pTol2Dest_nos2:nos2 CDS-nos2,3'utr_pBH
(Plasmid #226800)

• Purpose: Transgenic expression of the cloned nanos2 CDS+3'utr region from Danio rerio. Includes a red heart (pBH; cmlc2:RFP) marker.
• Depositing Lab: Florence Marlow
• Depositing Organization: Icahn School of Medicine at Mount Sinai
  Located in the United States of America
• Publication: Primordial germ cells differentially contribute to the germline in zebrafish. (unpublished)

Backbone

• Vector backbone: pTol2Dest_pBH
• Backbone manufacturer: Florence Marlow, Amanda Heim
• Backbone size w/o insert (bp): 6946
• Total vector size (bp): 11130
• Vector type: Zebrafish expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert 1

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

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: nanos2
• Species: D. rerio (zebrafish)
• Insert Size (bp): 1538
• Entrez Gene: nanos2

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: EBV

Terms and Licenses

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

Depositor Comments

Cloned by Maya Pahima.
The nos2 CDS+3'utr was cloned from zebrafish ovary gDNA using primers (3'-CACACCTAAAACACACACCTGGAC-5' and 3'-AAATAAAATATATCAGGAGGAAACAAATATATCAGG-5') and inserted into the pCR8 cloning vector using the TOPO™ TA Cloning Kit (Thermo Fisher, K250020) for stabilization and sequencing (pCR8-nos2-CDS-3'utr).
pTol2-nos2L:nos2-CDS-nos2,3’utr, cmlc2:dsred (pBH) was generated by combining p5E-nos2, and pCR8-nos2CDS-nos2,3’utr into the pTol2-R4/R2_cmlc2:dsred (pBH) backbone vector (Heim et al., 2014).