pRS41K-mScarlet-I-mNeonGreen-ADH1term-TEFpr-hphΔC (pKBJ011)
(Plasmid
#173462)
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PurposeC-SWAT donor for Selection Reconstitution tagging, contains 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 | 173462 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepRS41K-CEN ARS kanMX (type II donor template) (pMaM484)
- Backbone size w/o insert (bp) 6711
- Total vector size (bp) 8148
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Modifications to backboneVector pRS41K-CEN ARS kanMX (type II donor template) (pMaM484) was digested with BamHI-HF and SpeI-HF, a fragment of 6711bp was gel-purified. mScarlet-I-mNeonGreen insert was PCR amplified from pRS423-mScarletI-mNeonGreen-CYC1ter (pKBJ003.1). Vector and insert were assembled by Gibson and transformed into DH5α E. coli cells.
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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 numberLow Copy
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 ttaccctgttatccctac
- 3′ sequencing primer ttaaaacctaagagtcac (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:
pRS41K-mScarlet-I-mNeonGreen-ADH1term-TEFpr-hphΔC (pKBJ011) was a gift from Anton Khmelinskii (Addgene plasmid # 173462 ; http://n2t.net/addgene:173462 ; RRID:Addgene_173462) -
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