pE1.2

* Shuttle vector for inserting heterologous sequences in place of the E1 region of the Ad5 genome

* E1- and E3- substituted adenovirus vectors with E4 region WT or deleted (bipartite, dicistronic adenovirus vectors)

* "Armed" conditionally replicative adenovirus vectors (CRAds)

* AdenoQuick cloning system

pE1.2 is a shuttle vector designed for inserting expression cassettes in place of the E1 region of the Ad5 genome, in combination with the pE3.1 and AdenoQuick13.1 or pAd363 plasmids. It contains the first 353 base pairs (map unit 0-1) from the Ad5 genome (including the left ITR and packaging signal), preceded by PacI and SwaI sites, and followed by a multiple cloning site. Expression cassettes inserted into this site should contain a promoter, coding sequence and a polyA signal. The sequences encompassing the kanamycin-resistance gene, the adenovirus 0-1 map units and the multiple cloning site are flanked by two sets of restriction sites AlwNI, BstAPI, DraIII and PflMI which generate incompatible and non-symmetrical sticky ends. pE1.2 can also be used in combination with plasmids pAd328 and pE3.1 to construct "armed" conditionally replicative adenoviruses (CrAds) containing a heterologous promoter in front of the E1a TATA box, and a transgene in place of the E3 region.