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miR-206-1/miR-133b promoter GFP
(Plasmid #20793)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 20793 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBSII SK+ modified to introduce GFP and more cloning sites
  • Backbone manufacturer
    Bartel Lab
  • Vector type
    zebrafish expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    mir-206-1/mir-133b
  • Alt name
    miR-206
  • Alt name
    miR-133
  • Species
    D. rerio (zebrafish)
  • Insert Size (bp)
    4500
  • Entrez Gene
    mir206-1 (a.k.a. dre-mir-206-1)
  • Tag / Fusion Protein
    • GFP (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site ApaI (unknown if destroyed)
  • 3′ cloning site SmaI (destroyed during cloning)
  • 5′ sequencing primer CTGCCCCATTTAGACTCTTTCTTGACG
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

For polycistronic mir-206-1/mir-133b, we were not able to identify the transcriptional start precisely by 5′-RACE due to the repetitive sequence in this region. Analyses of EST clones (BM259633, CF999541, CF348604, DR729161) mapped the approximate start of the mir-206-1/mir-133b primary transcript to 1.4 kb from the mir-206 hairpin. A 4.5-kb upstream fragment that includes ∼0.9 kb of the primary transcript was amplified from genomic DNA using primers 3 and 4 (5'-CTGCCCCATTTAGACTCTTTCTTGACG-3' and 5'-CCAGAAGAACCCAGACGAAACCCAC-3'). The promoter fragment was fused to a GFP reporter gene. The miRNA promoter fusion was flanked by I-SceI meganuclease recognition sites to increase the efficiency of transgenensis.

Please note that Addgene's sequencing results show some differences from the published sequence. The depositing lab is aware of the differences.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    miR-206-1/miR-133b promoter GFP was a gift from David Bartel (Addgene plasmid # 20793 ; http://n2t.net/addgene:20793 ; RRID:Addgene_20793)
  • For your References section:

    Coherent but overlapping expression of microRNAs and their targets during vertebrate development. Shkumatava A, Stark A, Sive H, Bartel DP. Genes Dev. 2009 Feb 15. 23(4):466-81. 10.1101/gad.1745709 PubMed 19240133