pmalc4x_I499F_TEV
(Plasmid
#47703)
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PurposeExpresses MBP myocilin olfactomedin domain fusion protein with a TEV protease recognition sequence and the mutation I499F
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 47703 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepmalC4X
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Backbone manufacturerNEB
- Backbone size w/o insert (bp) 6645
- Total vector size (bp) 7458
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)XL1 Blue
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameOlfactomedin-like domain with I499F mutation
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Alt nameI499F
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Alt nameOLF
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SpeciesH. sapiens (human)
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Insert Size (bp)837
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Mutationchanged I499F
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Entrez GeneMYOC (a.k.a. GLC1A, GPOA, JOAG, JOAG1, TIGR)
- Promoter Ptac
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Tag
/ Fusion Protein
- MBP (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Unknown (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer MBP-F
- 3′ sequencing primer M13-F20; M13_pUC_Fwd (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
The Lieberman Lab recommends E.coli Rosetta Gami 2 competent cells for protein expression.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pmalc4x_I499F_TEV was a gift from Raquel Lieberman (Addgene plasmid # 47703 ; http://n2t.net/addgene:47703 ; RRID:Addgene_47703) -
For your References section:
The stability of myocilin olfactomedin domain variants provides new insight into glaucoma as a protein misfolding disorder. Burns JN, Turnage KC, Walker CA, Lieberman RL. Biochemistry. 2011 Jul 5;50(26):5824-33. doi: 10.1021/bi200231x. Epub 2011 Jun 9. 10.1021/bi200231x PubMed 21612213