CmCitrine Lck w/ NosUTRs
(Plasmid #66785)

• Purpose: expression of fluorescent membranes in sea urchin primordial germ cells
• Depositing Lab: Amro Hamdoun
• Publication: Campanale et al Dev Dyn. 2014 Jul;243(7):917-27. doi: 10.1002/dvdy.24133. Epub 2014 Apr 30.

Backbone

• Vector backbone: pGEM-T
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 3000
• Vector type: sea urchin, zebrafish, xenopus

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: Lck
• Alt name: LCK proto-oncogene, Src family tyrosine kinase
• Species: H. sapiens (human)
• Insert Size (bp): 2144
• Mutation: Tandem repeat of LCK the N-terminal membrane targeting sequence: MGCIKSKRKDNLNDDEAAMGCIKSKRKDNLNDDEAPVAT
• Entrez Gene: LCK (a.k.a. IMD22, LSK, YT16, p56lck, pp58lck)
• Promoter: T7
• Tags / Fusion Proteins:
  • 5' UTR from sea urchin Nanos (N terminal on insert)
  • mCitrine (C terminal on insert)
  • 3' UTR from sea urchin Nanos (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: ApaI (not destroyed)
• 3′ cloning site: NdeI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: SP6

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Lck was taken from Addgene plasmid #53749

Terms and Licenses

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

Depositor Comments

The membrane anchoring of mCitrine in sea urchin germ cells uses the sea urchin (S. purpuratus) 5' and 3' UTRs Nanos2 UTRs flanking this insert.