|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||14839||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4000
Vector typeYeast Expression
Growth in Bacteria
Insert Size (bp)3860
- Cloning method Restriction Enzyme
- 5′ cloning site BglII (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer n/a
- 3′ sequencing primer pBRrevBam primer (Common Sequencing Primers)
pNKY51 (hisG direct repeats separated by URA3): pNKY5 1 was built in four steps. (1) The backbone, pNKY3 was made by deleting the 2micron DNA from YEP24 with EcoRI and by inserting a BglII linker at the remaining EcoRI site (EcoR1 site is regenerated). (2) A 1.1kb BglII-BamHI fragment of pNK294 bearing Salmonella hisG DNA was inserted into the BglII site of pNKY3 to form pNKY49. (3) The same 1.1-kb hisG fragment as in (2) was inserted at the BamHI site of pNKY49 to form pNKY50. (4) The EcoRI site at the 5’ end of URA3 in pNKY50 was destroyed by fill in and ligation reactions. The resulting plasmid, pNKY51, contains the 3.8 kb hisG-URA3-hisG fragment that can be gel isolated by a BglII and BamHI digest.
Sequence deposited by author is for the hisG-URA3-hisG insert.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pNKY51 was a gift from Nancy Kleckner (Addgene plasmid # 14839 ; http://n2t.net/addgene:14839 ; RRID:Addgene_14839)
For your References section:A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Alani E, Cao L, Kleckner N. Genetics. 1987 Aug . 116(4):541-5. 10.1534/genetics.112.541.test PubMed 3305158
Map uploaded by the depositor.