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DNA Constructs

ACCESS RESOURCES

DNA Constructs

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DNA Constructs

Below is a list of dysferlin DNA constructs that are available to anyone in the research community.   The majority of these constructs are available via Addgene.  These constructs include both mouse and human dysferlin full length transcripts, as well as the various spliced isoforms, all possible deletion constructs, and full length human constructs containing various dysferlin missense mutations.  If there is a construct that you need that isn’t listed here, please reach out to the Jain Foundation at admin@jain-foundation.org to see if it is something we can help you obtain.

Dysferlin cDNA Clones

Eighteen (18) full length dysferlin cDNAs have been deposited to Addgene by the Jain Foundation and are available through that site.

The first 4 cDNAs (#1-4) are provided in the pDONR221 plasmid, which can be used as an entry vector for Invitrogen’s gateway system.  There are also several restriction sites available for traditional cloning.  Please view the example restriction digest confirming the identity of the plasmids. Additional information about the known dysferlin splice variants is also available.

The remaining 14 cDNAs (#5-18) are provided in the pUC57 plasmid and are all the possible spliced isoforms of human dysferlin through the alternative splicing of exon 1/1a, exon 5a, exon 17, and exon 40a.

*Note: NCBI Reference Sequence NM_003494.2 has been updated to NM_003494.3 since the creation of these plasmids. There is a one nucleotide base pair change between the two sequences, but the protein sequence is identical.

  1. Human dysferlin – original (accession #NM_003494*)
    View plasmid map and sequence information human_original.
  2. Human dysferlin – v1 (accession #DQ267935)
    Exon 1 variant of Human dysferlin.
    View plasmid map and sequence information human_v1.
  3. Mouse dysferlin – transcript variant 2 (accession #NM_001077694)
    Similar to Human dysferlin – original.
    View plasmid map and sequence information mouse_variant2.
  4. Mouse dysferlin – transcript variant 1 (accession #NM_021469)
    Similar to Human dysferlin – v1.
    View plasmid map and sequence information mouse_variant1
  5. pUC57 DYSF, alt exon 1a
    Full length DYSF containing the alternatively spliced exon 1a without exons 1, 5a, 17, or 40a
    View plasmid map and sequence information here
  6. pUC57 DYSF, alt exon 1, 5a, 17
    Full length DYSF containing alternatively spliced exon 1, 5a, 17, but not 1a or 40a
    View plasmid map and sequence information here 
  7. pUC57 DYSF, alt exon 1, 5a, 40a
    Full length DYSF containing alternatively spliced exons 1, 5a, 40a, but not 1a or 17
    View plasmid map and sequence information here
  8. pUC57 DYSF, alt exon 1, 5a
    Full length DYSF containing alternatively spliced exon 1, 5a, but not 1am 17 or 40a
    View plasmid map and sequence information here
  9. pUC57 DYSF, alt exon 1, 17, 40a
    Full length DYSF containing alternatively spliced exon 1, 17, 40a, but not 1a or 5a
    View plasmid map and sequence information here 
  10. pUC57 DYSF, alt exon 1, 17
    Full length DYSF containing alternatively spliced exons 1, 17 but not 1a, 5a or 40a
    View plasmid map and sequence information here 
  11. pUC57 DYSF, alt exon 1, 40a
    Full length DYSF containing alternatively spliced exons 1, 40a, but not 1a, 17 or 5a
    View plasmid map and sequence information here
  12. pUC57 DYSF, alt exon 1, 5a, 17, 40a
    Full length DYSF containing alternatively spliced exons 1, 5a, 17, and 40a, but not 1a
    View plasmid map and sequence information here
  13. pUC57 DYSF, alt exon 1a, 5a, 17
    Full length DYSF containing alternatively spliced exons 1a, 5a, 17 but not 1 or 40a
    View plasmid map and sequence information here 
  14. pUC57 DYSF, alt exon 1a, 5a, 40a
    Full length DYSF containing the alternatively spliced exon 1a, 5a, 40a without exon 1 
    View plasmid map and sequence information here 
  15. pUC57 DYSF, alt exon 1a, 5a
    Full length DYSF containing the alternatively spliced exon 1a, 5a without exons 1, 17, or 40a
    View plasmid map and sequence information here  
  16. pUC57 DYSF, alt exon 1a, 17, 40a
    Full length DYSF containing the alternatively spliced exon 1a, 17, 40a without exons 1 or 5a
    View plasmid map and sequence information here 
  17. pUC57 DYSF, alt exon 1a, 17
    Full length DYSF containing the alternatively spliced exon 1a, 17 without exons 1, 5a or 40a
    View plasmid map and sequence information here 
  18. pUC57 DYSF, alt exon 1a, 40a
    Full length DYSF containing the alternatively spliced exon 1a, 40a without exons 1, 5a or 17
    View plasmid map and sequence information here 

