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Adenovirus Shuttle Plasmids

AdenovirusThe University of Iowa VVC provides shuttle plasmids at no cost when intended for vector production. (A $25 FedEx overnight shipping fee will apply if no FedEx account is provided).

Interested in Ordering?

Please view our ordering information for shuttle plasmids to learn how to register for an account, place orders for in-stock vectors and make payment through our online system.

Plasmid Backbone

The pacAd5 backbone was developed in the laboratory of Dr. Beverly L. Davidson and is patented as the RAPAdTM system (U.S. Patent No. 6,830,920 B2). A description of the system can be found in "A simple method for the rapid generation of recombinant adenovirus vectors," published in Gene Therapy 7:1034-1038, 2000. This system eliminates many of the problems of standard adenoviral production. The RAPAdTMconstruction plasmids have left-hand ITR, E1a, and partial E1b sequence deletions, which greatly reduces wild type occurrence in the final vector preparations. 

Cloning Adenovirus Plasmids

The RapAdTM System uses a two plasmid transfection system to generate recombinant adenovirus particles, the shuttle plasmid and the viral backbone plasmid:

  • The Shuttle Plasmid: The shuttle plasmid is usually cloned by the investigator and contains the gene of interest driven by the desired promoter and any other elements needed for a single or bicistronic construct. Reporter genes such as eGFP may be cloned into this plasmid or may come from the viral backbone plasmid. See the advantages and disadvantages below. The VVC offers shuttle plasmids with a number of common promoters already prepared and are available free of charge. The VVC also offers limited cloning services for investigators.
  • The Viral Backbone Plasmid: This plasmid contains most of the adenoviral genome, including the partially (or fully) deleted E3 region. With a portion of the E3 deleted, an additional gene cassette can be added. The VVC offers several very convenient viral backbone plasmids with and without reporters for investigators to choose from. No cloning of these plasmids is necessary unless you have a custom design in mind. Simply choose the desired backbone when ordering the virus construction.

Insert Size

Adenovirus vectors can accommodates inserts of up to 7.5kb, this includes promoters, gene(s) of interest, and other elements. Larger inserts can be accommodated though, provided that an equivalent part of the viral genome has been properly deleted. The VVC currently has two backbones with additional portions of the E3 region deleted (known as the G0685 pacAd5 (9.2-100)∆1.6KbE3 and G0686 pacAd5 (9.2-100)∆3.1KbE3).


E1 Shuttle Plasmids

G0687 pacAd5mcspA

Cat: G0687

pacAd5 backbone, multiple cloning site, SV40 polyadenylation signal


G0688 pacAd5CMVmcspA

Cat: G0688

pacAd5 backbone, CMV (Cytomegalovirus) Promoter, multiple cloning site, SV40 polyadenylation signal


G0689 pacAd5RSVmcspA

Cat: G0689

pacAd5 backbone, RSV (Rous sarcoma virus) Promoter, multiple cloning site, SV40 polyadenylation signal


G0407 pacAd5CAGmcspA

Cat: G0407

pacAd5 backbone, CAG Promoter (CMV enhancer fused to the chicken beta-actin promoter), multiple cloning site, SV40 polyadenylation signal


G0796 pacAd5TREtightmcspA

Cat: G0796

pacAd5 backbone, TRE tight (TET Response Element) Promoter, multiple cloning site, SV40 polyadenylation signal


G0864 pacAd5hSyn1mscpA

Cat: G0864

pacAd5 backbone, tissue-specific human synapsin I promoter, multiple cloning site, SV40 polyadenylation signal


G0951 pacAd5mcswtIRESeGFPpA

Cat: G0951

pacAd5 backbone, multiple cloning site, wild-type IRES (internal ribosome entry site), eGFP (enhanced green fluorescent protein), SV40 polyadenylation signal. The IRES-eGFP functions as a non-fusion protein reporter without the addition of a second promoter.


G0950 pacAd5CMVmcswtIRESeGFPpA

Cat: G0950

pacAd5 backbone, multiple cloning site, wild-type IRES (internal ribosome entry site), eGFP (enhanced green fluorescent protein), SV40 polyadenylation signal. The IRES-eGFP functions as a non-fusion protein reporter without the addition of a second promoter.


G1059 pacAd5mcswtIRESmCherrypA

Cat: G1059

pacAd5 backbone, multiple cloning site, wild-type IRES (internal ribosome entry site), mCherry, SV40 polyadenylation signal. The IRES-mCherry functions as a non-fusion protein reporter without the addition of a second promoter.


G1071 pacAd5CMVmcswtIRESmCherrypA

Cat: G1071

pacAd5 backbone, CMV (Cytomegalovirus) Promoter, multiple cloning site, wild-type IRES (internal ribosome entry site), mCherry, SV40 polyadenylation signal. The IRES-mCherry functions as a non-fusion protein reporter without the addition of a second promoter.


G1072 pacAd5CAGmcswtIRESmCherrypA

Cat: G1072

pacAd5 backbone, CAG (CMV enhancer fused to the chicken beta-actin Promoter) Promoter, multiple cloning site, wild-type IRES (internal ribosome entry site), mCherry, SV40 polyadenylation signal. The IRES-mCherry functions as a non-fusion protein reporter without the addition of a second promoter.


G0996 pacAd5mU6miSafeSV40pA

Cat: G0996

pacAd5 backbone, Mouse U6 Promoter driving a scrambled miRNA control miSafe, Poly T termination signal, and SV40 polyadenylation signal.

Please reference: Boudreau, R. L., et al. (2011). "Rational Design of Therapeutic siRNAs: Minimizing Off-targeting Potential to Improve the Safety of RNAi Therapy for Huntington's Disease." Mol Ther 19(12): 2169-2177.


G0997 pacAd5mU6shSafeSV40pA

Cat: G0997 (scrambled control for use with G0455)

Ad5 backbone, Mouse U6 Promoter driving a scrambled shRNA control shSafe, Poly T termination signal, and SV40 polyadenylation signal.

Please reference: Boudreau, R. L., et al. (2011). "Rational Design of Therapeutic siRNAs: Minimizing Off-targeting Potential to Improve the Safety of RNAi Therapy for Huntington's Disease." Mol Ther 19(12): 2169-2177.

G1272 pacAd5CMVCas9SV40pA

Cat: G1272

Ad5 backbone, CMV Promoter driving human spCas9, and SV40 polyadenylation signal. MluI or AgeI estriction sites for gRNA cassette addition.

Manual 
Sequence