pBABE-puro-NLS-LSSmKate2
(Plasmid #34586)

• Depositing Lab: Vladislav Verkhusha
• Publication: Piatkevich et al Proc Natl Acad Sci U S A. 2010 Mar 23;107(12):5369-74. Epub 2010 Mar 8.

Backbone

• Vector backbone: pBABE-puro
• Backbone size w/o insert (bp): 5169
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: NLS-LSS-mKate2
• Alt name: Nuclear localization signal-LSS-mKate2
• Insert Size (bp): 795
• Mutation: K69Y/P131T/S148G/M167D/T183S/M196V compared to mKate but numbering is relative to EGFP.

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pBABE5'
• 3′ sequencing primer: pBABE3'

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

There is an 18bp deletion in Addgene's sequencing results relative to the full plasmid sequence (at position 2174-2192bp). This region is immediately after the LSS-mKate2 gene and is not critical for plasmid performance.

There is also a single nucleotide mismatch that results in an F22S change in the linker between the NLS and LSS-mKate2. It should not have any effect on the fusion performance.

Two other mismatches do not alter amino acid sequence.