pCaSpeR3/ncd-gfp #10
(Plasmid #66082)

• Purpose: stable germline transformation of Drosophila to produce Ncd-GFP expression
• Depositing Lab: Sharyn Endow
• Publication: Endow et al J Cell Biol. 1997 Jun 16;137(6):1321-36.

Backbone

• Vector backbone: pCaSpeR3
• Backbone size w/o insert (bp): 7814
• Modifications to backbone: Plasmid contains 3.1kb ncd promoter, 2.1kb ncd coding region and 0.7kb GFP
• Vector type: Insect Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Medium copy number plasmid; Preferred growth temperature 34-35C
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Ncd kinesin-14 motor protein
• Alt name: Ca[nd]
• Species: D. melanogaster (fly)
• Insert Size (bp): 5900
• Mutation: D699L
• GenBank ID: X52814
• Entrez Gene: ncd (a.k.a. Dmel_CG7831, CG7831, DmNCD, DmNcd, Dmel\CG7831, KIF C1, KIFC1, NCD, Ncd, ca, ca[nd], cd, l(3)05884, ndc)
• Promoter: ncd promoter (3.1kb upstream region of ncd start site)
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (destroyed during cloning)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pCasper-F (5'-GGGTTTTATTAACTTACAT-3')
• 3′ sequencing primer: Cas3 Rev (5'-AACTCATAATATTAATTAGACG-3')

Terms and Licenses

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

Depositor Comments

Other references describing Ncd-GFP expression:

Endow SA & Komma DJ 1996 J Cell Sci 109, 2429-2442 - PubMed ID: 8923204

Skold HN, Komma DJ & Endow SA 2005 J Cell Sci 118, 1745-1755 - PubMed ID: 15797926