pFRT/TO/HIS/FLAG/HA-MYEF2
(Plasmid #38069)

• Depositing Lab: Markus Landthaler
• Publication: Baltz et al Mol Cell. 2012 Jun 8;46(5):674-90.

Backbone

• Vector backbone: pFRT/TO/HIS/FLAG/HA-DEST
• Backbone size w/o insert (bp): 5508
• Total vector size (bp): 7311
• Vector type: Mammalian Expression
• Selectable markers: 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: MYEF2 myelin expression factor 2
• Alt name: MEF-2; MST156; MSTP156; HsT18564
• Species: H. sapiens (human)
• Insert Size (bp): 1803
• GenBank ID: NM_016132.3
• Entrez Gene: MYEF2 (a.k.a. HsT18564, MEF-2, MST156, MSTP156, myEF-2)
• Tag / Fusion Protein:
  • HIS/FLAG/HA (N terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV
• 3′ sequencing primer: BGH

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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

Depositor Comments

Digest and NGS results suggest that the original Gateway entry vector is still present in the DNA stock from which the bacterial stocks are created. Because the entry vector does not contain a promoter for the insert or selection marker, it should not interfere with function.

To attempt to remove the entry vector, recipient scientists could try to cure the plasmid from isolated colonies. Alternatively, they can linearize the entry vector with BmtI, HpaI or NheI digest, which will not cut the expression vector, and retransform into bacteria.