pTSlb-R756K
(Plasmid
#60731)
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PurposeExpresses both fragments of T7 RNAP split at position 179. The N-terminal fragment is driven by PLac while the C-terminal fragment is driven by PBAD. C-terminal fragment has the point mutations R756S
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 60731 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepBAD33
- Backbone size w/o insert (bp) 5500
- Total vector size (bp) 7600
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Modifications to backboneIPTG inducible promoter PLac drives expression 1-179 of split T7 RNAP while the arabinose promoter PBAD drives expression 180-880.
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Vector typeBacterial Expression, Synthetic Biology
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol, 25 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH10B
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Copy numberHigh Copy
Gene/Insert 1
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Gene/Insert nameResidues 1-179 of split T7 RNAP
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SpeciesSynthetic; T7 Phage
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Insert Size (bp)540
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MutationResidues 1-180 of split T7 RNAP
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GenBank IDGI:216012
Gene/Insert 2
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Gene/Insert nameResidues 180-880 of split T7 RNAP
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SpeciesSynthetic
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Insert Size (bp)2118
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MutationResidues 180-880 of split T7 RNAP. Contains mutation R756K
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GenBank IDGI:216012
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:
pTSlb-R756K was a gift from Matthew Bennett (Addgene plasmid # 60731 ; http://n2t.net/addgene:60731 ; RRID:Addgene_60731) -
For your References section:
Library of synthetic transcriptional AND gates built with split T7 RNA polymerase mutants. Shis DL, Bennett MR. Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5028-33. doi: 10.1073/pnas.1220157110. Epub 2013 Mar 11. 10.1073/pnas.1220157110 PubMed 23479654