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Addgene

CAG sHRPa-FRB-pre mGRASP
(Plasmid #73145)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 73145 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pCAG
  • Backbone size w/o insert (bp) 4247
  • Total vector size (bp) 6173
  • Vector type
    Mammalian Expression

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
    sHRPa-FRB-pre mGRASP
  • Species
    Synthetic
  • Insert Size (bp)
    1926
  • Promoter CAG
  • Tag / Fusion Protein
    • Flag tag

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer gctaaccatgttcatgccttc
  • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

sHRPa is the large split HRP fragment. It consists of amino acids 1-213 of horseradish peroxidase (HRP) with the following 4 mutations: T21I, P78S, R93G, N175S.

The insert contains the following features:
EcoRI-β integrin ss-AgeI-sHRPa-10 aa linker-XhoI-FRB-MfeI-flag-CD4-2(25-242 aa)-NRX1β(414-468 aa)-NotI-stop-HindIII

CAG promoter
10 aa linker: KGSGSTSGSG
FRB: we used a 94 aa sequence that matches residues 5-98 of chain A in PDB 1AUE
Nematode β integrin ss: MPPSTSLLLLAALLPFALPASDWKTGEVTAS

This construct is identical to the previously reported “pre-mGRASP” (Addgene ID 34910) except AgeI and NotI restriction sites are added here, and the 5 aa linker here is shorter than the previously reported 10 aa linker.

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:

    CAG sHRPa-FRB-pre mGRASP was a gift from Alice Ting (Addgene plasmid # 73145 ; http://n2t.net/addgene:73145 ; RRID:Addgene_73145)
  • For your References section:

    A split horseradish peroxidase for the detection of intercellular protein-protein interactions and sensitive visualization of synapses. Martell JD, Yamagata M, Deerinck TJ, Phan S, Kwa CG, Ellisman MH, Sanes JR, Ting AY. Nat Biotechnol. 2016 May 30. doi: 10.1038/nbt.3563. 10.1038/nbt.3563 PubMed 27240195