pJET-col2a1a-KI-donor
(Plasmid
#184876)
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PurposeDonor template for knock-in of p2a-CreERT2 into the medaka col2a1a locus via CRISPR/Cas9-mediated homology directed repair
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 184876 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepJET1.2/blunt
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Backbone manufacturerThermoFisher Scientific
- Backbone size w/o insert (bp) 2974
- Total vector size (bp) 7699
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Vector typeCre/Lox, CRISPR
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 numberHigh Copy
Gene/Insert 1
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Gene/Insert nameCollagen2a1a 5' homology arm
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Alt nameCol2a1a 5' homology arm
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SpeciesOryzias latipes
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Insert Size (bp)256
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GenBank ID
Cloning Information for Gene/Insert 1
- Cloning method Gibson Cloning
- 5′ sequencing primer CGACTCACTATAGGGAGAGCGGC
- 3′ sequencing primer GGCGATCCCTGAACATGTCCAT (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert nameCollagen2a1a 3' homology arm
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Alt nameCol2a1a 3' homology arm
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SpeciesOryzias latipes
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Insert Size (bp)198
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GenBank ID
Cloning Information for Gene/Insert 2
- Cloning method Gibson Cloning
- 5′ sequencing primer AGTTACAACTGGCTATGCCTGA
- 3′ sequencing primer AGAACATCGATTTTCCATGGCAG (Common Sequencing Primers)
Gene/Insert 3
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Gene/Insert namep2a-CreERT2
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SpeciesSynthetic
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Insert Size (bp)2040
Cloning Information for Gene/Insert 3
- Cloning method Gibson Cloning
- 5′ sequencing primer AATGAAAACCTCACGCTTGC
- 3′ sequencing primer CTAGTAACGGCCGCCAGTGT (Common Sequencing Primers)
Gene/Insert 4
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Gene/Insert nameMyosin light chain 2 promoter
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Alt nameCmlc2 promoter
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SpeciesD. rerio (zebrafish)
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Insert Size (bp)899
Cloning Information for Gene/Insert 4
- Cloning method Gibson Cloning
- 5′ sequencing primer GTCGCCGTCCAGCTCGACCA
- 3′ sequencing primer GTGCCCTTAAGTGCCAACAT (Common Sequencing Primers)
Gene/Insert 5
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Gene/Insert nameEGFP
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SpeciesSynthetic
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Insert Size (bp)718
Cloning Information for Gene/Insert 5
- Cloning method Gibson Cloning
- 5′ sequencing primer TCTGGATCTACGTAATACGACTCA
- 3′ sequencing primer TGCTGGAATCTGAGCACTTG (Common Sequencing Primers)
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:
pJET-col2a1a-KI-donor was a gift from Christoph Winkler (Addgene plasmid # 184876 ; http://n2t.net/addgene:184876 ; RRID:Addgene_184876) -
For your References section:
A novel non-disruptive and efficient knock-in allows fate tracing of resident osteoblast progenitors during repair of vertebral lesions in medaka. Hui TW, Winkler C. Development. 2022 May 20. pii: 275483. doi: 10.1242/dev.200238. 10.1242/dev.200238 PubMed 35593425