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pGEM-WingGFP-tan
(Plasmid #83473)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 83473 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pGEM T-easy
  • Backbone manufacturer
    Promega
  • Backbone size w/o insert (bp) 3015
  • Total vector size (bp) 7290
  • Vector type
    Bacterial Expression, Insect Expression, CRISPR

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    WingGFP-tan
  • Insert Size (bp)
    4275
  • Promoter Hsp70 and enhancer from D. melanogaster yellow gene for pupal wing expression

Cloning Information

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

To be used as a selectable marker in two-stage CRISPR-mediated allele replacement strategies as described in Lamb, et al 2016. This donor contains a selectable green fluorescent protein driven by an enhancer from the D. melanogaster yellow gene that can be screened in the developing pupal wing. The current insert contains homology arms targeting the D. americana tan gene, which can be replaced with homology arms targeting the experimenter's locus of choice.
Construct contains:
5’ homology arm
5’ target PAM
Unique portion of 5’ target site for reporter excision (restriction sites: BglII, BsiWI, Acc65I)
Pupal wing enhancer from D. melanogaster yellow gene
EGFP with Hsp70 promoter and SV40 polyadenylation signal
Unique portion of 3’ target site for reporter excision (restriction sites: NarI, Bsu36I, ClaI)
3’ homology arm
3’ target PAM

Restriction sites within pGEM backbone for homology arm removal: 5’ end – ApaI, SphI; 3’ end – SbfI, MluI

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:

    pGEM-WingGFP-tan was a gift from Patricia Wittkopp (Addgene plasmid # 83473 ; http://n2t.net/addgene:83473 ; RRID:Addgene_83473)
  • For your References section:

    Tools and strategies for scarless allele replacement in Drosophila using CRISPR/Cas9. Lamb AM, Walker EA, Wittkopp PJ. Fly (Austin). 2016 Aug 5:1-12. 10.1080/19336934.2016.1220463 PubMed 27494619