pEcRNAP5
(Plasmid
#128939)
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PurposepET21a derivative encoding E. coli core RNA polymerase subunits: RpoAppx234-241-(His)10, B, C, Z
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 128939 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET21a
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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 numberUnknown
Gene/Insert 1
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Gene/Insert nameRpoAppx234-241-(His)10
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SpeciesEscherichia coli
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Insert Size (bp)1025
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MutationResidues 234-241 are substituted for a PreScission protease site (GE Healthcare: LEVLFQGP).
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GenBank ID947794
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Entrez GenerpoA (a.k.a. b3295, ECK3282, pez, phs, sez)
- Promoter T7
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Tag
/ Fusion Protein
- His (C terminal on insert)
Cloning Information for Gene/Insert 1
- Cloning method Restriction Enzyme
- 5′ cloning site NdeI (unknown if destroyed)
- 3′ cloning site NcoI (unknown if destroyed)
- 5′ sequencing primer T7 Promotor (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert nameRpoB
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SpeciesEscherichia coli
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GenBank ID948488
Gene/Insert 3
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Gene/Insert nameRpoC
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SpeciesEscherichia coli
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GenBank ID948487
Gene/Insert 4
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Gene/Insert nameRpoZ
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SpeciesEscherichia coli
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GenBank ID948160
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:
pEcRNAP5 was a gift from Seth Darst (Addgene plasmid # 128939 ; http://n2t.net/addgene:128939 ; RRID:Addgene_128939) -
For your References section:
A novel method for the production of in vivo-assembled, recombinant Escherichia coli RNA polymerase lacking the alpha C-terminal domain. Twist KA, Husnain SI, Franke JD, Jain D, Campbell EA, Nickels BE, Thomas MS, Darst SA, Westblade LF. Protein Sci. 2011 Jun;20(6):986-95. doi: 10.1002/pro.622. Epub 2011 Apr 26. 10.1002/pro.622 PubMed 21416542