Shuttle vector for the construction of adenovirus vectors using the AdenoQuick2.0 system (WT E4 region)

pAd1130 is a 10,062 bp-long plasmid designed for constructing recombinant adenovirus vectors, in combination with the AdenoQuick2.0 plasmids (pAd1127, pAd1128, pAd1129, and their derivatives). It contains the sequence encompassing psn 32796-right end in the Ad5 genome, including the right ITR and the entire E4 region. The right ITR is flanked with PacI and SwaI sites.  The E4 region is terminated with two SfiI sites, which generate non-symmetrical sticky ends suitable for directional cloning. The plasmid contains a 5 kb stuffer made from scrambled phage λ DNA.  This stuffer increases the size of the ligation product of pAd1127, pAd1128, pAd1129, and pAd1130 so that it can be packaged efficiently into phage λ.

The entire sequence of pAd1130 was verified by sequencing.  It is identical to the sequence of the Adenovirus Type 5 Reference Material ARM, with one discrepancy in the stretch of T's immediately downstream from the E4 ORF3 stop codon.  That homopolymer is 11 nt-long in pAd1130, pAd5, and also in Ad5 GenBank # M73260.    It was reported to be 12-nt long in the Adenovirus Type 5 Reference Material ARM (Genbank # AY339865), and 14-nt long in Ad2 GenBank #NC_001405.