pT7PI-Sce ESARC D326A
(Plasmid
#195609)
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PurposeExpresses PI-SceI (VDE) homing endonuclease/intein from the T7 promoter that contains the D326A active site mutation
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 195609 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepT7-7
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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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namePI-SceI
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Alt nameVDE
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SpeciesS. cerevisiae (budding yeast)
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MutationT3593C, A3601C, T3602G
- Promoter T7
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Unknown (unknown if destroyed)
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
Expresses the excised intein of the VMA1 gene product including amino-terminal methionine-cysteine residues. Contains silent AvrII and RsrII sites near the borders of the N-terminal LAGLIDADG motif and silent EaeI and SnaBI sites near the borders of the C-terminal motif. Contains a D326A mutation in the second LAGLIDADG motif that inactivates the protein.
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:
pT7PI-Sce ESARC D326A was a gift from Frederick Gimble (Addgene plasmid # 195609 ; http://n2t.net/addgene:195609 ; RRID:Addgene_195609) -
For your References section:
Substitutions in conserved dodecapeptide motifs that uncouple the DNA binding and DNA cleavage activities of PI-SceI endonuclease. Gimble FS, Stephens BW. J Biol Chem. 1995 Mar 17;270(11):5849-56. doi: 10.1074/jbc.270.11.5849. 10.1074/jbc.270.11.5849 PubMed 7890714