pMT- eef1a1l1:sfGFP
(Plasmid #254856)

• Purpose: This plasmid is designed for Tol2-mediated transgenesis in zebrafish and drives ubiquitous cytoplasmic expression of superfolder GFP (sfGFP), under the control of an eef1a1l1 derived promoter.
• Depositing Lab: Dan Wagner
• Publication: eef1a1l1:h2b-mCherry2 (unpublished)

Backbone

• Vector backbone: pMTB Addgene #112225
• Backbone manufacturer: Sean Megason
• Backbone size w/o insert (bp): 4314
• Total vector size (bp): 6470
• Modifications to backbone: This pMT vector (mini-tol2) contains a ~2.2 kb genomic region upstream of the predicted eef1a1l1 translation start site, amplified from purified zebrafish genomic DNA, with the pMTB vector containing sfGFP (Addgene #112225) as a backbone. The actb2 promoter in pMTB-mem-sfGFP was replaced with the regulatory sequence upstream of eef1a1l1. This sequence includes a eef1a1l1 promoter region, non-coding exon 1, intron 1, and exon 2 up to the start codon. Membrane tags present in pMTB-mem-sfGFP were removed, resulting in cytoplasmic sfGFP. The plasmid can be co-injected with Tol2 transposase mRNA for transgenesis in zebrafish.

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: eef1a1l1
• Alt name: ef1a
• Species: D. rerio (zebrafish)
• Insert Size (bp): 2200
• Entrez Gene: eef1a1l1 (a.k.a. EFL1-alpha, chunp6927, eef1a, ef1a, ik:tdsubc_2a3, ik:tdsubc_2b3, tdsubc_2a3, wu:fa91c07, wu:fa94b03, wu:fi13b09, xx:tdsubc_2a3, xx:tdsubc_2b3)
• Promoter: eef1a1l1

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T3
• 3′ sequencing primer: T7

Terms and Licenses

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