pUASTLOTattB_vhhGFP4::Nrv1::tagBFP
(Plasmid #163925)

• Purpose: Expression of GrabFP-B-extracellular under control of UAS or LOP enhancer
• Depositing Lab: Markus Affolter
• Publication: Harmansa et al Elife. 2017 Apr 11;6. doi: 10.7554/eLife.22549.

Backbone

• Vector backbone: pUASTLOTattB
• Backbone manufacturer: Kanca, O. et.al., Raeppli: a whole-tissue labeling tool for live imaging of Drosophila development. Development (2014)
• Backbone size w/o insert (bp): 8900
• Total vector size (bp): 11000
• Vector type: Insect Expression, Cre/Lox

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: Fusion of vhhGPF4 (extracellular) nanobody with Nrv1 protein and tagBFP fluorophor
• Alt name: GrabFP-B extracellular
• Species: D. melanogaster (fly)
• Tag / Fusion Protein:
  • tagBFP (N terminal on insert)

Terms and Licenses

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

Depositor Comments

The TagBFP (Evrogen) coding sequence was inserted between the first and the second exon of the nervana 1 (Nrv1, FlyBase ID: FBgn0015776) cDNA (BDGP DGC clone LD02379). The vhhGFP4 coding fragment (Saerens et al., 2005) was inserted at the C-terminal end of Nrv1::TagBFP. A Drosophila Kozak sequence (CAAA) was added and subsequently vhhGFP4::Nrv1::TagBFP was inserted into the multiple cloning site (MCS) of the pUASTLOTattB vector(Kanca et al., 2014).

pUASTLOTattB vector:
Kanca, O., Caussinus, E., Denes, A. S., Percival-Smith, A. & Affolter, M. Raeppli: a
whole-tissue labeling tool for live imaging of Drosophila development.
Development 141, 472–480 (2014).

vhhGFP4 plasmid:
Saerens D, Pellis M, Loris R, Pardon E, Dumoulin M, Matagne A, Wyns L, Muyldermans S, Conrath K
J Mol Biol. 2005 Sep 23; 352(3):597-607.

tagBFP: Evrogen