pSCW01
(Plasmid
#72300)
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Purpose(Empty Backbone) Starting substrate for in vitro DNA mismatch repair substrate monitored by PstI cleavage. Derived from pUC19CPDrev H. HWang and J.B. Hays 2002 JBC 277:26136.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 72300 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepUC19CPDrev
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Backbone manufacturergift from Dr. John Hays
- Backbone size (bp) 2025
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Modifications to backbonepSCW01 were derived from pUC19CPDrev and follow the original numbering system with the four Nt.BstNBI endonuclease sites residing on the sense strand as defined by the pUC19 bla gene in pSCW01(Fig. 1A). A BbvC1 restriction endonuclease site was introduced at position 55 (Stratagene QuikchangeXL) in pUC19CPDrev and an existing BbvC1 site removed between positions 331 and 348. The resulting plasmid was digested with AatII and SapI, gel purified to remove the 76-bp intervening sequence, and ligated to synthetic 76-bp duplexes to create a 2030-bp plasmid, pSCW01, that served as homoduplex DNA controls in MMR assays. The plasmid was used to create circular DNA mismatch substrates for mismatch repair protein research
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Vector typeBacterial Expression ; derivative of pUC19
- Promoter lac
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
Resource Information
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Articles Citing this Plasmid
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.
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:
pSCW01 was a gift from Peggy Hsieh (Addgene plasmid # 72300 ; http://n2t.net/addgene:72300 ; RRID:Addgene_72300) -
For your References section:
In vitro studies of DNA mismatch repair proteins. Geng H, Du C, Chen S, Salerno V, Manfredi C, Hsieh P. Anal Biochem. 2011 Jun 15;413(2):179-84. doi: 10.1016/j.ab.2011.02.017. Epub 2011 Feb 15. 10.1016/j.ab.2011.02.017 PubMed 21329650