pAdZ5-mIR155
(Plasmid
#38291)
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Purpose(Empty Backbone)
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Depositing Lab
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 38291 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
This material is available to academics and nonprofits only.
Backbone
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Vector backbonepBeloBAC11
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Backbone manufacturerNEB
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Vector typeMammalian Expression, Adenoviral, RNAi
- Promoter CMV
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 50 μg/mL
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Growth Temperature30°C
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Growth Strain(s)SW102
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Growth instructionsUse low salt media (10g/L tryptone; 5g/L yeast extract; 5g/L salt). The depositing laboratory recommends that the bacteria be grown at 32C.
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Copy numberLow Copy
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer AATGTCGTAACAACTCCG
- 3′ sequencing primer ACCTGATGGTGATAAGAAG (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.
Depositor Comments
Please see the attached protocol or http://adz.cf.ac.uk/ for information on using this plasmid, including cloning, PCR, screening, minipreps, maxipreps, restriction digest, sequencing and virus preparation.
To clone an shRNA into pAdZ-mIR155 design two oligos, a ‘forward’ and ‘reverse’ oligo that overlap in the centre by 25bp. Each oligo will have 40bp homology to the vector, followed by either sense or antisense sequence to your target, then a loop sequence. They will overlap through the loop sequence. Please see the attached protocol for an example of oligo design.
To view the sequence of the vector backbone please see Addgene plasmid 38286 www.addgene.org/38286 Note that the vector numbers are different to those in the original publication - the original work used vectors in which the genome was derived from the AdEasy vectors. The new vectors contain a newly derived & sequenced Ad5 genome, but the expression cassettes etc remain identical to those in the original paper.
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:
pAdZ5-mIR155 was a gift from Richard Stanton (Addgene plasmid # 38291) -
For your References section:
Re-engineering adenovirus vector systems to enable high-throughput analyses of gene function. Stanton RJ, McSharry BP, Armstrong M, Tomasec P, Wilkinson GW. Biotechniques. 2008 Dec;45(6):659-62, 664-8. 10.2144/000112993 PubMed 19238796