pZAP1.2

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

* E3 and/or E4 regions WT or deleted

* Ad5 or Ad35 knob

* AdenoZap cloning system - ligation or in vitro recombination

pZAP1.2 is a shuttle vector designed for inserting expression cassettes in place of the E1 region of the Ad5 genome, in combination with RightZAP1.1 (WT E3), RightZAP1.2 (ΔE3), RightZap1.3 (ΔE3 + Ad35 knob) and other vectors from the AdenoZAP1.X cloning system. In contrast to pZAP1.1 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 (psn 1-353) and a 400 bp-long 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. When 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 one side, and either PflMI, DraIII, SfiI, BstAPI or AlwNI on the other side. When recombination in helper cells is used, the shuttle vector needs to be linearized using PacI or SwaI only.