Dat - EGFP - Tol2
(Plasmid #85418)

• Purpose: for making transgenic zebrafish expressing GFP under the control of cis-regulatory elements of the dopamine transporter (dat) gene
• Depositing Lab: Marc Ekker
• Publication: Xi et al Dev Dyn. 2011 Nov;240(11):2539-47. doi: 10.1002/dvdy.22742. Epub 2011 Sep 19.

Backbone

• Vector backbone: modified pGEM vector with Tol2 arms
• Vector type: zebrafish transgenesis

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Electroporation is recommended for transformation due to plasmid size.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: dopamine transporter
• Alt name: dat
• Species: D. rerio (zebrafish)
• Insert Size (bp): 27000
• Entrez Gene: slc6a3 (a.k.a. da, dat)
• Promoter: dat
• Tag / Fusion Protein:
  • EGFP

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: M13F20
• 3′ sequencing primer: M13-R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • National Institute of Genetics Limited Use Label License for Tol2 System
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Contains the dat genomic sequences, including approximately 13 kb of its 50-flanking region, but excluding the last coding exon and 30-flanking region. The EGFP coding sequence is cloned in frame into dat exon 1.

Due to large size, transformation and DNA prep efficiencies may be low. Electroporation is recommended for transformation and low copy prep protocols using large starting volumes are recommended for purifying the DNA.