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GenBank/EMBL accession number X06403.
Not available from Fermentas
The 4245 bp plasmid, pACYC184, is compatible with pMB1- or ColE1-related plasmids and can therefore be used together with a pMB1- or ColE1-derivative within the same cell. pACYC184 contains: (1) the replicon rep responsible for the replication of plasmid (source - plasmid p15A); (2) tet gene, encoding tetracycline resistance protein (source - plasmid pSC101); (3) cat gene, coding for chloramphenicol acetyl transferase that confers resistance to chloramphenicol (source - transposon Tn9).
The circular sequence is numbered such that 1 is the first G of the unique EcoRI site GAATTC and the count increases first through the 5'-terminal part of cat gene, the p15A material, the tet gene and finally through the 3'-terminal part of cat gene. The map shows enzymes that cut pACYC184 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 indicated rep region is sufficient to promote replication. DNA replication initiates at position 846 (+/- 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. Due to the presence of the cat gene on pACYC184, amplification of this plasmid should be performed using 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.
Rose, R.E., The nucleotide sequence of pACYC184, Nucleic Acids Res., 16, 355, 1988
Enzymes which cut pACYC184 DNA once:
AasI 3696, BamHI 1870, BclI 3542, BfmI 1633, BseJI 3442, BseMI 4234, BseSI 3779, BseYI 2444, Bsp68I 2467, BssSI 960, Bst1107I 594, BstAPI 2540, Bsu15I 1518, BtsI 3620, BveI 2558, Cfr42I 832, Eam1104I 1447, Eam1105I 3204, Eco32I 1680, Eco52I 2434, Eco81I 3592, Eco88I 2920, EcoRI 1, FalI 1395, HindIII 1524, NcoI 3945, PaeI 2057, PagI 1984, PdmI 635, PfoI 788, PsyI 3699, SalI 2146, ScaI 3831, TatI 3831, VspI 1406, XagI 2117, XbaI 1425.
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Updated liepos 25, 2008 10:22
