G., Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322, Proc. Natl. Acad. Sci. U.S.A., 75, 3737-3741, 1978.
Sutcliffe, J.G., Complete nucleotide sequence of the Escherichia coli plasmid pBR322, Cold Spring Harb. Symp. Quant. Biol., 43, Pt 1, 77-90,1979.
Watson, N., A new revision of the sequence of plasmid pBR322, Gene 70, 399-403, 1988.
Enzymes which cut pBR322 DNA once:
AatII 4284, AflIII 2473, BamHI 375, BoxI 712, Bpu10I 1580, BsaAI 2225, BseJI 1668, BsgI 1650, Bsp68I 972, Bst1107I 2244, Bsu15I 23, BveI 1063, CaiI 2884, Eam1105I 3361, Eco31I 3433, Eco32I 185, Eco52I 939, Eco88I 1425, Eco130I 1369, EcoRI 4359, Esp3I 2122, HindIII 29, Kpn2I 1664, MlsI 1444, Mva1269I 1353, NdeI 2295, NheI 229, PaeI 562, PfoI 2117, PscI 2473, PstI 3607, PsyI 2217, PvuI 3733, PvuII 2064, SalI 651, SapI 2350, ScaI 3844, SgrAI 409, SspI 4168, TstI 389, VspI 3537, XagI 622, XapI 4359.
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Updated liepos 25, 2008 10:10
