CA2259460C

CA2259460C - Recombinant canine adenovirus (cav) containing exogenous dna

Info

Publication number: CA2259460C
Application number: CA002259460A
Authority: CA
Inventors: Laurent Fischer
Original assignee: Merial LLC
Current assignee: Boehringer Ingelheim Animal Health USA Inc
Priority date: 1996-07-03
Filing date: 1997-06-30
Publication date: 2007-03-13
Anticipated expiration: 2017-06-30
Also published as: CA2259460A1

Abstract

Disclosed and claimed are recombinant adenoviruses, methods of making them, uses for them (including in immunological, immunogenic, vaccine or therapeutic compositions, or, as a vector for cloning, replicating or expressing DNA and methods of using the compositions and vector), expression products from them, and uses for the expression products. More particularly, disclosed and claimed are recombinant canine adenoviruses (CAV) and methods of making them, uses for them, expression products from them, and uses for the expression products, including recombinant CAV2 viruses. Additionally, disclosed and claimed are truncated promoters, expression cassettes containing the promoters, and recombinant viruses and plasmids containing the promoters or expression cassettes.

Description

RECOMBINANT CANINE ADENOVIRUS (CAV) CONTAINING EXOGENOUS DNA
FIELD OF THE INVENTION
This invention relates to recombinant adenoviruses, methods of making them, uses for them (including as a vector for replicating DNA), expression products from them, and uses for the expression products.
This invention also relates to promoters and expression cassettes, especially truncated promoters and expression cassettes containing the promoters.
More particularly, this invention relates to recombinant canine adenoviruses (CAV) and methods of making them, uses for them (including as a vector for replicating DNA), expression products from them, and uses for the expression products. Recombinant CAV2 viruses, especially those wherein the exogenous DNA has been inserted into the CAV2 E3 and/or into the right end of the genome between the right ITR and the E4 transcription unit, and methods of making them, uses for them (including in immunological, immunogenic, vaccine or therapeutic compositions, or as a vector for cloning, replicating or expressing DNA and methods of using the compositions or vector), expression products from them, and uses for the expression products are preferred.
However, the invention.broadly relates to a CAV
synthetically modified to contain therein exogenous DNA, wherein a non-essential region of the CAV genome or a portion thereof has been deleted from the CAV. The CAV
is preferably packaged as an infectious CAV with respect to cells in which CAV naturally replicates. The non-essential region of the CAV gename or portion thereof deleted from the CAV is preferably the E3 region or a portion thereof. The exogenous DNA is preferably present in the E3 region, the E1 region, the E4 region, or the

2 region located between the right ITR and the E4 region.
And, the CAV can be a CAV2.
The recombinant CAV can be a vector for expression or cloning of heterologous DNA. The heterologous DNA can encode any desired expression product. Preferred expression products include: an epitope of interest, a biological response modulator, a growth factor, a recognition sequence, a therapeutic gene, or a fusion protein. Thus, the heterologous DNA
can encode any or all of these products. Accordingly, the heterologous DNA can be a transgene.
The epitope of interest can be antigen or immunogen or epitope thereof of a human or veterinary pathogen or toxin. Therefore, the invention further relates to immunological, antigenic or vaccine compositions, containing the expression products.
Further, since the CAV vector, in certain instances, can be administered directly to a suitable host, the invention relates to compositions containing the CAV
vector. The compositions can be immunological, antigenic, vaccine, or therapeutic (e. g., compositions for stimulating an immunological response - local or systemic - including, but not limited to a protective response, or for gene therapy). The invention therefore futher relates to methods of inducing an immunological response, or of transferring genetic information (e. g., gene therapy) comprising administering the composition to a suitable vertebrate host (animal or human).
Additionally, since the expression product can be isolated from the CAV vector in vitro or from cells infected or transfected by the CAV vector in vitro, the invention relates to methods for expressing a product, e.g., comprising inserting the exogenous DNA into a CAV
as a vector to obtain a recombinant CAV, e.g., by recombination or by cleaving and ligating and obtaining recombinant CAV therefrom, followed by infection or transfection of suitable cells in vitro with the

3 recombinant CAV, and optionally extracting, purifying or isolating the expression product from the cells.
As the expression products can provide an antigenic, immunological or protective (vaccine) response, the invention further relates to products therefrom; namely, antibodies and uses thereof. More in particular, the expression products can elicit antibodies. The antibodies can be formed into monoclonal antibodies; and, the antibodies or expression products can be used in kits, assays, tests, and the like involving binding, so that the invention relates to these uses too.
Additionally, since the recombinants of the invention can be used to replicate DNA, the invention relates to recombinant CAV as a vector and methods for replicating DNA by infecting cells with the recombinant and harvesting DNA therefrom. The resultant DNA can be used as probes or primers or for amplification.
The invention still further relates to promoters and expression cassettes containing the promoters, for use in recombinant viruses or plasmids.
In this aspect, the invention specifically relates to a truncated transcriptionally active promoter for a recombinant virus or plasmid which comprises a region transactivated with a transactivating protein provided by the virus or a system into which the plasmid is inserted and the minimal promoter region of the promoter. The invention also relates to an expression cassette comprising the promoter, and to viruses or plasmids containing the promoter or expression cassette.
The expression cassette can include a functional truncated polyadenylation signal.
Several publications are cited in the following text, with full citation of each set forth in the section headed References or with full citation occurring where cited.

4 BACKGROUND OF THE INVENTION
The patent and scientific literature includes various viral vector systems, uses therefor, and exogenous DNA for expression of protein by such systems, as well as uses for such proteins and uses for products -from such proteins.
For instance, recombinant poxvirus (e. g., l0 vaccinia, avipox virus) and exogenous DNA for expression in viral vector systems can be found in U.S. Patent Nos.

5,174,993 and 5,505,941 (e. g., recombinant avipox virus, vaccinia virus; rabies glycopratein (G), gene, turkey influenza hemagglutinin gene, gp51,30 envelope gene of bovine leukemia virus, Newcastle Disease Virus (NDV) antigen, Felt/ envelope gene, RAV-1 env gene, NP
(nudeoprotein gene of Chicken/Pennsylvania/1/83 influenza virus), matrix and preplomer gene of infectious bronchitis virus; HSV gD; entomopox promoter, inter olio), U.S. Patent No. 5,338,683, e.g., recombinant vaccinia virus, avipox virus; DNA encoding Herpesvirus glycoproteins, inter olio; U.S. Patent No. 5,494,807 (e.g., recombinant vaccinia, avipox; exogenous DNA
encoding antigens from rabies, Hepatitis B, JEV, YF, Dengue, measles, pseudorabies, Epstein-Barr, HSV, HIV, SIV, EHV, BHV, HCMV, canine parvovirus, equine influenza, FeLV, FHV, Hantaan, C. tetani, avian influenza, mumps, NDV, inter olio); U.S. Patent No. 5,503,834 (e. g., recombinant vaccinia, avipox, Morbillivirus [e. g., measles F, hemagglutinin, inter olio]); U.s. Patent No.
4,722,848 (e.g., recombinant vaecinia virus; HSV tk, glycoproteins [e.g., gB, gD], influenza HA, Hepatitis B
[e.g., HBsAg], inter olio); U.K. Patent GB 2 269 820 B
and U.S. Patent No. 5,514,375 (recombinant poxvirus;
flavivirus structural proteins); WO 92/22641 (e. g., recombinant poxvirus; immunodeficiency virus, inter olio); WO 93/03145 (e. g., recombinant poxvirus; IBDV, inter olio); WO 94/16716 and U.S. Patent No. 5,833,975, filed January 19, 1994 (e. g., recombinant poxvirus; cytokine and/or tumor associated antigens, inter olio); and PCT/US94/06652 (Plasmodium antigens such 5 as from each stage of the Plasmodium life cycle).
Baculovirus expression systems, exogenous DNA
' for expression therein, and purification of recombinant proteins therefrom can be found in Richardson, C.D.
(Editor), Methods in Molecular Biology 39, "Baculovirus Expression Protocols" (1995 Humana Press Inc.) (see, e.a., Ch.l8 for influenza HA expression, Ch.l9 for recombinant protein purification techniques), Smith et al., "Production of Huma Beta Interferon in Insect Cells Infected with a Baculovirus Expression Vector," Molecular and Cellular Biology, Dec., 1983, Vol. 3, No. 12, p.
2156-2165; Pennock et al., "Strong and Regulated Expression of Escherichia coli B-Galactosidase in Infect Cells with a Baculovirus vector," Molecular and Cellular Biology Mar. 1984, Vol. 4, No. 3, p. 399-406; EPA 0 370 573 (Skin test and test kit for AIDS, discussing baculovirus expression systems containing portion of HIV-1 env gene, and citing U.S. application Serial No.
920,197, filed October 16, 1986 and EP Patent publication No. 265785).
U.S. Patent No. 4,769,331 relates to herpesvirus as a vector.
There are also poliovirus and adenovirus vector systems (see, e.a., Kitson et al., J. Virol. 65, 3068-3075, 1991; Grunhaus et al., 1992, "Adenovirus as cloning vectors," Seminars in Virology (Vol. 3) p. 237-52, 1993;
Ballay et al. EMBO Journal, vol. 4, p. 3861-65; Graham, Tibtech 8, 85-87, April, 1990; Prevec et al., J. Gen Virol. 70, 429-434).
PCT W091/11525 relates to CAV2 modified to contain a promoter-gene sequence within the region from the Smal site close to the end of the inverted terminal

6 PCT/US97/11486 repeat region up to the promoter for the early region 4 (E4).
CAV, and particularly CAV2, has numerous problems. Several of these problems are discussed below.
A significant problem is that the CAV genome can only accept a limited amount of exogenous DNA. That is, only a limited amount of exogenous DNA can be inserted into the CAV genome. Thus, CAV is "insert size limited" and therefore presents a significant problem which must be addressed if CAV is to be a useful vector for a constellation of cloning and expression applications.
The efficient transmission of many viral infections via the oronasal route has provided the impetus for assessing the efficacy of viral vector-based vaccine candidates via the same route. However, since the spread of most live replicating vaccines within the vaccinee and their spread to or contacts with the general environment are well documented (for examples see Schwartz et al., 1974, Mueller et al., 1989, Oualikene et al., 1994), the choice of an adequate viral vector is not obvious.
To address legitimate safety concerns, vector selection preferably involves consideration of characterized live attenuated vaccines as the apparent safety thereof is established. For vaccination of humans, various vectors based on replicating live attenuated viruses are under consideration. To date, there are documented approaches based on human adenoviruses (HAVs) serotype 4 and 7 (Lubeck et al., 1989, Chanda et al., 1990, Chengalvala et al., 2991, 1994, Hsu et al., 1994 ), influenza viruses (for a review Garcia-Sastre and Palese, 1995) and poliovirus and related viruses (for a review Girard et al., 1995).
In the field of veterinary medicine, several vectors based on replicating live attenuated viruses are currently being analyzed with the objective to apply those recombinant vectors as vaccines either parenterally

7 or via the natural route of infection, thereby stimulating local protection. Among the best characterized at this point are members of the poxviridae family [e.g., fowlpox-based vectors (Edbauer et al.
1990, Taylor et al., 1995 and ref. therein)], herpesviridae family [e. g., pseudorabies virus-based ' vectors (Sedegah et al. 1992, Mettenleiter et al. 1994, Hooft van Iddekinge et al., 1996 and ref. therein), turkey herpes virus-based vectors (Ross et al. 1993, to Darteil et al. 1995 and ref. therein), feline herpes virus-based vectors (Cole et al., 1990, Wardley et al., 1992, Willense et al., 1996), infectious laryngotracheitis virus-based vectors (Guo et al., 1994), bovine herpes virus-based vectors (Kit et al. 1991)] and to a lesser extent members of the Adenoviridae family [bovine adenovirus 3-based vectors (Mittal et al., 1995)].
The canine species provides an appropriate model for oronasal immunizations. As such, the canine adenovirus serotype 2 (CAV2) for which attenuated vaccinal strains exist that can be safely administrated either parenterally or via oronasal route, provides a viable immunization vehicle for canine vaccination.
Canine distemper virus (CDV) infection of dogs provides a good example of a respiratory infection in this target species. Further, a relatively direct experimental CDV
challenge system is accessible and allows a direct comparison between CAV2 based-vaccine candidates and previously developed classical CDV vaccines.
CAV2 was first isolated from an outbreak of upper respiratory tract infection in dogs by Ditchfield et al. (1962). Since then, the virus has been isolated from the respiratory tract of dogs with respiratory diseases both in the US and in Europe (Binn et al. 1967, Appel and Percy, 1970, Assaf et al. 1978, Danskin 1973).
Experimental studies have resulted in mild respiratory disease following aerosol inoculation of CAV2 (Swango et

8 al. 1970, Appel , 1970). Several CAV2-based vaccines have been developed and extensively used worldwide for the vaccination of puppies and adult dogs. Immunization with CAV2 has even been shown to protect against an experimental challenge exposure with a serologically related strain of CAV1, which is fatal to non-vaccinated dogs (Fairchild and Cohen, 1969, Appel et al. 1973, Bass et al. 1980). The apparent safety of CAV2 as a vaccine has been well evidenced by the lack of vaccine-induced and vaccine-associated complications in dogs and other animal species including man during its 30 years of utility. Further, results from field serological surveys indicate that many wild animals (foxes, raccoons, skunks and mongooses) are asymptomatically exposed to CAV2 or to an antigenically related virus infection (Summer et ai., 1988). A vaccinal strain of canine adenovirus serotype 2 (CAV2), therefore, provides a unique example of a safe replication-competent, host-restricted virus which can be considered for the derivation of effective vector-based vaccine candidate for vaccination, especially of dogs.
HAVs have been shown to be valuable mammalian cell expression vectors (for a review see Graham et al.
1988) and are currently being evaluated both as recombinant viral vaccine candidates (for reviews see Randrianarison-Jewtoukoff and Perricaudet 1995, Imler 1995) and as vectors for gene therapy (for reviews see Perricaudet and Perricaudet 1995). There are two major groups of HAVs, and a third, less explored, group of recombinant HAVs.
The first group of these adenovirus vectors corresponds to replication-incompetent recombinant adenoviruses which are based on viruses deleted of their E1 region. The E1 region encodes proteins which are essential for virus replication in tissue culture. It has, however, been demonstrated that replication-incompetent recombinant adenoviruses deleted of their E1

9 region can be propagated in the 293 cell line (Graham et al., 1977) which constitutively expresses the E1 region (Haj-Ahmad et al., 1986).
Deletion of the E1 region not only increases the amount of foreign DNA which can be inserted into HAVs, but also limits their replication in human cells and thus considerably improves the safety characteristics of the corresponding recombinant HAVs in humans. Most of the HAV-based vaccine candidates against veterinary and humans pathogens are currently based on E1-deleted vectors. Despite their limited replicative capacity, protection data in challenge experiments have been described (Prevec et al., 1989, McDermott et al., 1989, Lubeck et al., 1989, Eloit et al., 1990, Ragot et al., 1993, Wesseling et al., 1993, Both et al., 1993, Gallichan et al., 1993, Hsu et al., 1994, Breker-Klasser et al., 1995). The property of inducing a protective immune response even in the absence of vector replication is shared by other host restricted viral vectors, the most promising of which being the canarypox virus-based vector ALVAC (Taylor et al., 1991, see Perkus et al., 1995 for a review).
When the goal is a replication competent adenovirus vector, the use of the E1 region as an insertion site is thus not desirable; and, the E1 region therefore has heretofore had deficiencies and presented problems. These deficiencies and problems are compounded when a replication competent adenovirus displaying safety characteristics with respect to humans is desired. In particular, while the E1 region deletion in HAVs may limit replication in human cells and improve safety characteristics with respect to humans, as discussed below, the possibility of recombination between E1 transformed cell lines and E1 deleted recombinant adenoviruses has been documented and thus the safety profile of E1 transformed cell lines appears questionable, thereby rendering any benefit from using E1 region deleted adenoviruses potentially illusory and exascerbating deficiencies and problems heretofore in the use of E1 region deleted adenoviruses (since propagation of E1 region deleted adenoviruses is in cells which 5 constitutively express the E1 region).
The second group of adenovirus vectors corresponds to recombinant adenoviruses which are replication-competent in human cells but replication-incompetent in most non-human animal cells. Those

10 viruses are characterized by a substitution of part of the E3 region with foreign gene expression cassettes.
The E3 region has been shown to be non-essential both in vitro and in vivo for infectious virus formation (Kelly and Lewis 1973, Kapoor et al., 1981, Morin et al., 1987, Lubeck et al., 1989). Numerous recombinant HAVs have therefore been generated by replacement of part of the E3 region (Morin et al., 1987, Chengalvala et al., 1991, 1994, Prevec et al., 1989, Johnson et al., 1988, Lubeck et al., 1989, Dewar et al., 1989, Natuk et al., 1993, Hsu et al., 1994).
However, since proteins encoded by the E3 region have been shown to alter various aspects of the host immune responses (for a review see Wold and Gooding 1991), E3 deletion may have some impact on the pathogenic profile of corresponding recombinant viruses. Indeed, it has been demonstrated in a cotton rat model that deletion of the E3 region from HAV serotype 5 increases virus pulmonary pathogenicity (Ginsberg et al., 1989).
However, it has also been demonstrated that a recombinant bovine Ad3, partially deleted within its E3 region, produces lesions in cotton rats similar to those observed with the parental wt bovine Ad3, therefore suggesting that safety of bovine Ad3-based vectors may be sufficient for the derivation of live recombinant virus vaccines for cattle (Mittal et al., 1996). These results also show that the impact of deletions within the E3 region of

11 any specific adenovirus should be considered on a case-by-case approach.
The CAV2 E3 region has been identified and characterized previously (Linne, 1992). However, based on the available published data (Linne 1992), the precise definition of an insertion site in the CAV2 E3 region is - not obvious. DNA sequence analysis revealed that the organization of the CAV2 E3 region differs significantly from that described for HAVs. The human adenovirus E3 region corresponds to a stretch of at least 3 kbp containing at least 8 open reading frames (orf) whereas the CAV2 E3 region is only 1.5 kbp long and contains only 3 orfs. None of these orfs have a significant level of homology with HAV E3 orfs. From such preliminary comparative analyses, it appears reasonable to speculate that human and canine adenoviruses genomes have evolved differently.
The definition of an insertion site within the CAV2 E3 region is further complicated by the complex splicing and polyadenylation pattern which characterizes the adenovirus family (for a review Imperiale et al., 1995). RNA splicing donor and aceptor sites localized within the E3 region may be important for the maturation of several essential mRNAs even though their coding sequences are localized outside of the E3 region.
Further, since the E3 region is located within a genome region of high transcriptional activity (for a review Sharp et al., 1984), the insertion of foreign DNA
at this site has a potential detrimental impact on the biology of the recombinant virus. Additionally, the E3 region is located downstream of the major late promoter (MLP), where interference between transcription of recombinant gene and transcription initiated at the MLP
has been demonstrated (Zu et al., 1995).
Problems in the art to be addressed therefore include: minimizing phenotypic alterations of the recombinant virus, and the definition of an insertion

12 site in a less transcriptionnally active region. And, in general, it can be said that the E3 region presents problems in the art which should be addressed.
The less explored third group of recombinant HAVs is based on the insertion of recombinant DNA between the right inverted terminal repeat (ITR) and the E4 promoter. The ITRs contain sequences which are essential for viral DNA replication and efficient packaging of the viral genomic DNA. While a region between the right to inverted terminal repeat (ITR) and the E4 promoter may accommodate exogenous DNA sequences (Saito et al., 1985, Chanda et al., 1990), adenoviruses-based vectors have severe limitations in the amount of foreign DNA they can carry, as the packaging capacity of recombinant hAd5 is limited to a genome of approximatively 1050 of the wild-type genome (Bett et al. 1993); thus presenting a problem in the art.
While the region between the right ITR and the E4 region may represent an additional insertion site candidate for the generation of CAV2 recombinant viruses, and PCT WO 91/11525 may relate to a SmaI site close to the leftward extremity of the ITR as a potential insertion site. Contrary to the teachings of W091/11525, there appears to be an upper limit for insertion at this site as Applicant attempted insertions at this site and was able to insert a 400 by DNA fragment, but larger insertions such as a fragment approximately 1 kbp repeatedly failed to be introduced into the site. Hence, a problem in the art is the utility of this site.
Therefore, the E4 promoter region has heretofore had deficiencies and presented problems.
Initial characterization of the CAV2 genome at the molecular level has been described in the literature.
Restriction analysis of several strains of both CAV2 and CAV1 (Jouvenne et al., 1987, Macartney et al., 1988, Spibey and Cavanagh 1989) and sequence analysis of the corresponding E1, E3 and ITRs regions have been reported

13 (Cavanagh et al., 1991, Linne 1992). Although the overall genomic organization of canine adenoviruses is similar to those described for other Adenoviridae family members, the precise organisation of CAV2 genomic E3 region is unique.
Accordingly, one cannot merely extrapolate from " one member to another member of the Adenoviridae family, thereby providing yet another problem in the art.
Further still, when addressing any or all of the aforementioned deficiencies or problems, it would be preferred to avoid any dependence on an endogenous promoter like the E3 or the MLP promoters. However, the pattern of expression of the recombinant gene may be a critical parameter in the overall expression and ergo in the efficacy of the recombinant in a vaccine or immunological composition (Darteil et al., 1995, Xu et al., 1995, Hooft van Iddekinge et al., 1996).
Several cellular and viral promoters have been involved in the derivation of recombinant HAVs. Among the best characterized are b-actin, SV40 early, SV40 late, hAD MLP, and hCMV-IE (Zu et al., 1995). The hCMV-IE promoter may have promise as an upstream regulatory region, since it is associated with the highest level and the longest persistence of recombinant protein expression in tissue culture. This promoter also appears to operate in almost every cell line tested thus far. A potential for cell type independent promoter activity can be regarded as a clear advantage.
It has been demonstrated that the hCMV-IE
promoter can be transactivated by HAV infection (Gorman et al., 1989). The large size of this promoter (approximately 850 bp) is a problem with respect to the size limitations of recombinant CAV vector. Thus, one cannot merely extrapolate from past successes with this promoter to a recombinant CAV vector.
Adenoviruses are known to strongly repress the synthesis of cellular proteins after the onset of viral

14 DNA replication (for a review Zhang and Schneider, 1993).
Thus, replication-competent recombinant adenoviruses have heretofore had a potential for a strong limitation of the recombinant protein expression after the onset of DNA
replication.
Similarly, Saito et al. (1985) demonstrated that a recombinant human adenovirus serotype 5 can produce high amounts of recombinant mRNA but that almost no recombinant protein is obtained.
Late adenovirus mRNAs are characterized by the presence of a tripartite leader (TPL) sequence in their 5' untranslated region (5'UTR). The presence of the TPL
can be an important component of the translatability of late adenovirus mRNAs. Further, it has been demonstrated that in an hAd5 background, the presence of the TPL is a feature of the translational control of a recombinant SV40 T antigen expressed from adenovirus late promoter (Thummel et al. 1983).
Another important problem to address in the design of an expression cassette is the size of the polyadenylation signal. Even still further, the problems in the art include establishing conditions to transfect CAV2 DNA into monolayers. The infectivity of purified naked adenovirus DNA is low. Using a calcium phosphate-based procedure, Graham and Van der Berg (1973) report a yield of 1 pfu/mg of purified DNA. This is not an efficient process for isolating recombinant viruses.
Several approaches have been proposed to attempt to address this problem; but, none heretofore have fully addressed the problem, and particularly without raising additional issues such as safety.
For instance, DNA protein complexes have been purified and are reported to have an increased infectivity (5x103 pfu/mg) (Sharp et al., 1976) over naked DNA. Similarly, covalently closed circles of adenovirus DNA have also been shown to be infectious (Graham, 1984).

A widely used procedure to derive recombinant HAVs is based on the utilization of the 293 cell line which has been transformed with the HAV E1 region (Graham et al., 1977). Previously, it has been reported that the 5 derivation of bovine and canine adenovirus recombinants was dependent on the utilization of cell lines transformed with the corresponding adenovirus E1 region (PCT WO 91/11525, Mittal et al. , 1995a). However, since the genes encoded by the E1 region of some 10 adenoviruses have been shown to contribute to the transformation of rodent cells (reviewed by Grand, 1987), the safety profile of E1 transformed cell line appear questionable. The presence of potent transactivators within the adenovirus E1 region (for a review Nevins,

15 1993) is also well established and further extends safety concerns which can be raised regarding E1 transformed cell lines.
Thus, transfection conditions independent of use of an E1 transformed cell line, especially with good yields, would be a significant advance in the art.
Accordingly, it is believed that a recombinant CAV, preferably a recombinant CAV2, having exogenous DNA
inserted therein and a non-essential region or portion thereof deleted therefrom, especially such a CAV which is packaged as an infectious CAV with respect to cells in which CAV naturally replicates, or a CAV containing exogenous DNA within the E3 and/or the right end of the genome between the right ITR and the E4 transcription unit, and methods for making such recombinants, and uses for such recombinants, as described herein (above and below), has not been taught or suggested. Further, it is believed that a truncated transcriptionally active promoter for a recombinant virus or plasmid which comprises a region transactivated with a transactivating protein provided by the virus or a system into which the plasmid is inserted and the minimal promoter region of the promoter, an expression cassette comprising the

16 promoter, and viruses or plasmids containing the promoter or expression cassette, have not been heretofore described or suggested. And, such a recombinant CAV and methods of making and using such a recombinant CAV, and such a promoter, expression cassette and viruses and plasmids containing the promoter or expression cassette present an advancement over prior recombinants, especially since as to humans CAV is a non-replicating vector and the promoter and expression cassette address insert size limits of recombinant viruses.
OBJECTS AND SUMMARY OF THE INVENTION
it is an object of the invention to provide a recombinant adenovirus, preferably a recombinant canine adenovirus (CAV), such as a recombinant canine adenovirus-2 (CAV2).
It is a further object of the invention to provide such a recombinant which contains exogenous DNA, preferably in a non-essential region, and which has had a non-essential region of the CAV genome, or a portion 2o thereof, deleted therefrom; and, preferably to provide such a recombinant which is packaged as an infectious CAV
with respect to cells in which CAV naturally replicates.
It is also an object of the invention to provide such a recombinant CAV containing exogenous DNA
wherein the exogenous DNA is inserted into the E3 or both the E3 and the region located between the right ITR and the E4 transcription unit.
It is another object of the invention to provide a transcritionally active truncated promoter, an expression cassette containing the promoter, and viruses and plasmids containing the promoter or the expression cassette; including to provide such an expression cassette containing a truncated polyadenylation signal.
Further objects of the invention include any or all of: to provide expression products from such recombinants, methods for expressing products from such recombinants, compositions containing the recombinants or

17 the expression products, methods for using the expression products, methods for using the compositions, DNA from the recombinants, and methods for replicating DNA from the recombinants.
Another object of the invention is an adenovirus-based, e.g., CAV-based, preferably CAV2-based, vector, or compositions containing the vector, or methods for making or using the vector with consideration of any, any combination, or all, of the earlier-discussed deficiencies and/or problems in the art.
Accordingly, the invention surprisingly provides a CAV synthetically modified to contain therein exogenous DNA, wherein a non-essential region of the CAV
genome or a portion thereof has been deleted from the CAV. The CAV is preferably packaged as an infectious CAV
with respect to cells in which CAV naturally replicates.
Any non-essential region or portion thereof can be deleted from the CAV genome, and the viability and stability of the recombinant CAV resulting from the deletion can be used to ascertain whether a deleted region or portion thereof is indeed non-essential. The non-essential region of the CAV genome or portion thereof deleted from the CAV is preferably the E3 region or a portion thereof. The exogenous DNA is present in any non-essential region (and viability and stability of the recombinant CAV resulting from the insertion of exogenous DNA can be used to ascertain whether a region into which exogenous DNA is inserted is non-essential). The E3 region, the E1 region, the E4 region, or a region located between the right ITR and the E4 region, are presently preferred as non-essential regions for insertion of exogenous DNA into the CAV genome.
Additionally, the invention surprisingly provides a recombinant CAV comprising heterologous DNA in a non-essential region of the CAV genome, wherein the heterologous DNA is in the E3 or both the E3 and the

18 region located between the right ITR and the E4 transcription unit.
The CAV of these embodiments is preferably a CAV2.
The invention further provides a vector for cloning or expression of heterologous DNA comprising the recombinant CAV.
The heterologous DNA encodes an expression product comprising: an epitope of interest, a biological response modulator, a growth factor, a recognition sequence, a therapeutic gene, or a fusion protein.
An epitope of interest is an antigen or immunogen or immunologically active fragment thereof from a pathogen or toxin of veterinary or human interest.
An epitope of interest can be an antigen of a veterinary pathogen or toxin, or from an antigen of a veterinary pathogen or toxin, or another antigen or toxin which elicits a response with respect to the pathogen, of from another antigen or toxin which elicits a response with respect to the pathogen, such as, for instance: a Morbillivirus antigen, e.g., a canine distemper virus or measles or rinderpest antigen such a HA or F; a rabies glycoprotein, e.g., rabies glycoprotein G; an avian influenza antigen, e.g., turkey influenza HA, Chicken/Pennsylvania/1/83 influenza antigen such a nudeoprotein (NP); a bovine leukemia virus antigen, e.g., gp51,30 envelope; a Newcastle Disease Virus (NDV) antigen, e.g., HN or F; a feline leukemia virus antigen (FeLV), e.g., FeLV envelope protein; RAV-1 env; matrix and/or preplomer of infectious bronchitis virus; a Herpesvirus glycoprotein, e.g., a glycoprotein from feline herpesvirus, equine herpesvirus, bovine herpesvirus, pseudorabies virus, canine herpesvirus, or cytomegalovirus; a flavivirus antigen, e.g., a Japanese encephalitis virus (JEV) antigen; an immunodef iciency virus antigen, e.g., a feline immunodeficiency virus (FIV) antigen or a simian immunodeficiency virus (SIV)

19 antigen; a parvovirus antigen, e.g., canine parvovirus;
an equine influenza antigen; a Marek~s Disease virus antigen; an poxvirus antigen, e.g., an ectromelia antigen, a canarypox virus antigen or a fowlpox virus antigen; or an infectious bursal disease virus antigen, e.g., VP2, VP3, VP4.
' An epitope of interest can be an antigen of a human pathogen or toxin, or from an antigen of a human pathogen or toxin, or another antigen or toxin which to elicits a response with respect to the pathogen, or from another antigen or toxin which elicits a response with respect to the pathogen, such as, for instance: a Morbillivirus antigen, e.g., a measles virus antigen such as HA or F; a rabies glycoprotein, e.g., rabies virus glycoprotein G; an influenza antigen, e.g., influenza virus HA or N; a Herpesvirus antigen, e.g., a glycoprotein of a herpes simplex virus (HSV), a human cytomegalovirus (HCMV), Epstein-Barr; a flavivirus antigen, a JEV, Yellow Fever virus or Dengue virus antigen; a Hepatitis virus antigen, e.g., HBsAg; an immunodeficiency virus antigen, e.g., an HIV antigen such as gp120, gp160; a Hantaan virus antigen; a C. tetani antigen; a mumps antigen; a pneumococcal antigen, e.g., PspA; a Borrelia antigen, e.g., OspA, OspB, OspC of Borrelia associated with Lyme disease such as Borrelia burgdorferi, Borrelia afzelli and Borrelia garinii; a chicken pox (varicella zoster) antigen; or a Plasmodium antigen.
Of course, the foregoing lists are intended as exemplary, as the epitope of interest can be an antigen of any veterinary or human pathogen or from any antigen of any veterinary or human pathogen.
Since the heterologous DNA can be a growth factor or therapeutic gene, the recombinant CAV can be used in gene therapy. Gene therapy involves transferring . genetic information; and, with respect to gene therapy and immunotherapy, reference is made to U.S. Patent No.

, CA 02259460 2005-03-29 5, 252, 4'19, and to WO 94/16716 and U.S. Patent No. 5,833,975, filed January 29, 1994, 5 together with the documents cited therein. The growth factor or therapeutic gene, for example, can encode a disease-fighting protein, a molecule for treating cancer, a tumor suppressor, a cytokine, a tumor associated antigen, or 10 interferon; and, the growth factor or therapeutic gene can, for example, be selected from the group consisting of a gene encoding alpha-globin, beta-globin, gamma-globin, granulocyte macrophage-colony stimulating factor, tumor necrosis factor, an interleukin, macrophage colony 15 stimulating factor, granulocyte colony stimulating factor, erythropoietin, mast cell growth factor, tumor suppressor p53, retinoblastoma, interferon, melanoma associated antigen or B7.
The invention still further provides an

20 immunogenic, immunological or vaccine composition containing the recombinant CAV virus or vector, and a pharmaceutically acceptable carrier or diluent. An immunological composition containing the recombinant CAV
virus or vector (or an expression product thereof) elicits an immunological response - local or systemic.
The response can, but need not be, protective. An immunogenic composition containing the recombinant CAV
virus or vector (or an expression product thereof) likewise elicits a local or systemic immunological response which can, but need not be, protective. A
vaccine composition elicits a local or systemic , protective response. Accordingly, the terms "immunological composition" and "immunogenic composition"
include a "vaccine composition" (as the two former terms-can be protective compositions).
The invention therefore also provides a method of inducing an immunological response in a host

21 vertebrate comprising administering to the host an immunogenic, immunological or vaccine composition comprising the recombinant CAV virus or vector and a pharmaceutically acceptable carrier or diluent. For purposes of this specification, "animal" includes all vertebrate species, except humans; and "vertebrate"
' includes all vertebrates, including animals (as "animal"
is used herein) and humans. And, of course, a subset of "animal" is "mammal", which for purposes of this specification includes all mammals, except humans.
For human administration, recombinant CAV, especially CAV2, provides the advantage of expression without productive replication. This thus provides the ability to use recombinants of the invention in immunocompromised individuals; and, provides a level of safety to workers in contact with recombinants of the invention. Therefore, the invention comprehends methods for amplifying or expressing a protein by administering or inoculating a host with a recombinant CAV virus or vector, e.g., CAV2, whereby the host is not a canine or not a natural host of the recombinant virus or vector, and there is expression without productive replication.
Furthermore, since CAV, and especially CAV2, is used as vaccinial strains in dogs, the present invention provides a means for introducing additional epitope(s) of interest of antigens) of a canine pathogens) or toxins) into the vaccinial CAV,.e.g., CAV2, strains for a recombinant CAV expressing those additional epitope(s) of interest and thereby providing a means to elicit in vivo responses to those epitope(s) of interest and canine adenovirus by inoculating a dog or pup with the vaccinial recombinant CAV. The additional epitope(s) of interest can be an antigen of a canine pathogen (other than adenovirus) or toxin, from an antigen of a canine pathogen (other than adenovirus) or toxin, another . antigen which elicits a response in dogs or pups to the canine pathogen (other than adenovirus) or toxin, or from . , CA 02259460 2005-03-29

22 another antigen which elicits a response in dogs or pups to the canine pathogen (other than adenovirus) or toxin (an example of the latter two epitopes of interest are measles HA and F and epitopes thereon which elicit a protective response against canine distemper virus in dogs or pups; see U.S. Patent No. 5,503,834).
Accordingly the present invention provides that the recombinant vaccinial CAV can contain heterologous DNA encoding an epitope of interest from any antigen of a canine pathogen or toxin, for instance: rabies, canine herpesvirus, canine distemper virus, canine parvovirus and the like. In this regard, reference is made to copending U.S. Patent Nos. 5,688,920, filed March 29, 1995 (canine herpesvirus DNA), 5,756,102, filed April 6, 1994 (canine distemper), and U.S. Patent No. 5,843,456, filed June 7, 1995 (rabies combination compositions) and U.S. Patent No. 5,529,780 (canine herpesvirus DNA), together with the documents cited therein.
Thus, the invention envisions CAV recombinants containing exogenous DNA coding for more than one protein, e.g., coding for two or more epitopes such as antigens of canine pathogens. The invention also envisions compositions containing CAV recombinants in combination with other antigens.
The invention even further provides a therapeutic composition containing the recombinant CAV
virus or vector and a pharmaceutically acceptable carrier or diluent. The therapeutic composition is useful in the gene therapy and immunotherapy embodiments of the invention, e.g., in a method for transferring genetic information to an animal or human in need of such comprising administering to the host the composition;
and, the invention accordingly includes methods for transferring genetic information.
In yet another embodiment, the invention provides a method of expressing a protein or gene product

23 or an expression product which comprises infecting or transfecting a cell in vitro with a recombinant CAV virus or vector of the invention and optionally extracting, purifying or isolating the protein, gene product or S expression product or DNA from the cell. And, the invention provides a method for cloning or replicating a - heterologous DNA sequence comprising infecting or transfecting a cell in vitro or in vivo with a recombinant CAV virus or vector of the invention and optionallly extracting, purifying or isolating the DNA
from the cell or progeny virus The invention in another aspect provides a method for preparing the recombinant CAV virus or vector of the invention comprising inserting the exogenous DNA
into a non-essential region of the CAV genome.
The method can further comprise deleting a non-essential region from the CAV genome, preferably prior to inserting the exogenous DNA.
The method can comprise in vivo recombination (even though CAV DNA is infectious). Thus, the method can comprise transfecting a cell with CAV DNA in a cell-compatible medium in the presence of donor DNA comprising the exogenous DNA flanked by DNA sequences homologous with portions of the CAV genome, whereby the exogenous DNA is introduced into the genome of the CAV, and optionally then recovering CAV modified by the in vivo recombination.
The method can also comprise cleaving CAV DNA
to obtain cleaved CAV DNA, ligating the exogenous DNA to the cleaved CAV DNA to obtain hybrid CAV-exogenous DNA, tranfecting a cell with the hybrid CAV-exogenous DNA, and optionally then recovering CAV modified by the presence of the exogenous DNA.
Since in vivo recombination is comprehended, the invention accordingly also provides a plasmid comprising donor DNA not naturally occurring in CAV
encoding a polypeptide foreign to CAV, the donor DNA is

24 within a segment of CAV DNA which would otherwise be co-linear with a non-essential region of the CAV genome such that DNA from a non-essential region of CAV is flanking the donor DNA.
The exogenous DNA can be inserted into CAV to generate the recombinant CAV in any orientation which yields stable integration of that DNA, and expression thereof, when desired.
The exogenous DNA in the recombinant CAV virus or vector of the invention can include a promoter. The promoter can be from a herpesvirus. For instance, the promoter can be a cytomegalovirus (CMV) promoter, such as a human CMV (HCMV) or murine CMV promoter.
The promoter is preferably a truncated transcriptionally active promoter which comprises a region transactivated with a transactivating protein provided by the virus and the minimal promoter region of the full-length promoter from which the truncated transcriptionally active promoter is derived. For purposes of this specification, a "promoter" is composed of an association of DNA sequences corresponding to the minimal promoter and upstream regulatory sequences; a "minimal promoter" is composed of the CAP site plus TATA
box (minimum sequences for basic level of transcription;
unregulated level of transcription); and, "upstream regulatory sequences" are composed of the upstream elements) and enhancer sequence(s). Further, the term "truncated" indicates that the full-length promoter is not completely present, i.e., that some portion of the full-length promoter has been removed. And, the truncated promoter can be derived from a herpesvirus such as MCMV or HCMV, e.g., HCMV-IE or MCMV-IE.
The promoter can truncated so that there is up to a 40~ and even up to a 90% reduction in size, from a full-length promoter based upon base pairs; for instance, with the murine CMV-IE promoter, and HCMV-IE promoter, respectively. Indeed, a truncated promoter of the invention can consist essentially of an enhancer region which is transactivated by a transactivating protein provided by a virus or system into which the truncated promoter is inserted, and the mimimal promoter. Thus, as 5 little as 60% and even as little as 10~ of the original base pairs of the full-length promoter can be present in a truncated promoter of the invention.
Given that nature provided so many more base pairs for promoters than now has been discovered 10 necessary, the promoters, and expression cassettes, viruses and plasmids containing the truncated promoters of the invention are indeed surprising. Indeed, the promoters of the invention obtain superior performance in comparison with full-length promoters, and, without 15 necessarily wishing to be bound by any one particular theory, it is believed that this superior performance is due to the truncation. Further, truncation of promoters addresses the insert size limit problem of recombinant viruses and plasmids, particularly CAV.
20 Thus, the invention even still further provides, a truncated transcriptionally active promoter for a recombinant virus or plasmid which comprises a region transactivated with a transactivating protein provided by the virus or a system into which the plasmid

25 is inserted and the minimal promoter region of a full-length promoter from which the truncated transcriptionally active promoter. is derived.
Like the aforementioned promoter, the inventive promoter is preferably a herpesvirus, e.g., a MCMV or HCMV such as MCMV-IE or HCMV-IE promoter; and, there can be up to a 40% and even up to a 90% reduction in size, from a full-length promoter, based upon base pairs.
The invention thus also provides an expression cassette for insertion into a recombinant virus or plasmid comprising the truncated transcriptionally active promoter. The expression cassette can further include a functional truncated polyadenylation signal; for instance

26 an SV40 polyadenylation signal which is truncated, yet functional. Considering that nature provided a larger signal, it is indeed surprising that a truncated polyadenylation signal is functional; and, a truncated polyadenylation signal addresses the insert size limit problems of recombinant viruses such as CAV. The expression cassette can also include exogenous or heterologous DNA with respect to the virus or system into which it is inserted; and that DNA can be exogenous or heterologous DNA as described herein.
Even further surprisingly, the present invention provides a recombinant CAV, preferably CAV2, wherein at least one non-essential loci, such as the E3 region, is employed for generation of the recombinant.
Based on data derived from HAVs and bovine Ad3, part of this region may be non-essential both in vitro and in vivo for infectious virus formation and thus can be considered as an insertion region. Accordingly, in an aspect, the present invention provides the generation of a CAV E3 deletion or partial deletion mutant (e.g., E3 ORF1 and/or ORF2); and, this mutant additionally demonstrates that the entire CAV E3 region is not necessary in tissue culture and thus can be used as an insertion site in the generation of recombinant CAV. And therefore, the present invention encompasses a recombinant CAV wherein endogenous DNA is deleted and/or exogenous DNA introduced in the E3 region; preferably one or more non-essential domains within the E3 region, e.g., ORF2.
A deletion within the E3 region can also provide additional capacity for insertion of heterologous sequences into the CAV genome. For example, such deletions can compensate for the introduction of a large expression cassette into the right end of the genome. In this regard, by the methods herein taught, without undue experimentation, the skilled artisan can readily identify

