GgVcl 1-1066 FRET tail probe 1
(Plasmid #46276)

• Depositing Lab: Susan Craig
• Depositing Organization: Johns Hopkins University and School of Medicine
  Located in United States of America
• Publication: Chen et al J Cell Biol. 2005 May 9;169(3):459-70.

Backbone

• Vector backbone: pECFP-N3
• Backbone manufacturer: Craig lab
• Modifications to backbone: pECFP-N3 vector was constructed as an intermediate vector for generating tail probe. ECFP was PCR amplified with 5-GGTACCatggtgagcaagggc-3 and 5-GCGGCCGCTttacttgtacagctc-3 to generate 5' KpnI and 3' NotI. The product was subcloned into TOPO pCRII and sequenced. The KpnI and NotI fragment of pCRII/ECFP was used to replace the EGFP fragment of pEGFP-N3 to generate pECFP-N3.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Vcl 1-1066 + EYFP
• Alt name: Vinculin
• Alt name: Vcl
• Species: G. gallus (chicken)
• Mutation: addition of 9 bp linker and EYFP at aa 883
• Entrez Gene: VCL (a.k.a. VINC1)
• Promoter: CMV
• Tag / Fusion Protein:
  • ECFP (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: ?

Terms and Licenses

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

Depositor Comments

A 9-bp fragment encoding a NotI site was introduced by mutagenesis into pEGFP-C1/vinculin cDNA (chicken) immediately after the codon for aa 883 (Coutu and Craig, 1988) using
the Quick Change kit (Stratagene). The cDNA of EYFP (CLONTECH Laboratories, Inc.) minus the stop codon and flanked by NotI was inserted into NotI-digested vinculin.

DNA (Catalog # 46276-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 46276-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.