ninaE-stops m3-myc
(Plasmid #26855)

• Depositing Lab: Craig Montell
• Publication: Wang et al Neuron. 2008 Jan 10. 57(1):56-68.

Backbone

• Vector backbone: pCNX
• Vector type: Insect Expression

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: Slow Termination of Phototransduction
• Alt name: STOPS
• Alt name: CG31006
• Species: D. melanogaster (fly)
• Mutation: Amino acids 296-299 (SLRH in wild-type) are mutated to GPGD
• GenBank ID: CG31006
• Entrez Gene: stops (a.k.a. CG15557, CG15558, CG15559, CG31006, Dmel\CG31006, su(rdgB)69)
• Tag / Fusion Protein:
  • myc (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (unknown if destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: n/a

Terms and Licenses

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

Depositor Comments

To express the stops cDNA under the control of the ninaE promoter, the cDNA
(EST clone GH07253, DGRC) was subcloned between the NotI and XbaI sites
of pCNX. To express Myc-tagged wild-type STOPS in flies, the sequence encoding a Myc tag was fused to the 3' end
of the wild-type stops cDNA. The wild-type stops cDNAs was
subsequently subcloned between the NotI and XbaI sites of pCNX.

The pCNX vector was constructed as follows. First, the pNX vector (provided by Dr. J. O'Tousa, University of Notre Dame, Notre Dame, IN) was digested with KpnI and NotI, which liberated a 3 kb ninaE promoter sequence, and this fragment was subsequently introduced between the same sites of pCaspeR4. Second, to provide the signals for polyadenylation and 3' end cleavage of RNAs, a simian virus 40 3' region (0.5 kb) was excised from pCaspeR–heat shock protein 70 promoter (hs) and introduced between the XbaI and XhoI sites of the vector