EF06R
(Plasmid #42940)

• Depositing Lab: Eline Luning Prak
• Depositing Organization: University of Pennsylvania
  Located in United States of America
• Publication: Farkash et al Nucleic Acids Res. 2006 Feb 28;34(4):1196-204. Print 2006.
• Citations: 8 Articles

Backbone

• Vector backbone: pCEP4
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 10410
• Total vector size (bp): 23135
• Modifications to backbone: Blunt ligation of nt 1-1392 of pPur (BD/Clontech) into the NruI site of pCEP4 to insert PuroR
• Vector type: Mammalian Expression
• Selectable markers: Puromycin, Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Human L1 element
• Alt name: LINE-1
• Alt name: Long Interspersed Element 1
• Alt name: L1RT
• Species: H. sapiens (human)
• Insert Size (bp): 7904
• GenBank ID: M80343.1
• Entrez Gene: L1RE1 (a.k.a. L1.2, LRE1)
• Promoter: CMV MIE, L1 5'UTR
• Tag / Fusion Protein:
  • EGFP (opposite strand) (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: pCEP_fwd; LNCX
• 3′ sequencing primer: EGFP-C

Terms and Licenses

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

Depositor Comments

These plasmids consist of the L1RP element tagged with an enhanced green fluorescent protein (EGFP) cassette (pL1RP-EGFP) in a pCEP4 backbone (Invitrogen). The L1 element is driven by its 5′-UTR and an upstream CMV MIE promoter. L1-EGFP (EF06R, Addgene plasmid #42940) was derived from L1RP-EGFP by blunt ligation of 1–1392 nt of pPur (BD/Clontech) into the NruI site of pCEP4.

A negative control ‘dead’ L1-EGFP (EF05J, Addgene plasmid #42941) was similarly derived from pL1RP(JM111)-EGFP, which contains disabling mutations in ORF1.

An EN− plasmid (EF13E, Addgene plasmid #42942) was created by swapping 1927–3708 nt (Age I–Bcl I) from JM102D205A, which contains an endonuclease function-abrogating D205A point mutation in the EN domain, into EF06R.

A control plasmid (EF12J, Addgene plasmid #42943) was similarly derived from JM102. JM102 and JM102D205A are derived from L1.3, which differs from L1RP by 10 base changes in the region swapped.

I->T substitution at the end of ORF 2 should not affect plasmid function.

DNA (Catalog # 42940-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 # 42940-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.