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pT2-eab2-[GTR]
(Plasmid #46144)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46144 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBluescript II KS+
  • Backbone manufacturer
    stratagene
  • Total vector size (bp) 8831
  • Vector type
    Cre/Lox ; Tol2 zebrafish transgenic expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    EGFP/mCherry
  • Alt name
    EGFP
  • Alt name
    mCherry
  • Insert Size (bp)
    6500

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site KpnI (not destroyed)
  • 3′ cloning site SpeI (not destroyed)
  • 5′ sequencing primer M13R
  • 3′ sequencing primer M13F
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

This plasmid can be used to make a transgene that expresses EGFP but can be switched to express mCherry if inverted.

This reporter is controlled by a 2.3 kb regulatory sequence consisting of 0.2 kb Xenopus EF1alpha enhancer and 2.1 kb zebrafish
beta-actin 2 sequence including the 1.5 kb enhancer/promoter sequence, the 1st exon (86 bp) and part of the 1st intron (490 bp). To generate a complete intron, the splice acceptor from the rabbit beta-globin gene was placed upstream of the EGFP reporter and a synthetic splice acceptor was placed upstream of the mCherry reporter. The synthetic splice acceptor contains the zebrafish consensus sequence along with intronic and exonic splice enhancers.

There are some discrepancies between Addgene's and the depositor's sequence--these do not affect plasmid function.

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:

    pT2-eab2-[GTR] was a gift from Wenbiao Chen (Addgene plasmid # 46144 ; http://n2t.net/addgene:46144 ; RRID:Addgene_46144)
  • For your References section:

    FlEx-based transgenic reporter lines for visualization of Cre and Flp activity in live zebrafish. Boniface EJ, Lu J, Victoroff T, Zhu M, Chen W. Genesis. 2009 Jul;47(7):484-91. doi: 10.1002/dvg.20526. 10.1002/dvg.20526 PubMed 19415631