27 additional non-essential domains, preferably in the E3 region, and additional non-essential regions.
In another aspect, the invention surprisingly provides a recombinant CAV, preferably CAV2, wherein deletions within non-essential regions are relative to insertion of heterologous DNA. For instance, deletions ' within non-essential regions can be substantially similiar, e.g., compensatory, to the insertion of heterologous DNA in another region, such as, without l0 limitation, the E4/right ITR region.
Nucleotide sequence comparisons between the ITRs from various CAV2 strains indicate some variability immediately upstream of the right ITR (Cavanagh et al., 1991, Spibey, 1991). Applicants' engineered a novel and nonobvious insertion site within this region; and therefore, the present invention in a further aspect encompasses CAV recombinants having exogenous DNA
inserted therein. Further, the E4/right ITR region, as herein demonstrated, can surprisingly accept much larger fragments of heterologous DNA than the previously described SmaI site, further addressing the insert size limit of CAV .
Since the E4/right ITR site is localized in a region of the CAV genome with little transcriptional activity (for a review see Sharp et al., 1984), insertion thereinto does not significantly impact the biology of the CAV recombinant virus.
As discussed above, in an embodiment, the present invention provides novel and nonobvious expression cassettes) for insertion of exogenous DNA
into CAV; the cassettes) comprising appropriate heterologous eukaryotic regulatory sequences. In a preferred embodiment, the invention provides expression cassettes) rationally designed with consideration of packaging limitations and biological characteristics associated with viruses and plasmids such as adenovirus-based vectors. The ability to truncate MCMV and HCMV

28 promoters to as small as an enhancer region which is transactivated with a transactivating protein provided by the virus or system into which the promoter is inserted and the mimimal promoter demonstrates that promoters from other eukaryotic viruses, and especially from other herpesviruses, can be similarly truncated, without undue experimentation from this disclosure and the knowledge in the art; and, the invention comprehends truncated promoters from such other viruses.
In a more specific aspect, the present invention encompasses CAV, preferably CAV2, recombinants comprising the HCMV-IE or MCMV-IE promoter, preferably a truncated promoter therefrom. Preferably, the HCMV-IE or MCMV-IE promoter or a truncated promoter therefrom is transactivated by CAV-induced gene products.
In the aspects of the present invention which include a truncated transcriptionally active (or competent) promoter (preferably a truncated transcriptionally active eukaryotic virus promoter such as a herpesvirus promoter, e.g., a HCMV or MCMV
promoter), by "active" (or "competent"), the truncated transcriptionally active promoter should exhibit at least 80%, preferably at least 85%, more preferably at least 90%, and most preferably at least 95% of the transcriptional activity of the pristine or full length promoter. Deletion of nucleotides or of portions or of regions of the full length promoter can be done from the herein teachings, without undue experimentatin, for generation of active fragments in addition to those exemplified. The degree truncation, i.e., amount of base pairs deleted, from the original full length or pristine promoter, in terms of percentage, can be any amount up to 90%, so long as the truncated promoter remains "active" or "competent". Thus, a truncated transcriptionally active promoter can be, in terms of base pairs with respect to the full length or pristine promoter, about 5% to about 95%, preferably about 10% to

29 about 90%, more peferably about 10% to about 60% and most preferably about 10% to about 40% of the full length or pristine promoter, with specific embodiments being about 10% and about 40% of the full length or pristine promoter (i.e., deletions from the full length or pristine promoter, in terms of base pairs, of about 95% to about - 5%, preferably about 90% to about 10%, more preferably about 90% to about 40%, and most preferably about 90% to about 60% of the base pairs of the full length or pristine promoter, with deletions of about 90% and about 60% of base pairs of the full length or pristine promoter being specific embodiments). Indeed, all that need be retained of the original, full length or pristine promoter, at a minimum, is the minimal promoter and a region which is transactivated with a transactivating protein provided by the virus or system into which the promoter is inserted.
The deletion of portions of a promoter such as the HCMV-IE, is to reduce its size so as to address the deficiencies and/or problems of the size of promoters such as the HCMV-IE promoter and the packing limitations of adenoviruses.
In a particular aspect, the present invention provides an active fragment of the HCMV-IE having a size of 91 by or an active fragment of the MCMV-IE having a size of 466 bp, i.e., a truncated transcriptionally active HCMV-IE of about 91 by or.a truncated transcriptionally active MCMV-IE of about 466 bp. (The present invention can encompass HCMV-IE or MCMV-IE
fragments having substantial base pair size and/or homology with respect to the 91 by or 466 by fragment, e.g., as to base pair size and/or homology, at least 80%, preferably at least 85%, more preferably at least 90%, and most preferably at least 950 of the 91 by or 466 by fragment.) The fragment can be inserted into a CAV such as CAV2; and therefore, the invention encompasses a recombinant CAV such as CAV2 comprising an active fragment of HCMV-IE or MCMV-IE, i.e., a truncated transcriptionally active promoter derived from HCMV-IE or MCMV-IE, and preferably, the 91 by or 466 by fragment or an active fragment having substantial base pair size 5 and/or homology to the 91 by or 466 by fragment.
Size reduction considerations for preparing the particular 91 by or 466 by fragment, or any other active fragment of the HCMV-IE or MCMV-IE promoter, can, as discussed above, be from the known molecular organization 10 of the HCMV-IE or MCMV-IE promoter (Boshart et al., 1985) .
It is surprising that such small versions of the full length or pristine promoter, such as the 91 by or 466 by fragment, are still able to be "active" (as the 15 term is discussed above), and even drive an equivalent high level of transcription activity in CAV, particularly CAV2, infected cells as the 850 by version of HCMV-IE and the 766 by version of MCMV-IE, respectively.
The 91 by fragment or an active fragment having 20 substantial base pair size to the 91 by fragment is especially surprising as it is believed to be the smallest promoter element which has been used in an adenovirus-based recombinant virus.
By following the herein considerations applied 25 to the HCMV-IE and MCMV-IE promoter for generation of "active" fragments thereof, "active" fragments of promoters other than HCMV-IE or MCMV-IE, e.g., from other eukaryotic viruses such as other herpesviruses which are exogenous to adenovirus, e.g., CAV2, can be produced,

30 without undue experimentation; and therefore, the present invention provides a fragment of a promoter exogenous to an adenovirus, i.e., a truncated transcriptionally active promoter, which is active like the full length promoter in the adenovirus when introduced into the adenovirus.
The adenovirus is preferably CAV such as CAV2.
Thus, in another aspect the present invention provides a fragment of the murine CMV-IE (MCMV-IE)

31 promoter (Dorsh-Hasler et al., 1985), i.e., a truncated transcriptionally active promoter derived from MCMV-IE, which is active in adenovirus, e.g., CAV2. Indeed, in adenovirus such as CAV2 infected cells the 466 by MCMV-IE
promoter element exhibits activity like the HCMV-IE 91 by promoter element.
In yet another aspect, the invention provides a promoter which is active in adenovirus, e.g., CAV2, which has extended the translation of recombinant mRNAs into the late phase of the viral cycle; and, recombinants comprising the promoter, as well as compositions comprising the recombinants and methods for making and using the promoter, the recombinants and the compositions. Such a promoter can comprise an HCMV-IE
promoter or active fragment thereof wherein the 5'UTR has been replaced with the human Ad2 TPL.
In still another aspect, the invention provides an insertion cassette for generating recombinant adenoviruses, e.g., CAV2, and to recombinants comprising the cassette, as well as compositions comprising the recombinants and methods for making and using the cassette, the recombinants and the compositions. This cassette preferably comprises a minimizd polyadenylation sequence ("minimized poly-A"), such as a minimized polyadenylation sequence from SV40 ("minimized SV40 poly-A"). The minimized SV40 poly-A can be any length less than the full length or native or.pristine SV40 poly-A to as small as about 153 by (plus or minus 10~).
It is demonstrated herein that such a minimized SV40 poly-A is still associated with the same high level of steady stable mRNA as the wild-type element in adenovirus, e.g., CAV2, infected cells. The minimized SV40 poly-A cassette can be used to minimize DNA inserted into adenovirus; and, this addresses the capacity deficiencies and problems of adenoviruses. Further, from the minimization of the SV40 polyadenylation signal,

32 other similar sequences can be derived, from other sources, without undue experimentation.
Indeed, it is believed that heretofore an expression cassette having size and components which have been optimized for the expression of a recombinant protein by an adenovirus-based vector has not been described in the literature.
In an even further aspect, the present invention provides conditions and ergo methods to transfect purified adenovirus, e.g., CAV, preferably CAV2, DNA into canine monolayers.
In preferred embodiments of the invention, transfection conditions are independent of the utilization of a E1 transformed canine cell line. This procedure provides good yields, including yields of approximately 5x103 pfu/ug of purified CAV DNA. And, this procedure avoids the utilization of E1 transformed cells for the derivation and propagation of CAV recombinant viruses, thereby avoiding the safety issues surrounding E1 transformed cells.
The present invention thus provides recombinant adenoviruses, preferably CAV, more preferably CAV2, and methods for making and using them, and compositions containing them or expression products from them. Any suitable non-essential region can be used for insertion into the genome or deletion from the genome. Such sites include E4, E1, and E3. Two insertion sites are presently preferred: the first is within the E3 region and the second located between the right ITR and the E4 transcription unit (preferably the SmaI site); the former site or both sites {combined) are preferred. The CAV E3 ORF2, e.g., CAV2 E2 ORF2, is presently most preferred.
The results herein also demonstrate that the CAV E3 is non-essential for replication in tissue culture. This represents the first successful attempt to derive recombinant CAV viruses and thus constitutes a basis for products based upon recombinant CAV such as r CA 02259460 2005-03-29

33 CAV2, e.g., immunological, antigenic or vaccine compositions containing the recombinant CAV or expression products therefrom.
Accordingly, the present invention comprehends a CAV such as CAV2 synthetically modified to contain therein exogenous DNA (DNA not naturally occurring in CAV, or not naturally occurring in CAV at the insertion site) in a non-essential region of the CAV2 genome. The non-essential region is preferably the CAV E3 or both the 20 CAV E3 and the right end of the genome such as the SmaI
site.
The invention further comprehends antibodies elicited by the inventive compositions and/or recombinants and uses for such antibodies. The antibodies, or the product (epitopes of interest) which elicited them, or monoclonal antibodies from the antibodies, can be used in binding assays, tests or kits to determine the presence or absence of an antigen or antibody.
Flanking DNA used in the invention can be from the site of insertion or a portion of the genome adjacent thereto (wherein "adjacent" includes contiguous sequences, e.g., codon or codons, as well as up to as many sequences, e.g., codon or codons, before there is an intervening insertion site).
The exogenous or heterologous DNA (or DNA
foreign to CAV, or DNA not naturally occurring in CAV) can be DNA encoding any of the aforementioned epitopes of interest, as listed above. In this regard, with respect to Borrelia DNA, reference is made to U.S. Patent No.
5,523,089, W093/08306, PCT/US92/08697, Molecular Microbiology (1989), 3(4), 479-486, and PCT publications WO 93/04175 and WO 96/06165.
With respect to pneumococcal epitopes of interest, reference is made to Briles et al. WO 92/14488, with respect to tumor viruses reference is made to Molecular Bioloay of Tumor a CA 02259460 2005-03-29

34 Viruses, RNA TUMOR VIRUSES.(Second Edition, Edited by Weiss et al., Cold Spring Harbor Laboratory 1982) (e. a_.
page 44 et seq. - Taxonomy of Retroviruses).
With respect to DNA encoding other epitopes of interest, attention is directed to the documents cited in the BACKGROUND OF THE INVENTION, for instance: U.S. Patent Nos. 5,174,993 and 5,505,941 (e. g., recombinant avipox virus, vaccinia virus; rabies glycoprotein (G), gene, turkey influenza hemagglutinin gene, gp51,30 envelope gene of bovine leukemia virus, Newcastle Disease Virus (NDV) antigen, Felt/ envelope gene, RAV-1 env gene, NP (nudeoprotein gene of Chicken/Pennsylvania/1/83 influenza virus), matrix and preplomer gene of infectious bronchitis virus; HSV gD;
entomopox promoter, inter alia), U.S. Patent No.
5,338,683, e.g., recombinant vaccinia virus, avipox virus; DNA encoding Herpesvirus glycoproteins, inter alias U.S. Patent No. 5,494,807 (e. g., recombinant vacc.inia, avipox; exogenous DNA encoding antigens from rabies, Hepatitis B, JEV, YF, Dengue, measles, pseudorabies, Epstein-Barr, HSV, HIV, SIV, EHV, BHV, HCMV, canine parvovirus, equine influenza, FeLV, FHV, Hantaan, C. tetani, avian influenza, mumps, NDV, inter alia); U.S. Patent No. 5,503,834 (e. g., recombinant vaccinia, avipox, Morbillivirus [e.g., measles F, hemagglutinin, inter alia]); U.S. Patent No. 4,722,848 (e. g., recombinant vaccinia virus; HSV tk, glycoproteins [e.g., gB, gD], influenza HA, Hepatitis B [e.g., HBsAg], inter alia); U.K. Patent GB 2 269 820 B and U.S. Patent No. 5,514,375 (recombinant poxvirus; flavivirus structural proteins); WO 92/22641 (e. g., recombinant poxvirus; immunodeficiency virus, inter aZia); WO
93/03145 (e. g., recombinant poxvirus; IBDV. inter alia);
WO 94/16716 and U.S. Patent No. 5,833,975, filed January 19, 1994 (e. g., recombinant poxvirus;
cytokine and/or tumor associated antigens, inter alia);

and PCT/US94/06652 (Plasmodium antigens such as from each stage of the Plasmodium life cycle).
In particular, since the tag and other exogenous DNA had been incorporated into CAV2, as in the 5 recombinants described in the Examples, other exogenous DNA can be incorporated into CAV2. Therefore, instead of the exogenous DNA used to generate vCAl, vCA2, vCA3, vCA4, vCA5, vCA6, vCA7, vCA8, and vCA-CDVF1-@l2bp-up-SmaI, the exogenous DNA of the above-listed documents 10 and/or those otherwise cited herein are used to generate additional CAV2 recombinants with the exogenous DNA in regions as in vCA2 through vCA8 and vCA-CDVF1-@l2bp-up-SmaI and deletions as in vCA2 through vCA8 and vCA-CDVF1-@l2bp-up-SmaI (e.g., insertions in the E3 or at the 15 region between the right ITR and the E4 transcription unit ar at both sites and deletions in the E3 region) including recombinants containing coding for multiple antigens, as herein described (including with subfragment promoters, reduced or modified polyadenylation cassettes, 20 and promoters with 5' UTR replaced). Analysis demonstrates expression. Compositions are prepared by admixture with a carrier or diluent for administration to a vertebrate (animal or human) hosts for generating responses, including antibody responses.
25 The exogenous DNA can include a marker, e.g., a color or light marker. The exogenous DNA can also code for a product which would be detrimental to an insect host such that the expression product can be a pesticide or insecticide. The exogenous DNA can also code for an 30 ahti-fungal polypeptide; and, for information on such a polypeptide and DNA therefor, reference is made to U.S.
Patent No. 5,421,839 and the documents cited therein.
In addition, the present invention provides a

35 method for mapping a non-essential region in the adenovirus, preferably CAV, e.g., CAV2, genome, comprising preparing donor DNA comprising DNA not

36 naturally occurring in CAV present within a segment of CAV DNA otherwise co-linear with a portion of the CAV genome such that by in vivo recombination the donor DNA can be introduced into a region of the CAV genome, introducing said donor DNA into the CAV genome by in vivo recombination, recovering recombinants, and determining stability and viability thereof and expression or presence of the DNA not naturally occurring in CAV and/or absence of endogenous CAV DNA in the recombinants, whereby viability and stability of recombinants and expression or presence of the DNA not naturally occurring in CAV and/or absence of endogenous CAV DNA indicates that the region into which the donor DNA
was introduced is non-essential. This method is employed in the Examples below. The donor DNA can be marker DNA such that by hybridization one can determine whether it has been incorporated into the genome, e.g., hybridization to the marker DNA or failure to hybridize to endogenous DNA
replaced by the marker.
According to another aspect of the present invention, there is provided a recombinant canine adenovirus type 2 (CAV2) containing a deletion in the E3 region of the CAV2 genome and an insertion of heterologous DNA in the E3 region or in the region located between the E4 region and the right ITR region of the CAV2 genome, wherein the CAV2 replicates in a host.
According to still another aspect of the present invention, there is provided an immunogenic or vaccine composition containing the CAV2 as described herein, and a pharmaceutically acceptable carrier or diluent.

36a These and other objects and embodiments within the present invention are described or are obvious from the following detailed description.
BRIEF DESCRIPTION OF DRAWINGS
In the following Detailed Description, reference will be made to the accompanying drawings, wherein:
Figure 1 shows a complete DNA sequence of pLF027 ((6,995 bp) (SEQ ID N0: 1) CAV2 HindIII A fragment starts at nucleotide #689 and ends at nucleotide #4,725.
CAV2 E3 region starts at nucleotide #1,414 and ends at nucleotide #2,945. CAV2 E3 ORFl starts at nucleotide #8 and ends at nucleotide #346. CAV2 E3 ORF2 starts at nucleotide #384 and ends at nucleotide #1,478. CAV2 E3 ORF3 starts at nucleotide #1,019 and ends at nucleotide #483.
The~remaining nucleotides correspond to pBSSK+);
Figure 2 shows a restriction map of pLF027;

WO 98!00166 PCT/US97/11486

37 Figure 3 shows a complete DNA sequence of pLF047A ((6,959 bp) (SEQ ID NO: 2) The 23 by B_lqII/MluI
linker starts at nucleotide #1,485 and ends at nucleotide #1,508. The remaining sequences correspond to pLF027);
Figure 4 shows a restriction map of pLF047A;
Figure 5 shows a complete DNA sequence of ' pLF049A ((7,002 bp) (SEQ ID N0: 3) The 63 by BlqII/MluI
linker starts at nucleotide #2,138 and ends at nucleotide #2,201. The remaining sequences correspond to pLF047A);
Figure 6 shows a restriction map of pLF049A;
Figure 7 shows a complete DNA sequence of pLF086 ((6,581 bp) (SEQ ID NO: 4) The 63 by B1.q11/MluI
linker starts at nucleotide #2,295 and ends at nucleotide #2,358. The remaining sequences correspond to pLF047A);
Figure 8 shows a restriction map of pLF086;
Figure 9 shows a complete DNA sequence of pLF056 ((6,196 bp) (SEQ ID NO: 5) CAV2 SalI B fragment starts at nucleotide #1 and ends at nucleotide #3,274.
The right ITR (196 bp) starts at nucleotide #3,078 and ends at nucleotide #3,274. The SmaI site is localized at position #3,088. The remaining nucleotides correspond to pBSSK+) ;
Figure 10 shows a restriction map of pLF056;
Figure 11 shows a complete DNA sequence of pLF061 ((6,503 bp) (SEQ ID NO: 6) The 306 by heterologous DNA tag starts at nucleotide #3,091 and ends at nucleotide #3,397. The remaining nucleotides correspond to pLF056);
Figure 12 shows a restriction map of pLF061;
Figure 13 shows a complete DNA sequence of pLF022 ((4,504 bp) (SEQ ID NO: 7) The hCMV-IE (145 bp) promoter starts at nucleotide #2 and ends at nucleotide #147. All other nucleotides correspond to pCAT basic sequences and include: the CAT reporter gene which starts at nucleotide #209 and ends at nucleotide #868, the SV40 small t antigen and polyadenylation signal (856 bp) which starts at nucleotide #958 and ends at nucleotide #1,814

38 and the ampicillin resistance gene which starts at nucleotide #2,467 and ends at nucleotide #3,327);
Figure 14 shows a restriction map of pLF022;
Figure 15 shows a complete DNA sequence of pLF062 ((3,812 bp} (SEQ ID NO: 8} The hCMV-IE (145 bp) promoter starts at nucleotide #2 and ends at nucleotide #147. The CAT reporter gene starts at nucleotide #209 and ends at nucleotide #868. The SV40 polyadenylation signal (241 bp) starts at nucleotide #881 and ends at l0 nucleotide #1,122. The ampicillin resistance gene starts at nucleotide #1,775 and ends at nucleotide #2,635);
Figure 16 shows a restriction map of pLF062;
Figure 17 shows a complete DNA sequence of pLF066 {{4,009 bp) (SEQ ID NO: 9) The hCMV-IE (145 bp) promoter starts at nucleotide #2 and ends at nucleotide #147. The Ad2 TPL (202 bp) starts at nucleotide #154 and ends at nucleotide #356. The CAT reporter gene starts at nucleotide #406 and ends at nucleotide #1,065. The SV40 polyadenylation signal (241 bp} starts at nucleotide #1,077 and ends at nucleotide #1,319. The ampicillin resistance gene starts at nucleotide #1,972 and ends at nucleotide #2,832);
Figure 18 shows a restriction map of pLF066;
Figure 19 shows a complete DNA sequence of pLF069 ({3,955 bp) {SEQ ID NO: 10) The hCMV-IE (91 bp) promoter starts at nucleotide #2 and ends at nucleotide #93. The Ad2 TPL (202 bp) starts at nucleotide #100 and ends at nucleotide #302.The CAT reporter gene starts at nucleotide #352 and ends at nucleotide #1,011. The SV40 polyadenylation signal (241 bp) starts at nucleotide #1,024 and ends at nucleotide #1,265. The ampicillin resistance gene starts at nucleotide #1,918 and ends at nucleotide #2,778);
Figure 20 shows a restriction map of pLF069;
Figure 21 shows a complete DNA sequence of pLF077 {(3,861 bp) (SEQ ID NO: 11) The hCMV-IE (91 bp) promoter starts at nucleotide #2 and ends at nucleotide

39 #93. The Ad2 TPL (202 bp) starts at nucleotide #100 and ends at nucleotide #302. The CAT reporter gene starts at nucleotide #352 and ends at nucleotide #1,011. The SV40 polyadenylation signal (153 bp) starts at nucleotide #1018 and ends at nucleotide #1,171. The ampicillin resistance gene starts at nucleotide #1,824 and ends at nucleotide #2,684);
Figure 22 shows a restriction map of pLF077;
Figure 23 shows a complete DNA sequence of pLF091 ((3,888 bp) (SEQ ID NO: 12) The hCMV-IE (91 bp) promoter starts at nucleotide #2 and ends at nucleotide #93. The Ad2 TPL (202 bp) starts at nucleotide #100 and ends at nucleotide #302. The CAT reporter gene starts at nucleotide #352 and ends at nucleotide #1,011. The SV40 polyadenylation signal (153 bp) starts at nucleotide #1,018 and ends at nucleotide #1,164. The CAV2 12 nucleotides inserted at the 3' end of the SV40 polyadenylation signal are starting at nucleotide #1,165 and are finishing at nucleotide #1,176. The ampicillin resistance gene starts at nucleotide #1,851 and ends at nucleotide #2,711);
Figure 24 shows a restriction map of pLF091;
Figure 25 shows a complete DNA sequence of pLF092 ((7,379 bp) (SEQ ID NO: 13) The CAT expression cassette (as defined in pLF091) starts at nucleotide #1 and ends at nucleotide #1,179. The CAV2 left flanking arm (182 bp) starts at nucleotide #1,.180 and ends at nucleotide #1,362. The CAV2 right flanking arm (3,090 bp) starts at nucleotide #4,285 and ends at nucleotide #7,375. The remaining nucleotides corresponds to pBSSK+) ;
Figure 26 shows a restriction map of pLF092;
Figure 27 shows a complete DNA sequence of . pLF105 ((6,243 bp) (SEQ ID NO: 14) The polylinker starts at nucleotide #3,092 and ends at nucleotide #3,123. The CAV2 left flanking arm (182 bp) starts at nucleotide #3,123 and ends at nucleotide #3,321. The CAV2 right flanking arm (3,090 bp) starts at nucleotide #1 and ends at nucleotide #3,091. The remaining nucleotides correspond to pBSSK+);
Figure 28 shows a restriction map of pLF105;
5 Figure 29 shows a complete DNA sequence of pLF102 ((6,615 bp) (SEQ ID NO: 15) The 305 by BZaII/MluI
linker starts at nucleotide #1,471 and ends at nucleotide #1,776. The remaining sequences correspond to pLF086;
Figure 30 shows a restriction map of pLF102);
10 Figure 31 shows a complete DNA sequence of pLF1116A ((6,450 bp) (SEQ ID NO: 16) The 311 by MluI/MluI
linker starts at nucleotide #1,092 and ends at nucleotide #1,403. The remaining sequences correspond to pLF086);
Figure 32 shows a restriction map of pLF1i16A;
15 Figure 33 shows a complete DNA sequence of pLF100 ((6,247 bp) (SEQ ID NO: 17) The 302 by DraIII/MluI
linker starts at nucleotide #898 and ends at nucleotide #1,200. The remaining sequences correspond to pLF086);
Figure 34 shows a restriction map of pLF100;
20 Figure 35 shows a complete DNA sequence of pLF120 ((6,048 bp) (SEQ ID NO: 18) The 311 by DraIII/MluI
linker starts at nucleotide #898 and ends at nucleotide #1,209. The remaining sequences correspond to pLF086);
Figure 36 shows a restriction map of pLF120;
25 Figure 37 shows a complete DNA sequence of pLF043 ((5,109 bp) (SEQ ID NO: 19) CDV HA coding sequence starts at nucleotide #35 and ends. at nucleotide #2175.
CDV HA ORF stop codon is #1847. The partial vaccinia H6 promoter starts at nucleotide #7 and ends at nucleotide 30 #35. The remaining sequences correspond to pBSSK+);
Figure 38 shows a restriction map of pLF043;
Figure 39 shows a complete DNA sequence of pLF098 ((5,070 bp) (SEQ ID NO: 20) CDV HA expression cassette starts at nucleotide #1 and ends at nucleotide 35 #2372. The remaining sequences correspond to pLF069);
Figure 40 shows a restriction map of pLF098;

WO 98!00166 PCT/US97/11486 Figure 41 shows a complete DNA sequence of pLF099A ((8,618 bp) (SEQ ID NO: 21) CDV HA expression cassette starts at nucleotide #3120 and ends at nucleotide #5,494. The remaining sequences correspond to pLF105) ;
Figure 42 shows a restriction map of pLF099A;
Figure 43 shows a complete DNA sequence of pLF108 ((4,965 bp) (SEQ ID NO: 22) the 3' most region of the vaccinia virus H6 promoter is located between position#1 and 29; the CDV F1 coding sequence begins at position#30 and terminates at position#2,018; the remaining sequences correspond to pBSSK+);
Figure 44 shows a restriction map of pLF108;
Figure 45 shows a complete DNA sequence of pLFlll ((5,241 bp) (SEQ ID NO: 23) CDV F1 expression cassette begins at position #1 and terminates at position #2,556; the remaining sequences correspond to pLF069);
Figure 46 shows a restriction map of pLFlll;
Figure 47 shows a complete DNA sequence of pLF128 ((5,147 bp) (SEQ ID NO: 24) CDV F1 expression cassette begins at position #1 and terminates at position #2,452; the remaining sequences correspond to pLF077);
Figure 48 shows a restriction map of pLF128;
Figure 49 shows a complete DNA sequence of pLF130A ((8,792 bp) (SEQ ID NO: 25) CDV F1 expression cassette begins at position #3,126 and terminates at nucleotide #5,669; the CAV2 SalI.B left flanking arm (3,091 bp) is located between position #1 and 3,091; the CAV2 SalI.B right flanking arm (182 bp) is located between position #5,688 and 5,870; the remaining sequences correspond to pLF105); and, Figure 50 shows a restriction map of pLF130A.
DETAILED DESCRIPTION
As mentioned earlier, the present invention relates to recombinant adenovirus, such as CAV, preferably CAV2, methods for making and using them, and WO 98/00166 PCT/iJS97111486 to compositions containing them or their expression products; and, to promoters and expression cassettes .
More specifically, this invention relates to recombinant CAV such as CAV2, especially those wherein exogenous DNA has been inserted into a non-essential region and/or a non-essential region is deleted and methods of making them, uses for them (including as a vector for replicating DNA), expression products from them, and uses for the expression products. The CAV E3 region, preferably ORF2, is preferred for insertion and/or deletion.
The uses for recombinant viruses, and for products therefrom can be determined without undue experimentation from the documents set forth in the BACKGROUND OF THE INVENTION and the discussion under the SUMMARY OF THE INVENTION.
The heterologous or exogenous DNA in recombinants of the invention preferably encodes an expression product comprising: an epitope of interest, a biological response modulator, a growth factor, a recognition sequence, a therapeutic gene, or a fusion protein. With respect to these terms, reference is made to the following discussion, and generally to Kendrew, THE ENCYCLOPEDIA OF MOLECOLAR BIOLOGY (Blackwell Science Ltd 1995) and Sambrook, Fritsch, Maniatis, Molecular Cloning, A LABORATORY MANUAL (2d Edition, Cold Spring Harbor Laboratory Press, 1989).
As to antigens for use in vaccine or immunological compositions, reference is made to the documents and discussion set forth in the BACKGROUND OF
THE INVENTION and the discussion under the SUMMARY OF THE
INVENTION; see also Stedman's Medical Dictionary (24th edition, 1982, e.g., definition of vaccine (for a list of antigens used in vaccine formulations; such antigens or epitopes of interest from those antigens can be used in the invention, as either an expression product of the inventive recombinant virus, or in a multivalent composition containing an inventive recombinant virus or an expression product therefrom).
As to epitopes of interest, one skilled in the art can determine an epitope or immunodominant region of a peptide or polypeptide and ergo the coding DNA therefor from the knowledge of the amino acid and corresponding - DNA sequences of the peptide or polypeptide, as well as from the nature of particular amino acids (e. g., size, charge, etc.) and the codon dictionary, without undue experimentation.
A general method for determining which portions of a protein to use in an immunological composition focuses on the size and sequence of the antigen of interest. "In general, large proteins, because they have more potential determinants are better antigens than small ones. The more foreign an antigen, that is the less similar to self configurations which induce tolerance, the more effective it is in provoking an immune response." Ivan Roitt, Essential Immunolocry, 1988.
As to size: the skilled artisan can maximize the size of the protein encoded by the DNA sequence to be inserted into the viral vector (keeping in mind the packaging limitations of the vector). To minimize the DNA inserted while maximizing the size of the protein expressed, the DNA sequence can exclude introns (regions of a gene which are transcribed but which are subsequently excised from the primary RNA transcript).
At a minimum, the DNA sequence can code for a peptide at least 8 or 9 amino acids long. This is the minimum length that a peptide needs to be in order to stimulate a CD4+ T cell response (which recognizes virus infected cells or cancerous cells). A minimum peptide length of 13 to 25 amino acids is useful to stimulate a CD8+ T cell response (which recognizes special antigen presenting cells which have engulfed the pathogen). See - Kendrew, supra. However, as these are minimum lengths, these peptides are likely to generate an immunological response, i.e., an antibody or T cell response; but, for a protective response (as from a vaccine composition), a longer peptide is preferred.
With respect to the sequence, the DNA sequence preferably encodes at least regions of the peptide that generate an antibody response or a T cell response. One method to determine T and B cell epitopes involves epitope mapping. The protein of interest "is fragmented into overlapping peptides with proteolytic enzymes. The individual peptides are then tested for their ability to bind to an antibody elicited by the native protein or to induce T cell or B cell activation. This approach has been particularly useful in mapping T-cell epitopes since the T cell recognizes short linear peptides complexed with MHC molecules. The method is less effective for determining B-cell epitopes" since B cell epitopes are often not linear amino acid sequence but rather result from the tertiary structure of the folded three dimensional protein. Janis Kuby, Immunoloay, (1992) pp.
79-80.
Another method for determining an epitope of interest is to choose the regions of the protein that are hydrophilic. Hydrophilic residues are often on the surface of the protein and are therefore often the regions of the protein which are accessible to the antibody. Janis Kuby, Immunoloay, (1992) p. 81.
Yet another method for. determining an epitope of interest is to perform an X-ray crystallographic analysis of the antigen (full length)-antibody complex.
Janis Kuby, Immunoloay, (1992) p. 80.
Still another method for choosing an epitope of interest which can generate a T cell response is to identify from the protein sequence potential HLA anchor binding motifs which are peptide sequences which are known to be likely to bind to the MHC molecule.
The peptide which is a putative epitope of interest, to generate a T cell response, should be presented in a MHC complex. The peptide preferably contains appropriate anchor motifs for binding to the MHC
molecules, and should bind with high enough affinity to generate an immune response. Factors which can be 5 considered are: the HLA type of the patient (vertebrate, animal or human) expected to be immunized, the sequence of the protein, the presence of appropriate anchor motifs and the occurance of the peptide sequence in other vital cells.
10 An immune response is generated, in general, as follows: T cells recognize proteins only when the protein has been cleaved into smaller peptides and is presented in a complex called the "major histocompatability complex MHC" located on another cell's surface. There are two 15 classes of MHC complexes - class I and class II, and each class is made up of many different alleles. Different patients have different types of MHC complex alleles;
they are said to have a 'different HLA type.' Class I MHC complexes are found on virtually 20 every cell and present peptides from proteins produced inside the cell. Thus, Class I MHC complexes are useful for killing cells which when infected by viruses or which have become cancerous and as the result of expression of an oncogene. T cells which have a protein called CD4 on 25 their surface, bind to the MHC class I cells and secrete lymphokines. The lymphokines stimulate a response; cells arrive and kill the viral infected cell.
Class II MHC complexes are found only on antigen- presenting cells and are used to present 30 peptides from circulating pathogens which have been endocytosed by the antigen- presenting cells. T cells which have a protein called CD8 bind to the MHC class II
cells and kill the cell by exocytosis of lytic granules.
Some guidelines in determining whether a 35 protein is an epitopes of interest which will stimulate a T cell response, include: Peptide length - the peptide should be at least 8 or 9 ammino acids long to fit into the MHC class I complex and at least 13-25 amino acids long to fit into a class II MCH complex. This length is a minimum for the peptide to bind to the MHC complex. It is preferred for the peptides to be longer than these lengths because cells may cut the expressed peptides.
The peptide should contain an appropriate anchor motif which will enable it to bind to the various class I or class II molecules with high enough specificity to generate an immune response (See Bocchia, M. et al, Specific Binding of Leukemia Oncoaene Fusion Protein Peptides to HLA Class I Molecules, Blood 85:2680-2684;
Englehard, VH, Structure of peptides associated with class I and class II MHC molecules Ann. Rev. Immunol.
12:181 (1994)). This can be done, without undue experimentation, by comparing the sequence of the protein of interest with published structures of peptides associated with the MHC molecules. Protein epitopes recognized by T cell receptors are peptides generated by enzymatic degradation of the protein molecule and are prestnted on the cell surface in association with class I
or class II MHC molecules.
Further, the skilled artisan can ascertain an epitope of interest by comparing the protein sequence with sequences listed in the protein data base. Regions of the protein which share little or no homology are better choices for being an epitope of that protein and are therefore useful in a vaccine or immunological composition. Regions which share great homology with widely found sequences present in vital cells should be avoided.
Even further, another method is simply to generate or express portions of a protein of interest, generate monoclonal antibodies to those portions of the protein of interest, and then ascertain whether those antibodies inhibit growth in vitro of the pathogen from which the from which the protein was derived. The skilled artisan can use the other guidelines set forth in ~
~ CA 02259460 2005-03-29 this disclosure and in the art for generating or expressing portions of a protein of interest for analysis as to whether antibodies thereto inhibit growth in vitro.
For example, the skilled artisan can generate portions of a protein of interest by: selecting 8 to 9 or 13 to 25 amino acid length portions of the protein, selecting hydrophylic regions, selecting portions shown to bind from X-ray data of the antigen (full length)-antibody complex, selecting regions which differ in sequence from lO other proteins, selecting potential HLA anchor binding motifs, or any combination of these methods or other methods known in the art.
Epitopes recognized by antibodies are expressed on the surface of a protein. To determine the regions of a protein mast likely to stimulate an antibody response one skilled in the art can preferably perform an epitope map, using the general methods described above, or other mapping methods known in the art.
As can be seen from the foregoing, without undue experimentation, from this disclosure and the knowledge in the art, the skilled artisan can ascertain the amino acid and corresponding DNA sequence of an epitope of interest for obtaining a T cell, B cell and/or antibody response. In addition, reference is made to Gefter et al., U.S. Patent No. 5,019,384, issued May 28, 1991, and the documents it cites, (Note especially the "Relevant Literature"
section of this patent, and column 13 of this patent which discloses that: "A large number of epitopes have been defined for a wide variety of organisms of interest.
Of particular interest are those epitopes to which neutralizing antibodies are directed. Disclosures of - such epitopes are in many of the-references cited in the Relevant Literature section.") With respect to expression of a biological response modulator, reference is made to Wohlstadter, "Selection Methods," WO 93/19170, published 30 September 48 ' 1993, and the documents cited therein.
For instance, a biological response modulator modulates biological activity; for instance, a biological response modulator is a modulatory component such as a high molecular weight protein associated with non-NMDA
excitatory amino acid receptors and which allosterically -regulates affinity of AMPA binding (See Kendrew, supra).
The recombinant of the present invnention can express such a high molecular weight protein.
More generally, nature has provided a number of precedents of biological response modulators. Modulation of activity may be carried out through mechanisms as complicated and intricate as allosteric induced quaternary change to simple presence/absence, e.g., expression/degradation, systems. Indeed, the repression/activation of expression of many biological molecules is itself mediated by molecules whose activities are capable of being modulated through a variety of mechanisms.
Table 2 of Neidhardt et al Physiology of the Bacterial CeI1 (Sinauer Associates Inc., Publishers, 1990), at page 73, lists chemical modifications to bacterial proteins. As is noted in that table, some modifications are involved in proper assembly and other modifications are not, but in either case such modifications are capable of causing modulation of function. From that table, analogous chemical modulations for proteins of other cells can be determined, without undue experimentation.
In some instances modulation of biological functions may be mediated simply through the proper/improper localization of a molecule. Molecules may function to provide a growth advantage or disadvantage only if they are targeted to a particular location. For example, a molecule may be typically not taken up or used by a cell, as a function of that WO 98100166 PCTJUS9'7/11486 molecule being first degredaded by the cell by secretion of an enzyme for that degradation. Thus, production of the enzyme by a recombinant can regulate use or uptake of the molecule by a cell. Likewise, the recombinant can express a molecule which binds to the enzyme necessary for uptake or use of a molecule, thereby similarly ' regulating its uptake or use.
Localization targeting of proteins carried out through cleavage of signal peptides another type of modulation or regulation. In this case, a specific endoprotease catalytic activity can be expressed by the recombinant.
Other examples of mechanisms through which modulation of function may occur are RNA virus poly proteins, allosteric effects, and general covalent and non-covalent steric hindrance. HIV is a well studied example of an RNA virus which expresses non-functional poly-protein constructs. In HIV "the gag, pol, and env poly-proteins are processed to yield, respectively, the viral structural proteins p17, p24, and p15--reverse transcriptase and integrase--and the two envelope proteins gp41 and gp120" (Kohl et al., PNAS USA 85:4686-90 (1988)). The proper cleavage of the poly-proteins is crucial for replication of the virus, and virions carrying inactive mutant HIV protease are non-infectious (Id.). This is another example of the fusion of proteins down-modulating their activity. .Thus, it is possible to construct recombinant viruses which express molecules which interfere with endoproteases, or which provide endoproteases, for inhibiting or enhancing the natural expression of certain proteins (by interfering with or enhancing cleavage).
The functional usefulness of enzymes may also . be modulated by altering their capability of catalyzing a reaction. Illustrative examples of modulated molecules are zymogens, formation/disassociation of multi-subunit functional complexes, RNA virus poly-protein chains, allosteric interactions, general steric hindrance (covalent and non-covalent) and a variety of chemical modifications such as phosphorylation, methylation, acetylation, adenylation, and uridenylation (see Table 1 5 of Neidhardt, supra, at page 315 and Table 2 at page 73).
Zymogens are examples of naturally occurring protein fusions which cause modulation of enzymatic activity. Zymogens are one class of proteins which are converted into their active state through limited i0 proteolysis. See Table 3 of Reich, Proteases and Biological Control, Vol. 2, (1975) at page 54). Nature has developed a mechanism of down-modulating the activity of certain enzymes, such as trypsin, by expressing these enzymes with additional "leader" peptide sequences at 15 their amino termini. With the extra peptide sequence the enzyme is in the inactive zymogen state. Upon cleavage of this sequence the zymogen is converted to its enzymatically active state. The overall reaction rates of the zymogen are "about 105-106 times lower than those 20 of the corresponding enzyme" (See Table 3 of Reich, supra at page 54).
It is therefore possible to down-modulate the function of certain enzymes simply by the addition of a peptide sequence to one of its termini. For example, 25 with knowledge of this property, a recombinant can express peptide sequences containing additional amino acids at one or both terminii.
The formation or disassociation of multi-subunit enzymes is another way through which modulation 30 may occur. Different mechanisms may be responsible for the modulation of activity upon formation or disassociation of multi-subunit enzymes.
Therefore, sterically hindering the proper specific subunit interactions will down-modulate the 35 catalytic activity. And accordingly, the recombinant of the invention can express a molecule which sterically WO 98/OOlb6 PCT/US97I1I486 hinders a naturally occurring enzyme or enzyme complex, so as to modulate biological functions.
Certain enzyme inhibitors afford good examples of functional down-modulation through covalent steric hindrance or modification. Suicide substrates which irreversibly bind to the active site of an enzyme at a catalytically important amino acid in the active site are examples of covalent modifications which sterically block the enzymatic active site. An example of a suicide substrate is TPCK for chymotrypsin (Fritsch, Enzyme Structure and Mechanism, 2d ed; Freeman & Co. Publishers, 1984)). This type of modulation is possible by the recombinant expressing a suitable suicide substrate, to thereby modulate biological responses (e. g., by limiting enzyme activity).
There are also examples of non-covalent steric hindrance including many repressor molecules. The recombinant can express repressor molecules which are capable of sterically hindering and thus down-modulating the function of a DNA sequence by preventing particular DNA-RNA polymerase interactions.
Allosteric effects are another way through which modulation is carried out in some biological systems. Aspartate transcarbamoylase is a well characterized allosteric enzyme. Interacting with the catalytic subunits are regulatory domains. Upon binding to CTP or UTP the regulatory subunits are capable of inducing a quaternary structural change in the holoenzyme causing down-modulation of catalytic activity. In contrast, binding of ATP to the regulatory subunits is capable of causing up-modulation of catalytic activity (Fritsch, supra). Using methods of the invention, molecules can be expressed which are capable of binding and causing modulatory quaternary or tertiary changes.
In addition, a variety of chemical modifications, e.g., phosphorylation, methylation, acetylation, adenylation, and uridenylation may be carried out so as to modulate function. It is known that modifications such as these play important roles in the regulation of many important cellular components. Table 2 of Neidhardt, supra, at page 73, lists different bacterial enzymes which undergo such modifications. From that list, one skilled in the art can ascertain other enzymes of other systems which undergo the same or similar modifications, without undue experimentation. In addition, many proteins which are implicated in human disease also undergo such chemical modifications. For example, many oncogenes have been found to be modified by phosphorylation or to modify other proteins through phosphorylation or dephosphorylation. Therefore, the ability afforded by the invention to express modulators which can modify or alter function, e.g., phosphorylation, is of importance.
From the foregoing, the skilled artisan can use the present invention to express a biological response modulator, without any undue experimentation.
With respect to expression of fusion proteins by inventive recombinants, reference is made to Sambrook, Fritsch, Maniatis, Molecular Cloning, A LABORATORY MANUAL
(2d Edition, Cold Spring Harbor Laboratory Press, 1989) (especially Volume 3), and Kendrew, supra.
The teachings of Sambrook a al.;
can be suitably modified, without undue experiementation, from this disclosure, for the skilled artisan to generate recombinants expressing fusion proteins.
With regard to gene therapy and immunotherapy, reference is made to U.S. Patent Nos. 4,b90,915 and .
5,252,479, together with the documents cited therein it and on their face, and to WO 94/16716 and U.S. Patent No. 5,833,975, filed January 19, 1994, together with the documents cited therein:

