Appendix A2 . 2 Sequence of Ad^CFTR.10 
The sequence data for the genome of AdcvCFTR.10 is shown. The equivalent of 
bases 1-6100 in Ad^CFTR.10 were sequenced in the construction plasmid 
pAdCMVl/CFTR 1 by dideoxy chain- termination DNA sequencing (Sanger, F., et al. 
Proc . Nat. Acad. Sci. 1977; 74:5463-5467). Base 6100 is equivalent to Ad5 base 
3373. 
The remainder of the molecule (Ad5 3374-35935) is from Genbank [accession No. 
M73260]. The sequence has been annotated to remove the middle part of E3 , the 
Xbal fragment of Ad5 that is deleted in the Ad5 -based deletion mutant Addl324 
used to construct Ad^CFTR.10 (bases 28593- 30470 of Ad5). The minor 2 bp 
deletion in the VA-I region of the Ad5-based deletion mutant Addl324 (T.Shenk, 
personal communication) at Ad5 residues 10594-10595 [residues 13321-13322 of 
Ad^CFTR.10 (AC on the upper strand)] is still present in the sequence to al- 
low direct comparison between Ad^CFTR.10 and the AdCFTR vector used by the PI 
in the NY/NIH CF gene therapy study. This two base pair deletion interrupts a 
minor alternative transcriptional start site for VA-I; this mutation is 
present in Addl324 and Addl327 (personal communication T. Shenk) . 
In the annotated sequence of Ad^CFTR.10, sequences are indicated which com- 
prise the left inverted terminal repeat (ITR), Ela enhancer/promoter (Ela 
core, E2F) and encapsidation signal (AI-AVII, indicated as <£) . The expression 
cassette is oriented leftward with respect to the Ad5 genome and contains the 
CMV enhancer/promoter along with splicing signals directing expression of the 
human CFTR cDNA. Within the intron is an Sp6 RNA promoter. Following the cDNA 
is the SV40 early polyadenylation signal. The CMV promoter TATA box, CCAAT 
box, consensus splice donor and acceptor signals Sp6 promoter, intervening 
plasmid derived sequences along with the SV40 sequences are indicated. 
The CFTR cDNA sequence of AdsvCFTR.10 differs from the published sequence at 
five locations. The coordinates of the mutations are those of Riordan e£ al. 
Science 1989; 245:1066-1073). The differences found are at nucleotides 1990 (C 
instead of A) (a typographical error in the publication) and 2629 (T instead 
of C) as communicated to us by L-C Tsui, U. Toronto, from whom we obtained the 
cDNA. Three silent mutations at position 930, 933, 936 result in the con- 
version of the T, A, and T nucleotides to nucleotides C, G, C respectively. 
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