pCN50 (ATCC® 77099)

Applications: shuttle vector  /  Depositors: R Diaz Orejas

Permits and Restrictions

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Designations pCN50
Depositors R Diaz Orejas
Biosafety Level 1
Vector Information
Size (kb): 8.5000000000000000
Vector: pCN50 (plasmid)
Construction: pCN38, pBR322
Marker(s):ampR,kanR,tetR
Construct size (kb): 8.5
Features: marker(s): kanR, ampR, tetR
replicon: repA, pMB1
Applications
shuttle vector
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--6.2, 3.1; HindIII/PstI--4.5, 3.4, 0.8; PvuII--4.4 (doublet); PvuII/KpnI--4.4, 2.5, 1.6.
Contains the basic replicon of pPS10 which has been sequenced (EMBL accession number X58896). The repA portion is flanked by EcoRI sites (nt 536 and 1500), with the site at nt 536 between the repA promoter and coding sequences.
Multiple selectable markers permit insertional activation to screen for recombinants (PstI site in ampR).
Retains the mobilization site of pBR322-derived vectors and can be mobilized if suitable functions are provided in trans.
Constructed by ligating pCN38 (ATCC 77095) and pBR322 which had been linearized with PvuII.
The order of the major features in this plasmid is: PvuII - pBR322 (pMB1 ori - bla - tetR) - PvuII - BamHI - kanR - HindIII - repA.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Nieto C, et al. Cloning vectors, derived from a naturally occurring plasmid of Pseudomonas savastanoi, specifically tailored for genetic manipulations in Pseudomonas. Gene 87: 145-149, 1990. PubMed: 2110095

Ramon Diaz Orejas, personal communication

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