. CA 02259460 2005-03-29 A growth factor can be defined as multifunctional, locally acting intercellular signalling peptides which control both ontogeny and maintenance of tissue and function (see Kendrew, especially at page 4 5 5 et seq. ) .
The growth factor or therapeutic gene, for example, can encode a disease-fighting protein, a molecule for treating cancer, a tumor suppressor, a cytokine, a tumor associated antigen, or interferon; and, to the growth factor or therapeutic gene can, for example, be selected from the group consisting of a gene encoding alpha-globin, beta-globin, gamma-globin, granulocyte macrophage-colony stimulating factor, tumor necrosis factor, an interleukin (e. g., an interleuk in selected from interleukins 1 to 14, or l to 11, or any combinat ion thereof), macrophage colony stimulating f actor, granulocyte colony stimulating factor, erythropoietin, mast cell growth factor, tumor suppressor p53, retinoblastoma, interferon, melanoma associated antigen or B7. U.S. Patent No. 5,252,479 provides a list of proteins which can be expressed in an adenovirus system for gene therapy, and the skilled artisan is directed to that disclosure. WO 94/16716 and U.S. Patent No. 5,833,975, filed January 19, 1994, provide genes for cytokines and tumor associated antigens and immunotherapy methods, including ex vivo methods, and the skilled artisan is directed to those disclosures.
Thus, one skilled in the art can create recombinants expressing a growth factor or therapeutic gene and use the recombinants, from this disclosure and the knowledge in the art, without undue experimentation.
Moreover, from the foregoing and the knowledge in the art, no undue experimentation is required for the skilled artisan to construct an inventive recombinant which expresses an epitope of interest, a biological . response modulator, a growth factor, a recognition sequence, a therapeutic gene, or a fusion protein; or for the skilled artisan to use such a recombinant.
It is noted that the exogenous or heterologous DNA can itself include a promoter for driving expression in the recombinant CAV, or the exogenous DNA can simply be coding DNA and appropriately placed downstream from an endogenous promoter to drive expression. Further, multiple copies of coding DNA or use of a strong or early promoter or early and late promoter, or any combination thereof, can be done so as to amplify or increase expression. Thus, the exogenous or heterologous DNA can be suitably positioned with respect to an endogenous promoter Like the E3 or the MLP promoters, or those promoters can be translocated to be inserted at another location, with the exogenous or heterologous DNA. The coding DNA can be DNA coding for more than one protein so as to have expression of more than one product from the recombinant CAV.
The expression products can be antigens, immunogens or epitopes of interest; and therefore, the invention further relates to immunological, antigenic or vaccine compositions containing the expression products.
Further, since the CAV vector, in certain instances, can be administered directly to a suitable host, the invention relates to compositions containing the CAV, preferably CAV2, vector. Additionally, since the expression product can be isolated from the CAV, preferably CAV2, vector in vitro or from cells infected or transfected by the CAV vector in vitro, the invention relates to methods for expressing a product, e.g., comprising inserting the exogenous DNA into a CAV as a vector, e.g., by restriction/ligation or by recombination followed by infection or transfection of suitable cells in vitro with a recombinant CAV, and optionally extracting, purifying or isolating the expression product from the cells. Any suitable extraction, purification or isolation techniques can be employed; and reference is made to the discussion and documents in the BACKGROUND OF
THE INVENTION and SUMMARY OF THE INVENTION.
In particular, after infecting cells with the recombinant CAV, the proteins) from the expression of 5 the exogenous DNA are collected by known techniques such as chromatography (see Robbins, EPA 0162738A1; Panicali, r EPA 0261940A2); Richardson, su ra; Smith et al., supra;
Pennock et al., supra; EP Patent Publication No.
0265785). The collected proteins) can then be employed 10 in a vaccine, antigenic or immunological composition which also contains a suitable carrier.
Thus, the recombinant CAV can be used to prepare proteins such as antigens, immunogens, epitopes of interest, etc. which can be further used in 15 immunological, antigenic or vaccine compositions. It is noted that a recombinant CAV expressing a product detrimental to growth or development of insects can be used to prepare an insecticide, and a recombinant CAV
expressing a product detrimental to growth of plants can 20 be used to prepare a herbicide (by isolating the expression product and admixing it with an insecticidally or herbicidally acceptable carrier or diluent) and a recombinant CAV expressing an anti-fungal polypeptide can be used to prepare an anti-fungal preparation (by 25 isolating the expression product and admixing it with a suitable carrier or diluent).
As the expression products can provide an antigenic, immunological or protective (vaccine) response, the invention further relates to products 30 therefrom; namely, antibodies and uses thereof. More in particular, the expression products can elicit antibodies. The antibodies can be formed into monoclonal antibodies; and, the antibodies or expression products can be used in kits, assays, tests, and the like 35 involving binding, so that the invention relates to these uses too. Additionally, since the recombinants of the invention can be used to replicate DNA, the invention WO 98/00166 PCTlUS97/11486 relates to recombinant CAV as a vector and methods for replicating DNA by infecting or transfecting cells with the recombinant and harvesting DNA therefrom. The resultant DNA can be used as probes or primers or for amplification.
The administration procedure for recombinant CAV or expression product thereof, compositions of the invention such as immunological, antigenic or vaccine compositions or therapeutic compositions can be via a parenteral route (intradermal, intramuscular or subcutaneous). Such an administration enables a systemic immune response. The administration can be via a mucosal route, e.g., oral, nasal, genital, etc. Such an administration enables a local immune response.
More generally, the inventive antigenic, immunological or vaccine compositions or therapeutic compositions (compositions containing the CAV, preferably CAV2, recombinants of the invention or expression products) can be prepared in accordance with standard techniques well known to those skilled in the pharmaceutical or vetinary arts. Such compositions can be administered in dosages and by techniques well known to those skilled in the medical arts taking into consideration such factors as the breed or species, age, sex, weight, and condition of the particular patient, and the route of administration. The compositions can be administered alone, or can be co-administered or sequentially administered with other compositions of the invention or with other immunological, antigenic or vaccine or therapeutic compositions. Such other compositions can include purified native antigens or epitopes or antigens or epitopes from the expression by a recombinant CAV or another vector system; and are administered taking into account the aforementioned factors.
Examples of compositions of the invention include liquid preparations for orifice, e.g., oral, nasal, anal, genital, e.g., vaginal, etc., administration such as suspensions, syrups or elixirs; and, preparations for parenteral, subcutaneous, intradermal, intramuscular or intravenous administration (e. g., injectable administration) such as sterile suspensions or emulsions.
In such compositions the recombinant may be in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose or the like.
Antigenic, immunological or vaccine compositions typically can contain an adjuvant and an amount of the recombinant CAV or expression product to elicit the desired response. In human applications, alum (aluminum phosphate or aluminum hydroxide} is a typical adjuvant. Saponin and its purified component Quil A, Freund's complete adjuvant and other adjuvants used in research and veterinary applications have toxicities which limit their potential use in human vaccines.
Chemically defined preparations such as muramyl dipeptide, monophosphoryl lipid A, phospholipid conjugates such as those described by Goodman-Snitkoff et al. J, fmmunol. 147:410-415 (199I), encapsulation of the protein within a proteoliposome as described by Miller et al., ,7. Exp.
Med. 176:1739-1744 (1992), and encapsulation of the protein i:n lipid vesicles such as NovasomeTM lipid vesicles (Micro Vescular Systems, Inc., Nashua, NH) can also be used.
The composition may be packaged in a single dosage form for immunization by parenteral (i.e., intramuscular, intradermal or subcutaneous) administration or orifice administration, e.g., perlingual (i.e., oral), intragastric, mucosal including intraoral, intraanal, intravaginal, and the like administration. And again, the effective dosage and route of administration are determined by the nature of the composition, by the nature of the expression product, by expression level if recombinant CAV2 is directly used, and by known factors, such as breed or species, age, sex, weight, condition and nature of host, as well as LDSO and other screening procedures which are known and do not require undue experimentation. Dosages of expressed product can range from a few to a few hundred micrograms, e.g., 5 to 500 ,gig. The inventive recombinant can be administered in any suitable amount to achieve expression at these dosage levels. The vaccinal CAV2 is administered in an amount of about 103'5 pfu; thus, the inventive recombinant is preferably administered in at least this amount; more preferably about 104 pfu to about 106 pfu. Other suitable carriers or diluents can be water or a buffered saline, with or without a preservative. The expression product or recombinant CAV
may be lyophilized for resuspension at the time of administration or can be in solution.
The carrier may also be a polymeric delayed release system. Synthetic polymers are particularly useful in the formulation of a composition having controlled release. An early example of this was the polymerization of methyl methacrylate into spheres having diameters less than one micron to form so-called nano particles, reported by Kreuter, J., Microcapsules and Nanoparticles in Medicine and Pharmacology, M. Donbrow (Ed). CRC Press, p. 125-148.
Microencapsulation has been applied to the injection of microencapsulated pharmaceuticals to give a controlled release. A number of factors contribute to the selection of a particular polymer for microencapsulation. The reproducibility of polymer synthesis and the microencapsulation process, the cost of the microencapsulation materials and process, the toxicological profile, the requirements for variable release kinetics and the physicochemical compatibility of the polymer and the antigens are all factors that must be considered. Examples of useful polymers are polycarbonates, polyesters, polyurethanes, polyorthoesters and polyamides, particularly those that are biodegradable.
A frequent choice of a carrier for ' S pharmaceuticals and more recently for antigens is poly (d,l-lactide-co-glycolide) (PLGA). This is a ~ biodegradable polyester that has a long history of medical use in erodible sutures, bone plates and other temporary prostheses where it has not exhibited any toxicity. A wide variety of pharmaceuticals including peptides and antigens have been formulated into PLGA
microcapsules. A body of data has accumulated on the adaption of PLGA for the controlled release of antigen, for example, as reviewed by Eldridge, J.H., et al.
Current Topics in Microbiology and Immunology 1989, 146:59-66. The entrapment of antigens in PLGA
microspheres of 1 to 10 microns in diameter has been shown to have a remarkable adjuvant effect when administered orally. The PLGA microencapsulation process uses a phase separation of a water-in-oil emulsion. The compound of interest is prepared as an aqueous solution and the PLGA is dissolved in a suitable organic solvents such as methylene chloride and ethyl acetate. These two immiscible solutions are co-emulsified by high-speed stirring. A non-solvent for the polymer is then added, causing precipitation of the polymer around the aqueous droplets to form embryonic microcapsules. The microcapsules are collected, and stabilized with one of an assortment of agents (polyvinyl alcohol (PVA), gelatin, alginates, polyvinylpyrrolidone (PVP), methyl cellulose) and the solvent removed by either drying is vacuo or solvent extraction.
Thus, solid, including solid-containing-liquid, liquid, and gel (including "gel caps") compositions are envisioned. Additionally, the inventive vectors, e.g., recombinant CAV2, and the expression products therefrom can stimulate an immune or antibody response in animals.

From those antibodies, by techniques well-known in the art, monoclonal antibodies can be prepared and, those monoclonal antibodies, can be employed in well known antibody binding assays, diagnostic kits or tests to 5 determine the presence or absence of antigens) and therefrom the presence or absence of the natural causative agent of the antigen or, to determine whether an immune response to that agent or to the antigens) has simply been stimulated.
10 Monoclonal antibodies are immunoglobulin produced by hybridoma cells. A monoclonal antibody reacts with a single antigenic determinant and provides greater specificity than a conventional, serum-derived antibody. Furthermore, screening a large number of 15 monoclonal antibodies makes it possible to select an individual antibody with desired specificity, avidity and isotype. Hybridoma cell lines provide a constant, inexpensive source of chemically identical antibodies and preparations of such antibodies can be easily 20 standardized. Methods for producing monoclonal antibodies are well known to those of ordinary skill in the art, e.g., Koprowski, H. et al., U.S. Pat. No.
4,196,265, issued Apr. 1, 1989.
25 Uses of monoclonal antibodies are known. One such use is in diagnostic methods, e.g., David, G. and Greene, H., U.S. Pat. No. 4,376,110, issued Mar. 8, 1983.
Monoclonal antibodies have also been used to 30 recover materials by immunoadsorption chromatography, e.g. Milstein, C., 1980, Scientific American 243:66, 70.
Furthermore, the inventive recombinant CAV or expression products therefrom can be used to stimulate a 35 response in cells in vitro or ex vivo for subsequent reinfusion into a patient. If the patient is seronegative, the reinfusion is to stimulate an immune response, e.g., an immunological or antigenic response such as active immunization. In a seropositive individual, the reinfusion is to stimulate or boost the immune system against a pathogen.
The recombinant CAV of the invention are also useful for generating DNA for probes or for PCR primers which can be used to detect the presence or absence of hybridizable DNA or to amplify DNA, e.g., to detect a pathogen in a sample or for amplifying DNA.
Furthermore, as discussed above, the invention comprehends promoters and expression cassettes which are useful in adenovirus systems, as well as in any viral or cell system which provides a transactivating protein.
The promoter is preferably a truncated transcriptionally active promoter for a recombinant virus or plasmid which comprises a region transactivated with a transactivating protein provided by the virus or a system into which the plasmid is inserted and the minimal promoter region of a full-length promoter from which the truncated transcriptionally active promoter is derived.
Like the inventive promoter is preferably a derived from a eukaryotic virus such as a herpesvirus, e.g., a MCMV or HCMV such as MCMV-IE or HCMV-IE promoter;
and, there can be up to a 40% and even up to a 90%
reduction in size, from a full-length promoter, based upon base pairs.
The expression cassette.of the invention can further include a functional truncated polyadenylation signal; for instance an SV40 polyadenylation signal which is truncated, yet functional. The expression cassette can contain exogenous or heterologous DNA (with respect to the virus or system into which the promoter or expression cassette is being inserted); for instance exogenous or heterologous coding DNA as herein described above, and in the Examples. This DNA can be suitably positioned and operably linked to the promoter for expression. The expression cassette can be inserted in 62 ' any orientation; preferably the orientation which obtains maximum expression from the system or virus into which the expression cassette is inserted.
While the promoter and expression cassette are specifically exemplified with reference to adenoviruses, the skilled artisan can adapt these embodiments of the invention to other viruses and to plasmids for cells such as eukaryotic cells, without undue experimentation, by simply ascertaining whether the virus, plasmid, cell or system provides the transactivating protein.
As to HCMV promoters, reference is made to U.S.
Patent Nos. 5,168,062 and 5,385,839.
As to transfecting cells with plasmid DNA
for expression therefrom, reference is made to Fei~ner et al. (1994), J. Biol. Chem. 269, 2550-2561.
And, as to direct injection of plasmid DNA as a simple and effective method of vaccination against a variety of infectious diseases reference is made to Science, 259:1745-49, 1993.
It is therefore within the scope of this invention that the inventive promoter and expression cassette be used in systems other than adenovirus; for example, in plasmids for the direct injection of plasmid DNA.
Other utilities also exist for embodiments of the invention.
The following non-limiting Examples are given by way of illustration only and are not to be considered a limitation of this invention.
EXAMPLES
EXAMPLE 1 - Virus and cell line identifications The described stock of canine adenovirus type 2 (CAV2) was produced at Rhone Merieux Inc. (Athens, Ga) under the reference CAV2 Lot # 0830 pool - 033093, with a titer of 10~~4 TCID50/ml. Madin and Darby canine kidney (MDCK) cell line was also provided by Rhone Meri.eux Inc.

CAV2 is commercially available from Rhone Merieux Inc. as a canine vaccine.
- EXAMPLE 2 - Virus culture and cloning MDCK cell suspensions were seeded in MEM
(Gibco, Grand Island, NY) supplemented with 7.5o fetal bovine serum (Sigma, St Louis, MO), sodium pyruvate ~ (Gibco, 1 mM final), glutamine (Gibco, 2 mM final), penicillin (Gibco, 50 U/ml), streptomycin (Gibco, 50 mg/ml) and non essential amino acids (NEA)(Gibco, 0.1 mM
final) and cultured at 37itch in 5~ C02. Confluent MDCK
cells were infected with serial dilutions of CAV2 and cultured under a 0.6% agarose overlay at 37itch in 5% C02.
CAV2 was subjected to several rounds of plaque purification. A plaque purified CAV2 was amplified in a T25 MDCK flask. When the culture CPE was complete, infected cells were collected and their CAV2 content was titrated on MDCK cell monolayers under agarose. The virus stock was further amplified by infecting a confluent T175 MDCK flask with a multiplicity of infection (MOI) of 0.1. The titre of the T175 MDCK flask amplified virus was established to be 108 p.f.u/ml.
EXAMPLE 3 - Viral DNA purification Roller bottles containing confluent MDCK cell monolayers (108 cells/bottle) were infected at a MOI of 0.1 pfu/cell with plaque purified CAV2 virus. Three days later the infected monolayer were harvested and subjected to low speed centrifugation (1K g., 15 minutes, 15°C).
The cell pellets were stored at -70°C. The frozen pellets were subsequently thawed at 37°C and carefully resuspended in lOmM Tris HC1 pH 8.0 and 10 mM EDTA buffer (35 ml/108 cells) to limit cellular DNA shearing. SDS
was added to the resuspended pellets to a final concentration of 1°s. After 15 minutes incubation at room temperature NaCl was added to a concentration of 1.25 M.
After 3 hours incubation at 4°C the material was centrifuged at 25K g for 20 minutes at 4°C. Dense white pellets containing salts and cellular DNA were discarded and supernatants were digested with Proteinase K (300 ~,g/ml final concentration) at 42°C for 4 hours and subsequently heated at 65°C for 30 minutes. Two cycles of phenol-chloroform and chloroform extractions were performed prior to recovery of viral DNA by ethanol precipitation in the presence of 0.3 M sodium acetate pH
6Ø The viral DNA pellet was washed with 70% ethanol before being air dried for 1 hour and subsequently resuspended in 2 ml of H20. This procedure typically yields approximatively 4 mg of purified CAV2 DNA.
Purified viral DNA was stored at -20°C until further utilization.
EXAMPLE 4 - Viral DNA restriction analysis Aliquots of purified CAV2 DNA were digested with a set of restriction enzymes purchased from Boehringer Mannheim Corp. (Indianapolis, IN) accordingly to the manufacturer's specifications. Restricted DNA
samples were fractionated by electrophoresis on a 1%
agarose gel and the corresponding restriction fragments were visualized under UV light after staining of the gel with ethidium bromide (4 ~.g/ml). Table 1 summarizes the size of the various restriction fragments.
EXAMPLE 5 -Identification and characterization of the restriction fragment containing the E3 2 5 region 1. Southern blot analysis of specific endonuclease restricted CAV2 DNA.
Four ~g aliquots of purified CAV2 DNA were digested with BamHI, Ball, HindII, HindIII and PstI, respectively, before being fractionated by electrophoresis through a 1% agarose gel. The gel was soaked in 0.25 M HCL for 30 minutes before being washed in H20 for 5 minutes. Viral DNA was subsequently denatured in 0.5 M NaOH and 0.9M NaCl solution for 30 minutes. After being rinsed with H20 for 5 minutes, DNA
was renatured by two subsequent baths in 0.5 M tris HC1 pH 7.5 containing 3 M NaCl. DNA was subsequently transferred overnight in lOxSSC (1.5M NaCl, 0.15M Na Citrate pH 7.4) buffer onto a nylon membrane (Hybond N, Amersham Life Sciences, Cleveland, OH). The nylon membrane was air dried for one hour before being 5 submitted to UV cross-linking for 3 minutes. A 6 hours prehybridization was performed at 65°C in 4xSSC, 25~
- Denhardt's solution (v/v), 0.1% SDS (v/v), 0.1% Na pyrophosphate and denatured hering sperm DNA (500 ~,g/ml) solution.
10 2. Preparation of the probes specific for CAV2 PVIII and Fiber genes.
Since in most adenoviruses the E3 region is comprised between the two structural genes, PVIII and fiber, Applicant took advantage of a previously published 15 partial sequence of the CAV2 (Manhattan strain) genome (Linne, 1992) to design two specific primers pairs for each of these genes. Oligonucleotides LF189 (5'-TCAGTCATAGCCATCGACAGA-3') (SEQ ID NO: 26) and LF190 (5'-GTGCTGGCTGGCACGGGCATT-3') (SEQ ID N0: 27) were designed 20 to correspond to sequences within the 3' end of the CAV2 PVIII gene whereas oligonucleotides LF191 (5'-ATGTCCACCAAAGTCCCCTCT-3') (SEQ ID NO: 28) and LF192 (5'-CCCGGGGCGTCGTATGGATAT3') (SEQ ID NO: 29) were designed to correspond to sequences within the 5' end of the CAV2 25 fiber gene.
A 302 by DNA PVIII specific probe was generated by mixing 10 ng of purified CAV2 DNA with 5 ~C1 of lOX PCR
buffer, 3.75 ~1 of 2 mM dNTPs, 26 ~,1 H20, 0.25 ~C1 of Taq polymerase (5.0 u/~1), 5 ~,1 of 5 ~,M 5'end primer LF189 30 and 5 ul of 5 ~M 3'end primer LF190. A 30 cycle PCR
amplification was performed in a 0.5 ml tube containing

40 ~,1 of mineral oil using the following profile: 94°C 1 minute, 55°C 1 minute and 72°C 1 minute.
A 190 by DNA Fiber specific probe was generated by PCR by 35 swapping primer LF189 with primer LF191 and primer LF190 with primer LF192 in the previously described protocol.
Both PCR reactions were electrophoresed through a 1%

agarose gel and the corresponding PCR products were isolated using the Gene Clean procedure according to the manufacturer (Bio 101, Inc., La Jolla, CA) specifications. 100 ng aliquots of each probe was labelled by mixing with l~,g of random hexamers (Pharmacia, Piscataway, NJ) in a total volume of 13 ~C1 and subsequently boiled for 3 minutes before being incubated with 2.5 ~,1 of a dCTP, dTTP and dGTP mixture (each at a concentration of 0.5M), 2.3 ~,1 Klenow lOX
buffer, 1.5 ~1 Klenow enzyme (2u/~,1) and 5~1 of 32P-a-dATP (3000 Ci/mmol, 10 mCi/ml, NEN, Boston, MA) at RT for 4 hours. The reaction was stopped by adding 100 ~,1 of Stop solution (IBI Prime Time kit). 25 ~1 of each probe was heat denatured (100°C) for 3 minutes before being incubated overnight at 65°C with the previously described nylon membrane in a total volume of 50 ml of prehybridization solution. The nylon membrane was subsequently washed at 65°C in 6xSSC, O.lo SDS and 50 mM
Na Pyrophosphate solution for 2 hours. Viral DNA
restriction fragments complementary to the radiolabelled DNA probes were identified by autoradiography.
3. Identification and cloning of the restriction fragment containing the E3 region.
The HindIII fragment A (4.0 Kbp) was identified as the shortest well isolated restriction fragment recognized by both PVIII and Fiber probes, suggesting that it may contain the entire CAV2 E3 region. This fragment was isolated using Gene Clean procedure as previously described and subsequently subcloned into the HindIII site of the vector pBluescript SK+ (Stratagene, La Jolla, CA) generating plasmid pLF027.
4. Characterization of the CAV2 E3 region.
The CAV2 E3 region was analyzed by restriction digestion of pLF027 and by sequencing pLF027 according to Sequenase 2.0 kit instructions (US Biochemical, Cleveland, OH). Sequence analysis was performed using the MacVector software (Eastman Kodak, Rochester, NY). The pLF027 restriction map is shown in Figure 2. The corresponding sequence of the pLF027 including the CAV2 ' E3 region [defined as the DNA stretch between the PVIII
stop codon (#1,413 in pLF027) and the fiber ATG
initiation codon (#2,945 in pLF027)] is represented in Figure 1. Analysis of sequencing data revealed that the CAV2 E3 1,533 bps were 100 homologous with the previously identified CAV2 (Manhattan strain) E3 region (Linne, 1992). Analysis of the amino acid sequence deduced from the nucleotide sequence revealed that the rightward coding strand of the CAV2 E3 region encodes two potential polypeptides (ORF1 and ORF2) whereas the leftward coding strand encodes a single potential polypeptide (ORF3). The characteristics of these ORFs are presented in Table 2.
EXAMPLE 6 - Generation of donor plasmid pLF086.
1. Introduction of BalII and MluI restriction sites in the middle of the CAV2 E3 sequence.
In order to facilitate further manipulations, a 24 by DNA linker (5'-GATACGCGTTCCATTAGCAGATCT-3') (SEQ ID
NO: 30) containing unique BglII and MluI restriction sites were introduced between nucleotide #1487 and #1966 of the CAV2 E3 region (as described in Figure 1) by a double round PCR amplification procedure. Initial PCR
amplifications was performed using pLF027 DNA as template and using the following primer couples [LF327(5'-GGACACCTTTCTGATCAGTTCATT-3')/LF324(5'-GATACGCGTTCCATTA
GCAGATCTTTGAGGGGCCTGGAAATAGGC-3') (SEQ ID NO: 31, 32)]
and [LF326(5'-GGTTGTGTGGAAGACCCGGGGGCG-3')/LF325(5'-AGATCTGCTAATGGAA CGCGTATCGCTGCCCCCACAGTACAGCAA-3') (SEQ
ID NO: 33, 34)], to generate two partially overlapping DNA fragments of 838 by and 956 bp, respectively. The second round of PCR amplification was performed in the presence of both partially overlapping purified DNA
fragments and both external primers LF327 and LF326. The resultant 1,794 by DNA fragment was digested with PstI
and AatI and the resultant 890 by PstI/AatII fragment was purified and ligated with the 6,069 by PstI/AatII DNA
fragment of pLF027, generating pLF047A (Figures 3 and 4).
All PCR amplifications were performed using the conditions previously described. The 6,944 by MluI/B.q111 pLF047A was subsequently ligated with preannealed oligonucleotides LF328 (5'-GATCTGTTAACCCTAAGGCCATGGCATATGTCGCGAGGCCATCGTGGCCGCGGCCGC
A-3 ') (SEQ ID NO: 35) and LF329 (5'-CGCGTGCGGCCGCGGCCACGATGGCCTCGCGACATATGCCATGGCCTTAGGGTTAAC
A-3') to (SEQ ID NO: 36) generate pLF049A (Figures 5 and 6) .
This manipulation results in the exchanging of 60 by of the CAV2 E3 region with a 60 by BglII/MluI polylinker DNA
fragment. The size of the E3 region has not been modified and E3 ORF1 remained unaffected. However, sequences corresponding to E3 ORF2 have been disrupted and those of the E3 ORF3 were completely eliminated.
2. Generation of donor plasmid pLF086.
In order to delete part of the CAV2 E3 region a 428 by deletion was engineered 3' of the pLF049A MluI
site. A 537 by DNA fragment was generated by PCR as previously described using the pLF027 template and the primers pair LF361(5'-CTAGTCATCTTAACGCGTGTCCTCAACATCACCCGCGA-3')/LF334(5'-CTT
GCTTGTTATTAAAAAAAG-3') (SEQ ID NO: 37, 38). This 551 by fragment was subsequently digested with MluI and AatI
before being purified and ligated with the 6,284 by MIuI/AatII DNA fragment of pLF049A, generating pLF086 (Figures 7 and 8). This manipulation, which introduces a 27~ (428 bp) deletion of the E3 region, further expands the deletion of E3 ORF2 towards its 3'end but does not interfere with E3 ORF1 coding sequence.
EXAMPLE 7 - Cloning and characterization of the restriction fragment containing the right end of the viral g~enome 1. Cloning of the restriction fragment containing the right end of the viral genome.

Previously published restriction maps of the CAV2 (Glasgow strain) genome indicated the presence of a unique SalI restriction site located at 84.0 map units (Spibey and Cavanagh 1989). SalI digestion of CAV2 DNA
(30 ~,g) generated the predicted 3.2 kbp and 29 kbp DNA
fragments. The CAV2 DNA SalI B fragment (3.2 kbp) was ' gel purified using Gene Clean procedure as previously described and resuspended in 20 ~1 of H20.
Approximatively 3 ~g of purified SalI B fragment was denatured by the addition of 2 ~l of 1 N NaOH in a total volume of 22 ~cl for 90 minutes at RT to eliminate the known protein moiety (Robinson et al., 1973) which is covalently linked to the 5' termini of adenovirus genome.
The DNA was subsequently renatured by the addition of 1.3 ~C1 of 2M Tris HC1 pH 7.5 and incubated successively at 65°C for 1 hour and at RT for 1 hour before being ligated with the 2.919 by SalI/SmaI fragment of pBluescript SK+
to generate pLF056.
2. Characterization of the restriction fragment containing the right end of the viral genome.
The 3.2K by right end of the CAV2 genome was analyzed by restriction digestion of pLF056 and by sequencing of the same plasmid according to Sequenase 2.0 kit instructions. Sequence analysis was performed using the MacVector software. The pLF056 restriction map is shown in Figure 10, and Figure 9 shows the DNA sequence.
Sequencing data revealed that the CAV2 DNA SalI B
fragment is 3,274 by in length. Two unique restriction sites within the CAV2 genome have been localized within the CAV2 DNA SalI B fragment: BglII at position #587 and _SpeI at position #2,133. The 196 by ITR (Figure 9) nucleotide sequence of CAV2 situated at the right termini is 100$ homologous with the CAV2 right and left ITR
sequences previously published for the CAV2 Vaxitas and Glasgow strains, respectively (Cavanagh et al. 1991).
Analysis of the remainder of the CAV2 SalI-B fragment DNA
versus the DNA sequence of the previously mentioned CAV2 strains shows significant divergence with only 45 homology.
EXAMPLE 8 - Generation of pLF061 A NruI/EcoRV 312 by tag DNA fragment (Figure 5 il) was ligated with SmaI linearized pLF056 to generate pLF061 (Figure 11; restriction map shown in Fig. 12).
EXAMPLE 9 - Transfection of purified viral DNA into MDCR
cells Solution A was prepared by mixing 5 ~g of 10 purified CAV2 DNA with serum free MEM, supplemented as previously described, to a final volume of 300 ul.
Solution B was prepared by adding 40 ~1 of Lipofectamine reagent (Gibco) to 260 ul of supplemented but serum free MEM medium. Solutions A and B were mixed together and 15 incubated at RT for 30 minutes. The CAV2 DNA/liposome complexes were gently mixed with 2.4 ml of supplemented MEM medium (serum free) before being added to MDCK cell monolayer that was 75% confluent. After 24 hour incubation at 37°C in presence of 5% C02, the serum free 2o medium was removed and replaced by 3 ml of supplemented MEM medium containing 5% C02. The culture was incubated at 37°C in presence of 5% C02 for 8 days with 2 ml of supplemented MEM medium being added to it on the third day. No CPE could be evidenced during this incubation.
25 On day 8 the transfected MDCK cells were scraped off and harvested in a total volume of 5 ml. After 2 rounds of 2 minutes sonication on ice, 2 ml o.f the transfected culture were used to infect a 100% confluent MDCK
monolayer in a 150 mm diameter tissue culture dish for 1 3o hour at 37°C in presence of 5~ C02. The culture was subsequently overlaid with medium containing 0.6%
agarose. Plaques are appearing after 5 days at 37°C in the presence of 5% C02. Typically, a yield of at least 2,000 pfu/10 ~g of purified DNA is observed.
35 EXAMPLE 10 - Generation of recombinant CAV2 virus yCAl 1. In vitro generation of a recombinant CAV2 genome.
20 ~Cg of purified CAV2 DNA was digested with 30 U of SalI overnight at 37°C. The digested DNA was phenol chloroform extracted and ethanol precipitated before being resuspended in H20 to a concentration of 370 ng/~1.
5 ~g of SalI digested CAV2 DNA were in vitro ligated with 5 ~,g of the 3,557 by SalI/SacI pLF051 DNA fragment overnight at 15°C in the presence of 400 U of ligase (NEB, Beverly, MA) in a total volume of 50 ~,1.
2. Isolation of CAV2 recombinant virus vCAl.
The whole ligation reaction was subsequently used to transfect a 75% confluent MDCK monolayer as previously described. 4 ml of the harvested transfected culture were used to infect two 150 mm diameter tissue culture dishes. A total of 8 plaques became apparent after 10 days of incubation. All plaques were picked and resuspended in 1 ml of supplemented MEM medium before being sonicated for 2 x 2~ on ice. The clarified culture medium was serially diluted and used to infect 100%
confluent MDCK cells monolayer in 60 mm diameter tissue culture dishes. After 6 days of culture the agarose overlay was discarded and the infected monolayer was blotted onto nitrocellulose filters following the procedure described in Perkus et al. 1993. The filters were processed and subsequently hybridized with a labelled NruI/EcorV 312 by tag DNA fragment following classical procedures previously described.
Autoradiography experiments demonstrated that five out the initially detected 8 plaques contain recombinant CAV2 viruses. one well isolated plaque identified by plaque hybridization was picked and submitted to four additional rounds of plaque purification on MDCK cells.
Hybridization with the probe was confirmed after each round of purification. The plaque purified recombinant CAV2 virus was named vCAl.

3. Characterization of vCAl.
To further characterize vCA1 a small scale DNA
purification was performed. Briefly, purified vCA1 recombinant virus was used to infect a 100% confluent MDCK monolayer (206 cells). After 5 days, when CPE were completed, the infected culture was scraped and harvest.
The sonicated and clarified culture medium was treated with proteinase K (500 ~Cg/ml final concentration) for 2 hours at 42°C. The enzyme was inactivated by heating the reaction at 65°C for 20 minutes and the total DNA was subsequently phenol chloroform extracted and ethanol precipitated before being resuspended in H20. Purified total DNA was subsequently treated with RNase T1, phenol chloroform extracted and ethanol precipitated before being resuspended in H20 to a final concentration of 1.2 ~g/ml. 5 ~,g aliquots of purified vCA1 were independently digested with B_qlII and SpeI. Since those two sites are unique within the CAV2 genome a 29 kbp and 3 kbp fragments are expected from the BctlII digestion, whereas a 30.5 kbp and a 1.5 kbp fragments are expected from the _SpeI digestion. These restriction fragments are indeed observed demonstrating that vCA1 is a recombinant CAV2 virus which has incorporated 300 by of heterologous DNA
within the right end of its genome.
To further demonstrate that vCAl has indeed incorporated the expected tag DNA fragment, the VCA1 DNA
was analyzed by Southern blotting; and, this confirmed that vCAl indeed incorporated the tag DNA fragment.
To confirm that the CAV2 SmaI has been used as the insertion site, a 1.9 kbp DNA fragment was amplified from purified vCAi DNA with the couple of primers LF379 (5'-TCACGCCCTGGTCAGGGTGTT-3') (SEQ ID NO: 39) and LF407 (5'-GCCATCGCGTCAACCTGA-3') (SEQ ID NO: 40) using the conditions previously described. A partial sequence analysis of 1.940 by DNA fragment conducted using primers LF63 (5'-ATGATGTCTGGGGACATG-3') (SEQ ID NO: 41), LF379 (5'-TCACGCCCTGGTCAGGGTGTT-3') (SEQ ID NO: 42) and LF384 5'-ACCACGCGCCCACATTTT-3') (SEQ ID NO: 43) confirmed that the heterologous tag DNA was indeed inserted into the - CAV2 SmaI site to yield vCAl.
EXAMPLE ti - Generation of recombinant CAV2 virus yCA2 Ten ug of pLF086 were digested with HindLII and - the resulting 3.6 kbp DNA fragment was isolated using Gene Clean procedure as previously described and resuspended in H20 to a concentration of 100 ng/~,1. MDCK
l0 cells were transfected using the Lipofectamine based procedure previously described. Solution A was prepared by mixing 0.5 ~Cg of 3.600 by HindIII DNA fragment with 3 ~g of purified CAV2 DNA. Solution A total volume was brought to 3001 with supplemented serum free MEM medium.
Transfected cells were harvested after 8 days and plate out on 150 mm diameter tissue culture dishes as previously described. Plaques were lifted as previously described and hybridized with 5' end labelled oligonucleotide LF328. Five viral plaques crossreacting with the probe were picked and subsequently submitted to 4 rounds of plaque purification as previously described.
The plaque purified recombinant CAV2 virus was named vCA2. (Note that plaque purification is a use of the recombinant for replication of the DNA, or for replication of the virus, i.e., a vector use of the recombinant, thereby showing that there is no restriction or limit on the exogenous DNA).
2. Characterization of vCA2.
To characterize vCA2, a small scale DNA
purification was performed as previously described for vCAl. Purified vCA2 DNA and wild-type CAV2 DNA were independently digested by HindIII and the restricted DNAs were subsequently fractionated by electrophoresis through a 1% agarose gei. A 3.6 kbp HindIII fragment was visualized in the vCA2 sample whereas a 4.0 kbp fragment was present in the wild-type CAV2 sample, proving that the E3 region has been deleted of 428 by in vCA2 genome.

To further demonstrate that the expected tag (oligonucleotides LF328/LF329) has indeed been incorporated into the vCA2 E3 region, Southern blot was performed and this confirmed incorporation of the tag.
This result indicates that the complete CAV2 E3 ORF2 is not necessary in tissue culture. It also demonstrates that part of the CAV2 E3 ORF2 sequences can be exchanged with heterologous DNA and thus validates a second insertion site within the CAV2 genome. This results also proves that part of the CAV2 E3 region can be deleted to compensate for the introduction of foreign DNA into the SmaI site previously described in the derivation of vCAl.
EXAMPLE 12 - Generation of subfragment promoters, reduced or modified polyadenylation cassettes, promoters with 5' UTR replaced, and plasmids and recombinants containing same 1.1 Generation of pLF022, an expression vector in which the CAT reporter gene has been placed under the control of a subfragment (145 bp) of the HCMV-IE
promoter:
DNA from human cytomegalovirus (hCMV) (Towne strain) was prepared as described in Lafemina et al.
(1989). Amplification of the 3' end of the human cytomegalovirus immediate early promoter (hCMV-IE) was performed by PCR as previously described, using the primers pair LF172 (5'-ATCGTAAAGCTTAATGTCGTAATAACCCCGC-3')/LF159 (5'-TCTACTGCAGCCGGTGTCTTCTATGGAGGTCA-3') and hCMV DNA (long) as template. The resulting 166 by DNA
fragment was subsequently digested with PstI and HindIII
before being purified using Gene Clean procedure and directly ligated with the 4,348 by PstI/HindIII DNA
fragment of pCAT-Basic Vector (Promega, Madison, WI), generating pLF022 (Figures 13, 14, SEQ ID NO: 7). The regulatory sequences present in the pLF022 expression cassette are a 145 by fragment of the hCMV IE promoter and a 856 by cassette containing the SV40 small t antigen and polyadenylation signal.
- 1.2 Generation of pLF062, a derivative of pLF022 in which the SV40 polyadenylation cassette has 5 been reduced to 241 bp:
In order to reduce the size of the SV40 small t - antigen and polyadenylation signal cassette (856 bp) of pLF022, the following manipulations were performed. A
170 by DNA fragment was amplified by PCR using primers 10 LF377 (5'-TCTTCGCCCCCGTTTTCACCATGG-3') and LF378 (5'-ATCACGCCGCGGCTTAAAAAAATTACGCCCCGCCCT-3') and pLF022 DNA
(long) as template. The purified amplified fragment was resuspended in 18 ml H20 and incubated with lU of Klenow enzyme (Boehringer Mannheim, Indianapolis, IN) for 30 15 minutes at room temperature in the presence of 800 ~M
dNTPs. The modified DNA fragment was phenol-chloroform extracted and recovered by ethanol precipitation before being digested with NcoI. The resulting 136 by fragment was ligated with the 3,655 by NcoI/BsaBI DNA fragment of 20 pLF022, generating pLF062 (Figures 15, 16, SEQ ID NO: 8).
pLF062 contains two repeats of the consensus polyadenylation signal AATAAA downstream of the CAT gene.
The size of the CAT expression cassette in pLF062 is 1,119 by as compared to 1,804 by in pLF022. Regulatory 25 sequences in pLF062 expression cassette are a 145 by fragment of the hCMV-IE promoter and a 241 by cassette containing the SV40 polyadenylation signal.
1.3 Generation of pLF066, a derivative of pLFO62 in which the Ad2 TPL has been cloned downstream of 30 the HCMV-IE promoter:
In order to allow the expression of the reporter gene after the onset of CAV2 replication, pLF062 CAT expression cassette was modified by cloning the human Ad2 tripartite leader (Ad2 TPL) downstream of the hCMV-IE
35 promoter transcription start site.
Oligonucleotides SPH6ETr1 (5'-AATTCGGTACCAAGCTTCTTTATTCTATACTTAAAAAGTGAAAATAAATACAAAGGT

TCTTGACTCTCTTC-3', SPH6ETr2 (5'-CGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCG
CGGTCTTTCCAGT-3'), SPH6ETr3 (5'-ACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGTACTCCGCCACCGAGGGACCTGA
GCGAGTCCGCATC-3'), SPH6ETr4 (5'-GACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACC
AGTCACAGTCGCAAGCCCGGGT-3'), SPH6ETr5 (5'-CTTTGTATTTATTTTCAC
TTTTTAAGTATAGAATAAAGAAGCTTGGTACCG-3'), SPH6ETr6(5'GAAGAGT
TTGTCCTCAACCGCGAGCCCAACAGCTGGCCCTCGCAGACAGCGATGCGGAAGAGAG
TCAAGAAC-3'), SPH6ETr7 (5'-GCTCAGGTCCCTCGGTGGCGGAGTACGTTCGGAGGCCGACGGGTTTCCGATCCAAGA
GTACTGGAAAGACCGC-3'), and SPH6ETr8 (5'-CTAGACCCGGGCTTGCGACTGTGACTGGTTAGACGCCTTTCTCGAGAGGTTTTCCGA
TCCGGTCGATGCGGACTC-3,) were kinased and annealed and the 271 by product was gel purified.
The complete Ad2 TPL was subsequently amplified by PCR using primers LF394 (5'ATCGTCCTGCAGACTCTCTTCCGCATCGCTGTCTGC-3') and LF395 (5'-GCTCTAGACTTGCGACTGTGACTGGTTAG-3') and the gel purified annealed oligonucleotides as template.
The resulting 220 by DNA fragment was subsequently digested by PstI and XbaI before being purified using Gene Clean procedure as previously described and directly ligated with the 3,800 by PstI/XbaI pLF062 fragment, generating pLF066 (Figures 17, 18, SEQ ID NO: 9). Regulatory sequences in pLF066 expression cassette are a 145 by fragment of the hCMV-IE
promoter in which the 5'UTR has been replaced by the 202 by Ad2 TPL and a 241 by cassette containing the SV40 polyadenylation signal.
1.4 Generation of pLF069, a derivative of pLF066 in which the HCMV-IE 5'UTR has been replaced by the Ad2 TPL:
The HCMV-IE promoter 5'UTR (54 bp) present in pLF062 was deleted using the following procedure.
Annealed oligonucleotides LF397 (5'-CGTTTAGTGAACCGTCTGCA-3') and LF398 (5'-GACGGTTCACTAAACGAGCT-3') were ligated with the 3,936 by DNA fragment of pLF062, generating - pLF069 (Figure 19, 20, SEQ ID NO: 10). Regulatory sequences in pLF069 expression cassette are a 91 by fragment of the HCMV-IE promoter in which the 5'UTR has been replaced by the 202 by Ad2 TPL and a 241 by cassette - containing the SV40 polyadenylation signal.
1.5 Generation of pLF077, a derivative of pLF069 in which the SV40 polyadenylation cassette has been reduced to 153 bp:
A 160 by subfragment of SV40 polyadenylation sequences was amplified by PCR using oligonucleotides M13R (5'-GTAAAACGACGGCCAGT-3') and LF409 (5'-ATCGTCCCGCGGAATTGTTGTTGTTAACTTGTT-3') and pCAT Basic DNA
(long) as template. The resulting 145 by DNA fragment was subsequently digested by KspI and BamHI before being purified using Gene Clean procedure and directly ligated with the 3,716 by KspI/BamHI DNA fragment of pLF069, generating pLF077 (Figure 21, 22, SEQ ID NO: 11). The CAT expression cassette size in pLF077 is 1,161 by as compared to 1,804 by in pLF022 (36% reduction).
Regulatory sequences in pLF069 expression cassette are a 91 by fragment of the HCMV-IE promoter in which the 5'UTR
has been replaced by the 202 by Ad2 TPL and a 153 by cassette containing part of the SV40 polyadenylation signal.
1.6 Generation of pLFa9l, a derivative of pLF077 in which the 3' end of the polyadenylation signal has been modified:
The 12 by (5'-TTTTTGGGCGTT-3') which are localised upstream of SmaI site at the 5' end of the right ITR sequence in the CAV2 genome were introduced downstream of the pLF077 polyadenylation cassette using the following procedure. A 1,000 by DNA fragment was amplified by PCR using oligonucleotides LF423 (5'-ACGACCCGTAGAGGGCGTTGGACAGCAACTTGGCCTCGCGGTTGAGGACAAACTCTT
-3') and LF432 (5'-ATCGTCCCCGGGTTTTTGGGCGTTATCCAGACATGATAAGATACA-3') and pLF077 DNA (10 ng) as template. The 1,000 by PCR DNA
fragment was Gene Clean purified and modified by Klenow treatment before being digested by Ncol. The PCR
reaction was electrophoresed through a 1.2% agarose gel and the 295 by fragment was subsequently isolated using Gene Clean procedure.
pLF077 was digested by BamHI and subsequently modified by the action of Klenov enzyme before being digested by NcoI. The digestion reaction was electrophoresed through a 1% agarose gel and the 3,567 by restriction fragment was isolated using Gene Clean procedure, before being ligated with the aforementionned 295 by DNA fragment, resulting in pLF091 (Figures 23, 24, SEQ ID NO: 12).
1.7 Generation of pLF092, a CAT expression cassette donor plasmid:
The 1,180 by HindIII/SmaI DNA fragment of pLF091, which contains the entire CAT expression cassette, was modified by the action of Klenov enzyme and subsequently ligated with the 6.2 kbp SmaI linearized pLF056 to generate pLF092 (Figures 25, 26, SEQ ID NO:
13). This plasmid corresponds to a donor plasmid for the insertion of the CAT expression cassette into an insertion site 12 by upstream of the Smal site at the CAV2 genome 5' end.
1.8 Generation of pLF1.05, a donor plasmid for the insertion of foreign DNA 12 by upstream of the SmaI
site at the 5' end of the right ITR sequence in the CAV2 genome:
A polylinker [Nrul-AgeI-EcoRI-MluI-SalI-SmaI]
constituted of preanneled oligonucleotides LF446 (5'-GGGTTTTTGGGCGTTTCGCGAACCGGTGAATTCACGCGTGTCGACCCC-3') and LF447 (5'-CCCAAAAACCCGCAAAGCGCTTGGCCACTTAAGTGCGCACAGCTGGGG-3') was ligated with the 6.2 kbp SmaI linearized pLF056 to generate pLF105 (Figures 27, 28, SEQ ID NO: 14).

