pTol2Dest_nos2:d2GFP-nos2,3'utr_pCY
(Plasmid #226801)

• Purpose: Transgenic expression of destabilized GFP (d2GFP) with the nanos2 3'utr control element under the nanos2 promoter. Includes a yellow eye (pCY; αcrystalin-a:Venus) marker.
• Depositing Lab: Florence Marlow
• Publication: Primordial germ cells differentially contribute to the germline in zebrafish. (unpublished)

Backbone

• Vector backbone: pTol2Dest_pCY
• Backbone manufacturer: Christian Mosimann
• Backbone size w/o insert (bp): 4974
• Total vector size (bp): 9545
• Vector type: Zebrafish expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: nanos2
• Species: D. rerio (zebrafish)
• Insert Size (bp): 2417
• Entrez Gene: nanos2
• Promoter: nanos2

Cloning Information for Gene/Insert 1

• Cloning method: Gateway Cloning
• 5′ sequencing primer: ACTGGCCTAAAGGGTGTCCT
• 3′ sequencing primer: CGTCCCTTGCCTTTAGTCTG

Gene/Insert 2

• Gene/Insert name: nanos2
• Species: D. rerio (zebrafish)
• Insert Size (bp): 1075
• Entrez Gene: nanos2
• Promoter: nanos2

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: ATCAACCAATGCTGAATCTTTTTATAAAAAATTGAA
• 3′ sequencing primer: AAATAAAATATATCAGGAGGAAACAAATATATCAGG

Gene/Insert 3

• Gene/Insert name: d2egfp
• Species: Synthetic
• Promoter: nanos2

Cloning Information for Gene/Insert 3

• Cloning method: Gateway Cloning
• 5′ sequencing primer: M13F
• 3′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

Cloned by Maya Pahima.
nos2 control elements were cloned from zebrafish ovary gDNA.
Gateway recombination: pTol2-nos2L:d2egfp-nos2,3’utr, cryaa:Venus (pCY) was generated by combining p5E-nos2promL, pME-d2GFP (Collery and Link, 2011), and pCR4-nos2,3’utr (Chapter 6) into the pTol2-R4/R3_cryaa:Venus backbone vector (Mosimann et al., 2013).