p3E-nanos2 3'utr_R2-R3
(Plasmid #226799)

• Purpose: 3'-entry vector with the nanos2 3'utr cloned from Danio rerio. For use in Gateway cloning.
• Depositing Lab: Florence Marlow
• Publication: Primordial germ cells differentially contribute to the germline in zebrafish. (unpublished)

Backbone

• Vector backbone: pDONR-P2R-P3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2640
• Total vector size (bp): 3718
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

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

Cloning Information

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

Terms and Licenses

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

Depositor Comments

Cloned by Maya Pahima.
The putative zebrafish nos2 3'utr was cloned from zebrafish ovary gDNA using primers (3’-ATCAACCAATGCTGAATCTTTTTATAAAAAATTGAA-5' and 3’-AAATAAAATATATCAGGAGGAAACAAATATATCAGG-5') and inserted into the pCR4 cloning vector using the TOPO™ TA Cloning Kit (Thermo Fisher, K457502) for stabilization and sequencing (pCR4-nos2,3'utr). To insert the nos2,3’utr sequence into a Gateway Cloning vector, the region was amplified with attB primers (3'-GGGGACAGCTTTCTTGTACAAAGTGGTATATCAACCAATGCTGAATCTTTTTATAAAAAATTGAA-5' and 3'-GGGGACAACTTTGTATAATAAAGTTGTAAATAAAATATATCAGGAGGAAACAAATATATCAGG-5') and a BP reaction was performed with the Gateway BP Clonase II Enzyme mix (Invitrogen, 11789020) to recombine the amplicon into pDONR-P2R-P3 (Invitrogen, 12537-023) and produce p3E-nos2,3’utr.