WO 98!00166 PCTIUS97111486 1.9 Generation of recombinant CAV2 virus vCA3, which contains a CAT expression cassette inserted into - the right terminal end of the CAV2 genome:
Ten(10) ~g of pLF092 were digested with HindIII
and BamHI and the resulting 4.3 kbp DNA fragment was isolated using Gene Clean procedure and resuspended in H20 to a concentration of 100 ng/~Cl. MDCK cells were transfected using the Lipofectamine based procedure.
Solution A was prepared by mixing 0.4 ~cg of 4.3 kbp HindIII/BamHI pLF092 fragment with 4.4 ~Cg of purified CAV2 DNA. Solution A total volume was brought to 300,1 with supplemented serum free MEM medium. Transfected cells were harvested after 8 days and plated out on 150 mm diameter tissue culture dishes as previously described. A probe specific for the CAT reporter gene was generated by PCR using pCAT Basic DNA (10 ng) as template and primers pair LF218 (5'-ATCGTACATATGGAGp,AAAAAATCACTGGATAT-3')/ LF231 (5'-ATCGTAGATATCCTCGAGTTACGCCCCGCCCTGCCACTC-3'). The resultant 660 by DNA fragment was labelled by random priming using a procedure previously described and subsequently hybridized with nitrocellulose membrane to lift viral plaques, as previously described. A plaque crossreacting with the probe was picked and subsequently submitted to 4 rounds of plaque purification, as previously described. The plaque purified recombinant CAV2 virus was named vCA3.
2. Characterization of vCA3.
2.1. Analysis of CAT gene expression by recombinant virus vCA3.
2.1.1. Detection of CAT enzymatique activity in vCA3 infected MDCK cells lysates.
Purified vCA3 recombinant virus and wild-type CAV2 were used to independently infect 100% confluent MDCK monolayer (106 cells) at a M.O.I. of 20. After 24 hours at 37°C in the presence of 5% C02, the infected cultures were scraped and harvested. Cells pellets were washed 3 times with prewarmed (37°C) PBS (Ca2+ and Mg2+
free) before being resuspended in lml of 40 mM Tris-HC1, pH 7.5, 1 mM EDTA, pH 8.0 and 150 mM NaCl and incubated for 5 minutes at room temperature. The cells were 5 subsequently centrifuged at 12 Kg for 30 seconds at 4°C
and the resulting pellet was resuspended in 100 ml of 0.25M Tris-HC1, pH 8.0 before being subjected to 3 rapid freeze/thaw cycles with vigorous vortexing after each thaw cycle. Endogenous deacetyl activity was inactivated 10 by incubating the lysates at 65°C for 10 minutes. The supernatants of a 12 Kg centrifugation for 2 minutes at RT were assayed in a chloramphenicol acetyltransferase (CAT) assay as follows. Twenty-five ml of cell lysate was incubated for 2 hours at 37°C with 3 ml of [14C]
15 chloramphenicol (0.005 mCi/ml)(NEN, Boston, MA), 5 ml of n-Butyryl Coenzyme A (5 mg/ml) and 92 ml of 0.25 M Tris-HC1, pH 8Ø The reaction was terminated by adding 500 ml of ethyl acetate (Sigma, St Louis, MO) per tube. The reaction was vortexed with the mixed xylenes for 30 20 seconds and subsequently centrifuged at 12 K g for 1 minute. The upper, organic phase was transferred to a fresh tube and evaporated to dryness. The residue was resuspended in 25 ml of n-Butyryl Coenzyme A (5 mg/ml) and 10 ml of the resuspended material was subsequently 25 dotted onto a silica gel thin layer chromatography (TCL) silica plate (Baker, Philisburg, NY). The slica plate chromatography was run in a closed chamber for approximately 1 hour, until the solvent was half-way up the plate. The silica plate was subsequently dried and 3o autoradiogramed. Butyrylated chloramphenicol was clearly detected in the vCA2 sample whereas no modified chloramphenicol could be evidenced in the control wild-type CAV2 sample. This result demonstrates that the recombinant virus vCA3 expresses a functional CAT
35 activity and thus validates both the expression cassette we have engineered and the insertion site we have selected.
2.1.2. Detection of CAT protein by radioimmnuprecipitation from vCA3 infected MDCK cells lysates.
Radioimmunoprecipitation analyses were - performed as previously described (Pincus et al., 1992) using (35SJ methionine (1000 Ci/mmol, NEN)-labelled lysates derived from vCA3-infected MDCK cells and CAT
rabbit polyclonal serum (5'3'Inc, Boulder, CO). The immunoprecipitated CAT polypeptide was resolved by SDS-PAGE and visualized by fluorography using sodium salicylate.
Analysis of vCA3 genomic organisation by restriction enzyme activity:
vCA3 DNA was purified as previously described.
Purified total DNA was subsequently resuspended in H20 to a final concentration of 1.3 ~g/ml. 2 ~g aliquots of purified vCA3 were independently digested with BcrlII and SalI. Since those two sites are unique within the CAV2 genome a 28.2 kbp and 3.8 kbp fragments are expected from the BglII digestion, whereas a 27.8 kbp and a 4.2 kbp fragments are expected from the SalI digestion.
These restriction fragments are indeed observed demonstrating that vCA3 is a recombinant CAV2 virus which has incorporated 1,000 by of the CAT expression cassette within the right end of its genome.
EXAMPLE 13 - Generation of donor ~lasmid pLF102 In order to delete the 3' end of the E3 ORF2 without modifying the E3 ORF1, the following procedure was developed.
A PCR amplification was set up using pLF027 DNA as a template and the primers pair LF437 _ (5'ATCTTAACGCGTCCCTCAGCCTTCTAATGGGAC 3') and LF334 (5' CTTGCTTGTTATTfi,AAAAAAG 3') as previously described. The 329 by amplified DNA fragment was purified using the previously described Gene Clean procedure before being digested by MluI and SmaI. The resultant 287 by MluI/Smal DNA fragment was gel purified before being ligated with the 6,079 by MluI/SmaI DNA fragment of pLF086, generating pLF095. The pLF095 63 by BalII/Mlul linker was subsequently swapped with a 305 by BQ1II/MluI
linker of unrelated foreign DNA using the following procedure. A 305 by DNA fragment [nucleotide sequence described in Figures 29 and 30, see below] was obtained by digesting an unrelated plasmid with MluI and BalII.
The MluI and BalII digested DNA fragment was gel purified and subsequently ligated with the 6,315 by MluI/BqlII
DNA fragment of pLF095, generating pLF102 (Figure 29, SEQ
ID NO: 15).
The engineering of pLF102 results in the exchange of a 688 by fragment of CAV2 E3 (which represents 45% of the total E3 size) with 305 by of foreign DNA and is useful to further define the limits of non-essential subdomains within CAV2 E3 region.
EXAMPLE 14 - Generation of donor plasmid pLF116A
In order to delete a pLF027 EcoRV/AatII 1.8 kbp DNA fragment which contains two SphI restriction sites [at positions #3,770 and #3,870], the pLF027 EcoRV/AatII
5,163 by fragment was gel purified and subsequently treated with Klenow enzyme before being religated on itself to generate pLF094.
A 24 by DNA linker (5'-GATACGCGTTCCATTAGCAGATCT-3') containing unique BalII and MluI restriction sites was introduced into the pLF094 intergenic sequence between E3 ORF1 and E3 ORF2 by a double round PCR amplification procedure. Initial PCR
amplifications were performed using pLF027 DNA as template and the following primer couples (LF243(5' CGCGCACAAACTGGTAGGTGC 3')/LF436(5' AGATCTGCTAATGGAACGCGTATCAAGTTTAATAATATTATC 3')] and [LF435(5' GATACGCGTTCCATTAGCAGATCTGTTTTACAGCTACCA
3')/LF277(5' GTACAGTTATGTTGAAGG 3')], to generate two partially overlapping DNA fragments of 487 by and 698 bp, respectively. The second round of PCR amplification was performed in the presence of both partially overlapping - purified DNA fragments and both external primers LF243 and LF277. The amplified 1,185 by DNA fragment was digested with SphI and PstI and the resultant 566 by PstI/SphI fragment was purified and ligated with the - 4,653 by SphI/PstI partial digest of pLF094, generating pLF093. All PCR amplifications were performed using the conditions previously described.
A deletion of the 5' end of E3 ORF2 without modifying E3 ORF1 was engineered by the following procedure. The pLF093 XhoI/MluI 1,062 by fragment was gel purified and subsequently ligated with the 5,081 by XhoI/MluI fragment of pLF086, generating pLF115. MluI
linearized pLF115 DNA was subsequently ligated with a 311 by MluI/MluI fragment of unrelated foreign DNA, generating pLF116A and B. The complete DNA sequence of pLF116A including the sequence of the unrelated 31I by MluI/MluI fragment of foreign DNA is presented in Figure 31 (SEQ ID NO: 16), with the restriction map shown in Figure 32.
The engineering of pLF116A results in the exchange of a 876 by fragment of CAV2 E3 (which represents 57% of the total E3 size) with 311 by of foreign DNA and is useful to further define the limits of non-essential subdomains within CAV2 E3 region.
EXAMPLE 14 - Generation of donor plasmid pLF100 In order to delete simultaneously the 5' end of the E3 ORF2, the 3' end of the E3 ORF1 and the complete E3 ORF3, a 634 by fragment was deleted between the MluI(#1529) and DraIII(#889) restriction sites of pLF086 (Figures 7 and 8) and subsequently exchanged with a 302 by fragment of unrelated foreign DNA using the following procedure.
The 302 by DNA fragment was obtained by digesting an unrelated plasmid with MluI and DraIII. The MluI and DraIII digested DNA fragment was gel purified WO 98/00166 PCTlUS97/11486 and subsequently ligated with the 5,946 by MluI/DraIII
DNA fragment of pLF086, generating pLF100 (Figures 33, 34 SEQ ID NO: 17). The nucleotide sequence of the 302 by fragment is shown in Figure 33, and the restriction map is shown in Figure 34.
The engineering of pLF100 results in the exchange of a 1,060 by fragment of CAV2 E3 (which represents 690 of the total E3 size) with 302 by of foreign DNA and is useful to further define the limits of non-essential subdomains within CAV2 E3 region.
EXAMPLE Z5 - Generation of donor plasmid pLF120 In order to delete simultaneously the 3' end of the E3 ORF1, the almost complete E3 ORF2 and the complete E3 ORF3, a 882 by fragment was deleted between the MluI(#/,771) and DraIII(#889) restriction sites of pLF102 and subsequently exchanged with a 311 by fragment of unrelated foreign DNA using the following procedure.
pLF102 DNA was linearized by MluI and subsequently partially digested with DraIII. The resultant 5,733 by MluI/DraIII was subsequently ligated with a 311 by MluI/DraIII fragment of unrelated foreign DNA, generating pLF120 (Figures 35, 36, SEQ ID N0: 18).
The nucleotide sequence of the 311 by MluI/DraIII
fragment of unrelated foreign DNA is shown in Figure 35, and the restriction map is shown in Figure 36.
The engineering of pLFI20 results in the exchange of a 1,261 by fragment of CAV2 E3 (which represents 820 of the total E3 size) with 311 by of foreign DNA and is useful to further define the limits of non-essential subdomains within CAV2 E3 region. This is the largest deletion and indicates that practically all of the E3 region, e.g., about 80% to about 100%, such as up to about 80 to about 95% or up to about 80% to 90% or up to about 80o to 85% of the E3 region can be deleted.
EXAMPLE 16 - Generation of pLF043, a pB88K+ which contains the canine distemper virus (CDV) hemaaalutinin fliA) codina sequence 1. Generation of plasmid pSDCDVHA.
' The Onderstepoort strain of canine distemper virus (CDV) was obtained from Dr. M. Appel (Cornell 5 University, Ithaca, NY). RNA was harvested from CDV
infected Vero cells and cDNA was prepared in the - following manner.
RNA from CDV infected Vero cells was isolated by the guanidium isothiocyanate-cesium chloride method of 10 Chirgwin, et al., (1979). First strand cDNA was synthesized with AMV reverse transcriptase (Life Sciences, St. Petersburg, FL), the oligonucleotide primer CDVFSP (SEQ ID NO: 44) (5'-CCAGGACATAGCAAGCCAACAGGTC-3'), and RNA from CDV infected cells. CDVFSP (SEQ ID NO: 44) 15 primes 80 by upstream of the CDV fusion (F) start codon, yielding a positive sense single stranded cDNA product which contains the F and hemagglutinin (HA) coding sequences.
The HA-specific open reading frame (ORF) was 20 amplified from the first strand cDNA product by polymerase chain reaction (PCR) as previously described.
Oligonucleotide primers CDVHAI (SEQ ID NO: 45) (5'-CGATATCCGTTAAGTTTGTATCGTAATGCTCCCCTACCAAGAC-3') and CDVHA2 (SEQ ID NO: 46) (5'-25 GGGATAAAAATTAACGGTTACATGAGAATCTTATACGGAC-3'} were used in a PCR with the CDVFSP derived first strand cDNA as template. CDVHA1 contains the 3' most region of the vaccinia virus H6 promoter (Perkus, et al., 1989) followed by a sequence which primes from the translation 30 initiation codon into the CDV HA ORF. CDVHA2 (SEQ ID NO:
46) primes from the stop codon of the HA ORF toward the CDV HA 5' end. The resultant 1.8 kbp PCR product was treated with the Klenow fragment from the E. coli DNA
polymerase, in the presence of 20mM dNTPs, to blunt end 35 the fragment. The 1.8 kbp blunt-ended fragment was inserted between the NruI site within the H6 promoter, and the SmaI site 3' of the H6 promoter in pSD554 (see below). The resultant plasmid pCDVHA should have contained the H6 promoted CDV HA ORF, but there was an unexpected deletion at the CDV HA 5' end. Repair of the deletion is described below.
Plasmid pSD554 contains the vaccinia K1L host range gene (Gillard et al., 1986) and vaccinia H6 promoter followed by insertion sites, within flanking vaccinia arms. The flanking vaccinia arms replace the ATI region: open reading frames A25L and A26L (Goebel et al., 1990a,b). pSD554 was prepared in the following manner.
Left and right vaccinia flanking arms were constructed by PCR using the template pSD414 which contains vaccinia SalI B (Goebel et al., 1990a,b). The left arm was synthesized using oligonucleotide primers MPSYN267 (SEQ ID NO: 47) (5'-GGGCTGAAGCTTGCTGGCCGCTCATTAGACAAGCGAATGAGGGAC-3') and MPSYN268 (SEQ ID NO: 48) (5'-AGATCTCCCGGGCTCGAGTAATTAATTAATTTTTATTACACCAGAAAAGACGGCTTG
AGA T C-3') in a PCR with template pSD414. The right arm was synthesized using oligonucleotide primers MPSYN269 (SEQ ID NO: 49) (5'-TAATTACTCGAGCCCGGGAGATCTAATTTAATTTAATTTATATAACTCATTTTTTGA
ATA T ACT-3') and MPSYN270 (SEQ ID NO: 50) (5'-TATCTCGAATTCCCGCGGCTTTAAATGGACGGAACTCTTTTCCCC-3') in a PCR with template pSD414. The two PCR-derived fragments containing the left and right arms were combined in a PCR. The resultant PCR product was digested with EcoRI
and HindIII and a 0.9kbp fragment was isolated. The 0.9kb fragment was inserted between the pUC8 EcoRI and HindIII sites. The resultant plasmid pSD541 received the K1L gene, and additional insertion sites, in the following manner.
Plasmid pSD541 was digested with BalII and XhoI
and ligated with annealed complementary oligonucleotides MPSYN333 (SEQ ID NO: 51) (5'-GATCTTTTGTTAACAAAAACTAATCAGCTATCGCGAATCGATTCCCGGGGGATCCGG

TALC C-3') and MPSYN334 (SEQ ID NO: 52) (5'-TCGAGGGTACCGGATCCCCCGGGAATCGATTCGCGATAGCTGATTAGTTTTTGTTAA
- CAA A A-3'), generating plasmid pSD552. pSD452 (Perkus et al., 1990) contains the K1L gene. pSD452 was digested with F_ipal and partially digested with BcxlII and the resultant lkbp fragment containing the K1L gene was - inserted between the pSD552 BalII and HpaI sites. The resultant plasmid pSD553 was digested with NruI and a SmaI/NruI fragment containing the vaccinia H6 promoter (Perkus et al., 1989) was inserted. The resultant plasmid, pMP553H6, contains the vaccinia H6 promoter downstream from the K1L gene within the A26L insertion locus.
Plasmid pMP553H6 was digested with NruI and BamHI and ligated with annealed synthetic oligonucleotides MPSYN347 (SEQ ID NO: 53) (5'-CGATATCCGTTAAGTTTGTATCGTAATCTGCAGCCCGGGGGGG-3') and MPSYN348 (SEQ ID NO: 54) (5'-GATCCCCCGGGCTGCAGATTACGATACAAACTTAACGGATATCG-3'). The resultant plasmid pSD554 contains the K1L gene and the H6 promoter followed by insertion sites, within flanking vaccinia sequences which replace the ATI region.
The vaccinia virus H6 promoter and 5' end of the CDV HA ORF were added to pCDVHA as a PCR derived fragment. The ATG of the regulatory region H6 overlaps the CDV HA translation initiation codon in the PCR
derived fragment. The vaccinia virus H6 promoter has been described in Perkus, et al., 1989.
pEIVC5L contains the modified H6 promoter and a nonpertinent gene. pEIVCSL was used in a polymerase - 30 chain reaction with oligonucleotide primers H65PH (SEQ ID
NO: 55) (5'-ATCATCAAGCTTGATTCTTTATTCTATAC-3') and CDVHAH6 (SEQ ID NO: 56) (5'-GTCTTGGTAGGGGAGCATTACGATACAAACTTAACG-3') to generate a 156bp fragment. CDVHAH6 contains the - 5' 18 base pairs of CDV HA followed by a sequence which primes from the translation initiation codon toward the H6 promoter 5' end. H65PH (SEQ ID NO: 55) contains a HindIII site followed by a sequence which primes from the H6 promoter 5' end toward the 3' end. The 156 base pair PCR-derived H65PH/CDVHAH6 (SEQ ID NO: 55/SEQ ID NO: 56) product contains the H6 promoter and the 5' 18 base pairs of the CDV HA coding sequence.
The CDVFSP (SEQ ID NO: 44) first strand cDNA
product was used in a PCR with oligonucleotide primers CDVHAATG (SEQ ID NO: 57) (5'-ATGCTCCCCTACCAAGAC-3') and CDVHAECO (SEQ ID NO: 58) (5'-GTAATTAGTAAAATTCACCTTG-3') to generate a 459 base pair fragment. CDVHAATG (SEQ ID
NO: 57) primes from the translation initiation codon toward the CDV HA 3' end. CDVHAECO (SEQ ID NO: 58) primes from position 583 of the following H6 promoted CDV
HA sequence toward the CDV HA 5' end. The 156 base pair and 459 base pair PCR-derived fragments were pooled and used in a PCR with H65PH (SEQ ID NO: 55) and CDVHAECO
(SEQ ID NO: 58) to generate a 597 base pair fragment.
The PCR-derived product was digested with HindIII and EcoRI, generating a 520 base pair fragment which contains the H6 promoter and 5' most 387 base pairs of the CDV HA
coding sequence. The 520 base pair HindIII/EcoRI
digested PCR fragment was inserted between the HindIII
and EcoRI sites of pBSSK+ , yielding pBSCDVHASS. Plasmid pBSCDVHASS contains the H6 promoted 5' end of the CDV HA
ORF in pBSSK+, and the 3' end of the CDV HA ORF was added in the following manner.
Plasmid pCDVHA was digested with SmaI followed by partial digestion with EcoRI to generate a l.4kbp fragment containing the 3' end of the CDV HA ORF. The l.4kbp pCDVHA EcoRI/SmaI fragment was inserted between the EcoRI and SmaI sites of pBSCDVHASS. The resultant plasmid pBSCDVHA was digested with BamHI and partially digested with XhoI to generate a l.9kbp fragment containing the H6 promoted CDV HA open reading frame.
The l.9kbp BamHI/XhoI pBSCDVHA fragment was inserted between the BamHI and XhoI sites of pSD553 (see above).
The resultant plasmid pSDCDVHA contains the H6 promoted CDV HA gene in the ATI insertion site.

2. Generation of pLF043.
The pSDCDVHA 1,975 by HindIII/BamHI which - contains the CDV HA coding sequence and the 3' most region of the vaccinia virus H6 promoter, was gel purified and subsequently inserted between the corresponding restrictions sites of pBSSK+, generating pLF043 (Figures 37 and 38) (SEQ ID NO: 19).
EXAMPLE 17 - Generation of pLF098, which contains a complete CDV HA expression cassette A XbaI restriction site was engineered immediately upstream of the CDV HA initiation codon (ATG) in the following manner. A 409 by DNA fragment was amplified by PCR using pLF043 DNA as a template and the primers pair LF412 (5' CTGATCTCTAGAATGCTCCCCTACCAAGACAAG
3') (SEQ ID NO: 59) and LF413 (5' TGGAGATCGCGGAAGTCG 3') (SEQ ID NO: 60) as previously described. The PCR
amplified fragment was isolated using the Gene Clean procedure as previously described before being treated with the Klenow fragment from the E. coli DNA polymerase, in the presence of 20mM dNTPs and digested by St~eI and EcoRI. The resultant blunt-ended/SpeI 192 by DNA fragment was subsequently ligated with the 4,891 by NruI/Spel fragment of pLF043, generating pLF096.
A KspI restriction site was engineered immediately downstream of pLF096 CDV HA stop codon (TAA) in the following manner. A 204 by DNA fragment was amplified by PCR using pLF043 as.a template and the primers pair LF438 (5' TGTTTATGACCCAATCG 3') (SEQ ID NO:
61) and LF439 (5' ATGCTCCCGCGGTTAACGGTTACATGAGAATCT 3') (SEQ ID NO: 62) as previously described. The PCR
amplified fragment was isolated using the Gene Clean procedure as previously described before being digested with KsbI and AccI. The resultant 143 by DNA fragment was gel purified and subsequently ligated with the 4,594 by KS,~7I/AccI fragment of pLF096, generating- pLF097.
The 1,821 by pLF097 KSpI/XbaI fragment which contains the CDV HA coding sequence was subsequently WO 98!00166 PCTIUS97111486 ligated with the 3,246 by KSpI/XbaI fragment of pLF069, generating pLF098 (Figures 39, 40) (SEQ ID NO: 20).
EXAMPLE 18 - Generation of pLF099A, a donor plasmid for the insertion of CDV IiA expression 5 cassette 12 by upstream of the SmaI site at the CAV2 q~enome riqht end The 2,372 by BamHI/HindIII pLF098 fragment which contains the CDV HA coding sequence coupled to the regulatory sequences defined in pLF069 was treated with 10 the Klenow fragment from the E. coli DNA polymerase before being ligated with the 6,243 by NruI linearized pLF105, generating pLF099A and pLF099B. pLF099A
corresponds to the rightward orientation of the expression cassette (Figures 41, 42) (SEQ ID NO: 21).
15 EXAMPLE I9 - Generation and characterization of recombinant CAV2 virus vCA4 1. Generation of recombinant CAV2 virus vCA4 Ten ~,g of pLF102 were digested with HindIII and the resulting 3,652 by DNA fragment was isolated using 20 Gene Clean procedure as previously described and resuspended in H20 to a concentration of 100 ng/~,1. MDCK
cells were transfected using the Lipofectamine based procedure previously described. Solution A was prepared by mixing 0.5 ~,g of 3.6 kbp HindIII DNA fragment with 3 25 ~,g of purified vCA2 DNA. Solution A total volume was brought to 300,1 with supplemented serum free MEM medium.
Transfected cells were harvested after 8 days and plate out on 150 mm diameter tissue culture dishes as previously described. A probe specific for the 305 by 30 fragment of foreign DNA inserted into pLF102 was generated by PCR using pLF102 DNA (10 ng) as template and primers pair LF440 (5'-ATCAGTACGCGTATGGGCCACACACGGAGG-3')/ LF441 (5'-ATCAGTAGATCTGTTATTAGTGATATCAAA-3'). The resultant 305 by DNA fragment was labelled by random 35 priming using a procedure previously described and subsequently hybridized with nitrocellulose membrane used to lift viral plaques as previously described. Five viral plaques crossreacting with the probe were picked and subsequently submitted to 4 rounds of plaque - purification as previously described. The plaque purified recombinant CAV2 virus was named vCA4.
2. Characterization of vCA4 vCA4 DNA was purified as previously described.
- Purified total DNA was subsequently resuspended in H20 to a final concentration of 1.9 ~Cg/ml. 2 ~g aliquots of purified vCA4 were digested with HindIII. The expected 3, 667 by HindIII fragment was visualized in the vCA4 sample whereas a 4.0 kbp fragment was present in the wild-type CAV2 sample, proving that vCA4 genomic DNA
contains the partially deleted E3 region described in pLF102. VCA4 DNA was analyzed by Southern Blot which indicated that vCA4 has an E3 region 371 by shorter than the wild-type E3 region.
This result futher demonstrates non-essential subdomains of CAV2 E3 region. More specifically, the derivation of vCA4 demonstrates that the CAV2 E3 sequences comprised between position #1,470 and position #2,157 [ie 45% of the E3 region], as in pLF027 (see Figure 1, SEQ ID NO: 1) can be exchanged with heterologous DNA. It also further validates the CAV2 E3 as an insertion site within the CAV2 genome. This results also proves that part of the CAV2 E3 region can be deleted to compensate for the introduction of foreign DNA into the right end of CAV2 genome as previously described in the derivation of vCA3.
EXAMPLE 20 - Generation and characterization of recombinant CAV2 virus vCA5 1. Generation of recombinant CAV2 virus vCAS
Ten ~,g of pLF116A were digested with HindIII
and the resulting 3,487 by DNA fragment was isolated using Gene Clean procedure as previously described and resuspended in H20 to a concentration of 100 ng/~,1. MDCK
cells were transfected using the Lipofectamine based procedure previously described. Solution A was prepared by mixing 0.5 ~Cg of 3.5 kbp HindIII DNA fragment with 3 ~,g of purified vCA2 DNA. Solution A total volume was brought to 3001 with supplemented serum free MEM medium.
Transfected cells were harvested after 8 days and plate out on 150 mm diameter tissue culture dishes as previously described. A probe specific for the 311 by fragment of foreign DNA inserted into pLF216A was generated by PCR using pLF116A DNA (10 ng) as template and primers pair LF453(5'-ATCGTCATTGCCACGCGTATGGCAGAAGGATTTGCAGCCAAT-3')/ LF454 (5'-ATCGTCATTGCCACGCGTAACCAGGGACAATACTTGTTCATC-3'). The resultant 311 by DNA fragment was labelled by random priming using a procedure previously described and subsequently hybridized with nitrocellulose membrane used to lift viral plaques as previously described. Five viral plaques crossreacting with the probe were picked and subsequently submitted to 4 rounds of plaque purification as previously described. The plaque purified recombinant CAV2 virus was named vCAS.
2. Characterization of vCA5 vCA5 DNA is purified as previously described.
Purified total DNA is subsequently resuspended in H20 to a final concentration of 1.9 ~,g/ml. 2 ,ug aliquots of purified vCA5 are digested with HindIII. The expected 3,487 by HindIII fragment is visualized in the vCAS
sample whereas a 4.0 kbp fragment is present in the wild-type CAV2 sample, proving that vCAS genomic DNA contains the partially deleted E3 region described in pLF116A.
This result futher demonstrates non-essential subdomains of CAV2 E3 region. More specifically, the derivation of vCA5 demonstrates that the CAV2 E3 sequences comprised between position #1,088 and position #1,964 [ie 57% of the E3 region], as described in pLF027 (see figure 1, SEQ ID NO: I) can be exchanged with heterologous DNA. It also further validates the CAV2 E3 as an insertion site within the CAV2 genome. This result also proves that part of the CAV2 E3 region can be deleted to compensate for the introduction of foreign DNA
into the right end of CAV2 genome as previously described - in the derivation of vCA3.
ERAMPLE 21 - Generation and characterization of recombinant CAV2 virus vCA6 Z. Generation of recombinant CAV2 virus vCA6 - Ten ~g of pLF100 were digested with HindIII and the resulting 3,284 by DNA fragment was isolated using Gene Clean procedure as previously described and resuspended in H20 to a concentration of 100 ng/~,1. MDCK
cells were transfected using the Lipofectamine based procedure previously described. Solution A was prepared by mixing 0.5 ~g of 3.3 kbp HindIII DNA fragment with 3 ~Cg of purified vCA2 DNA. Solution A total volume was brought to 3001 with supplemented serum free MEM medium.
Transfected cells were harvested after 8 days and plate out on 150 mm diameter tissue culture dishes as previously described. A probe specific for the 311 by fragment of foreign DNA inserted into pLF100 was generated by PCR using pLF100 DNA (10 ng) as template and primers pair LF442(5~-ATCAGTCACGGTGTGTAAATGGGCCACACACGGAGG-3~)/ LF443 (5~-ATCAGTACGCGTGTTATTAGTGATATCAAA-3~). The resultant 302 by DNA fragment was labelled by random priming using a procedure previously described and subsequently hybridized with nitrocellulose membrane used to lift viral plaques as previously described. Five viral plaques crossreacting with the probe were picked and subsequently submitted to 4 rounds of plaque purification as previously described. The plaque purified recombinant CAV2 virus was named vCA6.
2. Characterization of vCA6 vCA6 DNA is purified as previously described.
- Purified total DNA is subsequently resuspended in H20 to a final concentration of 1.9 ~.g/ml. 2 ~,g aliquots of - purified vCA6 are digested with HindIII. The expected 3,284 by HindIII fragment was visualized in the vCA6 sample whereas a 4.0 kbp fragment is present in the wild-type CAV2 sample, proving that vCA6 genomic DNA contains the partially deleted E3 region described in pLF100.
This result futher demonstrates non-essential subdomains of CAV2 E3 region. More specifically, the derivation of vCA6 demonstrates that the CAV2 E3 sequences comprised between position #898 and position #1,949 [ie 69~ of the E3 region], as described in pLF027 (see Figure 1, SEQ ID NO: 1) can be exchanged with heterologous DNA. It also further validates the CAV2 E3 as an insertion site within the CAV2 genome. This results also proves that part of the CAV2 E3 region can be deleted to compensate for the introduction of foreign DNA into the right end of CAV2 genome as previously described in the derivation of vCA3.
EXAMPLE 22 - Generation and characterization of recombinant CAV2 virus vCA7 1. Generation of recombinant CAV2 virus vCA7 Ten ~Cg of pLF120 were digested with HindIII and the resulting 3,085 by DNA fragment was isolated using Gene Clean procedure as previously described and resuspended in H20 to a concentration of 100 ng/~C1. MDCK
Cells were transfected using the Lipofectamine based procedure previously described. Solution A was prepared by mixing 0.5 ~,g of 3.3 kbp HindIII DNA fragment with 3 ~g of purified vCA2 DNA. Solution A total volume was brought to 300~c1 with supplemented serum free MEM medium.
Transfected cells were harvested after 8 days and plate out on 150 mm diameter tissue culture dishes as previously described. A probe specific for the 311 by fragment of foreign DNA inserted into pLF100 was generated by PCR using pLF100 DNA (10 ng) as template and primers pair LF458(5'-ATCCGTACGCGTTAGAGGGCAAAGCCCGTGCAGCAGCGC-3')/ LF459 (5'-ATCCGTCACGGTGTGTAGATGGGTTGTTTTGTGGAGAAT-3'). The resultant 311 by DNA fragment was labelled by random priming using a procedure previously described and WO 98/001b6 PCT/US97/11486 subsequently hybridized with nitrocellulose membrane used to lift viral plaques as previously described. Cross reacivity between the probe and viral DNA has been evidenced.
5 This result indicates that a deletion of 1,259 by between position #898 and position #2,157, as - described in pLF027 (see Figure 1, SEQ ID NO: 1) is compatible with viral replication in tissue culture, further showing that essentially all of the E3 region can 10 be deleted.
EXAMPLE 23 - Generation of vCA8 Ten ~,g of pLF099A were digested with BalII and NotI and the resulting 5,131 by DNA fragment was isolated using Gene Clean procedure as previously described and 15 resuspended in H20 to a concentration of 100 ng/~,1. MDCK
cells were transfected using the Lipofectamine based procedure previously described. Solution A was prepared by mixing 0.5 ~g of 5.1 kbp BqlII/NotI DNA fragment with 3 ~,g of purified vCA2 DNA. Solution A total volume was 20 brought to 300u1 with supplemented serum free MEM medium.
Transfected cells were harvested after 8 days and plate out on 150 mm diameter tissue culture dishes as previously described. The 440 by EcoRI fragment of pSDCDVHA was labelled by random priming using a procedure 25 previously described and subsequently hybridized with nitrocellulose membrane used to lift viral plaques as previously described. Two viral.plaques cross-reacting with the probe were picked and are currently submitted to a plaque purification process as previously described.
30 The plaque purified recombinant CAV2 virus is named vCA8.
2. Characterization of vCA8 vCA8 DNA purification, restriction digestion, Southern Blot, and CDV HA expression analysis by radioimmunoprecipitation confirm insertion and 35 expression.
EXAMPLE 24 - Generation of pLF108, a pBSSK+ derived plasmid which contains the canine WO 98!00166 PCTIUS97111486 8istemper virus (CDV) fusion (F1) coding sequence 1. Generation of pATICDVF1 The CDV fusion (F) specific open reading frame (ORF) was amplified from cDNA by PCR using oligonucleotide primers CDVATGF1 (SEQ ID NO: 63) (5'-CATAAATTATTTCATTATCGCGATATCCGTTAAGTTTGTATCGTAATGCACAAGGGA
ATCCCCAA AAGC-3') and CDVFT (SEQ ID NO: 64) (5'-ATCATCGGATCCATAAAAATCAGTGTGATCTCACATAGGATTTCGAAG-3') with CDVFSP (SEQ ID NO: 44) derived first strand cDNA as the template. CDVATGF1 (SEQ ID NO: 63) contains the 3' most region of the vaccinia virus H6 promoter (Perkus, et al., 1989) followed by a sequence which primes from the CDV F
translation initiation codon into the CDV F ORF. CDVFT
(SEQ ID NO: 64) contains a BamHI site followed by a sequence which primes from the CDV F stop codon toward the CDV F 5' end. The resultant PCR product was digested with NruI and BamHI, yielding a 2 kbp fragment which was inserted into pSD554 between the NruI and BamHI sites.
The resultant plasmid pATICDVF1 contains the H6 promoted CDV F ORF in the vaccinia virus ATI insertion locus.
2. Generation of HC5LSP28 The C5 vector plasmid HC5LSP28 was constructed to remove the C5 ORF in the following manner.
Oligonucleotide primers C5A (SEQ ID NO: 65) (5'-ATCATCGAATTCTGAATGTTAAATGTTATACTTTG-3') and C5B (SEQ ID
NO: 66) (5'-GGGGGTACCTTTGAGAGTACCACTTCAG-3') were used in a PCR with genomic canarypox DNA as the template. The resultant 1.5 kbp fragment was digested at the C5A end with EcoRI and the other end remained blunt for insertion between the EcoRI and SmaI sites of pUC8, yielding plasmid C5LAB. Oligonucleotide primers C5C (SEQ ID NO:
67) (5'-GGGTCTAGAGCGGCCGCTTATAAAGATCTAAAATGCATAATTTC-3') and CSDA (SEQ ID NO: 68) (5'-ATCATCCTGCAGGTATTCTAAACTAGGAATAGATG-3') were used in a PCR with genomic canarypox DNA as template. The resultant 400 base pair fragment was digested at the CSDA

