KG#707
(Plasmid
#110911)
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PurposeExpresses the C. elegans sup-1(e995) minigene in ventral nerve cord cholinergic motor neurons
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 110911 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepUC
- Backbone size w/o insert (bp) 2124
- Total vector size (bp) 3236
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namesup-1 minigene
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SpeciesC. elegans (nematode)
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Insert Size (bp)1112
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer Unknown (Common Sequencing Primers)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Made by IDT Custom Gene Synthesis. This sup-1 minigene can be inserted in C. elegans introns as a gene editing marker. The minigene begins with the 491 bp unc-17beta promoter. This is an engineered derivative of the full unc-17 promoter that drives expression specifically in cholinergic motor neurons of the ventral nerve cord (AS, DA, DB, VA, VB neuron classes) and not in cholinergic neurons elsewhere in the animal (Jim Rand, personal communication). The sup-1 portion of the minigene contains a single, small native intron (we removed a second, much larger intron) and includes the e995 suppressor mutation that rescues the unc-17(e245) mutant phenotype. The minigene finishes with the C. briggsae unc-119 3’ UTR, previously used in the unc-119 marker gene (FROKJAER-JENSEN et al. 2008). After searching the reverse orientation of the minigene sequence with the consensus spice acceptor sequence WTNYAG, we replaced 11 potential splice acceptors in the coding region, intron, and 3’ UTR with silent mutations. The AT content of the minigene is 63%, which approaches the 70% average for introns (BLUMENTHAL AND STEWARD 1997). The total size of the sup-1 minigene is 1112 bp.
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.
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For your Materials & Methods section:
KG#707 was a gift from Kenneth Miller (Addgene plasmid # 110911 ; http://n2t.net/addgene:110911 ; RRID:Addgene_110911) -
For your References section:
Local microtubule organization promotes cargo transport in C. elegans dendrites. Harterink M, Edwards SL, de Haan B, Yau KW, van den Heuvel S, Kapitein LC, Miller KG, Hoogenraad CC. J Cell Sci. 2018 Oct 22;131(20). pii: jcs.223107. doi: 10.1242/jcs.223107. 10.1242/jcs.223107 PubMed 30254025