Shuttle plasmid for the construction of adenovirus vectors using the AdenoQuick2.0 system (WT E2 region and late genes)
- Easy manipulation of sequences coding for DNA polymerase, terminal protein, DNA-binding protein, IVa2, penton, hexon, 100K, IIIa, pVII, pVIII, ...
pAd1128 is a 25,657 bp-long plasmid that contains the Ad5 E2 region and most of the late genes. It designed for constructing recombinant adenovirus vectors, in combination with the AdenoQuick2.0 plasmids (pAd1127, pAd1129, pAd1130, and their derivatives).
pAd1128 includes among others the complete ORFs encoding IVa2, DNA polymerase, pre-terminal protein, pIIIa, penton, pV, pX, pVI, hexon, protease, DNA-binding protein, 100K, and pVIII. These adenovirus sequences are flanked by two SfiI sites, which generate non-symmetrical sticky ends suitable for directional cloning.
The entire sequence of the pAd1128 was confirmed by sequencing. It contains the sequence encompassing psn 4032-27851 in the Ad5 genome (Adenovirus Type 5 Reference Material, ARM, GenBank AY339865), with the following exceptions:
- The two SfiI sites naturally present in WT Ad5 DNA were mutated by substituting A for G and C at positions 16291 and 16294 in the Ad5 genome, and C and G for respectively G and C at positions 18979 and 18982 in the pAd1128, introducing silent mutations in the adenovirus pVII and DNA-binding protein coding sequences.
- At position ~10060, the stretch of “A” has 14 residues in pAd1128, while 13 in ARM (GenBank AY339865), 12 in Ad5 (GenBank M73260) and 17 in Ad2 (GenBank NC_001405). This homopolymer is located immediately downstream from (and includes partially) the TAA stop codon of pIIIa coding sequence.
|IVa2 (orf, unspliced)||1692-65||Ad5|
|DNA pol (orf)||4341-1171||Ad5|
|origin of replication||24497-23910||pUC19|
|Vector Map||Sequence||Product Information Sheet|
|Vector Map||pAd1128.txt (25.68 KB)||Product_Information_pAd1128.pdf (96.67 KB)|