end with PstI and the other end remained blunt for insertion between the SmaI and PstI sites of CSLAB, - yielding plasmid pCSL. Annealed complementary oligonucleotides CP26 (SEQ ID NO: 69) (5'-- 5 GTACGTGACTAATTAGCTATAAAA.AGGATCCGGTACCCTCGAGTCTAGAATCGATCC
CGGGTTTT TATGACTAGTTAATCAC-3') and CP27 (SEQ ID NO: 70) - (5r-GGCCGTGATTAACTAGTCATAAAAACCCGGGATCGATTCTAGACTCGAGGGTACCGG
ATCCTTTT TATAGCTAATTAGTCAC-3') were inserted between the pCSL AS~718 and NotI sites. The resultant- plasmid HC5LSP28 is a locus C5 vector plasmid.
3. Generation of pBSCDVHAVQ
Oligonucleotides RW132 (SEQ ID NO: 71) (5'-AGCTTCCCGGGTTAATTAATTAGTCATCAGGCAGGGCGAGAACGAGACTATCTGCTC
GTTAATTA ATTAG-3') and RW133 (SEQ ID NO: 72) (5'-AGCTCTAATTAATTAACGAGCAGATAGTCTCGTTCTCGCCCTGCCTGATGACTAATT
AATTAACC CGGGA-3') were annealed to form a double-stranded linker sequence. The RW132/RW133 (SEQ ID NO:
71/SEQ ID NO: 72) double-stranded sequence was inserted into the HindIII site 5' of the H6 promoted CDV HA ORF in pBSCDVHASS, generating plasmid pBSCDVHAVQ.
4. Generation of pCSCDVHAF1 The 2 kbp pBSCDVHAVQ SmaI fragment, which contains the H6 promoted CDV HA ORF, was inserted into the HC5LSP28 SmaI site, generating plasmid pCSLCDVHA.
The 2.1 kbp pATICDVFl HpaI/BamHI fragment, containing the H6 promoted CDV F ORF, was ligated with the pCSLCDVHA
SmaI/BamHI 6.5 kbp DNA fragment, generating plasmid pCSLCDVHAF1 which contains the H6 promoted CDV F and H6 promoted CDV HA ORFs, with their transcripts directed away from each other, in the C5 locus.
6. Generation of vector plasmid pC6L
The C6 vector pC6L was constructed to remove the C6 ORF in the following manner. Oligonucleotide primers C6A1 (SEQ ID NO: 73) (5'-ATCATCGAGCTCGCGGCCGCCTATCAAAAGTCTTAATGAGTT-3'), C6B1 (SEQ
ID NO: 74) (5'-GAATTCCTCGAGCTGCAGCCCGGGTTTTTATAGCTAATTAGTCATTTTTTCGTAAGT
AAGTATTT TTATTTAA-3'), C6C1 (SEQ ID NO: 75) (5'-CCCGGGCTGCAGCTCGAGGAATTCTTTTTATTGATTAACTAGTCAAATGAGTATATA
TAATTGAA AAAGTAA-3') and C6D1 (SEQ ID NO: 76) (5'-GATGATGGTACCTTCATAAATACAAGTTTGATTAAACTTAAGTTG-3') were used to construct pC6L. Oligonucleotide primers C6A1 (SEQ ID NO: 73) and C6B1 (SEQ ID NO: 74) were used in a PCR with canarypox DNA template to generate a 380 base pair fragment. A second PCR reaction with the canarypox ZO DNA template, and oligonucleotide primers C6C1 (SEQ ID
NO: 75) and C6D1 (SEQ ID N0: 76), generated a 1,155 base pair fragment. The two PCR reaction products were pooled and primed for a final PCR with C6A1 (SEQ ID NO: 73) and C6D1 (SEQ ID NO: ?6), yielding a 1,613 base pair fragment. The final PCR product was digested with SacI
and KpnI, and inserted between the SacI and Kt~nI sites of pBSSK+. The resultant C6 insertion plasmid was designated as pC6L.
7. Generation of pMM103 pCSLCDVHAF1 was digested with BamHI and treated with the Klenow fragment from the E. coli DNA polymerase, in the presence of 20 ~M dNTPs to blunt end the BamHI
site, followed by digestion with SmaI. The 4.2 kbp blunt ended BamHI to SmaI fragment, containing the H6 promoted CDV F and H6 promoted CDV HA ORFs, was inserted into the SmaI site of pC6L, generating plasmid pMM103.
8. Generation of pLF108 The pMM103 HindIII/BamHI 1,961 by DNA fragment which contains the CDV F1 coding sequence and the 3' most region of the vaccinia virus H6 promoter, was gel purified and subsequently inserted between the corresponding restrictions sites of pBSSK+, generating pLF108 (Figures 43, 44, SEQ ID NO: 22).
EXAMPLE 25 - Generation of pLFili, which contains a complete CDV F1 expression cassette A pLF108 XbaI restriction site was engineered immediately upstream of the CDV F1 initiation codon (ATG) in the following manner. A 473 by DNA fragment was amplified by PCR using pLF108 DNA as a template and - LF448A (5' ACTGTACTCGAGTCTAGAATGCACAAGGGAATCCCCAAAAGC 3') (SEQ ID NO: 77) and RW830 (5' ATTCCAATGTATCTGAGC 3') (SEQ
ID NO: 78) as primers. The PCR amplified fragment was isolated using the Gene Clean procedure as previously described before being digested by XhoI and CelII. The resultant XhoI/CelII 136 by DNA fragment was subsequently ligated with the 4,783 by XhoI/CelII fragment of pLF108, generating pLF109.
The XbaI (#2,035) was deleted and a KSpI
restriction site was engineered immediately downstream of pLF108 CDV F1 stop codon (TGA#2,016) in the following manner. A 431 by DNA fragment was amplified by PCR using pLF109 as a template and LF449 (5'ACTGTACCGCGGTCAGTGTGATCTCACATAGGATTTCGA 3') (SEQ ID
NO: 79) and CDV-FG (5' GGTTGAAATAGATGGTG 3') (SEQ ID NO:
80) as the primers. The PCR amplified fragment was isolated using the Gene Clean procedure as previously described before being digested with KspI and BfrI. The resultant 255 by DNA fragment was gel purified and subsequently ligated with the 4,631 by KspI/BfrI fragment of pLF109, generating pLF110.
The 1,997 by pLF110 KspI/XbaI fragment which contains the CDV F1 coding sequence was subsequently ligated with the 3,244 by K~I/XbaI fragment of pLF069, generating pLF111 (Figures 45, 46, SEQ ID NO: 23).
EXAMPLE 26 - Generation of pLF128, which contains a modified complete CDV F1 expression cassette In order to reduce the size of the polyadenylation cassette in the CDV F1 expression cassette from 241 by to 153 bp, the following manipulations were performed. The pLF077 KspI/BamHI 146 by fragment was gel purified as previously described and subsequently ligated with the pLF111 Kspl/BamHI 5,002 by WO 98/00166 PCTlUS97111486 fragment in order to generate pLF128 (Figures 47, 48, SEQ
ID NO: 24).
EXAMPLE 27 -Generation of pLF130A, a donor plasmid for insertion of CDV F1 expression cassette 12 by upstream of SmaI site at CAV2 genome right end Plasmid pLF128 was digested by BamHI and subsequently partially digested by HindIII. The BamHI/HindIII 2,451 by fragment contains the CDV Fl coding sequence coupled to the regulatory sequences in pLF077, and was treated with the Klenow fragment from the E. coli DNA polymerase before being ligated with the 6,243 by NruI linearized pLF105, generating pLF130A and pLF130B. pLF130A corresponds to the rightward orientation of the expression cassette (Figures 49, 50, SEQ ID NO: 25).
EXAMPLE 28 - Generation of vCA-CDVF1-@l2bp-up-SmaI
Ten ~g of pLF130A were digested with BalII and NotI and the resulting 5,305 by DNA fragment was isolated using the Gene Clean procedure as previously described and resuspended in H20 to a concentration of 100 ng/~,1.
MDCK cells were transfected using the Lipofectamine based procedure as previously described. Solution A was prepared by mixing 0.5 ~,g of 5.3 kbp BalII/NotI DNA
fragment with 3 ~g of purified vCA2 DNA. Solution A
total volume was brought to 300,1 with supplemented serum free MEM medium. Transfected cells were harvested after 8 days and plated out on 150 mm diameter tissue culture dishes as previously described. The 1.4 kbp EcoRI/BamHI
DNA fragment of pATICDVFl was labelled by random priming using the procedure previously described and subsequently hybridized with a nitrocellulose membrane to lift viral plaques, as previously described. Two viral plaques cross-reacting with the probe were picked and are subjected to a plaque purification process, as previously described to yield vCA-CDVF1-@l2bp-up-SmaI. This virus is characterized by restriction digestion (DNA analysis) and Southern Blot radioimmunoprecipitation (expression analysis).
EXAMPLE 29 - Additional Recombinants Since the tag and other exogenous DNA had been incorporated into CAV2, other exogenous DNA can be incorporated into CAV2. Therefore, instead of the exogenous DNA used to generate vCAl, vCA2, vCA3, vCA4, vCAS, vCA6, vCA7, vCA8, and vCA-CDVFl-@l2bp-up-SmaI, exogenous DNA as described in U.S. Patent Nos. 5,174,993 and 5,505,941 (e. g., recombinant avipox virus, vaccinia virus; rabies glycoprotein (G), gene, turkey influenza hemagglutinin gene, gp51,30 envelope gene of bovine leukemia virus, Newcastle Disease Virus (NDV) antigen, Felt/ envelope gene, RAV-1 env gene, NP (nudeoprotein gene of Chicken/Pennsylvania/1/83 influenza virus), matrix and preplomer gene of infectious bronchitis virus; HSV gD;
entomopox promoter, inter alia), U.S. Patent No.
5,338,683, e.g., recombinant vaccinia virus, avipox virus; DNA encoding Herpesvirus glycoproteins, inter alias U.S. Patent No. 5,494,807 (e. g., recombinant vaccinia, avipox; exogenous DNA encoding antigens from rabies, Hepatitis B, JEV, YF, Dengue, measles, pseudorabies, Epstein-Barr, HSV, HIV, SIV, EHV, BHV, HCMV, canine parvovirus, eguine influenza, FeLV, FHV, Hantaan, C. tetani, avian influenza, mumps, NDV, inter alia); U.S. Patent No. 5,503,834 (e. g., recombinant vaccinia, avipox, Morbillivirus [e.g., measles F, hemagglutinin, inter alia]); U.S. Patent No. 4,722,848 (e. g., recombinant vaccinia virus; HSV tk, glycoproteins [e.g., gB, gD], influenza HA, Hepatitis B [e.g., HBsAg], inter alia); U.K. Patent GB 2 269 820 B and U.S. Patent No. 5,514,375 (recombinant poxvirus; flavivirus structural proteins); WO 92/22641 (e. g., recombinant poxvirus; immunodeficiency virus, inter alia); WO
93/03145 (e. g., recombinant poxvirus; IBDV, inter alia);
WO 94/16716 and U.S. Patent No. 5,833,975, filed January 19, 1994 (e. g., recombinant poxvirus;

cytokine and/or tumor associated antigens, inter alia);
PCT/US94/06652 (Plasmodium antigens such as from each stage of the Plasmodium life cycle); U.S. Patent No.
5,523,089, W093/08306, PCT/US92/08697, Molecular Microbiology (1989), 3(4), 479-486, PCT publications WO
93/04175, and WO 96/06165 (Borrelia antigens and DNA
therefor); and Briles et al. WO 92/14488 (pneumococcal DNA), are used to generate additional CAV2 recombinants with the exogenous DNA in regions as in vCA2 through vCA8 and vCA-CDVF1-@l2bp-up-SmaI and deletions as in vCA2 through vCA8 and vCA-CDVF1-@l2bp-up-SmaI (e. g., insertions in the E3 or at the region between the right ITR and the E4 transcription unit or at both sites and deletions in the E3 region) including recombinants containing coding for multiple antigens, as herein described (including with subfragment promoters, reduced or modified polyadenylation cassettes, and promoters with 5' UTR replaced). Analysis demonstrates expression.
Compositions are prepared by admixture with a carrier or diluent for administration to a vertebrate (animal or human) hosts for generating responses, including antibody responses.

Table 1. Sizes of CAV2 DNA restriction fragments.
CAV2 DNA restriction fragments sizes - Fragment ~4~' A B C D E F G H I J K
BamHI
14 8.1 6.1 2.1 0.8 0.7 1 o EcoRI
20 8.2 3.8 Asp718 9.5 4.8 3.8 3.2 3.2 3 2.5 0.85 0.75 SaII
15 29 3.2 BgIII
29 2.8 BgII
6.1 S 4.1 3.2 2 1.7 1.5 1.5 1 0.7 ND

Table 2.
Characteristics of CAV2 E3 region ORFs ORFl ORF2 ORF3 MW {KDa.) 12.6 40.7 18.6 pI
6.48 7.45 9.68 Limits in Fig 3 Having thus described in detail preferred embodiments of the present invention, it is to be understood that the invention defined by the appended claims is not to be limited by particular details set forth in the above description as many apparent variations thereof are possible without departing from the spirit or scope thereof.

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SEQUENCE LISTING
(1) GENERAL INFORMATIOIJ:
(i) APPLICANT: MERIAL, INC.
(B) STREET: 1?Ø BOX 2999, STATION D
(C) CITY: OT'PAWA
(D) STATE: ONT
(E) COUNTRY: CANADA
(F) ZIP: K1P 5Y6 (v) COMPUTER READABLE FORM:
(A) MEDIUM T'.fPE: Floppy disk (B) COMPUTER:; IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE;; ASCII (text) (vi) CURRENT APPLICATION DATA:
(A) APPLICAT:=ON NUMBER: CA 2,259,460 (B) FILING DATE: 30-JUN-1997 (C) CLASSIFICATION:
(vii) PRIOR APPLICA7.'ION DATA:
(A) APPLICATION NUMBER: US 08/675,556 (B) FILING DATE: 03-JUL-1996 (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 08/675,566 (B) FILING DATE: 03-JUL-1996 (viii) ATTORNEY/AGEN7.' INFORMATION:
(A) NAME: SMART & BIGGAR
(B) REGISTRA7.'ION NUMBER:
(C) REFERENCF;/DOCKET NUMBER: 77354-34 (ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: (613)-232-2486 (B) TELEFAX: (613)-232-8440 (2) INFORMATION
FOR
SEc~
ID N0:1:

(i) S EQUENCE
CHARACTERISTICS:

(A) LENGTH:6994 base pairs (B) TYPE:
nucleic acid (C) STRANDEDIQESS:
single (D) TOPOLOGY:
linear (ii) MOLECULE
TYPE:
DNA
(genomic) (xi) SEQUENCE
DESCRIPTION:
SEQ
ID N0:1:

GGAAATTGTAAACGTTAATA'CTTTGTTAAA ATTCGCGTTAAATTTTTGTT AAATCAGCT~60 CAACGTCAAAGGGCGAAAAA(:CGTCTATCA GGGCGATGGCCCACTACGTG AACCATCACC240 CTAATCAAGTTTTTTGGGGTC:GAGGTGCCG TAAAGCACTAAATCGGAACC CTAAAGGGAG300 CACACCCGCCGCGCTTAATGC:GCCGCTACA GGGCGCGTCGCGCCATTCGC CATTCAGGCT480 CAACCGCAAACGGGACACGCC:GGCGCCTCC CAGGACTACTCCACCCAAAT GAATTGGTTT840 TTGATAACCCAGGCAGAAATAP.CCAAAACT CCCAGAACAATAATGGATCC GCCAATTTGG960 TGCCCCTCCCAAATAGGTAT.?~AAAAGCCCAGTGCTGGCTGGCACGGGCATTCAGCTTAGC 1140 GAAGACATCCCCAGCGCCTC~~TGGATCAGGCCCGACGGCATATTCCAGCTAGGAGGGGGG 1200 TCTCGCTCGTCCTTCAGCCC,~ACGCAAGCATTCCTCACCCTGCAACAGGCATCCTCGACG 1260 ATTGTAACCAACTCTGTCGA'PGGCTATGACTGAGGAGAGCATGGACCAGGTGGAGGTGAA 1440 TTTATGTGCTAACTGGTTTTi~CAACCCAGCACTTGCCTTTGAAGGGTTTGATATTCCAGA 1560 ACTGTGCCACAATGGCCATGi~TATGATCT'GCTCATACTCTCGCCTGGGATCCCACATTAA 1680 TATGTATAACCTTAACTAGA'~AATATTATTAAACTTGTTTTACAGCTACCACCATAATGC 1800 CCCTCATTAAGGAGAGCAAC'.CACGAGGGCGCTGAGCACTACTACCTTGTCTATATTTATG 2100 GCTTCCAAGCCCAAAAAATC':'CTAAAGTCAGAAGCGGAGGCAGAAAAGAGAACCTGCATC 2340 CCAACTGGGCCTTGGTTACC7.'ATACTGGAGACCTTCTTGTCTTGCATGTTTCGCCAAACA 2400 TCAACATAACTGTACCCAAC7.'GGCAACAAAATCTAGTAACCATATTTAATCAACACGAGC 2520 CCCCAAAAAAGGGCGATAAT7.'ATGAGGACAGTTTTATGGAATGGACTCTGTTTAAAAAGC 2580 AAGAAGACAC

GCCATTCCATGCCCGTCGCC.ATACCTGACACTGCAATGCCTATATATATTTCCATCATGT 2820 GGCCCATGTAGCTTGTCAAA'rAAACTTACCTAATTTTTGCTAAGACGTCTGGGTCCTGCG 2940 TTTCTATGTCCACCAAAGTC~~CCTCTTCCCAGCTTTGGTACTTCCACTTGTGCGCGCGAG 3000 CCAGCTTGCGGATGTGCTTG;~AAGATAATGTGGTCTCTCCCAACAGCTTCCCGTTCACCA 3060 GCACCAGGGCCATGAAGCGG;~CACGAAGAGCTCTACCTGCAAATTATGACCCTGTATATC 3120 CATACGACGCCCCCGGGTCT'rCCACACAACCCCCTTTTTTTAATAACAAGCAAGGTCTCA 3180 ACAAGGTTTTGTCTTTTACC'rCCCCATTACATAAAAATGAAAACACTGTATCCCTAGCGC 3420 GGCAACCCCTCCCCCGCCTCTCACCTTTACATCACCCCTAGAAP.AAAATGAAAACACAGT 3840 TGGAAGTGGTTTAAGAATATC:TGGAGGCAGCCTCACGGTGGCCACTGGACCTGGCCTTTC 4020 ACCCCCAGCTGCCCCCATAA<:ACTGTGGACAGGGCCTGGCCTAGCATTAATGGCTTTATG 4200 TAATGACACTCCAGTAATTACiGTCTTTATATGCCTAACCAGAGACAGCAACTTAGTCACA 4260 AACTGTGCAT

GCTAATTATTAACAAACCAA.AAGGCGTTGCCACTTACACCCTTACCTTTAGGTTTTTAAA 4620 CTTTAACAGACTAAGCGGAG~~TACCCTGTTTAAAACTGATGTCTTAACCTTTACCTATGT 4680 AGGCGAAAATCAATAAAACC,?~GAAAAAAATAAGGGGAAAAGCTTGATATCGAATTCCTGC 4740 AGCCCGGGGGATCCACTAGT'rCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAGCTTTTGT 4800 TCCCTTTAGTGAGGGTTAAT'PCCGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTG 4860 TGAAATTGTTATCCGCTCAC:~ATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAA 4920 GTTCGGCTGCGGCGAGCGGTi~TCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAA 5160 TCAGGGGATAACGCAGGAAAc;AACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGT 5220 GACCGCTGCGCCTTATCCGG'.CAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTA 5580 TCGCCACTGGCAGCAGCCAC'.CGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCT 5640 TGCGCTCTGCTGAAGCCAGT'.PACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAA 5760 CAAACCACCGCTGGTAGCGG'..'GGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAA 5820 AAAGGATCTCAAGAAGATCC5:'TTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAA 5880 TTAAATTAAAAATGAAGTTT7.'AAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGAC 6000 ATAGTTGCCTGACTCCCCGTC:GTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGC 6120 AACGTTGTTGCCATTGCTAC.AGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCA 6360 TTCAGCTCCGGTTCCCAACG.ATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAA 6420 GCGGTTAGCTCCTTCGGTCCrCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCA 6480 CTCATGGTTATGGCAGCACT~~CATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTT 6540 TCTGTGACTGGTGAGTACTC,~.ACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGT 6600 TGCTCTTGCCCGGCGTCAAT;~CGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTG 6660 CTCATCATTGGAAAACGTTC'rTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGA 6720 TCCAGTTCGATGTAACCCAC'PCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACC 6780 GGTTATTGTCTCATGAGCGGi~TACATATTTGAATGTATTT AGAAAAATAA ACAAATAGGG6960 (2) INFORMATION FOR SE(,2 ID N0:2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6958 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:2:
TCGACGGTAT CGATAAGCTT ".'GCTCAACAA ATACTGTCAA GGACTCGAGT CCGGCTCTGA 60 CTGAGCAATG TCTAAAGAAA 7.'ACCAACCCC TTATATGTGG AGCTACCAAC CGCAAACGGG 120 TGTTCAGGAA GCCGCCCCAC C:CAAAACGGT CACTCTGCCC AGAAACCACA CCCTAGAACA 360 CGCCTCCTGGATCAGGCCCG.ACGGCATATTCCAGCTAGGAGGGGGGTCTCGCTCGTCCTT 540 CAGCCCAACGCAAGCATTCC'PCACCCTGCAACAGGCATCCTCGACGCCGCGCGCAGGAGG 600 CGTGGGCACCTACCAGTTTG'rGCGCGAATTTGTGCCAGAGGTATACCTTAACCCTTTTTC 660 AGCATGCCCAAACCTGCACGCGCCC'rCGCTGCTTTGCAAAGGAGGGTTTATGTGCTAACT 840 GCCATGATATGATCTGCTCA'PACTCTCGCCTGGGATCCCACATTAACATAAGATGTATTT 1020 GCAACAAGCCGCGGCCCCACi~TGAGCCTCATTGAGGCAGCCTGTTCTATGTATAACCTTA 1080 ACTAGATAATATTATTAAAC'PTGTTTTACAGCTACCACCATAATGCGCTTCAGCTTCTTC 1140 GGCCTGCAGTTTTCTGCAAA(:TTTTCTGAGGATGGCCTGTACATCGCCCTCATTAAGGAG 1380 ACTGCAAATGAGTCTGCCCA(:GGGCCTATTTCCAGGCCCCTCAAAGATCTGCTAATGGAA 1500 GACCCCAGCGGCTTCCAAGCC:CAAAAAATCTCTAAAGTCAGAAGCGGAGGCAGAAAAGAG 1620 AACCTGCATCCCAACTGGGC<:TTGGTTACCTATACTGGAGACCTTCTTGTCTTGCATGTT 1680 TTTAAAAAGCTCAAAAAAGG<:TTATTTAGAGTAACTTGCAGAGCCAAGTCAATATTCCCA 1920 GAGTGCGTCCTCAACATCACC:CGCGACGGAACTTTCCTGCTTATTGGGGATAGCAAAAAG 1980 TGCGCGCGAGCCAGCTTGCG~ATGTGCTTGAAAGATAATGTGGTCTCTCCCAACAGCTTC 2340 CCGTTCACCAGCACCAGGGC~~ATGAAGCGGACACGAAGAGCTCTACCTGCAAATTATGAC 2400 CCTGTATATCCATACGACGC~~CCCGGGTCTTCCACACAACCCCCTTTTTTTAATAACAAG 2460 TTTTCTACGTTAGGTGCCAT'PAAACTTTCCACAGGTCCCGGACTCACCCTCAACGAGGGC 2580 ACTCCCCCTCCCCCGCTACAi~TTCTCCCCTCCCCTCACAAAAACAGGTGGTACTGTTTCC 2820 ACGCACCTCCCTTGAAAAAAi~CTGACCAGCAAGTTAGCCTCCAAGTAGGCTCGGGTCTCA 2940 GACGTTTGGTGGCAACCCCT(:CCCCGCCTCTCACCTTTACATCACCCCTAGAAA:SAAATG3120 CTCTAAGTGCTGGAAGTGGT'.CTAAGAATATCTGGAGGCAGCCTCACGGTGGCCACTGGAC 3300 TGGCTTTATGTAATGACACTC:CAGTAATTAGGTCTTTATATGCCTAACCAGAGACAGCAA 3540 CCAGTCACAATTTAGCCTAA7.'TATGGAGTTTGATCAGTTTGGACAGCTTATGTCCACAGG 3660 AAACATTAACTCCACCACTAC:TTGGGGAGAAAAGCCCTGGGGCAATAACACTGTACAGCC 3720 AAGGCGTTGC

GAATTCCTGCAGCCCGGGGG'.ATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTC 4080 GTTTCCTGTGTGAAATTGTT.ATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCAT 4200 AAAGTGTAAAGCCTGGGGTG~~CTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTC 4260 GCATCACAAAAATCGACGCTc~AAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATA 4620 CGGATACCTGTCCGCCTTTC'rCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTG 4740 ATTTGGTATCTGCGCTCTGC'.CGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTG 5040 ATCCGGCAAACAAACCACCG(:TGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTAC 5100 GCGCAGAAAAAAAGGATCTCAP.GAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCA 5160 ACCATCTGGCCCCAGTGCTGC:AATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTT 5460 ATCAGCAATAAACCAGCCAGC:CGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATC 5520 GCGACCGAGTTGCTCTTGCC~GGCGTCAATACGGGATAATACCGCGCCACATAGCAGAAC 5940 TTTAAAAGTGCTCATCATTG~AAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACC 6000 GCTGTTGAGATCCAGTTCGA'rGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTT 6060 TACTTTCACCAGCGTTTCTG~~GTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGG 6120 CATTTATCAGGGTTATTGTC'TCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAA 6240 GCCGAAATCGGCAAAATCCC'PTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTT 6420 GTTCCAGTTTGGAACAAGAG'PCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGA 6480 AAAACCGTCTATCAGGGCGA'PGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTG 6540 GGGCGATCGGTGCGGGCCTC'.CTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCA 6840 (2) INFORMATION FOR SEQ ID N0:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7001 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:
linear (ii) M OLECULE
TYPE:
DNA (genomic) (xi) S EQUENCE
DESCRIPTION:
SEQ ID
N0:3:

TTTGTTAAAATTCGCGTTAA.ATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGA 60 AATCGGCAAAATCCCTTATA.AATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCC 120 CGTCTATCAGGGCGATGGCC~~ACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTC 240 GAGGTGCCGTAAAGCACTAA.?~TCGGAACC:CTAAAGGGAGCCCCCGATTTAGAGCTTGACG 300 GGCGCTGGCAAGTGTAGCGG'rCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGC 420 ATCGGTGCGGGCCTCTTCGC'rATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCG 540 ACCAAAACTCCCAGAACAATi~ATGGATCCGCCAATTTGGCCAGCTGCCATGCTTGTTCAG 960 AAAAGCCCAGTGCTGGCTGG(:ACGGGCATTCAGCTTAGCGAAGACATCCCCAGCGCCTCC 1140 GGCTATGACTGAGGAGAGCA7.'GGACCAGGTGGAGGTGAACTGCCTGTGTGCTCAGCATGC 1440 AATATTATTAAACTTGTTTT.ACAGCTACCACCATAATGCGCTTCAGCTTCTTCATCGCCG1800 CTTGCCTCCTACACCTAGAA~TGGGCTTGGGGGCCAATGTCAGTTGGATAAACTCTGACA1920 CAGGCCAGGCCCCGATTTGC~~TCTCCAATGGCATGTGCAACGCTACCCAGCAAGGCCTGC1980 AGTTTTCTGCAAACTTTTCT~~AGGATGGCCTGTACATCGCCCTCATTAAGGAGAGCAACT2040 ACGAGGGCGCTGAGCACTAC'rACCTTGTCTATATTTATGGAGACTGCTACCAAACTGCAA2100 ATGAGTCTGCCCACGGGCCT:~1TTTCCAGGCCCCTCAAAGATCTGTTAACCCTAAGGCCAT2160 GTTACCTATACTGGAGACCT'PCTTGTCTTGCATGTTTCGCCAAACACCCTTGGACTGTGG2400 CCCAACTGGCAACAAAATCTi~GTAACCATATTTAATCAACACGAGCCCCCAAAAAAGGGC2520 GATAATTATGAGGACAGTTT'PATGGAATGGACTCTGTTTAAAAAGCTCAAAAAAGGCTTA2580 GACGGAACTTTCCTGCTTAT'CGGGGATAGCAAAAAGACCCCCTATGTCATCCTGCTGCCC2700 AAAGTCCCCTCTTCCCAGCT':.'TGGTACTTCCACTTGTGCGCGCGAGCCAGCTTGCGGATG3000 TGCTTGAAAGATAATGTGGT(:TCTCCCAACAGCTTCCCGTTCACCAGCACCAGGGCCATG3060 GGGTCTTCCACACAACCCCC7.'TTTTTTAATAACAAGCAAGGTCTCACTGAGTCACCCCCA3180 TTTACCTCCCCATTACATAA.AAATGAAAACACTGTATCCCTAGCGCTAGGAGATGGGTTA 3420 TCCCCTCCCCTCACAAAAAC.AGGTGGTACTGTTTCCTTGCCCCTGCAAGACTCCATGCAA 3540 ACCAGCAAGTTAGCCTCCAA~~TAGGCTCGGGTCTCACCGTGATTAACGAACAGTTGCAAG 3660 CTGTCCAGCCTCCCGCAACC.~CCTACAACGAGCCTCTTTCCAAAACTGACAATTCTGTTT 3720 GCGCGGGCTTGTCTGTACAA:~ACAACGCCCTAGTAGCCACACCTCCCCCACCCTTAACCT 3900 CCATAACACTGTGGACAGGGc:CTGGCCTAGCATTAATGGCTTTATGTAATGACACTCCAG 9200 TAATTAGGTCTTTATATGCC'~AACCAGAGACAGCAACTTAGTCACAGTAAATGCTAGCTT 4260 GTGCATGCCTAACAGAGAAG'.L'TTACTCCACTCCCGCCGCCACCATCACCCGCTGTGGACT 4500 AACCAAAAGGCGTTGCCACT'.~ACACCCTTACCTTTAGGTTTTTAAACTTTAACAGACTAA 4620 ACTAGTTCTAGAGCGGCCGCC:ACCGCGGTGGAGCTCCAGCTTTTGTTCCCTTTAGTGAGG 4800 GCTCACAATTCCACACAACA7.'ACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTA 4920 AGCGGTATCAGCTCACTCAA.AGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGC 5160 AGGAAAGAACATGTGAGCAA.AAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTT 5220 TCAGAGGTGGCGAAACCCGA~AGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTC 5340 CCTCGTGCGCTCTCCTGTTC~~GACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCC 5400 TTCGGGAAGCGTGGCGCTTT~~TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGT 5460 ATCCGGTAACTATCGTCTTG:~1GTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGC 5580 TAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAA:9AGGATCTCAAGA 5820 TGCTACAGGCATCGTGGTGT(:ACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTC 6360 CCAACGATCAAGGCGAGTTA(:ATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTT 6420 AGCACTGCATAATTCTCTTAC:TGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGA 6540 GTCAATACGGGATAATACCG<:GCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAA 6660 GAGCGGATACATATTTGAAT;~TATTTAGAA AAATAAACAAATAGGGGTTCCGCGCACATT 6960 TCCCCGAAAAGTGCCACCTG~~GAAATTGTA AACGTTAATAT 7001 (2) INFORMATION
FOR
SEQ
ID N0:4:

(i) S EQUENCE S:
CHAR;~CTERISTIC

(A) LENGTH:0578 basepairs (B) TYPE:
nucleic acid (C) STRANDEDNESS: le sing (D) TOPOLOGY:
linear (ii) OLECULE
M TYPE:
DNA (genomic) (xi) EQUENCE
S DESCRIPTION:
SEQ ID
N0:4:

CGGATACATATTTGAATGTA'CTTAGAAAAATAAACAAATAGGGGTTCCGC GCACATTTCC120 TTTTGTTAAATCAGCTCATT'.PTTTAACCAATAGGCCGAAATCGGCAAAAT CCCTTATAAA240 TTAAAGAACGTGGACTCCAAC:GTCAAAGGGCGAAAAACCGTCTATCAGGG CGATGGCCCA360 CGGAACCCTAAAGGGAGCCCC:CGATTTAGAGCTTGACGGGGAAAGCCGGC GAACGTGGCG480 CATTCGCCATTCAGGCTGCGC:AACTGTTGGGAAGGGCGATCGGTGCGGGC CTCTTCGCTA660 GCGAATTGGGTACCGGGCCCC:CCCTCGAGGTCGACGGTATCGATAAGCTT TGCTCAACAA840 ATACTGTCAAGGACTCGAGTC:CGGCTCTGACTGAGCAATGTCTAAAGAAA TACCAACCCC900 CACTCTGCCCAGAAACCACA~~CCTAGAACAGGCTATGACCAACTCTGGGGCGCAGCTAGC 1200 GGGAGGACGACAGCTGTGCC~~CTCCCAAATAGGTATAAAAAGCCCAGTGCTGGCTGGCAC 1260 GGGCATTCAGCTTAGCGAAG,~1CATCCCCAGCGCCTCCTGGATCAGGCCCGACGGCATATT 1320 GCTTTGCAAAGGAGGGTTTA'PGTGCTAACTGGTTTTACAACCCAGCACTTGCCTTTGAAG 1680 GGTTTGATATTCCAGACTCT'PACCAAGAGGGACACGGTGTGGACATAGAAGTTAAGTGTT 1740 CCCACCACTCCAGCAAACTG'PGCCACAATGGCCATGATATGATCTGCTCATACTCTCGCC 1800 TTGAGGCAGCCTGTTCTATG'CATAACCTTAACTAGATAATATTATTAAACTTGTTTTACA 1920 GGCTTGGGGGCCAATGTCAG'.CTGGATAAACTCTGACACAGGCCAGGCCCCGATTTGCCTC 2100 GATGGCCTGTACATCGCCCTC:ATTAAGGAGAGCAACTACGAGGGCGCTGAGCACTACTAC 2220 CTTGTCTATATTTATGGAGAC:TGCTACCAAACTGCAAATGAGTCTGCCCACGGGCCTATT 2280 GGCCGCGGCCGCACGCGTGTC:CTCAACATCACCCGCGACGGAACTTTCCTGCTTATTGGG 2400 GATAGCAAAAAGACCCCCTA7.'GTCATCCTGCTGCCCTTTTTTGCAAACCCCAAAGAAGAC 2460 ATGGGACTAAACAACAAAATC:AGGCCCATGTAGCTTGTCAAATAAACTTACCTAATTTTT 2640 CCAACAGCTTCCCGTTCACC.AGCACCAGGGCCATGAAGCGGACACGAAGAGCTCTACCTG 2820 CAAATTATGACCCTGTATAT~CATACGACGCCCCCGGGTCTTCCACACAACCCCCTTTTT 2880 TTAATAACAAGCAAGGTCTC.ACTGAGTCACCCCCAGGAACCCTGGCTGTCAATGTTTCCC 2940 CTCCACTAACCTTTTCTACG'TTAGGTGCCATTAAACTTTCCACAGGTCCCGGACTCACCC 3000 AAATCACTGTTGAAAATGTC;~ACAAGGTTTTGTCTTTTACCTCCCCATTACATAAAAATG 3120 CTCGGGTCTCACCGTGATTAi~CGAACAGTTGCAAGCTGTCCAGCCTCCCGCAACCACCTA 3420 CGTGCAGAGCGGACGTTTGG'rGGCAACCCCTCCCCCGCCTCTCACCTTTACATCACCCCT 3540 AGAAAAAAATGAAAACACAG'PGTCGCTACAAGTAGGCGCGGGCTTGTCTGTACAAAACAA 3600 GGCCACTGGACCTGGCCTTT(:CCATCAAAATGGAACAATAGGGGCTGTAGTAGGTGCAGG 3780 CCTCAAGTTTGAAAACAATGC:CATTCTTGCAAAACTAGGCAACGGTCTAACCATTAGAGA 3840 ATGTCCACAGGAAACATTAAC:TCCACCACTACTTGGGGAGAAAAGCCCTGGGGCAATAAC 4140 ACTGTACAGCCACGCCCAAGC:CACACCTGGAAACTGTGCATGCCTAACAGAGAAGTTTAC 4200 TCCACTCCCGCCGCCACCATC:ACCCGCTGTGGACTAGACAGCATTGCAGTCGACGGTGCC 4260 CAGCAGAAGTATCGACTGCA7.'GCTAATTATTAACAAACCAAAAGGCGTTGCCACTTACAC 4320 GCCGGAAGCATAAAGTGTAA.AGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATT4680 ATCGGCCAACGCGCGGGGAG.?~GGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTC4800 ACTGACTCGCTGCGCTCGGT~~GTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCG4860 GTAATACGGTTATCCACAGA.~1TCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGC4920 CAGCAAAAGGCCAGGAACCG'rAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGC4980 CCCCCTGACGAGCATCACAA;~AATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGA5040 CTATAAAGATACCAGGCGTT'rCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACC5100 AGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCT~GGGCTGTGTG5220 AACCCGGTAAGACACGACTTi~TCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGA5340 GCGAGGTATGTAGGCGGTGC'CACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACT5400 GGTAGCTCTTGATCCGGCAAi~CAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAG5520 AGGATCTTCACCTAGATCCT'CTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATA5700 TATGAGTAAACTTGGTCTGAC:AGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCG5760 ATCTGTCTATTTCGTTCATC(:ATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATA5820 CGGGAGGGCTTACCATCTGGC:CCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCG5880 GCAACTTTATCCGCCTCCATC:CAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGT6000 TCGCCAGTTAATAGTTTGCGC:AACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGC6060 CATAGCAGAACTTTAAAAGT~~CTCATCATTGGAAAACGTTCTTCGGGGCG AAAACTCTCA6420 AGGATCTTACCGCTGTTGAG.?~TCCAGTTCGATGTAACCCACTCGTGCACC CAACTGATCT6480 (2) INFORMATION
FOR SEQ
ID N0:5:

(i) SEQUENCE
CHARACTERISTICS:

(A) LENGTH:6196 base pairs (B) TYPE:
nucleic acid (C) STRANDEDNESS:
single (D) TOPOLOGY:
linear (ii) MOLECULE
TYPE:
DNA (genomic) (xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:5:

AACAAAACCATCAACTGGAA':.'GCAAGAATAGTCCAGCACGGTGGGTTCAA TCTAAAAATG420 CAGCACAGTTTTTAAGAGAAC:AATAGTTTTGAAGGCTACAAGATTTACAC TTAAGCACCA660 GCCAGTAATTATAAGTGCTT7.'TAAGAACTACCCCTAGCTCAGGGTTAATG CACCTTTTAA720 TGGCCTCCATGCAGGCTTTA7.'GGACAGTTCTAAAAAAAGACAGTCTAAAA TAAATGTAGT780 GACAGTGTACTCGCCACGTTrGAGCCTCTGCCAGGCAGCAGTGCTTAGTTACTATCAACT 1020 CAATACCCGCATTGCATGTA.?~.ACCCCCCAAAGAGCAGTTTTTCATGCCTGTGTAGCACAT 1080 CATCCCACAAAATAGGAATT'rCATAGCATAAAGCAAAGCAATTACAATATTTAGGAACTC 1140 TATGAACAAAAACTAAACAC'PTCTAACAAAGATACAGTGACAATCTCCCTTCCTCTAAAA 1260 GCATTGTTTACATTAGGGTG;~TTATTAACAACGTCAGAAATTTCTTTAATTAAAGTGCCT 1320 AAAGTAATCCCAGGCTTGTT'PTTATCAACAGCCTTAAACATGCTTTCACAAAATATAGAA 1440 TATAGCTACAAAGACCTGCA'CCCCCTCCTTAGCAGACAGCTCTTGCACACACGCAGTAAC 1800 TATCCACCGCTTAAGAAAAG(:TTTAAGCCCAGCGCACATAACAGCTCCAATGTTTTTATC 1860 CAAGGAGAGCAAAATTTCAGC:AAGCGCAGGCTCAACAGTAATAGTGAAGCAGAGGCATTT 1920 CAGACGAGGCTCACTAGCTGC:AGTCGCCATTTATGAGGTCTGCAATAAAAAACAACTCAT 1980 CAGCAGCTGAAAAAGTGCAC'CTTGACCTCATTAAGCCACTGCATATGCAAGTCCTCATCT 2040 TCTTCTTTTAGCAAAGTACA<:ATGCTGTTTGGACTAGTATACACAATAGAAGTCACAATG 2160 AGCATTGAAACCCCGCGACA<:AGGTCAGTCTCGCGGTCTTGATCTCTTATTATAGCGACC 2280 AAAAGCAACACTTACTTATT<:AGCAGTCACAAGAATGTTGGGCTCCCAAGTGACAGACAA 2460 TCCTGAAGAGAAACGGCGGT.AGCCTGGATATCTGCAACGGACCCAAAACCTTCAGTGTCA 2580 CTTCCAATAAACAGATAAAA~~TCTAAATAGTCCCCACTTAAAACCGAAACAGCCGCGGCA 2640 AAGGTAGGACACGGACGCAC'rTCCTGAGCCCTAATAAGGCTAAACACCACACGGCGCAGT 2700 TCAGAAGGCAAAAAGTCTGT,~P.GCTCTAGCTGAGCACACACACTCTCCACTAGACACTTG 2760 TGAAGCCTCAGACAAAAACA'rGCTCCCATAGACACTCCTAAAGCTGCCATTGTACTCACG 2820 TGCCAAGTACAGTCATAAAA'rGTGGGCGCGTGGTAAATGTTAAGTGCAGTTTCCCTTTGG 3000 CCACACCTCTTTGTCCTGTA'CATTATTGATGATGGGGGGATCCACTAGTTCTAGAGCGGC 3300 CGTAATCATGGTCATAGCTG'CTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACA 3420 CATTAATTGCGTTGCGCTCAC:TGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGC 3540 CCTCGCTCACTGACTCGCTGC:GCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACT 3660 CAAAGGCGGTAATACGGTTA~~CCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAG 3720 GGCTCCGCCCCCCTGACGAGC:ATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACC 3840 CGACAGGACTATAAAGATACC:AGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTG 3900 TTGAGTCCAACCCGGTAAGAC:ACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGA 4140 GCTACACTAGAAGGACAGTA7.'TTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAA 4260 AAAGTATATATGAGTAAACT'PGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTA 4560 GCTCACCGGCTCCAGATTTA'TCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAA 4740 TTACATGATCCCCCATGTTG'rGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTG 4980 TCAGAAGTAAGTTGGCCGCAc;TGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTC 5040 TTACTGTCATGCCATCCGTAi~GATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCAT 5100 TCTGAGAATAGTGTATGCGGc:GACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATA 5160 CCGCGCCACATAGCAGAACT'CTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAA 5220 AATGTATTTAGAAAAATAAA(:AAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC 5520 CTCATTTTTTAACCAATAGGC:CGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGAC 5640 CGAGATAGGGTTGAGTGTTG'.'TCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGA 5700 GAAAGCGAAAGGAGCGGGCGC:TAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAAC 5940 ACGTTGTAAA ACGACGGCCA ~TGAATTGTA ATACGACTCA CTATAGGGCG AATTGGGTAC 6180 (2) INFORMATION
FOR SEQ
ID N0:6:

(i) SEQUENCE S:
CHAR:~CTERISTIC

(A) LENGTH:'0503 pairs base (B) TYPE:
nucleic acid (C) STRANDEDNESS: le sing (D) TOPOLOGY:
linear (ii) MOLECULE
TYPE:
DNA (genomic) (xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:6:

