Looking for small vector

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Fri Nov 20 19:36:01 EST 1992


LEES at SASK.USASK.CA writes:

>If does anyone have information about small vector which has 
>both ampicillin and kanamycin markers with less than 3Kb,
>please let me know.

On the plasmid pNH-Kan/oriT, there is a 1.4 kb kanamycin resistance
gene that can be easily subcloned onto an ampicillin resistant plasmid.
If you were to do this on any pUC vector, you would have a 1.4 + 2.7 kb
construct. That's more than you care for, but the omega fragment may
be useful to you. Is there a reason for it being less than 3 kb?

The reference is:

@ARTICLE{Hengen1992.oriT,
AUTHOR = "P. N. Hengen
        and V. N. Iyer",
TITLE = "DNA cassettes containing the origin of transfer (oriT) of two
        broad-host-range transfer systems",
JOURNAL = "Biotechniques",
YEAR = "1992",
VOLUME = "13",
PAGES = "57-62"}

Abstract:

Plasmid constructs are described which carry retrievable DNA cassettes
containing the origin of transfer region (oriT) from two broad-host-range
plasmids. Restriction of these high copy number plasmids with any one of a
variety of enzymes yields a linear DNA fragment of convenient size
containing the oriT region of either pCU1 or RK2. This DNA can be
ligated into any vector or recombinant plasmid containing a compatible
enzyme site and can be easily identified by size on an agarose gel.
Any plasmid can therefore be mobilized using a number of helper strains or
conjugative plasmids derived from the parental plasmids. In addition, the
cassettes can be used for a variety of genetic manipulations including
"selectable" linker mutagenesis.


Paul N. Hengen
National Cancer Institute
Frederick Cancer Research and Development Center
Frederick, Maryland 21702-1201 USA



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