myr-Abr
(Plasmid #38174)

• Depositing Lab: Nora Heisterkamp
• Publication: Cho et al Mol Cell Biol. 2007 Feb;27(3):899-911. Epub 2006 Nov 20.

Backbone

• Vector backbone: pcDNA3.1(+)
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 7800
• Vector type: Mammalian 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: ABR transcript variant 2
• Species: H. sapiens (human)
• Insert Size (bp): 2600
• Mutation: addition of a Src myristoylation sequence (see comments)
• Entrez Gene: ABR (a.k.a. MDB)

Cloning Information

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

Terms and Licenses

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

Depositor Comments

Constructed by Young Jin Cho. See Supplemental Fig. S3B in PMID 17116687 for membrane location.
Mutagenesis done using Stratagene Quick Change kit. This is ABR transcript 2/isoform B. Abr in pAcG2T (AddGene plasmid id 38162; this parent plasmid may contain an E24G mutation present -gag to ggg- introduced by Taq polymerase at an earlier cloning step) was used to introduce the Src myristoylation sequence at the N-temrnial end of Abr. We inserted 6 amino acids into the original sequence (BamHI-) GGATCC ATG GAG GAG GAA GAG GAG GCG…to get (KpnI-) GGTACC ATG GGC AGC AGC AAG AGC AAG GAG GAG GAA GAG etc. The first amino acids now are M-G-S-S-K-S-K-
The plasmid was digested with KpnIxEcoRI and the 2.8 kb insert ligated into pcDNA3.1(+) digested with KpnIxEcoRI. The construct includes the natural stop codon and 3’UT sequences but the protein does not contain a tag.