TTGCCACTTACACCCTTACC'CTTAGGTTTTTAAACTTTAACAGACTAAGC GGAGGTACCC120 AAAGTTACTCTGTAAACAGT'.CCTTTCACAACAGCACAAAACATAGGTATT AGTTAACAGT300 TCATTTGGGCTATAATAATA'.CACATTTTCTTGGGTGGCAAAGCAAGGGTC GGTAATCTCA360 AGAAACGCTGTTGAGGTTCAC:TAAGCACAGGTTTTGAATCTGTCGGCAGC GTCCATGCAT480 CACTACAGGTGTCTTTTCAAC:CTCTTTCAGCACCCGCTCTATTACAGATC TCACCCACAC600 CCAGTAATTATAAGTGCTTT7.'AAGAACTACCCCTAGCTCAGGGTTAATGC ACCTTTTAAT720 AGTGTTTCTAAATATAATAC7.'CCCCACATAGTTAATTTCATCAGGCCTGC TAGAATTTAC840 CACTTTCTGAACCACATGCT7.'GCTAGCCATGCATTGTAAAGACAAGCTGT TAGAGCAGTG960 AATACCCGCATTGCATGTAA.ACCCCCCAAAGAGCAGTTTTTCATGCCTGTGTAGCACATC 1080 CACCACAGCAGTCACGTGAC.ATGTTGTCTCAGCAGTGCAGTTGCCTTCCATCCTACAATT 1200 ATGAACAAAAACTAAACACT'PCTAACAAAGATACAGTGACAATCTCCCTTCCTCTAAAAG 1260 CATTGTTTACATTAGGGTGA'rTATTAACAACGTCAGAAATTTCTTTAATTAAAGTGCCTT 1320 TAAAATGTGCAAGAGCATCA'rCATACTCAAAACCAAGCTGAGAGTAAAAGACCACCTTAA 1380 AAGTAATCCCAGGCTTGTTT'PTATCAACAGCCTTAAACATGCTTTCACAAAATATAGAAG 1440 CAGTAACATCATCAATGGTG'rCGAAGAGAAACTCCATAGGAGACTCCAGCATTGATCCAA 1500 GCTCTCTAACAAAATCTTCC'rCAAAATGAATAATGCCCTTTACACAAACGCGGGGCAGAC 1560 ATCCACCGCTTAAGAAAAGC'CTTAAGCCCAGCGCACATAACAGCTCCAATGTTTTTATCC 1860 AGCAGCTGAAAAAGTGCACT'.CTGACCTCATTAAGCCACTGCATATGCAAGTCCTCATCTA 2040 AATGGTCCTTCAGAGTGATG"..'TGCACTCATAGAAGTAGGCAGCTCCGGCAGCCATTCTGC 2340 AGGTAGGACACGGACGCACTrCCTGAGCCCTAATAAGGCTAAACACCACACGGCGCAGTT 2700 CAGAAGGCAAAAAGTCTGTA.?~GCTCTAGCTGAGCACACACACTCTCCACTAGACACTTGT 2760 ACGGCTGGCTGTCAGAGGAG:~GCTATGAGGATGAAATGCCAAGCACAGCGTTTATATAGT 2880 CCTCAAAGTAGGGCGTGTGG;~1AAACGAAAAGGAATATAACGGGGCGTTTGAGGAAGTGGT 2940 GGTTGGCCCGGAAAGTTCAC:~AAAAGTACAGCACGTCCTTGTCACCGTGTCAACCACAAA 3060 CCATATATTCATGTCCCCAGi~CATCATAGTCAGCACCATTTTCTTCTCCTTTTGCCAGTA 3180 CACCAAGAGCTGAAAGAAAT'CGAGGTATGGACACTTGGATGGTGATGTTCCCTGCCTCCG 3360 TTTAAGCCACACCTCTTTGT(:CTGTATATTATTGATGATGGGGGGATCCACTAGTTCTAG 3600 AGCTGCATTAATGAATCGGC(:AACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTT 3900 CCGCTTCCTCGCTCACTGACi.'CGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAG 3960 CTCACTCAAAGGCGGTAATAC:GGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACA 4020 CTCCTGTTCCGACCCTGCCGC;TTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCG 4260 TGGCGCTTTCTCATAGCTCAC:GCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCA 4320 AGCTGGGCTGTGTGCACGAAC;CCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACT 4380 ATCGTCTTGAGTCCAACCCG~~TAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA 4940 ACAGGATTAGCAGAGCGAGG'rATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTA 4500 ACTACGGCTACACTAGAAGG,~CAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCT 4560 TCGGAAAAAGAGTTGGTAGC'rCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTT 4620 TGAGATTATCAAAAAGGATC'PTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAAT 4800 GCAGAAGTGGTCCTGCAACT'CTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAG 5100 TCGTGGTGTCACGCTCGTCG'CTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAA 5220 ATTCTCTTACTGTCATGCCA':CCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCA 5400 GGCGAAAACTCTCAAGGATC'.'TACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTG 5580 CACCCAACTGATCTTCAGCA~.'CTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAG 5640 TCTTCCTTTTTCAATATTAT7.'GAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACA 5760 ACCATCACCCTAATCAAGTT7.'TTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCC 6120 AGGGAAGAAA GCGAAAGGAG~~GGGCGCTAGGGCGCTGGCAAGTGTAGCGG TCACGCTGCG6240 CGTAACCACC ACACCCGCCG~~GCTTAATGCGCCGCTACAGGGCGCGTCGC GCCATTCGCC6300 (2) INFO1~MATION
FOR SEQ ID N0:7:

(i) SEQUENCE CHARR~CTERISTICS:

(A) LENGTH:4503 base pairs (B) TYPE:
nucleic acid (C) STRANDEDIJESS:
single (D) TOPOLOGY:
linear (ii) MOLECULE
TYPE: DNA (genomic) (xi) SEQUENCE
DESCRIPTION:
SEQ ID N0:7:

GTCTATATAA GCAGAGCTCG'.CTTAGTGAACCGTCAGATCGCCTGGAGACG CCATCCACGC120 GAGATTTTCA GGAGCTAAGGAP.GCTAAAATGGAGAAAAAAATCACTGGAT ATACCACCGT240 TAAGCACAAG TTTTATCCGGC:CTTTATTCACATTCTTGCCCGCCTGATGA ATGCTCATCC420 GGCCTATTTC CCTAAAGGGT7.'TATTGAGAATATGTTTTTCGTCTCAGCCA ATCCCTGGGT660 GAGTTTCACC AGTTTTGATT7.'AAACGTGGCCAATATGGACAACTTCTTCG CCCCCGTTTT720 CACCATGGGC AAATATTATAC:GCAAGGCGACAAGGTGCTGATGCCGCTGG CGATTCAGGT780 GCCTGGTGCTACGCCTGAAT:~AGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGAT 960 CTTTGTGAAGGAACCTTACT'rCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGAT 1020 TTAAAGCTCTAAGGTAAATA'rAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCT 1080 AATTGTTTGTGTATTTTAGA'rTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAA 1140 TGCCTTTAATGAGGAAAACC'rGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGC 1200 GGACTTTCCTTCAGAATTGC'CAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCT 1320 GGAAAAATATTCTGTAACCT'rTATAAGTAGGCATAACAGTTATAATCATAACATACTGTT 1440 TTATTGCAGCTTATAATGGT'PACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAG 1740 TCTGGATCCCCCGGAATTCA(:TGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTG 1860 GCGTTACCCAACTTAATCGCC:TTGCAGCACATCCCCCCTTCGCCAGCTGGCGTAATAGCG 1920 AAGAGGCCCGCACCGATCGCC:CTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCC 1980 ACTGGGTCATGGCTGCGCCCC:GACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTT 2160 GTCTGCTCCCGGCATCCGCT7.'ACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTC 2220 AGAGGTTTTCACCGTCATCAC:CGAAACGCGCGAGGCAGTTCTTGAAGACGAAAGGGCCTC 2280 GGTGCACGAGTGGGTTACAT~~GAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTT 2640 CGCCCCGAAGi~ACGTTTTCC.~ATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTA 2700 GACTTGGTTGAGTACTCACC;~GTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGA 2820 GAATTATGCAGTGCTGCCAT;~ACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACA 2880 ACGATGCCTGTAGCAATGGCi~ACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACT 3060 CTAGCTTCCCGGCAACAATTi3ATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTT 3120 CTGCGCTCGGCCCTTCCGGC'CGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGT 3180 GGGTCTCGCGGTATCATTGCi~GCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTT 3240 AAAAAACCACCGCTACCAGCC~GTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTT 3600 TAGTTAGGCCACCACTTCAAC~AACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATC 3720 CTGTTACCAGTGGCTGCTGC<:AGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGA 3780 AGCTTGGAGCGAACGACCTAC:ACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGC 3900 GGAGAGCGCACGAGGGAGCT7.'CCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGG 4020 TGAGCTGATACCGCTCGCCGC:AGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAA 4260 (2) INFORMATION
FOR
SEQ
ID N0:8:

(i) S EQUENCE
CHARACTERISTICS:

(A) LENGTH::3822 base pairs (B) TYPE:
nucleic acid (C) STRANDED1JESS:
single (D) TOPOLOGY:
linear (ii) MOLECULE
TYPE:
DNA
(genomic) (xi) EQUENCE
S DESCRIPTION:
SEQ ID
N0:8:

TAAGCACAAGTTTTATCCGGC:CTTTATTCACATTCTTGCCCGCCTGATGA ATGCTCATCC420 GGAATTCCGTATGGCAATGAP.AGACGGTGAGCTGGTGATATGGGATAGTG TTCACCCTTG480 GGCCTATTTCCCTAAAGGGT7.'TATTGAGAATATGTTTTTCGTCTCAGCCA ATCCCTGGGT660 CACCATGGGCAAATATTATAC:GCAAGGCGACAAGGTGCTGATGCCGCTGG CGATTCAGGT780 TACCACATTTGTAGAGGTTTTACTTGCTTTP.AAAAACCTCCCACACCTCC CCCTGAACCT960 TTGTGGTTTGTCCAAACTCA'rCAATGTATCTTATCATGTCTGGATCCCCCGGAATTCACT 1140 GGCCGTCGTTTTACAACGTCGTGAC'rGGGAAAACCCTGGCGTTACCCAACTTAATCGCCT 1200 TTCCCAACAGTTGCGCAGCC'rGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTAC 1320 GCATCTGTGCGGTATTTCACi~CCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGC 1380 ACACCCGCCAACACCCGCTGi~CGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTA 1500 GAAACGCGCGAGGCAGTTCT'CGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGG 1620 GCGGAACCCCTATTTGTTTA'~TTTTCTAAATACATTCAAATATGTATCCGCTCATGAGAC 1740 AATAACCCTGATAAATGCTTC:AATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATT 1800 TCCGTGTCGCCCTTATTCCC'.CTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAG 1860 TGATGAGCACTTTTAAAGTTC:TGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGC 2040 CACTGGGGCCAGATGGTAAGC:CCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG 2580 GAGCGCAGATACCAAATACT(iTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGA 3000 ACTCTGTAGCACCGCCTACA'PACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCA 3060 CAGGGGGAAACGCCTGGTAT(:TTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGC 3360 GTCGATTTTTGTGATGCTCG'.CCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGG 3420 CCCCTGATTCTGTGGATAACC:GTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCA 3540 GCCGAACGACCGAGCGCAGCCiAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCAATACGCAA 3600 CCAGGCTTTACACTTTATGC7.'TCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACA 3780 (2) INFORMATION FOR SEQ ID N0:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 41009 base pairs (B) TYPE: nucleic acid (C) STRANDEDDIESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE
DESC1~IPTION:
SEQ ID
N0:9:

GTCTATATF1AGCAGAGCTCG'PTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGC 120 GGCCAGCTGTTGGGCTCGCGc;TTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGG 240 ATCGACCGGATCGGAAAACC'CCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGTCTA 360 ACTGGATATACCACCGTTGA'CATATCCCAATGGCATCGTAAAGF1ACATTTTGAGGCATTT 480 GATAGTGTTCACCCTTGTTA(:ACCGTTTTCCATGAGCF1AACTGF1AACGTTTTCATCGCTC 720 TGGAGTGAATACCACGACGA~'TTCCGGCAGTTTCTACACATATATTCGCAAGATGTGGCG 780 TGTTACGGTGF1AAACCTGGC<:TATTTCCCTAAAGGGTTTATTGAGAATATGTTTTTCGTC 840 TTCTTCGCCCCCGTTTTCAC<:ATGGGCF1AATATTATACGCAAGGCGACAAGGTGCTGATG 960 F1ATGAATTACAACAGTACTGC;GATGAGTGGCAGGGCGGGGCGTAACCGCGGCGTGATTAA 1080 ATTCACTGGCCGTCGTTTTAC;AACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTA 1380 ATCGCCTTGCAGCACATCCCC;CCTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCG 1440 ATCGCCCTTCCCF1ACAGTTGC;GCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTC 1500 CTGATGCCGCATAGTTF:AGCC:AGTACACTCCGCTATCGCTACGTGACTGGGTCATGGCTG 1620 CGCCCCGACACCCGCCAACAC;CCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCAT 1680 CCGCTTACAGACAAGCTGTGi'~CCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGT 1740 CATCACCGAAACGCGCGAGGCAGTTCT'PGAAGACGAAAGGGCCTCGTGATACGCCTATTT 1800 TTATAGGTTAATGTCATGATi~ATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGA 1860 AATGTGCGCGGAACCCCTAT'PTGTTTA'rTTTTCTAAATACATTCAAATATGTATCCGCTC 1920 ATGAGACAATAACCCTGATAi~ATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATT 1980 CAACATTTCCGTGTCGCCCT'CATTCCC'rTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCT 2040 TACATCGAACTGGATCTCi~ACAGCGGTi~AGATCCTTGAGAGTTTTCGCCCCGAAGAACGT 2160 TTTCCAATGATGAGCACTTT'CAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGAC 2220 GCCGGGCAAGAGCAACTCGG'CCGCCGCi~TACACTATTCTCAGAATGACTTGGTTGAGTAC 2280 TCACCAGTCACAGAAAAGCA'PCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCT 2340 GCCATAACCATGAGTGATAA(;ACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCG 2400 AAGGAGCTAACCGCTTTTTT(~CACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGG 2460 GAACCGGAGCTGAATGAAGC(:ATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCA 2520 CAATTAATAGACTGGATGGACiGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTT 2640 CCGGCTGGCTGGTTTATTGC'.CGATAAA'CCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATC 2700 ATTGCAGCACTGGGGCCAGA'.CGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGG 2760 AAGCATTGGTAACTGTCAGA(:CAAGTT'.CACTCATATATACTTTAGATTGATTTAAAACTT 2880 CATTTTTAATTTAAAAGGAT(:TAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC 2940 CCTTAACGTGAGTTTTCGTT(:CACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCT 3000 TCTTGAGATCCTTTTTTTCT<~CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTA 3060 CCAGCGGTGGTTTGTTTGCCCiGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGC 3120 TTCAGCAGAGCGCAGATACC~1AATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCAC 3180 TTCAAGAACTCTGTAGCACCC~CCTACA'.CACCTCGCTCTGCTAATCCTGTTACCAGTGGCT 3240 GCTGCCAGTGGCGATAAGTCC~TGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGAT 3300 AAGGCGCAGCGGTCGGGCTG~~P.CGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACG 3360 ACCTACACCGAACTGAGATAC:CTACAGC:GTGAGCATTGAGAAAGCGCCACGCTTCCCGAA 3420 GGGAGAAAGGCGGACAGGTA'rCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGG 3480 AACGCGGCCTTTTTACGGTTCCTGGCC'rTTTGCTGGCCTTTTGCTCACATGTTCTTTCCT 3660 TACGCAAACCGCCTCTCCCCc~CGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGT 3840 TTCCCGACTGGAAAGCGGGCe~GTGAGCGCAACGCAATTAATGTGAGTTACCTCACTCATT 3900 AGGCACCCCAGGCTTTACAC'PTTATGC'rTCCGGCTCGTATGTTGTGTGGAATTGTGAGCG 3960 GATAACAATTTCACACAGGA~~ACAGCTATGACCATGATTACGCCAAGCT 4009 (2) INFORMATION FOR SEQ ID N0:10:
( i ) SEQUENCE CHARACTERIS'PICS
(A) LENGTH: :3955 baae pairs (B) TYPE: nucleic acid (C) STRANDEDtdESS: single (D) TOPOLOGY.; linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:10:
TAATGTCGTAACAACTCCGC(:CCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAG 60 TGCGAGGGCCAGCTGTTGGG(~TCGCGG'.CTGAGGACAAACTCTTCGCGGTCTTTCCAGTAC 180 TCTTGGATCGGAAACCCGTCC~GCCTCC(iAACGGTACTCCGCCACCGAGGGACCTGAGCGA 240 GTCCGCATCGACCGGATCGG~~AAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCA 300 AGTCTAGAGGATCTGAGCTTC~GCGAGA~PTTTCAGGAGCTAAGGAAGCTAAAATGGAGAAA 360 AAAATCACTGGATATACCAC<:GTTGATATATCCCAATGGCATCGTAAAGAACATTTTGAG 420 GCATTTCAGTCAGTTGCTCA~~.TGTACC'.CATAACCAGACCGTTCAGCTGGATATTACGGCC 480 GCCCGCCTGATGAATGCTCA7.'CCGGAA':CTCCGTATGGCAATGAAAGACGGTGAGCTGGTG 600 ATATGGGATAGTGTTCACCC'rTGTTACACCGTTTTCCATGAGCAAACTGAAACGTTTTCA 660 GACAACTTCTTCGCCCCCGT'PTTCACCATGGGCAAATATTATACGCAAGGCGACAAGGTG 900 GATTAATCAGCCATACCACA'PTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACC 1080 CTTATAATGGTTACAAATAAi~GCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTT 1200 CACTGCATTCTAGTTGTGGT'PTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATCC 1260 CCCGGAATTCACTGGCCGTCc~TTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCC 1320 AACTTAATCGCCTTGCAGCA(~ATCCCCCCTTCGCCAGCTGGCGTAATAGCGAAGAGGCCC 1380 GCACCGATCGCCCTTCCCAA(:AGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGT 1440 ATTTTCTCCTTACGCATCTG'CGCGGTA'PTTCACACCGCATATGGTGCACTCTCAGTACAA 1500 TCTGCTCTGATGCCGCATAG'CTAAGCCi~GTACACTCCGCTATCGCTACGTGACTGGGTCA 1560 TGGCTGCGCCCCGACACCCG(:CAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCC 1620 CACCGTCATCACCGAAACGC(~CGAGGCAGTTCTTGAAGACGAAAGGGCCTCGTGATACGC 1740 CTATTTTTATAGGTTAATGT(:ATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTT 1800 CGGGGAAATGTGCGCGGAAC(:CCTATT'CGTTTATTTTTCTAAATACATTCAAATATGTAT 1860 CCGCTCATGAGACAATAACC(:TGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATG 1920 AGTATTCAACATTTCCGTGT(:GCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTT 1980 TTTGCTCACCCAGAAACGCTC~GTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGA 2040 GTGGGTTACATCGAACTGGA'..'CTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAA 2100 GAACGTTTTCCAATGATGAGC:ACTTTT~~AAGTTCTGCTATGTGGCGCGGTATTATCCCGT 2160 ATTGACGCCGGGCAAGAGCA~3CTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTT 2220 GAGTACTCACCAGTCACAGA~~AAGCAT(:TTACGGATGGCATGACAGTAAGAGAATTATGC 2280 AGTGCTGCCATAACCATGAG7.'GATAACACTGCGGCCAACTTACTTCTGACAACGATCGGA 2340 GGACCGAAGGAGCTAACCGC'rTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGAT 2400 CGTTGGGAACCGGAGCTGAA'PGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCT 2460 GCCCTTCCGGCTGGCTGGTT'rATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGC 2640 CTGATTAAGCATTGGTAACT(~TCAGACCAAGTTTACTCATATATACTTTAGATTGATTTA 2820 AAACTTCATTTTTAATTTAAi~AGGATC'rAGGTGAAGATCCTTTTTGATAATCTCATGACC 2880 AAAATCCCTTAACGTGAGTT'PTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAA 2940 GGATCTTCTTGAGATCCTTT'CTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCA 3000 CCGCTACCAGCGGTGGTTTG'CTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTA 3060 ACTGGCTTCAGCAGAGCGCA(~ATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGC 3120 CACCACTTCAAGAACTCTGTi~GCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCA 3180 GTGGCTGCTGCCAGTGGCGA'PAAGTCG'PGTCTTACCGGGTTGGACTCAAGACGATAGTTA 3240 CCGGATAAGGCGCAGCGGTC(~GGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAG 3300 CGAACGACCTACACCGAACT(~AGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTT 3360 CCCGAAGGGAGAAAGGCGGA(:AGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGC 3420 ACGAGGGAGCTTCCAGGGGGAAACGCC'~GGTATCTTTATAGTCCTGTCGGGTTTCGCCAC 3480 CTCTGACTTGAGCGTCGATT'.PTTGTGA'CGCTCGTCAGGGGGGCGGAGCCTATGGAAAAAC 3540 GCCAGCAACGCGGCCTTTTTACGGTTC(:TGGCCTTTTGCTGGCCTTTTGCTCACATGTTC 3600 TTTCCTGCGTTATCCCCTGA'.~TCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGAT 3660 ACCGCTCGCCGCAGCCGAAC(~ACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAG 3720 CGCCAATACGCAAACCGCCT<:TCCCCG(:GCGTTGGCCGATTCATTAATGCAGCTGGCACG 3780 CTCATTAGGCACCCCAGGCT~:TACACT'.CTATGCTTCCGGCTCGTATGTTGTGTGGAATTG 3900 TGAGCGGATAACAATTTCAC~~CAGGAAACAGCTATGACCATGATTACGCCAAGCT 3955 (2) INFORMATION FOR SEQ ID NO:11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3861 base pairs (B) TYPE: nu~~leic acid (C) STRANDEDIVESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC1~IPTION: SEQ ID NO:11:
TAATGTCGTAACAACTCCGC(:CCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAG 60 GTCTATATAAGCAGAGCTCG'CTTAGTGAACCGTCTGCAGACTCTCTTCCGCATCGCTGTC 120 TGCGAGGGCCAGCTGTTGGG(~TCGCGG'PTGAGGACAAACTCTTCGCGGTCTTTCCAGTAC 180 TCTTGGATCGGAAACCCGTC(3GCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGA 240 AGTCTAGAGGATCTGAGCTT(~GCGAGATTTTCAGGAGCTAAGGAAGCTAAAATGGAGAAA 360 AAAATCACTGGATATACCAC(:GTTGATATATCCCAATGGCATCGTAAAGAACATTTTGAG 420 GCATTTCAGTCAGTTGCTCAATGTACC'~ATAACCAGACCGTTCAGCTGGATATTACGGCC 480 GCCCGCCTGATGAATGCTCA~CCCGGAA'CTCCGTATGGCAATGAAAGACGGTGAGCTGGTG 600 ATATGGGATAGTGTTCACCC'.CTGTTACACCGTTTTCCATGAGCAAACTGAAACGTTTTCA 660 TCGCTCTGGAGTGAATACCA(:GACGAT'.CTCCGGCAGTTTCTACACATATATTCGCAAGAT 720 GTGGCGTGTTACGGTGAAAA(:CTGGCC'CATTTCCCTAAAGGGTTTATTGAGAATATGTTT 780 TTCGTCTCAGCCAATCCCTGC~GTGAGT'.CTCACCAGTTTTGATTTAAACGTGGCCAATATG 840 GACAACTTCTTCGCCCCCGT'..'TTCACCATGGGCAAATATTATACGCAAGGCGACAAGGTG 900 CTGATGCCGCTGGCGATTCAC~GTTCATC:ATGCCGTCTGTGATGGCTTCCATGTCGGCAGA 960 TGTTGTTGTTAACTTGTTTA7.'TGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCAC 1080 AAATTTCACAAATAAAGCAT7.'TTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCAT 1140 CAATGTATCTTATCATGTCTC~GATCCCCCGGAATTCACTGGCCGTCGTTTTACAACGTCG 1200 TGACTGGGAAAACCCTGGCG'rTACCCAACTTAATCGCCTTGCAGCACATCCCCCCTTCGC 1260 CAGCTGGCGTAATAGCGAAG.~GGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCT 1320 GAATGGCGAATGGCGCCTGA'TGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACA 1380 CGCGCCCTGACGGGCTTGTC'rGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTC 1560 GAAGACGAAAGGGCCTCGTGi~TACGCC'rATTTTTATAGGTTAATGTCATGATAATAATGG 1680 TTTCTTAGACGTCAGGTGGCi~CTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTAT 1740 AATAATATTGAAAAAGGAAGi~GTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCT 1860 TTTTTGCGGCATTTTGCCTT(~CTGTTT'rTGCTCACCCAGAAACGCTGGTGAAAGTAAAAG 1920 ATGCTGAAGATCAGTTGGGTGCACGAG'PGGGTTACATCGAACTGGATCTCAACAGCGGTA 1980 AGATCCTTGAGAGTTTTCGC(:CCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTC 2040 TGCTATGTGGCGCGGTATTA'CCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCA 2100 TACACTATTCTCAGAATGAC'CTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGG 2160 ATGGCATGACAGTAAGAGAA'L'TATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGG 2220 TGGGGGATCATGTAACTCGC(:TTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAA 2340 ACGACGAGCGTGACACCACGATGCCTG'.CAGCAATGGCAACAACGTTGCGCAAACTATTAA 2400 CTGGCGAACTACTTACTCTA(~CTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATA 2460 AAGTTGCAGGACCACTTCTG(:GCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAAT 2520 CTGGAGCCGGTGAGCGTGGG~'CTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGC 2580 CCTCCCGTATCGTAGTTATC~.'ACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATA 2640 GACAGATCGCTGAGATAGGTC~CCTCAC'.CGATTAAGCATTGGTAACTGTCAGACCAAGTTT 2700 AGATCCTTTTTGATAATCTC~1TGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAG 2820 CGTCAGACCCCGTAGAAAAG~1TCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAA 2880 TCTGCTGCTTGCAAACAAAA~~AACCAC(:GCTACCAGCGGTGGTTTGTTTGCCGGATCAAG 2940 AGCTACCAACTCTTTTTCCG.AAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTG 3000 ACCTCGCTCTGCTAATCCTG'TTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTA 3120 CCGGGTTGGACTCAAGACGA'TAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGG 3180 GTTCGTGCACACAGCCCAGC'TTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC 3240 GTGAGCATTGAGAAAGCGCC;~CGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAA 3300 TTTATAGTCCTGTCGGGTTTCGCCACC'TCTGACTTGAGCGTCGATTTTTGTGATGCTCGT 3420 TTTGCTGGCCTTTTGCTCACATGTTCT'TTCCTGCGTTATCCCCTGATTCTGTGGATAACC 3590 AGTCAGTGAGCGAGGAAGCGc;AAGAGCGCCAATACGCAAACCGCCTCTCCCCGCGCGTTG 3660 GCCGATTCATTAATGCAGCTc~GCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCG 3720 CAACGCAATTAATGTGAGTT~~CCTCAC'rCATTAGGCACCCCAGGCTTTACACTTTATGCT 3780 TCCGGCTCGTATGTTGTGTGc~AATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTA 3840 TGACCATGATTACGCCAAGC'P 3861 (2) INFORMATION FOR SES2 ID N0:12:
( i ) SEQUENCE CHAR~~CTERIS'PICS
(A) LENGTH: 3888 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDLdESS: single (D) TOPOLOGY:: linear_ (ii) MOLECULE TYPE;. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:12:
TAATGTCGTA ACAACTCCGC C:CCGTTGACG CAAATGGGCG GTAGGCGTGT ACGGTGGGAG 60 GTCTATATAA GCAGAGCTCG 7.'TTAGTGAAC CGTCTGCAGA CTCTCTTCCG CATCGCTGTC 120 TGCGAGGGCC AGCTGTTGGG C:TCGCGGTTG AGGACAAACT CTTCGCGGTC TTTCCAGTAC 180 TCTTGGATCG GAAACCCGTC C~GCCTCCGAA CGGTACTCCG CCACCGAGGG ACCTGAGCGA 240 GTCCGCATCGACCGGATCGG.~AP.ACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCA 300 GCATTTCAGTCAGTTGCTCAATGTACC'TATAACCAGACCGTTCAGCTGGATATTACGGCC 480 TTTTTAAAGACCGTAAAGAA:~AATAAGCACAAGTTTTATCCGGCCTTTATTCACATTCTT 540 GCCCGCCTGATGAATGCTCA'TCCGGAA'TTCCGTATGGCAATGAAAGACGGTGAGCTGGTG 600 ATATGGGATAGTGTTCACCC'TTGTTACACCGTTTTCCATGAGCAAACTGAAACGTTTTCA 660 TCGCTCTGGAGTGAATACCA(~GACGAT'TTCCGGCAGTTTCTACACATATATTCGCAAGAT 720 GTGGCGTGTTACGGTGAAAAc~CTGGCC'TATTTCCCTAAAGGGTTTATTGAGAATATGTTT 780 TTCGTCTCAGCCAATCCCTGGGTGAGT'TTCACCAGTTTTGATTTAAACGTGGCCAATATG 840 GACAACTTCTTCGCCCCCGT'PTTCACCATGGGCAAATATTATACGCAAGGCGACAAGGTG 900 ATGCTTAATGAATTACAACA(~TACTGCGATGAGTGGCAGGGCGGGGCGTAACCGCGGAAT 1020 TGTTGTTGTTAACTTGTTTA'~TGCAGC'TTATAATGGTTACAAATAAAGCAATAGCATCAC 1080 AAATTTCACAAATAAAGCAT'.CTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCAT 1190 CAATGTATCTTATCATGTCT(~GATAACGCCCAAAAACCCGGGGACGATGATCCCCCGGAA 1200 TTCACTGGCCGTCGTTTTACAACGTCG'CGACTGGGAAAACCCTGGCGTTACCCAACTTAA 1260 TCGCCTTGCAGCACATCCCC(~CTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGA 1320 TCGCCCTTCCCAACAGTTGCC~CAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCT 1380 CCTTACGCATCTGTGCGGTATTTCACA(:CGCATATGGTGCACTCTCAGTACAATCTGCTC 1440 TGATGCCGCATAGTTAAGCCAGTACAC'.CCCGCTATCGCTACGTGACTGGGTCATGGCTGC 1500 GCCCCGACACCCGCCAACAC(:CGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATC 1560 CGCTTACAGACAAGCTGTGA(:CGTCTC(:GGGAGCTGCATGTGTCAGAGGTTTTCACCGTC 1620 ATCACCGAAACGCGCGAGGC~~GTTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTT 1680 ATGTGCGCGGAACCCCTATT7.'GTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCA 1800 TGAGACAATAACCCTGATAA~~.TGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTC 1860 AACATTTCCGTGTCGCCCTT~~.TTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTC 1920 ACCCAGAAACGCTGGTGAAAC~TAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTT 1980 ACATCGAACTGGATCTCAAC.AGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTT 2040 TTCCAATGATGAGCACTTTT.AAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACG 2100 CACCAGTCACAGAAAAGCAT~~TTACGG.ATGGCATGACAGTAAGAGAATTATGCAGTGCTG 2220 CCATAACCATGAGTGATAAC.~1CTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA 2280 AACCGGAGCTGAATGAAGCC;ATACCAA..~1CGACGAGCGTGACACCACGATGCCTGTAGCAA 2400 ATTTTTAATTTAAAAGGATC'PAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCC 2820 CTTAACGTGAGTTTTCGTTC(:ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTT 2880 CTTGAGATCCTTTTTTTCTG(;GCGTAA'PCTGCTGCTTGCAAACAAAAAAACCACCGCTAC 2940 CAGCGGTGGTTTGTTTGCCG(~ATCAAGi~GCTACCAACTCTTTTTCCGAAGGTAACTGGCT 3000 TCAGCAGAGCGCAGATACCAAATACTG'tCCTTCTAGTGTAGCCGTAGTTAGGCCACCACT 3060 TCAAGAACTCTGTAGCACCG(;CTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTG 3120 CTGCCAGTGGCGATAAGTCG'.CGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATA 3180 CCTACACCGAACTGAGATACC:TACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAG 3300 GGAGAAAGGCGGACAGGTAT(:CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGG 3360 AGCTTCCAGGGGGAAACGCCTGGTATC'PTTATAGTCCTGTCGGGTTTCGCCACCTCTGAC 3420 TTGAGCGTCGATTTTTGTGA~.'GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCA 3480 ACGCGGCCTTTTTACGGTTCC:TGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTG 3540 GCCGCAGCCGAACGACCGAG(:GCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCAAT 3660 ACGCAAACCGCCTCTCCCCGC:GCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTT 3720 TCCCGACTGGAAAGCGGGCA~TGAGCGCAACGCAATTAATGTGAGTTACC TCACTCATTA3780 GGCACCCCAGGCTTTACACTrTATGCTTCCGGCTCGTATGTTGTGTGGAA TTGTGAGCGG3840 ATAACAATTTCACACAGGAA.?~CAGCTATGACCATGATTACGCCAAGCT 3888 (2) INFORMATION FOR SEQ ID N0:13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '7379 base pairs (B) TYPE: nu~~leic acid (C) STRANDEDIVESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC1~IPTION: SEQ ID N0:13:
TAATGTCGTAACAACTCCGCc~CCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAG 60 GTCTATATAAGCAGAGCTCG'CTTAGTGAACCGTCTGCAGACTCTCTTCCGCATCGCTGTC 120 TGCGAGGGCCAGCTGTTGGG(:TCGCGG'PTGAGGACAAACTCTTCGCGGTCTTTCCAGTAC 180 GTCCGCATCGACCGGATCGG~~.AAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCA 300 AGTCTAGAGGATCTGAGCTT(~GCGAGA'PTTTCAGGAGCTAAGGAAGCTAAAATGGAGAAA 360 AAAATCACTGGATATACCAC(:GTTGATRTATCCCAATGGCATCGTAAAGAACATTTTGAG 420 GCATTTCAGTCAGTTGCTCA~~TGTACC'CATAACCAGACCGTTCAGCTGGATATTACGGCC 480 GCCCGCCTGATGAATGCTCA'.~.'CCGGAATTCCGTATGGCAATGAAAGACGGTGAGCTGGTG 600 ATATGGGATAGTGTTCACCC~.'TGTTACRCCGTTTTCCATGAGCAAACTGAAACGTTTTCA 660 TCGCTCTGGAGTGAATACCA<:GACGAT'.CTCCGGCAGTTTCTACACATATATTCGCAAGAT 720 GTGGCGTGTTACGGTGAAAA<:CTGGCC'.CATTTCCCTAAAGGGTTTATTGAGAATATGTTT 780 GACAACTTCTTCGCCCCCGT7.'TTCACCATGGGCAAATATTATACGCAAGGCGACAAGGTG 900 TGTTGTTGTTAACTTGTTTA'PTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCAC1080 AAATTTCACAAATAAAGCAT'TTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCAT1140 CAATGTATCTTATCATGTCT~~GATAACGCCCAAAAACCCGGGGCGCCGGCCAAAAGTCCG1200 CGGAACTCGCCCTGTCGTAA:?~ACCACGCCTTTGACGTCACTGGACATTCCCGTGGGAACA1260 AATTTAAGCCACACCTCTTTGTCCTGT.ATATTATTGATGATGGGGGGATCCACTAGTTCT1380 TCCACACAACATACGAGCCGGAAGCAT:~AAGTGTAAAGCCTGGGGTGCCTAATGAGTGAG1560 CTAACTCACATTAATTGCGT'rGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTG1620 TTCCGCTTCCTCGCTCACTGi~CTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATC1740 AGCTCACTCAAAGGCGGTAA'PACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAA1800 CATGTGAGCAAAAGGCCAGCi~AAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTT1860 TTTCCATAGGCTCCGCCCCC(:TGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTG1920 GCGAAACCCGACAGGACTATi~AAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCG1980 CTCTCCTGTTCCGACCCTGC(~GCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAG2040 CGTGGCGCTTTCTCATAGCT(~ACGCTG'rAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTC2100 CAAGCTGGGCTGTGTGCACG~~ACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAA2160 CTATCGTCTTGAGTCCAACC(:GGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGG2220 TAACAGGATTAGCAGAGCGA(~GTATGTi~GGCGGTGCTACAGAGTTCTTGAAGTGGTGGCC2280 TAACTACGGCTACACTAGAAGGACAGTi~TTTGGTATCTGCGCTCTGCTGAAGCCAGTTAC2340 CTTCGGAAAAAGAGTTGGTA(~CTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGG2400 CATGAGATTATCAAAAAGGA':.'CTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAA2580 ATCAATCTAAAGTATATATG~~GTAAAC'.PTGGTCTGACAGTTACCAATGCTTAATCAGTGA2640 GGCACCTATCTCAGCGATCTC~TCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGT2700 GTAGATAACTACGATACGGG.AGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCG 2760 AGACCCACGCTCACCGGCTC~~AGATTT.ATCAGCAATAAACCAGCCAGCCGGAAGGGCCGA 2820 CATCGTGGTGTCACGCTCGT~~GTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATC 3000 GATCGTTGTCAGAAGTAAGT'rGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCA 3120 GGATAATACCGCGCCACATAGCAGAAC'PTTAAAAGTGCTCATCATTGGAAAACGTTCTTC 3300 GGGGCGAAAACTCTCAAGGA'rCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCG 3360 TGCACCCAACTGATCTTCAGc:ATCTTT'rACTTTCACCAGCGTTTCTGGGTGAGCAAAAAC 3420 AGGAAGGCAAAATGCCGCAAi~AAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCAT 3480 ACTCTTCCTTTTTCAATATTi~TTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATA 3540 CATATTTGAATGTATTTAGAi~AAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAA 3600 AGTGCCACCTGGGAAATTGTi~AACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGT 3660 GAACCATCACCCTAATCAAG'CTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAAC 3900 CCTAAAGGGAGCCCCCGATT'.CAGAGCT'CGACGGGGAAAGCCGGCGAACGTGGCGAGAAAG 3960 GAAGGGAAGAAAGCGAAAGG~~GCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTG 4020 CGCGTAACCACCACACCCGC(:GCGCTTAATGCGCCGCTACAGGGCGCGTCGCGCCATTCG 4080 CCATTCAGGCTGCGCAACTG~.'TGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGC 4140 CAGCTGGCGAAAGGGGGATG!.'GCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCC 4200 CAGTCACGACGTTGTAAAACC~ACGGCCAGTGAATTGTAATACGACTCACTATAGGGCGAA 4260 TTGGGTACCGGGCCCCCCCTC:GAGGTCGACGGTGCCCCCAGCAGAAGTATCGACTGCATG 4320 CTAATTATTAACAAACCAAA~~GGCGTTGCCACTTACACCCTTACCTTTAGGTTTTTAAAC 4380 TTTAACAGACTAAGCGGAGG7.'ACCCTGTTTAAAACTGATGTCTTAACCTTTACCTATGTA 4440 CAAAACATAGGTATTAGTTA.ACAGTTCATTTGGGCTATAATAATATACATTTTCTTGGGT 4620 CACGGTGGGTTCAATCTAAA.zIATGAAGAi~ACGCTGTTGAGGTTCACTAAGCACAGGTTTT 4740 GAATCTGTCGGCAGCGTCCA'TGCATCATAGCTTGTCTCAAAGCAGATTGTCTTCTTTCCT 4800 CTGCCTTGGAAGTGGTTTGG'TGAAGCACTACAGGTGTCTTTTCAACCTCTTTCAGCACCC 4860 GCTCTATTACAGATCTCACC~~ACACAGCACAGTTTTTAAGAGAACAATAGTTTTGAAGGC 4920 TACAAGATTTACACTTAAGC;ACCAGCCAGTAATTATAAGTGCTTTTAAGAACTACCCCTA 4980 GCTCAGGGTTAATGCACCTT'PTAATGGCCTCCATGCAGGCTTTATGGACAGTTCTAAAAA 5040 AAGACAGTCTAAAATAAATG'PAGTGAG'TGTTTCTAAATATAATACTCCCCACATAGTTAA 5100 TTTCATCAGGCCTGCTAGAA'rTTACAAACTCTCGGTACCACATATACTTTTTATTCATAG 5160 CCCCACCCTTAATAAAGTCC'rCAATCACTTTCTGAACCACATGCTTGCTAGCCATGCATT 5220 AGCAGTGCTTAGTTACTATCi~ACTCAA'PACCCGCATTGCATGTAAACCCCCCAAAGAGCA 5340 GTTTTTCATGCCTGTGTAGCi~CATCATCCCACAAAATAGGAATTTCATAGCATAAAGCAA 5400 AGCAATTACAATATTTAGGAi~CTCTCACCACAGCAGTCACGTGACATGTTGTCTCAGCAG 5460 TGCAGTTGCCTTCCATCCTA(:AATTATGAACAAAAACTAAACACTTCTAACAAAGATACA 5520 GTGACAATCTCCCTTCCTCTi~AAAGCA'PTGTTTACATTAGGGTGATTATTAACAACGTCA 5580 GAAATTTCTTTAATTAAAGTt~CCTTTAAAATGTGCAAGAGCATCATCATACTCAAAACCA 5640 AGCTGAGAGTAAAAGACCAC(:TTAAAAGTAATCCCAGGCTTGTTTTTATCAACAGCCTTA 5700 AACATGCTTTCACAAAATAT~~GAAGCAGTAACATCATCAATGGTGTCGAAGAGAAACTCC 5760 ATAGGAGACTCCAGCATTGA'.CCCAAGC'CCTCTAACAAAATCTTCCTCAAAATGAATAATG 5820 CCCTTTACACAAACGCGGGGC:AGACGA'CGGTGGGCCATCGCGTCAACCTGi~AACACATTT5880 TACAGTAAACAAAGCTAGCT<:CGCAGTGGTAAAGTCATGCCCATGGGTGAGGCCAAAATC 5940 CTTAAAAAAGCTATCTAAGT~~GTTGGTCATCCCCTCAGTTAAAAAGTTTTGCAGCTGGGT 6000 GGTGCATACCACATAGTGCC~~.GCTTATAGCTACAAAGACCTGCATCCCCTCCTTAGCAGA 6060 CAGCTCTTGCACACACGCAG7.'AACTAT(:CACCGCTTAAGAAAAGCTTTAAGCCCAGCGCA 6120 AGTAATAGTGAAGCAGAGGC;~TTTCAGACGAGGCTCACTAGCTGCAGTCGCCATTTATGA 6240 GGTCTGCAATAAAAAACAAC'TCATCAGCAGCTGAAAAAGTGCACTTTGACCTCATTAAGC 6300 CACTGCATATGCAAGTCCTC;~TCTATGCCGCAGCCCAGACCCTCAATCCAGCCCCGAATG 6360 TACACTTTAATAAGAGATTC~~1ACCTCTTCTTTTAGCAAAGTACACATGCTGTTTGGACTA 6420 CGAAGGTTAAAAATGGACTG'PAACAGCATTGAAACCCCGCGACACAGGTCAGTCTCGCGG 6540 TCTTGATCTCTTATTATAGCGACCAAA'rGGTCCTTCAGAGTGATGTTGCACTCATAGAAG 6600 TAGGCAGCTCCGGCAGCCAT'PCTGCAAAATAACAAAACACCACTAAGCATAGCACCATCA 6660 CCAAGCATGAAAACAGGTAAi~AACAAAAGCAACACTTACTTATTCAGCAGTCACAAGAAT 6720 GTTGGGCTCCCAAGTGACAGi~CAAGCC'TAATGCAAGGTGGGCACAGTCTCCGGAATAAGT 6780 TGACAAAAGTCACGCCGCAAAGCTTCC'rGAAGAGAAACGGCGGTAGCCTGGATATCTGCA 6840 AGGCTAAACACCACACGGCG(:AGTTCAGAAGGCAAAAAGTCTGTAAGCTCTAGCTGAGCA 7020 CACACACTCTCCACTAGACA(:TTGTGAAGCCTCAGACAAAAACATGCTCCCATAGACACT 7080 CCTAAAGCTGCCATTGTACT(:ACGGACGGCTGGCTGTCAGAGGAGAGCTATGAGGATGAA 7140 ATGTTAAGTGCAGTTTCCCT'.CTGGCGG'PTGGCCCGGAAAGTTCACAAAAAGTACAGCACG 7320 (2) INFORMATION FOR SEQ ID N0:14:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6243 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDPdESS: single (D) TOPOLOGY; linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:14:
GTCGACGGTGCCCCCAGCAG.AAGTATCGACTGCATGCTAATTATTAACAA 60 ACCAAAAGGC

