KG#707
(Plasmid #110911)

• Purpose: Expresses the C. elegans sup-1(e995) minigene in ventral nerve cord cholinergic motor neurons
• Depositing Lab: Kenneth Miller
• Publication: Harterink et al J Cell Sci. 2018 Oct 22;131(20). pii: jcs.223107. doi: 10.1242/jcs.223107.

Backbone

• Vector backbone: pUC
• Backbone size w/o insert (bp): 2124
• Total vector size (bp): 3236
• Vector type: Bacterial 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: sup-1 minigene
• Species: C. elegans (nematode)
• Insert Size (bp): 1112

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: Unknown

Resource Information

• Supplemental Documents:
  • Annotated ApE and Strider formatted file--download free ApE plasmid editor to view

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.