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GenBank/EMBL accession number X06402.
Not available from Fermentas
The 3941 bp plasmid pACYC177 is compatible with pMB1- or ColE1-related plasmids and can therefore be used with a pMB1- or ColE1-derivative within the same cell. pACYC177 contains: (1) the replicon rep responsible for the replication of plasmid (source - plasmid p15A); (2) kan gene, coding for aminoglycoside 3'-phosphotransferase that confers resistance to kanamycin (source - transposon Tn903); (3) bla gene, coding for beta-lactamase that confers resistance to ampicillin (source - transposon Tn3).
The circular sequence is numbered such that 1 is the first G of the unique HincII site GTTGAC and the count increases first through the 3'-terminal part of bla gene, the p15A material, the Tn903 material and finally through the 5'-terminal part of bla gene. The map shows enzymes that cut pACYC177 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). In the bla gene, nucleotides 3699-3767 code for a signal peptide. The indicated rep region is sufficient to promote replication. DNA replication initiates at position 1330 (+/- 1) and proceeds in the direction indicated.
Derivatives of p15A are normally present in a somewhat lower copy number than pMB1 derivatives and cannot be amplified to the same extent as pMB1 derivatives. Plasmid pACYC177 can be amplified using chloramphenicol or spectinomycin.
References
Chang, A.C.Y. and Cohen, S.N., Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid, J. Bacteriol., 134, 1141-1156, 1978.
Mok, Y.K., Clark, D.R., Kam, K.M. and Shaw, P.C., BsiYI, a novel thermophilic restriction enzyme that recognizes 5' CCNNNNNNNGG 3' and the discovery of a wrongly sequenced site in pACYC177, Nucleic Acids Res., 19, 2321-2323, 1991.
Rose, R.E., The nucleotide sequence of pACYC177, Nucleic Acids Res., 16, 356, 1988.
Enzymes which cut pACYC177 DNA once:
AasI 3397, AatII 3563, AdeI 1818, AlfI 2024, Alw44I 3814, BamHI 3320, BcgI 18, BfiI 495, BfmI 299, BfuI 3638, BglI 421, BpiI 3496, BsgI 1506, BshNI 592, Bsp68I 2009, Bsp119I 852, Bst1107I 1577, Bsu15I 2044, CaiI 987, Cfr9I 2226, Cfr42I 1339, Eam1105I 540, Eco24I 2005, Eco31I 473, Eco57I 3799, Eco91I 3279, EcoO109I 3505, Esp3I 2364, FalI 771, FaqI 2791, GsuI 455, HincII 1, HindIII 2472, NheI 1588, NsbI 320, PfoI 1382, PsiI 3483, PstI 299, ScaI 62, SgfI 2350, SgrAI 1501, SmaI 2226, SmuI 3470, TatI 62, Van91I 2611, XagI 2262, XceI 1013, XhoI 1952, XmiI 1577.
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Updated liepos 25, 2008 10:21