CTGTTTAAAACTGATGTCTT.zIACCTTTACCTATGTAGGCGAAAATCAATAAAACCAGAAA 180 AAAATAAGTTTAAAAGCTTT.~TTTTTCATACACGCGAGCGGTAAGGCTGCCGCCTTCAGG 240 AAAAGTTACTCTGTAAACAG'TTCTTTCACAACAGCACAAAACATAGGTATTAGTTAACAG 300 TTCATTTGGGCTATAATAAT:?~TACATTTTCTTGGGTGGCAAAGCAAGGGTCGGTAATCTC 360 AACAAAACCATCAACTGGAA'PGCAAGAATAGTCCAGCACGGTGGGTTCAATCTAAAAATG 420 TCATAGCTTGTCTCAAAGCAGATTGTC'rTCTTTCCTCTGCCTTGGAAGTGGTTTGGTGAA 540 GCACTACAGGTGTCTTTTCAACCTCTT'PCAGCACCCGCTCTATTACAGATCTCACCCACA 600 CAGCACAGTTTTTAAGAGAACAATAGT'rTTGAAGGCTACAAGATTTACACTTAAGCACCA 660 GCCAGTAATTATAAGTGCTT'CTAAGAACTACCCCTAGCTCAGGGTTAATGCACCTTTTAA 720 TGGCCTCCATGCAGGCTTTA'PGGACAG'PTCTAAAAAAAGACAGTCTAAAATAAATGTAGT 780 CAAACTCTCGGTACCACATA'CACTTTT'PATTCATAGCCCCACCCTTAATAAAGTCCTCAA 900 TCACTTTCTGAACCACATGC'CTGCTAGCCATGCATTGTAAAGACAAGCTGTTAGAGCAGT 960 GACAGTGTACTCGCCACGTT'CGAGCCTCTGCCAGGCAGCAGTGCTTAGTTACTATCAACT 1020 CAATACCCGCATTGCATGTAP.ACCCCCCAAAGAGCAGTTTTTCATGCCTGTGTAGCACAT 1080 CATCCCACAAAATAGGAATT'CCATAGCATAAAGCAAAGCAATTACAATATTTAGGAACTC 1140 TCACCACAGCAGTCACGTGA(:ATGTTG'CCTCAGCAGTGCAGTTGCCTTCCATCCTACAAT 1200 TATGAACAAAAACTAAACAC'.CTCTAACAAAGATACAGTGACAATCTCCCTTCCTCTAAAA 1260 GCATTGTTTACATTAGGGTG~~TTATTAACAACGTCAGAAATTTCTTTAATTAAAGTGCCT 1320 TTAAAATGTGCAAGAGCATC~~TCATAC'.CCAAAACCAAGCTGAGAGTAAAAGACCACCTTA 1380 GCAGTAACATCATCAATGGTC~TCGAAGAGAAACTCCATAGGAGACTCCAGCATTGATCCA 1500 AGCTCTCTAACAAAATCTTCC:TCAAAATGAATAATGCCCTTTACACAAACGCGGGGCAGA 1560 CGATGGTGGGCCATCGCGTC~~ACCTGAAACACATTTTACAGTAAACAAAGCTAGCTCCGC 1620 CAGACGAGGCTCACTAGCTG~AGTCGCCATTTATGAGGTCTGCAATAAAAAACAACTCAT 1980 CAGCAGCTGAAAAAGTGCAC'PTTGACCTCATTAAGCCACTGCATATGCAAGTCCTCATCT 2040 ATGCCGCAGCCCAGACCCTC.?~ATCCAGCCCCGAATGTACACTTTAATAAGAGATTCAACC 2100 TCTTCTTTTAGCAAAGTACA~~ATGCTGTTTGGACTAGTATACACAATAGAAGTCACAATG 2160 AGGGGCCCGCTGTGGCTGGA;~AGCCTGCGCACAGCCCGAAGGTTAAAAATGGACTGTAAC 2220 TCCTGAAGAGAAACGGCGGTi~GCCTGGATATCTGCAACGGACCCAAAACCTTCAGTGTCA 2580 CTTCCAATAAACAGATAAAACTCTAAA'PAGTCCCCACTTAAAACCGAAACAGCCGCGGCA 2640 AAGGTAGGACACGGACGCAC'CTCCTGAGCCCTAATAAGGCTAAACACCACACGGCGCAGT 2700 TGAAGCCTCAGACAAAAACA'PGCTCCCATAGACACTCCTAAAGCTGCCATTGTACTCACG 2820 TCCTCAAAGTAGGGCGTGTG(~AAAACGi~AAAGGAATATAACGGGGCGTTTGAGGAAGTGG 2940 TGCCAAGTACAGTCATAAAA'.CGTGGGCGCGTGGTAAATGTTAAGTGCAGTTTCCCTTTGG 3000 CGGTTGGCCCGGAAAGTTCA(:AAAAAG'PACAGCACGTCCTTGTCACCGTGTCAACCACAA 3060 AACCACAAATAGGCACAACGC:CCAAAAACCCGGGTCGACACGCGTGAATTCACCGGTTCG 3120 CGAAACGCCCAAAAACCCGG(~GCGCCGGCCAAAAGTCCGCGGAACTCGCCCTGTCGTAAA 3180 ACCACGCCTTTGACGTCACTGGACATT(:CCGTGGGAACACCCTGACCAGGGCGTGACCTG 3240 AACCTGACCGTCCCATGACCC:CGCCCCTTGCAACACCCAAATTTAAGCCACACCTCTTTG 3300 TCCTGTATATTATTGATGATC~GGGGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTG 3360 ATAGCTGTTTCCTGTGTGAA.ATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGG 3480 GCGCTCACTGCCCGCTTTCC.AGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGG 3600 AAAGGCCAGGAACCGTAAAA:~GGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCC 3840 ACCCCCCGTTCAGCCCGACC(~CTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCC 4140 GGTAAGACACGACTTATCGCCACTGGC;~1GCAGCCACTGGTAACAGGATTAGCAGAGCGAG 4200 CTCTTGATCCGGCAAACAAA(:CACCGC'PGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCA 4380 GATTACGCGCAGAAAAAAAG(~ATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGA 4440 CGCTCAGTGGAACGAAAACT(:ACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGAT 4500 GTAAACTTGGTCTGACAGTT~~CCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTG 4620 TCTATTTCGTTCATCCATAG'('TGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGA 4680 GGGCTTACCATCTGGCCCCA(~TGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCC 4740 TTTATCCGCCTCCATCCAGT(:TATTAA'CTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCC 4860 GTTTGGTATGGCTTCATTCA(~CTCCGG'CTCCCAACGATCAAGGCGAGTTACATGATCCCC 4980 CATGTTGTGCAAAAAAGCGG~.'TAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTT 5040 GGCCGCAGTGTTATCACTCAi.'GGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCC 5100 TTGAAGCATTTATCAGGGTT.ATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAA 5520 AACGTTAATATTTTGTTAAA.~TTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAAC 5640 CAATAGGCCGAAATCGGCAA.~1ATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTG 5700 TTTTTGGGGTCGAGGTGCCG'rAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTT 5880 GCGCTTAATGCGCCGCTACAc;GGCGCGTCGCGCCATTCGCCATTCAGGCTGCGCAACTGT 6060 TGGGAAGGGCGATCGGTGCGc~GCCTCT'rCGCTATTACGCCAGCTGGCGAAAGGGGGATGT 6120 GCTGCAAGGCGATTAAGTTGc~GTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACG 6180 ACGGCCAGTGAATTGTAATAc~GACTCACTATAGGGCGAATTGGGTACCGGGCCCCCCCTC 6240 (2) INFORMATION FOR SE(,2 ID N0:15:
( i ) SEQUENCE CHAR~~.CTERI S'rICS
(A) LENGTH: t~612 base pairs (B) TYPE: nucleic acid (C) STRANDEDtdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE.. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:15:
AAGCTTTGCTCAACAAATAC'PGTCAAGGACTCGAGTCCGGCTCTGACTGAGCAATGTCTA 60 AAGAAATACCAACCCCTTAT;~TGTGGAGCTACCAACCGCAAACGGGACACGCCGGCGCCT 120 CCCAGGACTACTCCACCCAA;~1TGAATTGGTTTAGTGCTGGGCCATCAATGATTAGTCAAG 180 CAGTGCTGGCTGGCACGGGCATTCAGC'PTAGCGAAGACATCCCCAGCGCCTCCTGGATCA 480 CATTCCTCACCCTGCAACAGcsCATCCTCGACGCCGCGCGCAGGAGGCGTGGGCACCTACC 600 AGTTTGTGCGCGAATTTGTG(~CAGAGG'PATACCTTAACCCTTTTTCAGGACCACCGGACA 660 CCTTTCCTGATCAGTTCATT(JCTAACTACGACATTGTAACCAACTCTGTCGATGGCTATG 720 TGCACGCGCCCTCGCTGCTT'CGCAAAGGAGGGTTTATGTGCTAACTGGTTTTACAACCCA 840 GCACTTGCCTTTGAAGGGTT'CGATATTCCAGACTCTTACCAAGAGGGACACGGTGTGGAC 900 ATAGAAGTTAAGTGTTCCCA(:CACTCCAGCAAACTGTGCCACAATGGCCATGATATGATC 960 CCCCACATGAGCCTCATTGA(~GCAGCC'PGTTCTATGTATAACCTTAACTAGATAATATTA 1080 TTAAACTTGTTTTACAGCTA(:CACCATAATGCGCTTCAGCTTCTTCATCGCCGCCGTTCT 1140 TTTCTGCACCACAGGGGCCA(~CAATGACATTGTGACTTGCTGCGCCCACACACCTTGCCT 1200 CCTACACCTAGAAGTGGGCT~CGGGGGCCAATGTCAGTTGGATAAACTCTGACACAGGCCA 1260 GGCCCCGATTTGCCTCTCCA~~TGGCATGTGCAACGCTACCCAGCAAGGCCTGCAGTTTTC 1320 TGCAAACTTTTCTGAGGATGC~CCTGTACATCGCCCTCATTAAGGAGAGCAACTACGAGGG 1380 CGCTGAGCACTACTACCTTG~.'CTATAT'.PTATGGAGACTGCTACCAAACTGCAAATGAGTC 1440 TGCCCACGGGCCTATTTCCA<~GCCCCTCAAAGATCTGTTATTAGTGATATCAAAGATGGT 1500 CCGGTTCTTGTACTCGGGCC~1TATATTCATGTCCCCAGACATCATAGTCAGCACCATTTT 1560 CTTCTCCTTTTGCCAGTAGA7.'GCGAGTTTGTGCCAGCTCTTCAACAGAAACATTGTGACC 1620 1~~

TGATGTTCCCTGCCTCCGTGrGTGGCCCATACGCGTCCCTCAGCCTTCTAATGGGACTAA 1800 ACAACAAAATCAGGCCCATG'TAGCTTGTCAAATAAACTTACCTAATTTTTGCTAAGACGC 1860 GTGCGCGCGAGCCAGCTTGC~~GATGTGCTTGAAAGATAATGTGGTCTCTCCCAACAGCTT 1980 GCAAGGTCTCACTGAGTCACCCCCAGG,?~1ACCCTGGCTGTCAATGTTTCCCCTCCACTAAC 2160 CTTTTCTACGTTAGGTGCCA'rTAAACT'PTCCACAGGTCCCGGACTCACCCTCAACGAGGG 2220 TGAAAATGTCAACAAGGTTT'rGTCTTT'rACCTCCCCATTACATAAAAATGAAAACACTGT 2340 ATCCCTAGCGCTAGGAGATG(~GTTAGAAGATGAAAATGGCACCCTTAAAGTGACCTTCCC 2400 TACTCCCCCTCCCCCGCTACi~ATTCTCCCCTCCCCTCACAAAAACAGGTGGTACTGTTTC 2460 CTTGCCCCTGCAAGACTCCA'rGCAAGTGACAAATGGAAAACTGGGCGTTAAGCTACCACC 2520 TACGCACCTCCCTTGAAAAAi~ACTGACCAGCAAGTTAGCCTCCAAGTAGGCTCGGGTCTC 2580 ACCGTGATTAACGAACAGTTGCAAGCTc.;TCCAGCCTCCCGCAACCACCTACAACGAGCCT 2640 CTTTCCAAAACTGACAATTC'CGTTTCTCTGCAAGTAGGTGCCGGCCTTGCCGTGCAGAGC 2700 GGACGTTTGGTGGCAACCCC'CCCCCCGCCTCTCACCTTTACATCACCCCTAGAAAAAAAT 2760 GCTCTAAGTGCTGGAAGTGG'CTTAAGAATATCTGGAGGCAGCCTCACGGTGGCCACTGGA 2940 CCTGGCCTTTCCCATCAAAA'.CGGAACAATAGGGGCTGTAGTAGGTGCAGGCCTCAAGTTT 3000 GAAGCAACCCAACCCCCAGC'.iGCCCCCATAACACTGTGGACAGGGCCTGGCCTAGCATTA 3120 ATGGCTTTATGTAATGACAC~'CCAGTAATTAGGTCTTTATATGCCTAACCAGAGACAGCA 3180 ACTTAGTCACAGTAAATGCT~~GCTTTG'CGGGAGAGGGGGGGTATCGAATAGTCAGCCCTA 3240 CCCAGTCACAATTTAGCCTA~1TTATGGAGTTTGATCAGTTTGGACAGCTTATGTCCACAG 3300 CACGCCCAAGCCACACCTGG.~AACTGTGCATGCCTAACAGAGAAGTTTACTCCACTCCCG 3420 ATCGACTGCATGCTAATTAT'PAACAAACCAAAAGGCGTTGCCACTTACACCCTTACCTTT 3540 AGGTTTTTAAACTTTAACAG;~1CTAAGCGGAGGTACCCTGTTTAAAACTGATGTCTTAACC 3600 TTTACCTATGTAGGCGAAAA'PCAATAAAACCAGAAAAAAATAAGGGGAAAAGCTTGATAT 3660 CCAGCTTTTGTTCCCTTTAG'PGAGGGTTAATTCCGAGCTTGGCGTAATCATGGTCATAGC 3780 TGTTTCCTGTGTGAAATTGT'PATCCGC'rCACAATTCCACACAACATACGAGCCGGAAGCA 3840 CACTGCCCGCTTTCCAGTCGGGAAACC'PGTCGTGCCAGCTGCATTAATGAATCGGCCAAC 3960 TGCGCTCGGTCGTTCGGCTG(~GGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGT 4080 TATCCACAGAATCAGGGGATi~ACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGG 4140 CCAGGAACCGTAAAAAGGCC(~CGTTGC'PGGCGTTTTTCCATAGGCTCCGCCCCCCTGACG 4200 AGCATCACAAAAATCGACGC'CCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGAT 4260 ACCAGGCGTTTCCCCCTGGA~~GCTCCC'PCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTA 4320 CCGGATACCTGTCCGCCTTT(:TCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCT 4380 GTAGGTATCTCAGTTCGGTG'PAGGTCG'PTCGCTCCAAGCTGGGCTGTGTGCACGAACCCC 4440 GACACGACTTATCGCCACTG(~CAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATG 4560 TAGGCGGTGCTACAGAGTTC'.CTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAG 4620 TATTTGGTATCTGCGCTCTG(:TGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTT 4680 GATCCGGCAAACAAACCACC(~CTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTA 4740 CGCGCAGAAAAAAAGGATCT(:AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTC 4800 AGTGGAACGAAAACTCACGT~.'AAGGGA'CTTTGGTCATGAGATTATCAAAAAGGATCTTCA 4860 CTTGGTCTGACAGTTACCAA':.'GCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTAT 4980 TTCGTTCATCCATAGTTGCC"..'GACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCT 5040 TACCATCTGGCCCCAGTGCT(~CAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATT 5100 CCGCCTCCATCCAGTCTATT,~1ATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTA 5220 TGTGCAAAAAAGCGGTTAGC'rCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCG 5400 CAGTGTTATCACTCATGGTT:~TGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCG 5460 TAAGATGCTTTTCTGTGACTGGTGAGT:ACTCAACCAAGTCATTCTGAGAATAGTGTATGC 5520 TTACTTTCACCAGCGTTTCT(~GGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGG 5760 GAATAAGGGCGACACGGAAA'CGTTGAA'PACTCATACTCTTCCTTTTTCAATATTATTGAA 5820 GCATTTATCAGGGTTATTGT(~TCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA 5880 TAATATTTTGTTAAAATTCG(:GTTAAA'PTTTTGTTAAATCAGCTCATTTTTTAACCAATA 6000 GGCCGAAATCGGCAAAATCC(~TTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGT 6060 TGTTCCAGTTTGGAACAAGA(~TCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCG 6120 GGGGTCGAGGTGCCGTAAAG(:ACTAAA'PCGGAACCCTAAAGGGAGCCCCCGATTTAGAGC 6240 CGCTAGGGCGCTGGCAAGTG'.PAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCT 6360 TAATGCGCCGCTACAGGGCGC:GTCGCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGA 6420 AGGGCGATCGGTGCGGGCCT(:TTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGC 6480 AAGGCGATTAAGTTGGGTAA(:GCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGC 6540 CAGTGAATTGTAATACGACTC:ACTATAGGCGAATTGGGTACCGGGCCCCCCCTCGAGGTC 6600 (2) INFORMATION FOR SE(2 ID N0:16:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6447 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:16:
AAGCTTTGCTCAACAAATAC'rGTCAAGGACTCGAGTCCGGCTCTGACTGAGCAATGTCTA 60 AAGAAATACCAACCCCTTAT:~TGTGGAGCTACCAACCGCAAACGGGACACGCCGGCGCCT 120 CTCCCAGAACAATAATGGATCCGCCAA'rTTGGCCAGCTGCCATGCTTGTTCAGGAAGCCG 300 CTGGGGCGCAGCTAGCGGGAc~GACGACAGCTGTGCCCCTCCCAAATAGGTATAAAAAGCC 420 CAGTGCTGGCTGGCACGGGCATTCAGC'TTAGCGAAGACATCCCCAGCGCCTCCTGGATCA 480 GGCCCGACGGCATATTCCAG(~TAGGAGGGGGGTCTCGCTCGTCCTTCAGCCCAACGCAAG 540 CATTCCTCACCCTGCAACAG(~CATCCTCGACGCCGCGCGCAGGAGGCGTGGGCACCTACC 600 AGTTTGTGCGCGAATTTGTG(:CAGAGG'rATACCTTAACCCTTTTTCAGGACCACCGGACA 660 CCTTTCCTGATCAGTTCATT(:CTAACTACGACATTGTAACCAACTCTGTCGATGGCTATG 720 ACTGAGGAGAGCATGGACCA(~GTGGAGGTGAACTGCCTGTGTGCTCAGCATGCCCAAACC 780 TGCACGCGCCCTCGCTGCTT'CGCAAAGGAGGGTTTATGTGCTAACTGGTTTTACAACCCA 840 GCACTTGCCTTTGAAGGGTT'.CGATATTCCAGACTCTTACCAAGAGGGACACGGTGTGGAC 900 ATAGAAGTTAAGTGTTCCCA(:CACTCCAGCAAACTGTGCCACAATGGCCATGATATGATC 960 CCCCACATGAGCCTCATTGAC~GCAGCC'CGTTCTATGTATAACCTTAACTAGATAATATTA 1080 TTAAACTTGATACGCGTATGC~CAGAAGGATTTGCAGCCAATAGACAATGGATAGGACCAG 1140 AAGAAGCTGAAGAGTTATTA(~ATTTTGATATAGCAACACAAATGAGTGAAGAAGGACCAC 1200 TAAATCCAGGAGTAAACCCA'.:'TTAGGG'PACCTGGAATAACAGAAAAAGAAAAGCAAAACT 1260 TGGAAGAAGGAAATGCAGGT.~AGTTTAGAAGAGCAAGATTTTTAAGGTATTCTGATGAAC 1380 i'~AGACACTCCAATTTTAATGGCCCTTAGCCATTCCATGCCCGTCGCCATACCTGACACTG 1560 CAATGCCTATATATATTTCC;9TCATGTTTTTTATTGTGGCCATGCTAGCCACCCTCAGCC 1620 TTCTAATGGGACTAAACAAC;~AP.ATCAGGCCCATGTAGCTTGTCAAATi'~AACTTACCTAA 1680 CTCTCCCAACAGCTTCCCGT'PCACCAGCACCAGGGCCATGi~AGCGGACACGAAGAGCTCT 1860 ACCTGCAAATTATGACCCTG'TATATCCATACGACGCCCCCGGGTCTTCCACACAACCCCC 1920 TTTTTTTAATAACAAGCAAGGTCTCAC'PGAGTCACCCCCAGGAACCCTGGCTGTCAATGT 1980 CACCCTCAACGAGGGCAAGT'rACAAGCCAGCTTAGGGCCCGGCCTCATCACAAATACCGA 2100 GGGCCAAATCACTGTTGAAAi~TGTCAACAAGGTTTTGTCTTTTACCTCCCCATTACATAA 2160 AAATGAAAACACTGTATCCC'CAGCGCTAGGAGATGGGTTAGAAGATGAAAATGGCACCCT 2220 TAAAGTGACCTTCCCTACTC(~CCCTCCCCCGCTACAATTCTCCCCTCCCCTCACAAAAAC 2280 AGGTGGTACTGTTTCCTTGC(;CCTGCAAGACTCCATGCAAGTGACAAATGGAAAACTGGG 2340 CGTTAAGCTACCACCTACGCi~CCTCCC'PTGAAAAAAACTGACCAGCAAGTTAGCCTCCAA 2400 ACCTACAACGAGCCTCTTTC(:AAAACTGACAATTCTGTTTCTCTGCAAGTAGGTGCCGGC 2520 CTTGCCGTGCAGAGCGGACG'fTTGGTGGCAACCCCTCCCCCGCCTCTCACCTTTACATCA 2580 CCCCTAGAAAAAAATGAAAA(:ACAGTGTCGCTACAAGTAGGCGCGGGCTTGTCTGTACAA 2640 i~ACAACGCCCTAGTAGCCACi~CCTCCCCCACCCTTAACCTTTGCCTATCCCTTAGTi~AAA2700 ACGGTGGCCACTGGACCTGGC:CTTTCCCATCAAAATGGAACAATAGGGGCTGTAGTAGGT 2820 GCAGGCCTCAAGTTTGAAAA(:AATGCCi~TTCTTGCAAAACTAGGCAACGGTCTAACCATT 2880 AGAGATGGCGCTATTGAAGC1~ACCCAA(:CCCCAGCTGCCCCCATAACACTGTGGACAGGG 2940 CCTGGCCTAGCATTAATGGC'..'TTATGTAATGACACTCCAGTAATTAGGTCTTTATATGCC 3000 TAACCAGAGACAGCAACTTAC~TCACAGTAAATGCTAGCTTTGTGGGAGAGGGGGGGTATC 3060 AGCTTATGTCCACAGGAAAC.ATTAACTCCACCACTACTTGGGGAGAAAAGCCCTGGGGCA 3180 TTTACTCCACTCCCGCCGCC.~1CCATCACCCGCTGTGGACTAGACAGCATTGCAGTCGACG 3300 GTGCCCAGCAGAAGTATCGA~~TGCATGCTAATTATTAACAAACCAAAAGGCGTTGCCACT 3360 TACACCCTTACCTTTAGGTT'TTTAAACTTTAACAGACTAAGCGGAGGTACCCTGTTTAAA 3420 GGAAAAGCTTGATATCGAAT'rCCTGCAGCCCGGGGGATCCACTAGTTCTAGAGCGGCCGC 3540 CACCGCGGTGGAGCTCCAGC'rTTTGTTCCCTTTAGTGAGGGTTAATTCCGAGCTTGGCGT 3600 TACGAGCCGGAAGCATAAAG'rGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACAT 3720 TAATTGCGTTGCGCTCACTGCCCGCTT'TCCAGTCGGGAAACCTGTCGTGCCAGCTGCATT 3780 AATGAATCGGCCAACGCGCG(~GGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCT 3840 CGCTCACTGACTCGCTGCGC'PCGGTCG'TTCGGCTGCGGCGAGCGGTATCAGCTCACTCAA 3900 AAGGCCAGCAAAAGGCCAGGi~ACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGC 4020 TCCGCCCCCCTGACGAGCAT(:ACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGA 4080 CGACCCTGCCGCTTACCGGA~PACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTT 4200 CTCATAGCTCACGCTGTAGG'.CATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCT 4260 GTGTGCACGAACCCCCCGTT(;AGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTG 4320 AGTCCAACCCGGTAAGACAC(~ACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTA 4380 GCAGAGCGAGGTATGTAGGC(~GTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCT 4440 ACACTAGAAGGACAGTATTT(~GTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAA 4500 GAGTTGGTAGCTCTTGATCCC~GCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTT 4560 GCAAGCAGCAGATTACGCGC~~GAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTA 4620 CGGGGTCTGACGCTCAGTGG~~ACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTAT 4680 CAAAAAGGATCTTCACCTAG~~TCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAA 4740 GTATATATGAGTAAACTTGG7.'CTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCT 4800 GTCCTGCAACTTTATCCGCC'TCCATCC.AGTCTATTAATTGTTGCCGGGAAGCTAGAGTAA 5040 GTAGTTCGCCAGTTAATAGT'TTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGT 5100 CATGATCCCCCATGTTGTGC;~AAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCA 5220 GAAGTAAGTTGGCCGCAGTG'TTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTA 5280 CTGTCATGCCATCCGTAAGA'TGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCT 5340 GAGAATAGTGTATGCGGCGAt~CGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCG 5400 CGCCACATAGCAGAACTTTAi~AAGTGC'TCATCATTGGAAAACGTTCTTCGGGGCGAAAAC 5460 TCTCAAGGATCTTACCGCTG'TTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACT 5520 GATCTTCAGCATCTTTTACT'~TCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAA 5580 ATGCCGCAAAAAAGGGAATAe~GGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTT 5640 TTCAATATTATTGAAGCATT'CATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAAT 5700 GTATTTAGAAAAATAAACAA~~TAGGGG'TTCCGCGCACATTTCCCCGAAAAGTGCCACCTG 5760 GGAAATTGTAAACGTTAATA'CTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC 5820 ATTTTTTAACCAATAGGCCG~~AATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA 5880 GATAGGGTTGAGTGTTGTTC(:AGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC 5940 CAACGTCAAAGGGCGAAAAA(:CGTCTA'CCAGGGCGATGGCCCACTACGTGAACCATCACC 6000 CTAATCAAGTTTTTTGGGGT(:GAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG 6060 CCCCCGATTTAGAGCTTGAC(~GGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA 6120 AGCGAAAGGAGCGGGCGCTA(~GGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC 6180 CACACCCGCCGCGCTTAATGC:GCCGCTACAGGGCGCGTCGCGCCATTCGCCATTCAGGCT 6240 GCGCAACTGTTGGGAAGGGCC~ATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAA 6300 AGGGGGATGTGCTGCAAGGCC~ATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACG 6360 TTGTAAAACGACGGCCAGTG~~ATTGTAATACGACTCACTATAGGCGAATTGGGTACCGGG 6420 CCCCCCCTCGAGGTCGACGG7.'ATCGAT 6447 1~~
(2) INFORMATION FOR SEQ ID N0:17:
(i) SEQUENCE CHAR:~CTERISTICS:
(A) LENGTH: 6244 base pairs (B) TYPE: nucleic acid (C) STRANDED1VESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:17:
AAGCTTTGCTCAACAAATAC'CGTCAAGGACTCGAGTCCGGCTCTGACTGAGCAATGTCTA 60 AAGAAATACCAACCCCTTATi~TGTGGAGCTACCAACCGCAAACGGGACACGCCGGCGCCT 120 CCCAGGACTACTCCACCCAAe~TGAATTGGTTTAGTGCTGGGCCATCAATGATTAGTCAAG 180 TTTATGGCATTAGAGACTTG(:GCAACAAAGTTTTGATAACCCAGGCAGAAATAACCAAAA 240 CTCCCAGAACAATAATGGAT(:CGCCAA'1'TTGGCCAGCTGCCATGCTTGTTCAGGAAGCCG 300 CCCCACCCAAAACGGTCACT(:TGCCCAGAAACCACACCCTAGAACAGGCTATGACCAACT 360 CAGTGCTGGCTGGCACGGGC~~TTCAGC'PTAGCGAAGACATCCCCAGCGCCTCCTGGATCA 480 GGCCCGACGGCATATTCCAG(:TAGGAGGGGGGTCTCGCTCGTCCTTCAGCCCAACGCAAG 540 CATTCCTCACCCTGCAACAG(~CATCCTCGACGCCGCGCGCAGGAGGCGTGGGCACCTACC 600 AGTTTGTGCGCGAATTTGTG(:CAGAGG'CATACCTTAACCCTTTTTCAGGACCACCGGACA 660 CCTTTCCTGATCAGTTCATT(;CTAACTACGACATTGTAACCAACTCTGTCGATGGCTATG 720 ACTGAGGAGAGCATGGACCA(~GTGGAGGTGAACTGCCTGTGTGCTCAGCATGCCCAAACC 780 TGCACGCGCCCTCGCTGCTT~"GCAAAGGAGGGTTTATGTGCTAACTGGTTTTACAACCCA 840 GCACTTGCCTTTGAAGGGTT'..'GATATTCCAGACTCTTACCAAGAGGGACACGGTGTGTAA 900 ATGGGCCACACACGGAGGCAC~GGAACATCACCATCCAAGTGTCCATACCTCAATTTCTTT 960 CAGCTCTTGGTGCTGGCTGG~.'CTTTCTCACTTCTGTTCAGGTGTTATCCACGTGACCAAG 1020 CAAACTCGCATCTACTGGCA~~AAGGAGAAGAAAATGGTGCTGACTATGATGTCTGGGGAC 1140 ATGAATATATGGCCCGAGTAC:AAGAAC(:GGACCATCTTTGATATCACTAATAACACGCGT 1200 GTCCTCAACATCACCCGCGA~GGAACTTTCCTGCTTATTGGGGATAGCAAAAAGACCCCC 1260 ATGTTTTTTATTGTGGCCAT~~CTAGCCACCCTCAGCCTTCTAATGGGACTAAACAACAAA 1440 GAGCCAGCTTGCGGATGTGC'PTGAAAGATAATGTGGTCTCTCCCAACAGCTTCCCGTTCA 1620 ATCCATACGACGCCCCCGGG'rCTTCCACACAACCCCCTTTTTTTAATAACAAGCAAGGTC 1740 TCACTGAGTCACCCCCAGGA:~CCCTGGCTGTCAATGTTTCCCCTCCACTAACCTTTTCTA 1800 CGTTAGGTGCCATTAAACTT'PCCACAGGTCCCGGACTCACCCTCAACGAGGGCAAGTTAC 1860 CGCTAGGAGATGGGTTAGAAciATGAAAATGGCACCCTTAAAGTGACCTTCCCTACTCCCC 2040 CTCCCCCGCTACAATTCTCCcJCTCCCC'rCACAAAAACAGGTGGTACTGTTTCCTTGCCCC 2100 TCCCTTGAAAAAAACTGACCAGCAAGT'PAGCCTCCAAGTAGGCTCGGGTCTCACCGTGAT 2220 TAACGAACAGTTGCAAGCTG'CCCAGCC'rCCCGCAACCACCTACAACGAGCCTCTTTCCAA 2280 AACTGACAATTCTGTTTCTC'CGCAAGTAGGTGCCGGCCTTGCCGTGCAGAGCGGACGTTT 2340 GGTGGCAACCCCTCCCCCGC(:TCTCACCTTTACATCACCCCTAGAAAAAAATGAAAACAC 2400 AGTGTCGCTACAAGTAGGCG(~GGGCTTGTCTGTACAAAACAACGCCCTAGTAGCCACACC 2460 TCCCCCACCCTTAACCTTTG(:CTATCCCTTAGTAAAAAATGACAACCATGTAGCTCTAAG 2520 TGCTGGAAGTGGTTTAAGAA'.CATCTGGAGGCAGCCTCACGGTGGCCACTGGACCTGGCCT 2580 TTCCCATCAAAATGGAACAA'..'AGGGGC'CGTAGTAGGTGCAGGCCTCAAGTTTGAAAACAA 2640 TGCCATTCTTGCAAAACTAGC~CAACGG'CCTAACCATTAGAGATGGCGCTATTGAAGCAAC 2700 CCAACCCCCAGCTGCCCCCA':'AACACT(~TGGACAGGGCCTGGCCTAGCATTAATGGCTTT 2760 ATGTAATGACACTCCAGTAA".'TAGGTC'.CTTATATGCCTAACCAGAGACAGCAACTTAGTC 2820 ACAGTAAATGCTAGCTTTGTC~GGAGAGGGGGGGTATCGAATAGTCAGCCCTACCCAGTCA 2880 CAATTTAGCCTAATTATGGAC~TTTGAT(:AGTTTGGACAGCTTATGTCCACAGGAAACATT 2940 AACTCCACCACTACTTGGGG.AGAAAAGCCCTGGGGCAATAACACTGTACAGCCACGCCCA 3000 ATCACCCGCTGTGGACTA~GA~~AGCATTGCAGTCGACGGTGCCCAGCAGAAGTATCGACTG 3120 TGTAGGCGAAAATCAATAAA;~CCAGAAAAAAATAAGGGGAAAAGCTTGATATCGAATTCC 3300 TGCAGCCCGGGGGATCCACT;~GTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAGCTTT 3360 TGTTCCCTTTAGTGAGGGTT;AATTCCGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCT 3420 AAAGCCTGGGGTGCCTAATGAGTGAGC'rAACTCACATTAATTGCGTTGCGCTCACTGCCC 3540 AGAGGCGGTTTGCGTATTGGGCGCTCT'PCCGCTTCCTCGCTCACTGACTCGCTGCGCTCG 3660 GAATCAGGGGATAACGCAGGi~AAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAAC 3780 CGTAAAAAGGCCGCGTTGCTc~GCGTTT'PTCCATAGGCTCCGCCCCCCTGACGAGCATCAC 3840 AAAAATCGACGCTCAAGTCA(~AGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCG 3900 TTTCCCCCTGGAAGCTCCCT(:GTGCGC'rCTCCTGTTCCGACCCTGCCGCTTACCGGATAC 3960 CTGTCCGCCTTTCTCCCTTC(~GGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTAT 4020 CTCAGTTCGGTGTAGGTCGT'CCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAG 4080 CCCGACCGCTGCGCCTTATC(:GGTAAC'PATCGTCTTGAGTCCAACCCGGTAAGACACGAC 4140 TTATCGCCACTGGCAGCAGC(:ACTGGTe~ACAGGATTAGCAGAGCGAGGTATGTAGGCGGT 4200 GCTACAGAGTTCTTGAAGTG(~TGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGT 4260 ATCTGCGCTCTGCTGAAGCCAGTTACC'PTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGC 4320 AAACAAACCACCGCTGGTAG(:GGTGGT'PTTTTTGTTTGCAAGCAGCAGATTACGCGCAGA 4380 AAAAAAGGATCTCAAGAAGA'.CCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAAC 4440 GAAAACTCACGTTAAGGGAT'.CTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATC 4500 CTTTTAAATTAAAAATGAAG'"."TTTAAA'PCAATCTAAAGTATATATGAGTAAACTTGGTCT 4560 GACAGTTACCAATGCTTAATC:AGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCA 4620 TCCATAGTTGCCTGACTCCCC:GTCGTG'CAGATAACTACGATACGGGAGGGCTTACCATCT 4680 GGCCCCAGTGCTGCAATGAT.~1CCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCA 4740 CGCAACGTTGTTGCCATTGC'TACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCT 4920 TCATTCAGCTCCGGTTCCCA:ACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAA 4980 AAAGCGGTTAGCTCCTTCGG'rCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTA 5040 AGTTGCTCTTGCCCGGCGTCAP.TACGGGATAATACCGCGCCACATAGCAGAACTTTAAAA 5220 GTGCTCATCATTGGAAAACG'PTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTG 5280 ACCAGCGTTTCTGGGTGAGCi~AAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGG 5400 GCGACACGGAAATGTTGAATi~CTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTAT 5460 CAGGGTTATTGTCTCATGAGc~GGATACATATTTGAATGTATTTAGAAAAATAAACAAATA 5520 GGGGTTCCGCGCACATTTCCc~CGAAAAGTGCCACCTGGGAAATTGTAAACGTTAATATTT 5580 TGTTAAAATTCGCGTTAAAT'CTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAA 5640 TCGGCAAAATCCCTTATAAA'CCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAG 5700 TTTGGAACAAGAGTCCACTA'CTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCG 5760 TCTATCAGGGCGATGGCCCA(:TACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGA 5820 GGTGCCGTAAAGCACTAAAT(;GGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGG 5880 GAAAGCCGGCGAACGTGGCG~~GAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGG 5940 CGCTACAGGGCGCGTCGCGC(:ATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGAT 6060 CGGTGCGGGCCTCTTCGCTA'.CTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGAT 6120 TAAGTTGGGTAACGCCAGGG'L'TTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAAT 6180 TGTAATACGACTCACTATAG(~CGAATTGGGTACCGGGCCCCCCCTCGAGGTCGACGGTAT 6240 (2) INFORMATION FOR SE(2 ID N0:18:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6095 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:18:
AAGCTTTGCTCAACAAATAC'fGTCAAGGACTCGAGTCCGGCTCTGACTGAGCAATGTCTA 60 TTTATGGCATTAGAGACTTGCGCAACA;~AGTTTTGATAACCCAGGCAGAAATAACCAAAA 240 CTCCCAGAACAATAATGGAT(~CGCCAA'rTTGGCCAGCTGCCATGCTTGTTCAGGAAGCCG 300 CTGGGGCGCAGCTAGCGGGA(~GACGACAGCTGTGCCCCTCCCAAATAGGTATAAAAAGCC 420 CAGTGCTGGCTGGCACGGGCATTCAGC'rTAGCGAAGACATCCCCAGCGCCTCCTGGATCA 480 GGCCCGACGGCATATTCCAG(:TAGGAGGGGGGTCTCGCTCGTCCTTCAGCCCAACGCAAG 540 AGTTTGTGCGCGAATTTGTG(:CAGAGG'rATACCTTAACCCTTTTTCAGGACCACCGGACA 660 CCTTTCCTGATCAGTTCATT(;CTAACTACGACATTGTAACCAACTCTGTCGATGGCTATG 720 ACTGAGGAGAGCATGGACCA(~GTGGAGGTGAACTGCCTGTGTGCTCAGCATGCCCAAACC 780 TGCACGCGCCCTCGCTGCTT'.CGCAAAGGAGGGTTTATGTGCTAACTGGTTTTACAACCCA 840 GCACTTGCCTTTGAAGGGTT'.CGATATTCCAGACTCTTACCAAGAGGGACACGGTGTGTAG 900 ATGGGTTGTTCTGTGGAGAA'.CGTTGGACAGTGTAAAGTATGCTGCCAGGGGCGTCCGCGA 960 CTGACCAAGTGAAAACATCA'..'TGTAATAGGAGTTTGTTCTCCATGTCTCTTGTTGGTCTA 1020 CTGATGCTGTGGTGGCAGTGC:CCGCTGCTTCGCCTTGCGGCGCTGCACGGGCTTTGCCCT 1200 CTAACGCGTCCCTCAGCCTTC:TAATGGGACTAAACAACAAAATCAGGCCCATGTAGCTTG 1260 TCAAATAAACTTACCTAATT'PTTGCTAAGACGCTGGGTCCTGCGTTTCTATGTCCACCAA 1320 GCGGACACGAAGAGCTCTACCTGCAAA'rTATGACCCTGTATATCCATACGACGCCCCCGG 1500 GTCTTCCACACAACCCCCTT'PTTTTAATAACAAGCAAGGTCTCACTGAGTCACCCCCAGG 1560 TACCTCCCCATTACATi~AAAi~TGAAAACACTGTATCCCTAGCGCTAGGAGATGGGTTAGA 1800 AGATGAAAATGGCACCCTTAi~AGTGACCTTCCCTACTCCCCCTCCCCCGCTACAATTCTC 1860 CCCTCCCCTCACAAAAACAGGTGGTAC'TGTTTCCTTGCCCCTGCAAGACTCCATGCAAGT 1920 GACAAATGGAAAACTGGGCG'PTAAGCTACCACCTACGCACCTCCCTTGAAAAAAACTGAC 1980 CAGCAAGTTAGCCTCCAAGTi~GGCTCGGGTCTCACCGTGATTAACGAACAGTTGCAAGCT 2040 GTCCAGCCTCCCGCAACCAC(~TACAACGAGCCTCTTTCCAAAACTGACAATTCTGTTTCT 2100 CTGCAAGTAGGTGCCGGCCT'CGCCGTGCAGAGCGGACGTTTGGTGGCAACCCCTCCCCCG 2160 CCTCTCACCTTTACATCACC(:CTAGAAAAAAATGAAAACACAGTGTCGCTACAAGTAGGC 2220 GCGGGCTTGTCTGTACAAAA(:AACGCCCTAGTAGCCACACCTCCCCCACCCTTAACCTTT 2280 GCCTATCCCTTAGTAAAAAA'CGACAACCATGTAGCTCTAAGTGCTGGAAGTGGTTTAAGA 2340 GGCAACGGTCTAACCATTAG~~GATGGCGCTATTGAAGCAACCCAACCCCCAGCTGCCCCC 2520 ATAACACTGTGGACAGGGCC'CGGCCTAGCATTAATGGCTTTATGTAATGACACTCCAGTA 2580 AGTTTGATCAGTTTGGACAGC:TTATGTCCACAGGAAACATTAACTCCACCACTACTTGGG 2760 GCATGCCTAACAGAGAAGTT'.~ACTCCA(:TCCCGCCGCCACCATCACCCGCTGTGGACTAG 2880 ACAGCATTGCAGTCGACGGT(~CCCAGCAGAAGTATCGACTGCATGCTAATTATTAACAAA 2940 CCAAAAGGCGTTGCCACTTAC:ACCCTTACCTTTAGGTTTTTAAACTTTAACAGACTAAGC 3000 GGAGGTACCCTGTTTAAAAC'rGATGTCTTAACCTTTACCTATGTAGGCGA 3060 AAATCAATAA

