pZAP2.2

* Shuttle vector for inserting heterologous sequences in place of the E1 region of the Ad5 genome ("first-generation" vectors)

* E3 region WT or deleted

* AdenoZap2.x cloning system - ligation or homologous recombination in helper cells

pZAP2.2 is a shuttle vector appropriate for inserting expression cassettes in place of the E1 region of the Ad5 genome, in conjunction with AdenoZAP2.1 system.  In contrast to pZAP2.1 it can be used either in the in vitro DNA ligation approach or in the approach based on DNA recombination in helper cells. It contains a multiple cloning site located between the first map unit (mu) of the Ad5 genome and a 400 bp-long Ad5 sequence corresponding to mu 9.8-10.9 (psn 3504-3907) in the Ad5 genome.  Expression cassettes inserted into this site should contain a promoter-cDNA-polyA signal.  In case the in vitro DNA ligation approach is used, the left arm DNA (which corresponds to the left ITR, packaging signal and expression cassette) can be excised from the vector with either PacI or SwaI on the left side, and I-SceI on the right side.  In case the recombination in helper cells is used, the shuttle vector needs to be linearized using PacI or SwaI only.