Appendix A2 . 1 . 1 Expression Plasmid pAdCMVl/CFTR-1 
Ad5 1-454 
The expression plasmid contains the CMV promoter with splice donor and ac- 
ceptor sequences directing the expression of human CFTR cDNA. Within the in- 
tron is an Sp6 promoter. The SV40 early poly A signal is located immediately 
3' of the CFTR cDNA. Flanking the CFTR expression cassette are the left 454 
base pairs of Ad5 , which contain the ITR, origin of replication and packaging 
signals followed by Ad5 base pairs 3329-5788. These latter sequences are used 
for homologous recombination with Ad5 to generate the viral vector 
AdcvCFTR.10. CFTR expression is leftwards with respect to the viral genome. 
Not shown in the plasmid are the Bla gene (Amp r ) , 'Col El origin of DNA repli- 
cation, T7 and Sp6 RNA promoters derived from the pGEM 2 (Promega) vector 
backbone. The CFTR cDNA was cloned in the laboratory of L-C Tsui and provided 
to us in the plasmids pBQ4.7 and pBQ6 . 2 . Three differences from the published 
sequences have been noted by Tsui et al. pAdCMVl/CFTR- 1 is derived from pBQ6 . 2 
and contains the mutations indicated by Tsui et al. Three additional silent 
mutations were found in pBQ4 . 7 , pBQ6 . 2 and pAdCMVl/CFTR- 1 at amino acids 266 
to 268 (nucleotides 930, 933, 936 of the published sequence). The published 
sequence is 5' ATC GAG AAC while the sequence of our gene is 5' ATt GAa AAt. 
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