pDA171
(Plasmid
#74251)
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PurposeIntermediate cloning vector, with MCS2 containing inducible stable peptide driven by pSTL1 (Venus 2xNLSs SZ1).
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 74251 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepRS305
- Backbone size w/o insert (bp) 5053
- Total vector size (bp) 7281
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Modifications to backboneThe MCS was removed by PvuII digestion. The coding sequence is now flanked by AatII and SphI.
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Vector typeBacterial Expression, Yeast 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
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Gene/Insert nameInducible peptide of the dPSTR
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SpeciesS. cerevisiae (budding yeast)
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Insert Size (bp)2228
- Promoter pSTL1
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Tag
/ Fusion Protein
- Venus (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site AatII (not destroyed)
- 3′ cloning site SphI (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer T3 (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byThe SynZips were provided by the Keating lab.
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
This intermediate cloning vector is used to change the measured promoter (here pSTL1), by cloning it between SacI (or AatII) and XbaI (or SpeI). The entire inducible peptide has to be inserted into a pSIV by cloning AatII / SphI (into pDA183 or pDA200 for instance).
For more information, see Aymoz et al. Nat. Comm. 2016.
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:
pDA171 was a gift from Serge Pelet (Addgene plasmid # 74251 ; http://n2t.net/addgene:74251 ; RRID:Addgene_74251) -
For your References section:
Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters. Aymoz D, Wosika V, Durandau E, Pelet S. Nat Commun. 2016 Apr 21;7:11304. doi: 10.1038/ncomms11304. 10.1038/ncomms11304 PubMed 27098003