Tagged Dysferlin Plasmids

Tagged Dysferlin plasmids constructed by Dr. Steve Vogel are available from Addgene

C terminal HA tag: click here

C terminal Venus tag:  click here

Codon Optimized Dysferlin

The plasmid was deposited at Addgene by Dr. Matt Hirsch. It is available for $65 and is shipped as bacteria in an agar stab. click here

Human dysferlin missense mutations constructs

Dr. Mohan Viswanathan and Dr. Lenny Guarente have developed 115 full length human DYSF cDNA constructs with individual DYSF missense mutations.  Information about which missense mutations are available and how to acquire these constructs can be obtained from the Jain Foundation (admin@jain-foundation.org).

Human Dysferlin deletion constructs

Plasmids containing human dysferlin with every single exon that can be skipped without disrupting the open reading frame were developed by the Jain Foundation and are available through Addgene.  These human dysferlin deletion constructs are in the multiple cloning site of pUC57-Kan and can be easily cloned into an expression vector for analysis. All constructs were made using the full-length original human sequence which contains exons 1 and 17, but does not include 1a, 5a, or 40a (accession #NM_003494).

Deletion construct Requests: All of the deletion plasmids are available from Addgene.

Download sequence information:
When you click on the links below, you will download a zipped folder containing 4 files including: 1) A pdf with the vector map, full length sequence information, locations of unique restriction sites, and the location of the deletion, 2) the full sequence of the plasmid and insert in a word document, 3) the full sequence of the plasmid and insert in a fasta file and 4) a fully annotated sequence file that can be opened using DNAStar Lasergene software.

  1. Full-length human dysferlin – original in pUC57-Kan (accession #NM_003494)
    Download plasmid map and sequence information: Full-Length_Dysferlin
  2. Exon 7 deletion
    Download plasmid map and sequence information: DYSF_Exon_7_del
  3. Exon 8 deletion
    Download plasmid map and sequence information: DYSF_Exon_8_del
  4. Exon 9 deletion
    Download plasmid map and sequence information: DYSF_Exon_9_del
  5. Exon 14 deletion
    Download plasmid map and sequence information: DYSF_Exon_14_del
  6. Exon 17 deletion
    Download plasmid map and sequence information: DYSF_Exon_17_del
  7. Exon 19 deletion
    Download plasmid map and sequence information: DYSF_Exon_19_del
  8. Exon 24 deletion
    Download plasmid map and sequence information: DYSF_Exon_24_del
  9. Exon 25 deletion
    Download plasmid map and sequence information: DYSF_Exon_25_del
  10. Exon 30 deletion
    Download plasmid map and sequence information: DYSF_Exon_30_del
  11. Exon 32 deletion
    Download plasmid map and sequence information: DYSF_Exon_32_del
  12. Exon 34 deletion
    Download plasmid map and sequence information: DYSF_Exon_34_del
  13. Exon 35 deletion
    Download plasmid map and sequence information: DYSF_Exon_35_del
  14. Exon 36 deletion
    Download plasmid map and sequence information: DYSF_Exon_36_del
  15. Exon 37 deletion
    Download plasmid map and sequence information: DYSF_Exon_37_del
  16. Exon 38 deletion
    Download plasmid map and sequence information: DYSF_Exon_38_del
  17. Exon 41 deletion
    Download plasmid map and sequence information: DYSF_Exon_41_del
  18. Exon 42 deletion
    Download plasmid map and sequence information: DYSF_Exon_42_del
  19. Exon 43 deletion
    Download plasmid map and sequence information: DYSF_Exon_43_del
  20. Exon 45 deletion
    Download plasmid map and sequence information: DYSF_Exon_45_del
  21. Exon 49 deletion
    Download plasmid map and sequence information: DYSF_Exon_49_del