pET15b+PCNA3 T180C
(Plasmid
#66181)
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PurposeExpresses the T180C mutant of Sulfolobus solfataricus PCNA3
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 66181 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET-15b(+)
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 5698
- Total vector size (bp) 6485
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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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Growth instructionsSupplement with 1% glucose is recommended to suppress leaky expression.
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Copy numberLow Copy
Gene/Insert
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Gene/Insert nameDNA polymerase sliding clamp 3
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Alt namePCNA3
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SpeciesSynthetic; Sulfolobus solfataricus P2
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Insert Size (bp)798
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MutationT180C
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GenBank IDNP_341944
- Promoter T7 promoter
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Tag
/ Fusion Protein
- His6 tag (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NdeI (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer T7 Terminal (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 PCNA3 gene was subcloned from a synthetic gene whose codons are optimized for E. coli 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:
pET15b+PCNA3 T180C was a gift from Teruyuki Nagamune (Addgene plasmid # 66181 ; http://n2t.net/addgene:66181 ; RRID:Addgene_66181) -
For your References section:
Introduction of selective intersubunit disulfide bonds into self-assembly protein scaffold to enhance an artificial multienzyme complex's activity. Hirakawa H, Kakitani A, Nagamune T. Biotechnol Bioeng. 2013 Jul;110(7):1858-64. doi: 10.1002/bit.24861. Epub 2013 Feb 22. 10.1002/bit.24861 PubMed 23404255