pANG02
(Plasmid
#178084)
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Purpose(Empty Backbone) Gentamycin resistant backbone for in vivo Gateway reaction. Plasmid contains gateway cassette with restriction sites to add promoters or C and N Terminal tags. Compatible with pANG01 or pANG03.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 178084 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepGP704-gentR, pET28b, pDONR201
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Backbone manufacturerNA, Sigma, Invitrogen
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Modifications to backbonepANG02 was constructed with the aacC from a pGP704-gentR construct, oriTRP4 from pG704, and oriVpBM1 from pET28b. The Gateway sequence was obtained from commercial vector pDONR201 (Invitrogen) and inserted into the EcorV sites of minimal vectors by Gibson assembly to include unique PmeI and PacI restriction sites flanking the att sites.
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Vector typeBacterial Expression
- Promoter NA
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol and Gentamicin, 25 & 10 μg/mL
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Growth Temperature37°C
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Growth Strain(s)ccdB Survival
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Growth instructionsPlasmid contains ccdB toxin. Strain grows under standard laboratory conditions in LB media supplemented with antibiotics to maintain plasmid.
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Copy numberLow Copy
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer M13 Forward for Gateway site
- 3′ sequencing primer M13 Reverse for Gateway site (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.
Depositor Comments
The plasmids from this article are also used with plasmid pXint129 (www.addgene.org/178208)
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:
pANG02 was a gift from Sören Abel (Addgene plasmid # 178084 ; http://n2t.net/addgene:178084 ; RRID:Addgene_178084) -
For your References section:
A single step three-strain in vivo Gateway reaction. Gillman AN, Helleux A, Abel S. Plasmid. 2021 Nov;118:102608. doi: 10.1016/j.plasmid.2021.102608. Epub 2021 Nov 18. 10.1016/j.plasmid.2021.102608 PubMed 34801582