pFA6a-mScarlet-I-mNeonGreen-kanMX (pKBJ007)
(Plasmid
#173454)
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PurposeContains mScarletI-mNeonGreen tandem fluorescent protein timer
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 173454 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepFA6a-mCherry-sfGFP-kanMX (pMaM17)
- Backbone size w/o insert (bp) 5614
- Total vector size (bp) 5595
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Modifications to backbonePlasmid pFA6a-mCherry-sfGFP-kanMX (pMaM17) was digested with SalI-HF and BamHI-HF to cut out the fluorescent proteins; the 4150bp fragment was gel-purified. mScarletI-mNeonGreen insert was amplified by PCR from plasmid pRS423-mScarletI-mNeonGreen-CYC1ter (pKBJ003.1) and gel-purified. Digested-vector and amplified-insert were assembled by Gibson and transformed into E. coli Dh5a.
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Vector typeYeast Expression
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Selectable markersNeomycin (select with G418)
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 namemScarlet-I-mNeonGreen
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Insert Size (bp)1440
- Promoter no promoter
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer agagtgcaccatatggac
- 3′ sequencing primer ctgagaaagcaacctgac (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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:
pFA6a-mScarlet-I-mNeonGreen-kanMX (pKBJ007) was a gift from Anton Khmelinskii (Addgene plasmid # 173454 ; http://n2t.net/addgene:173454 ; RRID:Addgene_173454) -
For your References section:
High-Throughput Analysis of Protein Turnover with Tandem Fluorescent Protein Timers. Fung JJ, Blocher-Juarez K, Khmelinskii A. Methods Mol Biol. 2022;2378:85-100. doi: 10.1007/978-1-0716-1732-8_6. 10.1007/978-1-0716-1732-8_6 PubMed 34985695