pNKY51
(Plasmid #14839)

• Depositing Lab: Nancy Kleckner
• Publication: Alani et al Genetics. 1987 Aug . 116(4):541-5.

Backbone

• Vector backbone: YEP24
• Backbone size w/o insert (bp): 4000
• Vector type: Yeast Expression
• Selectable markers: URA3

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: hisG-URA3-hisG
• Insert Size (bp): 3860

Cloning Information

• 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

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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.