ppBAalbNixE1E2 OpIE2-GFP
(Plasmid #173505)

• Purpose: transgenesis vector for Aedes albopictus ectopic expression of the Nix gene
• Depositing Lab: Eric Marois
• Publication: Lutrat et al Commun Biol. 2022 Mar 7;5(1):210. doi: 10.1038/s42003-022-03165-7.

Backbone

• Vector backbone: pDONR221 P4r-P3r
• Backbone manufacturer: Invitrogen
• Total vector size (bp): 8700
• Modifications to backbone: addition of the Nix gene under control of its own promoter and a SV40 terminator, addition of an OpiE2-GFP-SV40 marker gene
• Vector type: Insect Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Nix masculinisation gene
• Species: Aedes albopictus
• Insert Size (bp): 3800
• Mutation: Nix isoforms with exons 1, 2 separated by intron 1
• Promoter: own

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (Golden Gate Cloning) (destroyed during cloning)
• 3′ cloning site: BsaI (Golden Gate Cloning)I (destroyed during cloning)
• 5′ sequencing primer: M13R
• 3′ sequencing primer: custom primer

Gene/Insert 2

• Gene/Insert name: EGFP
• Species: Synthetic
• Promoter: OpIE2

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (Golden Gate cloning) (destroyed during cloning)
• 3′ cloning site: BsaI (Golden Gate cloning) (destroyed during cloning)
• 5′ sequencing primer: custom primer
• 3′ sequencing primer: M13R

Resource Information

• Supplemental Documents:
  • ppBAalbNixIsof3-4 OpIE2GFP.gbk

Terms and Licenses

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

Depositor Comments

The OpIE2-GFP transgenesis marker may become detectable at late stages of mosquito development.

Note that the plasmid sequence spanning Nix was assembled from database sequence. The actual sequence in the plasmid may contain differences due to natural polymorphism, especially in the Nix promoter