AACCAGAAAAAAATAAGGGG;~AAAGCTTGATATCGAATTCCTGCAGCCCGGGGGATCCAC 3120 TAATTCCGAGCTTGGCGTAA'rCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGC 3240 TGTCGTGCCAGCTGCATTAA'PGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTG 3420 CGGTATCAGCTCACTCAAAG(~CGGTAA'rACGGTTATCCACAGAATCAGGGGATAACGCAG 3540 GAAAGAACATGTGAGCAAAAc~GCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGC 3600 TGGCGTTTTTCCATAGGCTCc~GCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTC 3660 AGAGGTGGCGAAACCCGACA(~GACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCC 3720 TCGTGCGCTCTCCTGTTCCGi~CCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTT 3780 CGGGAAGCGTGGCGCTTTCTI:ATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCG 3840 TTCGCTCCAAGCTGGGCTGT(~TGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTAT 3900 CCGGTAACTATCGTCTTGAG'PCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAG 3960 GGTGGCCTAACTACGGCTAC~~CTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGC 4080 GCGGTGGTTTTTTTGTTTGC~~AGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAG 4200 ATCCTTTGATCTTTTCTACG(~GGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGA 4260 TTTTGGTCATGAGATTATCARAAAGGA'PCTTCACCTAGATCCTTTTAAATTAAAAATGAA 4320 TCAGTGAGGCACCTATCTCA(~CGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCC 4440 TACCGCGAGACCCACGCTCAC:CGGCTC(:AGATTTATCAGCAATAAACCAGCCAGCCGGAA 4560 GGGCCGAGCGCAGAAGTGGT(:CTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTT 4620 GCCGGGAAGCTAGAGTAAGT~~GTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTG 4680 CTACAGGCATCGTGGTGTCA(:GCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCC 4740 GTCCTCCGATCGTTGTCAGA.?~GTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAG 4860 CACTGCATAATTCTCTTACT~~TCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGT 4920 ACTCAACCAAGTCATTCTGA~~AATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGT 4980 CAATACGGGATAATACCGCG~~CACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAAC 5040 GTTCTTCGGGGCGAAAACTC'TCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAAC 5100 CCACTCGTGCACCCAACTGA'PCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAG 5160 CCCGAAAAGTGCCACCTGGGAAATTGT;AAACGTTAATATTTTGTTAAAATTCGCGTTAAA 5400 TTTTTGTTAAATCAGCTCAT'rTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAA 5460 ATCAAAAGAATAGACCGAGA'PAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACT 5520 ATTAAAGAACGTGGACTCCA~~CGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCC 5580 ACTACGTGAACCATCACCCTi~ATCAAG'rTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAA 5640 TCGGAACCCTAAAGGGAGCCCCCGATT'rAGAGCTTGACGGGGAAAGCCGGCGAACGTGGC 5700 GAGAAAGGAAGGGAAGAAAG(~GAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGT 5760 CACGCTGCGCGTAACCACCA(:ACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCGCG 5820 CCATTCGCCATTCAGGCTGC(~CAACTG'PTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCT 5880 ATTACGCCAGCTGGCGAAAG(~GGGATG'PGCTGCAAGGCGATTAAGTTGGGTAACGCCAGG 5940 GTTTTCCCAGTCACGACGTT(~TAAAACGACGGCCAGTGAATTGTAATACGACTCACTATA 6000 GGCGAATTGGGTACCGGGCC(:CCCCTCGAGGTCGACGGTATCGAT 6045 (2) INFORMATION FOR SEQ ID N0:19:
(i) SEQUENCE CHARACTERIS'CICS:
(A) LENGTH: .'i109 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDPdESS: single (D) TOPOLOGY.. linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:19:
AAGCTTTCGCGATATCCGTT:AAGTTTGTATCGTAATGCTCCCCTACCAAGACAAGGTGGG 60 ATTTAGCAGATTGCTGAAAG:~GGATATGGAGAAATCAGAGGCCGTACATCACCAAGTCAT 300 AGATGTCTTGACACCGCTCT'PCAAGAT'rATTGGAGATGAGATTGGGTTACGGTTGCCACA 360 AAAGCTAAACGAGATCAAACi~ATTTATCCTTCAAAAGACAAATTTCTTCAATCCGAACAG 420 AGAATTCGACTTCCGCGATC'PCCACTGGTGCATTAACCCGCCTAGTACGGTCAAGGTGAA 480 TTTTACTAATTACTGTGAGTc:AATTGGGATCAGAAAAGCTATTGCATCGGCAGCAAATCC 540 TGGAGCTACTACTTCAGTAG(~CAAAGT'rTTCCCCCTATCAGTCTCATTATCCATGTCTTT 660 GATCTCAAGAACCTCAGAGG'CAATCAA'rATGCTGACCGCTATCTCAGACGGCGTGTATGG 720 CAAAACTTACTTGCTAGTGC(:TGATGA'PATAGAAAGAGAGTTCGACACTCGAGAGATTCG 780 AGTCTTTGAAATAGGGTTCA'CCAAAAGGTGGCTGAATGACATGCCATTACTCCAAACAAC 840 CAACTATATGGTACTCCCGA~~.GAATTCCAAAGCCAAGGTATGTACTATAGCAGTGGGTGA 900 GTTGACACTGGCTTCCTTGT(~TGTAGAi~GAGAGCACTGTATTATTATATCATGACAGCAG 960 TGGTTCACAAGATGGTATTC'CAGTAGTGACACTGGGGATATTTTGGGCAACACCTATGGA 1020 TCACATTGAGGAAGTGATAC(:TGTCGCTCACCCATCAATGAAGAAAATACATATAACAAA 1080 GAAACAAGAAGAACAAAAAG(~TTGTCTGGAGTCAGCTTGTCAAAGAAAAACCTACCCCAT 1200 GTGCAACCAAGCGTCATGGG~~ACCCTTCGGAGGAAGACAGTTGCCATCTTATGGGCGGTT 1260 GACATTACCTCTAGATGCAAC~TGTTGACCTTCAACTTAACATATCGTTCACATACGGTCC 1320 GGTTATACTGAATGGAGATG(iTATGGA'CTATTATGAAAGCCCACTTTTGAACTCCGGATG 1380 AGACCAGTTCACTGTACTCC(:CCATGTGTTAACATTTGCGCCCAGGGAATCAAGTGGAAA 1500 TTGTTATTTACCTATTCAAA(:ATCTCAAATTAGAGATAGAGATGTCCTCATTGAGTCCAA 1560 TATAGTGGTGTTGCCTACAC.AGAGTATTAGATATGTCATAGCAACGTATGACATATCACG 1620 AAGTGATCATGCTATTGTTT.ATTATGTTTATGACCCAATCCGGACGATTTCTTATACGCA 1680 CCCATTTAGACTAACTACCA.?~GGGTAGACCTGATTTCCTAAGGATTGAATGTTTTGTGTG 1740 GGATGACAATTTGTGGTGTC.~1CCAATTTTACAGATTCGAGGCTGACATCGCCAACTCTAC 1800 AACCAGTGTTGAGAATTTAG'rCCGTATRAGATTCTCATGTAACCGTTAAAATCCCTGACA 1860 GTATGATGATACACATCTCA.~TTGGCCTTAGGCATGATAACTGCGGTGAGAAATCCCTTA 1920 CAGACGATTGAATTAAACCA'rCTCTAGCATTATAAAAAAACTAAGGATCCAAGATCCTTT 1980 CAATTGTAACCAATAAGCTAGTATCTA'TTTTAGAATACGCACGAATTAGACATAACTATC 2100 CTTTTGTTCCCTTTAGTGAGGGTTAAT'PCCGAGCTTGGCGTAATCATGGTCATAGCTGTT 2280 GCCCGCTTTCCAGTCGGGAARCCTGTCcsTGCCAGCTGCATTAATGAATCGGCCAACGCGC 2460 GGGGAGAGGCGGTTTGCGTA'CTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCG 2520 CACAGAATCAGGGGATAACG(:AGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAG 2640 GAACCGTAAAAAGGCCGCGT'PGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCA 2700 GGCGTTTCCCCCTGGAAGCT(:CCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGG 2820 ATACCTGTCCGCCTTTCTCC(:TTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAG 2880 GTATCTCAGTTCGGTGTAGG'.CCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGT 2940 TCAGCCCGACCGCTGCGCCT'.CATCCGG'~AACTATCGTCTTGAGTCCAACCCGGTAAGACA 3000 CGACTTATCGCCACTGGCAG(:AGCCAC'~GGTAACAGGATTAGCAGAGCGAGGTATGTAGG 3060 TGGTATCTGCGCTCTGCTGA~~GCCAGT'.CACCTTCGGAAAAAGAGTTGGTAGCTCTTGATC 3180 CGGCAAACAAACCACCGCTGC~TAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCG 3240 CAGAAAAAAAGGATCTCAAG~~AGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTG 3300 GTCTGACAGTTACCAATGCT'rAATCAG'PGAGGCACCTATCTCAGCGATCTGTCTATTTCG 3480 TTCATCCATAGTTGCCTGAC'PCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACC 3540 ATCTGGCCCCAGTGCTGCAA'PGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATC 3600 AGCAATAAACCAGCCAGCCGt~AAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGC 3660 CTCCATCCAGTCTATTAATTt~TTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAG 3720 TTTGCGCAACGTTGTTGCCA'CTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTAT 3780 GGCTTCATTCAGCTCCGGTTt~CCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTG 3840 CAAAAAAGCGGTTAGCTCCT'CCGGTCC'PCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGT 3900 GTTATCACTCATGGTTATGGt~AGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAG 3960 ATGCTTTTCTGTGACTGGTG~~GTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCG 4020 ACCGAGTTGCTCTTGCCCGG(:GTCAATACGGGATAATACCGCGCCACATAGCAGAACTTT 4080 AAAAGTGCTCATCATTGGAAt~.ACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCT 4140 AAGGGCGACACGGAAATGTT(~AATACTCATACTCTTCCTTTTTCAATATTATTGAAGCAT 4320 TTATCAGGGTTATTGTCTCA'.CGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACA 4380 AATAGGGGTTCCGCGCACAT'.CTCCCCGAAAAGTGCCACCTGGGAAATTGTAAACGTTAAT 4440 ATTTTGTTAAAATTCGCGTT~~AATTTT'CGTTAAATCAGCTCATTTTTTAACCAATAGGCC 4500 GAAATCGGCAAAATCCCTTA'.PAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTT 4560 CCAGTTTGGAACAAGAGTCC~~CTATTA~'~AGAACGTGGACTCCAACGTCAAAGGGCGAAAA 4620 ACCGTCTATCAGGGCGATGG(:CCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGG 4680 AGGGCGCTGGCAAGTGTAGC(~GTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAAT 4860 GCGCCGCTACAGGGCGCGTC(~CGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGG 4920 CGATCGGTGCGGGCCTCTTC<~CTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGG 4980 CGATTAAGTTGGGTAACGCCAGGGTTT'.PCCCAGTCACGACGTTGTAAAACGACGGCCAGT 5040 GAATTGTAAT ACGACTCACT i~TAGGGCGAA TTGGGTACCG GGCCCCCCCT CGAGGTCGAC 5100 (2) INFORMATION FOR SES2 ID N0:20:
(i) SEQUENCE CHARACTERIS'PICS:
(A) LENGTH: 5067 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:20:
AAGCTTAATGTCGTAACAAC'.CCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGT 60 GGGAGGTCTATATAAGCAGA(~CTCGTT'CAGTGAACCGTCTGCAGACTCTCTTCCGCATCG 120 CTGTCTGCGAGGGCCAGCTG'.CTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCC 180 AGTACTCTTGGATCGGAAAC(:CGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTG 240 TCGCAAGTCTAGAATGCTCC(:CTACCAAGACAAGGTGGGTGCCTTCTACAAGGATAATGC 360 AAGAGCCAATTCAACCAAGC'.CGTCCTTAGTGACAGAAGGACATGGGGGCAGGAGACCACC 420 TTATTTGTTGTTTGTCCTTC'.CCATCTTATTGGTTGGTATCCTGGCCTTGCTTGCTATCAC 480 TGGAGTTCGATTTCACCAAG~CATCAAC'CAGTAATATGGAATTTAGCAGATTGCTGAAAGA 540 GGATATGGAGAAATCAGAGG(:CGTACA'.CCACCAAGTCATAGATGTCTTGACACCGCTCTT 600 CAAGATTATTGGAGATGAGA'~TGGGTTACGGTTGCCACAAAAGCTAAACGAGATCAAACA 660 CCACTGGTGCATTAACCCGCC:TAGTACGGTCAAGGTGAATTTTACTAATTACTGTGAGTC 780 AATTGGGATCAGAAAAGCTA'.'TGCATCGGCAGCAAATCCTATCCTTTTATCAGCCCTATC 840 TGGGGGCAGAGGTGACATAT"CCCACCACACAGATGCAGTGGAGCTACTACTTCAGTAGG 900 CAAAGTTTTCCCCCTATCAG'.'CTCATTATCCATGTCTTTGATCTCAAGAACCTCAGAGGT 960 AATCAATATGCTGACCGCTA~,'CTCAGACGGCGTGTATGGCAAAACTTACTTGCTAGTGCC 1020 TGATGATATAGAAAGAGAGT'..'CGACAC'.CCGAGAGATTCGAGTCTTTGAAATAGGGTTCAT 1080 CAAAAGGTGGCTGAATGACA'PGCCATTACTCCAAACAACCAACTATATGGTACTCCCGAA 1140 GAATTCCAAAGCCAAGGTAT(~TACTATAGCAGTGGGTGAGTTGACACTGGCTTCCTTGTG 1200 TGTAGAAGAGAGCACTGTAT'CATTATATCATGACAGCAGTGGTTCACAAGATGGTATTCT 1260 AGTAGTGACACTGGGGATAT'PTTGGGCAACACCTATGGATCACATTGAGGAAGTGATACC 1320 TTCAATTGCAACCTGGATGG'CGCCTGCCCTGGCCTCTGAGAAACAAGAAGAACAAAAAGG 1440 ACCCTTCGGAGGAAGACAGT'('GCCATC'PTATGGGCGGTTGACATTACCTCTAGATGCAAG 1560 TGTTGACCTTCAACTTAACA'CATCGTTCACATACGGTCCGGTTATACTGAATGGAGATGG 1620 TATGGATTATTATGAAAGCC(:ACTTTTGAACTCCGGATGGCTTACCATTCCCCCCAAAGA 1680 CGGAACAATCTCTGGATTGA'CAAACAAAGCAGGTAGAGGAGACCAGTTCACTGTACTCCC 1740 CCATGTGTTAACATTTGCGCCCAGGGAi~TCAAGTGGAAATTGTTATTTACCTATTCAAAC 1800 GAGTATTAGATATGTCATAG(;AACGTA'PGACATATCACGAAGTGATCATGCTATTGTTTA 1920 TTATGTTTATGACCCAATCCGGACGAT'PTCTTATACGCACCCATTTAGACTAACTACCAA 1980 CCAATTTTACAGATTCGAGG(~TGACATCGCCAACTCTACAACCAGTGTTGAGAATTTAGT 2100 ATGAATGCAATTGTTGTTGT'.CAACTTG'PTTATTGCAGCTTATAATGGTTACAAATAAAGC 2280 TCCAAACTCATCAATGTATC~PTATCATGTCTGGATCCCCCGGAATTCACTGGCCGTCGTT 2400 CCCCCCTTCGCCAGCTGGCG'CAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAG 2520 GGTATTTCACACCGCATATG(~TGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTA 2640 AGCCAGTACACTCCGCTATC(iCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCA 2700 ACACCCGCTGACGCGCCCTG~~CGGGCT'CGTCTGCTCCCGGCATCCGCTTACAGACAAGCT 2760 GTGACCGTCTCCGGGAGCTGC:ATGTGT(:AGAGGTTTTCACCGTCATCACCGAAACGCGCG 2820 i~GGGCCTCGT

GATAATAATGGTTTCTTAGAt:GTCAGG'rGGCACTTTTCGGGGAAATGTGCGCGGAACCCC 2940 TATTTGTTTATTTTTCTAAA'CACATTCAAATATGTATCCGCTCATGAGACAATAACCCTG 3000 CCTTATTCCCTTTTTTGCGGI:ATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGT 3120 TTTTAAAGTTCTGCTATGTG(~CGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACT 3300 CGGTCGCCGCATACACTATT(:TCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA 3360 TAACACTGCGGCCAACTTAC'CTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTT 3480 TTTGCACAACATGGGGGATCATGTAAC'rCGCCTTGATCGTTGGGAACCGGAGCTGAATGA 3540 AGCCATACCAAACGACGAGC(~TGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCG 3600 CAAACTATTAACTGGCGAAC'CACTTAC'PCTAGCTTCCCGGCAACAATTAATAGACTGGAT 3660 GGAGGCGGATAAAGTTGCAG(~ACCACT'PCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTAT 3720 TGCTGATAAATCTGGAGCCGGTGAGCG'PGGGTCTCGCGGTATCATTGCAGCACTGGGGCC 3780 AGATGGTAAGCCCTCCCGTA'CCGTAGT'PATCTACACGACGGGGAGTCAGGCAACTATGGA 3840 TGAACGAAATAGACAGATCG(~TGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTC 3900 AGACCAAGTTTACTCATATA'CACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAG 3960 GATCTAGGTGAAGATCCTTT'CTGATAA'PCTCATGACCAAAATCCCTTAACGTGAGTTTTC 4020 GTTCCACTGAGCGTCAGACC(~CGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTT 4080 TCTGCGCGTAATCTGCTGCT'CGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTT 4140 GCCGGATCAAGAGCTACCAAt:TCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGAT 4200 ACCAAATACTGTCCTTCTAG'CGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGC 9260 ACCGCCTACATACCTCGCTC'CGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAA 4320 CTGAACGGGGGGTTCGTGCAt:ACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAG 9440 GTATCCGGTAAGCGGCAGGG'CCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAA 4560 GTGATGCTCGTCAGGGGGGC (~GAGCCTATG GAAAAACGCCAGCAACGCGG CCTTTTTACG4680 GTTCCTGGCCTTTTGCTGGC t~TTTTGCTCA CATGTTCTTTCCTGCGTTAT CCCCTGATTC4740 TGTGGATAACCGTATTACCG (~CTTTGAGTG AGCTGATACCGCTCGCCGCA GCCGAACGAC4800 CGAGCGCAGCGAGTCAGTGA t~CGAGGAAGC GGAAGAGCGCCAATACGCAA ACCGCCTCTC4860 CCCGCGCGTTGGCCGATTCA 'CTAATGCAGC TGGCACGACAGGTTTCCCGA CTGGAAAGCG4920 GGCAGTGAGCGCAACGCAAT 'CAATGTGAGT TACCTCACTCATTAGGCACC CCAGGCTTTA4980 CACTTTATGCTTCCGGCTCG 'CATGTTGTGT GGAATTGTGAGCGGATAACA ATTTCACACA5040 GGAAACAGCTATGACCATGA 'CTACGCC 5067 (2) INFORMATION
FOR
SE(.2 ID N0:21:

(i) SEQUENCE
CHARACTERISTICS:

(A) LENGTH: 8618 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear ( ii ) MOLECULE
TYPE;
DNA
( genomic ) (xi) SEQUENCE
DESCRIPTION:
SEQ
ID N0:21:

TTGCCACTTACACCCTTACC'.CTTAGGT'CTTTAAACTTTAACAGACTAAGCGGAGGTACCC 120 AAATAAGTTTAAAAGCTTTA'.CTTTTCA'CACACGCGAGCGGTAAGGCTGCCGCCTTCAGGA 240 TCATTTGGGCTATAATAATA'.CACATTT'CCTTGGGTGGCAAAGCAAGGGTCGGTAATCTCA 360 ACAAAACCATCAACTGGAAT(~CAAGAA'CAGTCCAGCACGGTGGGTTCAATCTAAAAATGA 420 AGAAACGCTGTTGAGGTTCA(~TAAGCACAGGTTTTGAATCTGTCGGCAGCGTCCATGCAT 480 CACTACAGGTGTCTTTTCAA(:CTCTTTCAGCACCCGCTCTATTACAGATCTCACCCACAC 600 AGCACAGTTTTTAAGAGAACAATAGTT'~TGAAGGCTACAAGATTTACACTTAAGCACCAG 660 CCAGTAATTATAAGTGCTTT'CAAGAAC'rACCCCTAGCTCAGGGTTAATGCACCTTTTAAT 720 GGCCTCCATGCAGGCTTTAT(~GACAGTTCTAAAAAAAGACAGTCTAAAATAAATGTAGTG 780 AGTGTTTCTAAATATAATAC'CCCCCACATAGTTAATTTCATCAGGCCTGCTAGAATTTAC 840 CACTTTCTGAACCACATGCT'CGCTAGCCATGCATTGTAAAGACAAGCTGTTAGAGCAGTG 960 ACAGTGTACTCGCCACGTTTGAGCCTC'PGCCAGGCAGCAGTGCTTAGTTACTATCAACTC 1020 ATCCCACAAAATAGGAATTT(~ATAGCATAAAGCAAAGCAATTACAATATTTAGGAACTCT 1140 CACCACAGCAGTCACGTGAC~~TGTTGTCTCAGCAGTGCAGTTGCCTTCCATCCTACAATT 1200 ATGAACAAAAACTAAACACT'PCTAACAAAGATACAGTGACAATCTCCCTTCCTCTAAAAG 1260 CATTGTTTACATTAGGGTGA'.CTATTAACAACGTCAGAAATTTCTTTAATTAAAGTGCCTT 1320 TAAAATGTGCAAGAGCATCA'CCATACTCAAAACCAAGCTGAGAGTAAAAGACCACCTTAA 1380 AAGTAATCCCAGGCTTGTTT'CTATCAACAGCCTTAAACATGCTTTCACAAAATATAGAAG 1440 CAGTAACATCATCAATGGTG'CCGAAGAGAAACTCCATAGGAGACTCCAGCATTGATCCAA 1500 GTGGTAAAGTCATGCCCATG(~GTGAGGCCAAAATCCTTAAAAAAGCTATCTAAGTAGTTG 1680 ATAGCTACAAAGACCTGCAT(:CCCTCC'PTAGCAGACAGCTCTTGCACACACGCAGTAACT 1800 ATCCACCGCTTAAGAAAAGC'.L'TTAAGCCCAGCGCACATAACAGCTCCAATGTTTTTATCC 1860 AGCAGCTGAAAAAGTGCACT'.PTGACCTCATTAAGCCACTGCATATGCAAGTCCTCATCTA 2040 TGCCGCAGCCCAGACCCTCAATCCAGC(:CCGAATGTACACTTTAATAAGAGATTCAACCT 2100 CTTCTTTTAGCAAAGTACAC~~TGCTGT'CTGGACTAGTATACACAATAGAAGTCACAATGA 2160 GCATTGAAACCCCGCGACACAGGTCAG'CCTCGCGGTCTTGATCTCTTATTATAGCGACCA 2280 AATGGTCCTTCAGAGTGATG~CTGCACTCATAGAAGTAGGCAGCTCCGGCAGCCATTCTGC 2340 AAAGCAACACTTACTTATTCi~GCAGTCACAAGAATGTTGGGCTCCCAAGTGACAGACAAG 2460 CCTAATGCAAGGTGGGCACA(~TCTCCGGAATAAGTTGACAAAAGTCACGCCGCAAAGCTT 2520 CCTGAAGAGAAACGGCGGTAGCCTGGA'PATCTGCAACGGACCCAAAACCTTCAGTGTCAC 2580 TTCCAATAAACAGATAAAAC'CCTAAATAGTCCCCACTTAAAACCGAAACAGCCGCGGCAA 2640 AGGTAGGACACGGACGCACT'CCCTGAGCCCTAATAAGGCTAAACACCACACGGCGCAGTT 2700 CAGAAGGCAAAAAGTCTGTA~3GCTCTAGCTGAGCACACACACTCTCCACTAGACACTTGT 2760 GAAGCCTCAGACAAAAACAT(~CTCCCA'PAGACACTCCTAAAGCTGCCATTGTACTCACGG 2820 GCCAAGTACAGTCATAAAAT(~TGGGCGCGTGGTAAATGTTAAGTGCAGTTTCCCTTTGGC 3000 ACCACAAATAGGCACAACGC(:CAAAAACCCGGGTCGACACGCGTGAATTCACCGGTTCGA 3120 GCTTAATGTCGTAACAACTC(:GCCCCG'PTGACGCAAATGGGCGGTAGGCGTGTACGGTGG 3180 GAGGTCTATATAAGCAGAGC'.CCGTTTAGTGAACCGTCTGCAGACTCTCTTCCGCATCGCT 3240 GTCTGCGAGGGCCAGCTGTT(~GGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAG 3300 TACTCTTGGATCGGAAACCC(~TCGGCC'CCCGAACGGTACTCCGCCACCGAGGGACCTGAG 3360 CGAGTCCGCATCGACCGGAT(:GGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTC 3420 GAGCCAATTCAACCAAGCTG~CCCTTAG'CGACAGAAGGACATGGGGGCAGGAGACCACCTT 3540 ATTTGTTGTTTGTCCTTCTCATCTTAT'CGGTTGGTATCCTGGCCTTGCTTGCTATCACTG 3600 GAGTTCGATTTCACCAAGTA~PCAACTAGTAATATGGAATTTAGCAGATTGCTGAAAGAGG 3660 ATATGGAGAAATCAGAGGCC(~TACATCACCAAGTCATAGATGTCTTGACACCGCTCTTCA 3720 AGATTATTGGAGATGAGATT(~GGTTACGGTTGCCACAAAAGCTAAACGAGATCAAACAAT 3780 TTATCCTTCAAAAGACAAAT'.CTCTTCP.ATCCGAACAGAGAATTCGACTTCCGCGATCTCC 3840 ACTGGTGCATTAACCCGCCT~~GTACGG'CCAAGGTGAATTTTACTAATTACTGTGAGTCAA 3900 TTGGGATCAGAAAAGCTATTC~CATCGGCAGCAAATCCTATCCTTTTATCAGCCCTATCTG 3960 GGGGCAGAGGTGACATATTC(:CACCACACAGATGCAGTGGAGCTACTACTTCAGTAGGCA 4020 AAGTTTTCCCCCTATCAGTC'..'CATTATCCATGTCTTTGATCTCAAGAACCTCAGAGGTAA 4080 ATGATATAGA (;ACACTCGAGAGATTCGAGTCTTTGAAATAGGGTTCATCA 4200 AAGAGAGTTC

AAAGGTGGCTGAATGACATG(:CATTAC'CCCAAACAACCAACTATATGGTACTCCCGAAGA 4260 TAGAAGAGAGCACTGTATTA'.CTATATCATGACAGCAGTGGTTCACAAGATGGTATTCTAG 4380 TAGTGACACTGGGGATATTT'.CGGGCAACACCTATGGATCACATTGAGGAAGTGATACCTG 4440 CAATTGCAACCTGGATGGTG(:CTGCCCTGGCCTCTGAGAAACAAGAAGAACAAAAAGGTT 4560 CCTTCGGAGGAAGACAGTTG(:CATCTTATGGGCGGTTGACATTACCTCTAGATGCAAGTG 4680 TTGACCTTCAACTTAACATA'.CCGTTCACATACGGTCCGGTTATACTGAATGGAGATGGTA 4740 TGGATTATTATGAAAGCCCA(:TTTTGAACTCCGGATGGCTTACCATTCCCCCCAAAGACG 4800 CTCAAATTAGAGATAGAGAT(;TCCTCA'PTGAGTCCAATATAGTGGTGTTGCCTACACAGA 4980 GTAGACCTGATTTCCTAAGGATTGAAT(;TTTTGTGTGGGATGACAATTTGTGGTGTCACC 5160 GTATAAGATTCTCATGTAAC(:GTTAACCGCGGCGTGATTAATCAGCCATACCACATTTGT 5280 AGAGGTTTTACTTGCTTTAA~3AAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAAT 5340 CTTGCAACACCCAAATTTAA(;CCACACCTCTTTGTCCTGTATATTATTGATGATGGGGGG 5700 GAGGGTTAATTCCGAGCTTG(;CGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTT 5820 ATCCGCTCACAATTCCACACP.ACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTG 5880 CCTAATGAGTGAGCTAACTCi~CATTAA'rTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGG 5940 GTATTGGGCGCTCTTCCGCT'CCCTCGC'rCACTGACTCGCTGCGCTCGGTCGTTCGGCTGC 6060 GGCGAGCGGTATCAGCTCAC'CCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATA 6120 TCCCTTCGGGAAGCGTGGCG(:TTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGT 6420 AGGTCGTTCGCTCCAAGCTG(~GCTGTG'PGCACGAACCCCCCGTTCAGCCCGACCGCTGCG 6480 CCTTATCCGGTAACTATCGT(;TTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGG 6540 TGAAGTGGTGGCCTAACTACC~GCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGC 6660 TGAAGCCAGTTACCTTCGGAAAAAGAG'CTGGTAGCTCTTGATCCGGCAAACAAACCACCG 6720 AAGAAGATCCTTTGATCTTT'.CCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTT 6840 AATGAAGTTTTAAATCAATC'CAAAGTA'CATATGAGTAAACTTGGTCTGACAGTTACCAAT 6960 CAATGATACCGCGAGACCCA(:GCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAG 7140 GTTCCCAACGATCAAGGCGA(~TTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCT 7380 CCTTCGGTCCTCCGATCGTTC~TCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTA 7440 TGGCAGCACTGCATAATTCTC:TTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTG 7500 GTGAGTACTCAACCAAGTCA7.'TCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCC 7560 CGGCGTCAATACGGGATAAT~~.CCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTG 7620 TCATGAGCGGATACATATTT(~AATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCA 7920 ACTAAATCGGAACCCTAAAG(~GAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAA 8280 AGCGGTCACGCTGCGCGTAA(:CACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGC 8400 GTCGCGCCATTCGCCATTCA(~GCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTC 8460 TTCGCTATTACGCCAGCTGG(:GAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAAC 8520 GCCAGGGTTTTCCCAGTCAC(~ACGTTGTAAAACGACGGCCAGTGAATTGTAATACGACTC 8580 ACTATAGGGCGAATTGGGTAC:CGGGCCCCCCCTCGAGG 8618 (2) INFORMATION FOR SEQ ID N0:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 4965 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2dESS: single (D) TOPOLOGY;: linear (ii) MOLECULE TYPE;. DNA (genomic) S
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:22:

AGCTCCAAAA

GGACCTCCCGAGCACGACACi~GCACAACATCAGCTCAGCGATCCACGCACTACGATCCTC 180 GAACATCGGACAGACCCGTC'rCCTACACCATGAACAGGACCAGGTCCCGCAAGCAAACCA 240 ACTCAGGCTCTCATTGCACC'PGGTTAG'rCCTGTGGTGCCTCGGAATGGCCAGTCTCTTTC 420 ATAGTGTCCATTACAAGATCATGACTAtJGCCCAGTCACCAGTACTTGGTCATAAAACTGA 540 TGCCTAATGTTTCACTTATA(~AGAATTGTACCAAAGCAGAATTAGGTGAGTATGAGAAAT 600 TATTGAATTCAGTCCTCGAA(:CAATCAACCAAGCTTTGACTCTAATGACCAAGAATGTGA 660 AGCCCCTGCAGTCATTAGGG'CCAGGTAGGAGACAAAGGCGTTTTGCAGGAGTGGTACTTG 720 CAGGTGTAGCTTTAGGAGTG(~CTACAGCTGCACAAATCACTGCAGGAATAGCTTTACATC 780 AATCCAACCTCAATGCTCAA(~CAATCCAATCTCTTAGAACCAGCCTTGAACAGTCTAACA 840 AAGCTATAGAAGAAATTAGG(~AGGCTACCCAAGAAACCGTCATTGCCGTTCAGGGAGTCC 900 AGGACTACGTCAACAACGAA(:TCGTCCCTGCCATGCAACATATGTCATGTGAATTAGTTG 960 CGAGTTTACGTGACCCTATT'.CCAGCCGAGATATCAATTCAGGCACTGATTTATGCTCTTG 1080 GAGGAGAAATTCATAAGATAC:TTGGGAAGTTGGGATATTCTGGAAGTGATATGATTGCAA 1140 TCATCCTAAGTATCTCATAC(:CAACTTTATCAGAAGTCAAGGGGGTTATAGTCCACAGAC 1260 AGTCAGCCATTTGTAGCCAGAACTCCC't'GTATCCCATGAGCCCACTCTTACAACAATGTA 1440 TTAGGGGCGACACTTCATCT'..'GTGCTCGGACCTTGGTATCTGGGACTATGGGCAACAAAT 1500 GCACAAGCACAATTATTAAT<:AGAGTCCTGATAAGTTGCTGACATTCATTGCCTCCGATA 1620 CCTGCCCACTGGTTGAAATAC~ATGGTGCTACTATCCAAGTTGGAGGCAGGCAATACCCTG 1680 . 198 CCTCTAACCAGATCCTTGAG;~CGGTTAGGCGCTCTTCCTTCAATTTTGGCAGTCTCCTCA 1860 GCGTTCCTATATTAAGTTGTi'-1CAGCCCTGGCTTTGTTGTTGCTGATTTACTGTTGTAAAA 1920 GACGCTACCAACAGACACTCi'~AGCAGCATACTAAGGTCGATCCGGCATTTAAACCTGATC 1980 TAACTGGAACTTCGAAATCC'PATGTGAGATCACACTGACTCGAGATCCACTAGTTCTAGA 2040 CTTGGCGTAATCATGGTCATi~GCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCC 2160 ACACAACATACGAGCCGGAAt~CATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTA 2220 ACTCACATTAATTGCGTTGCcsCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCA 2280 GCTGCATTAATGAATCGGCCi~ACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTC 2340 CGCTTCCTCGCTCACTGACTc:GCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGC 2400 TCACTCAAAGGCGGTAATAC(~GTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACAT 2460 GTGAGCAAAAGGCCAGCAAAi3GGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTT 2520 CCATAGGCTCCGCCCCCCTGi~CGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCG 2580 TCCTGTTCCGACCCTGCCGC'PTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGT 2700 GGCGCTTTCTCATAGCTCAC(~CTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAA 2760 GCTGGGCTGTGTGCACGAAC(:CCCCGT'PCAGCCCGACCGCTGCGCCTTATCCGGTAACTA 2820 TCGTCTTGAGTCCAACCCGG~CAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAA 2880 CTACGGCTACACTAGAAGGAC:AGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTT 3000 CGGAAAAAGAGTTGGTAGCT(:TTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTT 3060 TTTTGTTTGCAAGCAGCAGA~'TACGCG(:AGAAAAAAAGGATCTCAAGAAGATCCTTTGAT 3120 CTTTTCTACGGGGTCTGACG(:TCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCAT 3180 GAGATTATCAAAAAGGATCT~.'CACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATC 3240 ACCTATCTCAGCGATCTGTC".'ATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTA 3360 CCCACGCTCACCGGCTCCAG;~TTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCG 3480 CAGAAGTGGTCCTGCAACTT'rATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGC 3540 TAGAGTAAGTAGTTCGCCAGT TAATAG'rTTGCGCAACGTTGTTGCCATTGCTACAGGCAT 3600 CGTGGTGTCACGCTCGTCGT'PTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAG 3660 GCGAGTTACATGATCCCCCA'PGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGAT 3720 TTCTCTTACTGTCATGCCATt~CGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAA 3840 GTCATTCTGAGAATAGTGTA'rGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGA 3900 GCGAAAACTCTCAAGGATCT'CACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGC 4020 ACCCAACTGATCTTCAGCAT(~TTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGG 4080 AAGGCAAAATGCCGCAAAAAi~GGGAATAAGGGCGACACGGAAATGTTGAATACTCATACT 4140 CTTCCTTTTTCAATATTATT(~AAGCAT'TTATCAGGGTTATTGTCTCATGAGCGGATACAT 4200 ATTTGAATGTATTTAGAAAAi~TAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGT 4260 GCCACCTGGGAAATTGTAAA(~GTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAA 4320 ATCAGCTCATTTTTTAACCAi~TAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAA 4380 TAGACCGAGATAGGGTTGAG'PGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAAC 4440 GTGGACTCCAACGTCAAAGG(~CGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAA 4500 GTAACCACCACACCCGCCGC(~CTTAATGCGCCGCTACAGGGCGCGTCGCGCCATTCGCCA 4740 TTCAGGCTGCGCAACTGTTG(JGAAGGG(:GATCGGTGCGGGCCTCTTCGCTATTACGCCAG 4800 CTGGCGAAAGGGGGATGTGC~"."GCAAGG(:GATTAAGTTGGGTAACGCCAGGGTTTTCCCAG 4860 TCACGACGTTGTAAAACGACC~GCCAGT(sAATTGTAATACGACTCACTATAGGGCGAATTG 4920 GGTACCGGGCCCCCCCTCGAC~GTCGACGGTATCGATAAGCTTGAT 4965 (2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5241 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:23:
AAGCTTAATGTCGTAACAAC'PCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGT 60 GGGAGGTCTATATAAGCAGAGCTCGTT')?AGTGAACCGTCTGCAGACTCTCTTCCGCATCG 120 CTGTCTGCGAGGGCCAGCTG~PTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCC 180 AGTACTCTTGGATCGGAAAC(:CGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTG 240 ACAAGACCGCCCCCCACAAC(:CAGCACCGAACTCGAAGAGACCAGGACCTCCCGAGCACG 420 CGTCTCCTACACCATGAACA(~GACCAGGTCCCGCAAGCAAACCAGCCACAGATTGAAGAA 540 AGGAGCGAGATCCCAGATCG~~AAGGCGGCAACCCAATGCAATCAACTCAGGCTCTCATTG 660 CACCTGGTTAGTCCTGTGGTC~CCTCGGAATGGCCAGTCTCTTTCTTTGTTCCAAGGCTCA 720 GATACATTGGAATAATTTGT<:AACTATTGGGATTATCGGGACTGATAGTGTCCATTACAA 780 TATAGAGAATTGTACCAAAG(:AGAATTAGGTGAGTATGAGAAATTATTGAATTCAGTCCT 900 CGAACCAATCAACCAAGCTT7.'GACTCTAATGACCAAGAATGTGAAGCCCCTGCAGTCATT 960 AGTGGCTACAGCTGCACAAA7.'CACTGCAGGAATAGCTTTACATCAATCCAACCTCAATGC 1080 TCAAGCAATCCAATCTCTTAC~AACCAGCCTTGAACAGTCTAACAAAGCTATAGAAGAAAT 1140 TAGGGAGGCTACCCAAGAAA(:CGTCATTGCCGTTCAGGGAGTCCAGGACTACGTCAACAA 1200 CGAACTCGTCCCTGCCATGCi'~ACATATGTCATGTGAATTAGTTGGGCAGAGATTAGGGTT 1260 TATTTCAGCCGAGATATCAA'rTCAGGCACTGATTTATGCTCTTGGAGGAGAAATTCATAA 1380 GATACTTGGGAAGTTGGGATi~TTCTGGAAGTGATATGATTGCAATCTTGGAGAGTCGGGG 1440 GATAAAAACAAAAATAACTCi3TGTTGATCTTCCCGGGAAATTCATCATCCTAAGTATCTC 1500 ATACCCAACTTTATCAGAAG'CCAAGGGGGTTATAGTCCACAGACTGGAAGCGGTTTCTTA 1560 CCAGAACTCCCTGTATCCCA'CGAGCCCACTCTTACAACAATGTATTAGGGGCGACACTTC 1740 ATCTTGTGCTCGGACCTTGG'PATCTGGGACTATGGGCAACAAATTTATTCTGTCAAAAGG 1800 TAATATCGTCGCAAATTGTG(:TTCTATACTATGTAAGTGTTATAGCACAAGCACAATTAT 1860 CAAAGTTGCCTTAGGCCCTG(:TATATCACTTGATAGGTTAGATGTAGGTACAAACTTAGG 2040 GAACGCCCTTAAGAAACTGGATGATGC'CAAGGTACTGATAGACTCCTCTAACCAGATCCT 2100 TGAGACGGTTAGGCGCTCTT(:CTTCAATTTTGGCAGTCTCCTCAGCGTTCCTATATTAAG 2160 ACTCAAGCAGCATACTAAGG'.PCGATCCGGCATTTAAACCTGATCTAACTGGAACTTCGAA 2280 GTTTTACTTGCTTTAAAAAAC:CTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAAT 2400 GCAATTGTTGTTGTTAACTTC~TTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGC 2460 CTCATCAATGTATCTTATCA"..'GTCTGGATCCCCCGGAATTCACTGGCCGTCGTTTTACAA 2580 CGTCGTGACTGGGAAAACCC7.'GGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCC 2640 TTCGCCAGCTGGCGTAATAG(:GAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGC 2700 AGCCTGAATGGCGAATGGCG(:CTGATGC:GGTATTTTCTCCTTACGCATCTGTGCGGTATT 2760 TCACACCGCATATGGTGCAC7.'CTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAG 2820 TACACTCCGCTATCGCTACG7.'GACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCC 2880 GCTGACGCGCCCTGACGGGC7.'TGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACC 2940 . 202 GTCTCCGGGAGCTGCATGTG'rCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAG 3000 TTCTTGAAGACGAAAGGGCC'PCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAAT 3060 AAAAGATGCTGAAGATCAGT'CGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAG 3360 CGGTAAGATCCTTGAGAGTT'PTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAA 3420 AGTTCTGCTATGTGGCGCGG'~ATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCG 3480 CCGCATACACTATTCTCAGA~~TGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCT 3540 TACGGATGGCATGACAGTAA(~AGAATTATGCAGTGCTGCCATAACCATGAGTGATAACAC 3600 TGCGGCCAACTTACTTCTGA(:AACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCA 3660 CAACATGGGGGATCATGTAA(:TCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCAT 3720 ACCAAACGACGAGCGTGACAC:CACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACT 3780 ATTAACTGGCGAACTACTTA(:TCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGC 3840 GGATAAAGTTGCAGGACCAC'.PTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGA 3900 TAAATCTGGAGCCGGTGAGCC~TGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGG 3960 TAAGCCCTCCCGTATCGTAG'.CTATCTA(:ACGACGGGGAGTCAGGCAACTATGGATGAACG 4020 AGTTTACTCATATATACTTTAGATTGA'CTTAAAACTTCATTTTTAATTTAAAAGGATCTA 4140 CTGAGCGTCAGACCCCGTAGAAAAGAT(:AAAGGATCTTCTTGAGATCCTTTTTTTCTGCG 4260 CGTAATCTGCTGCTTGCAAAC:AAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGA 4320 TCAAGAGCTACCAACTCTTT'."TCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAA 4380 TACTGTCCTTCTAGTGTAGCC:GTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCC 4940 TACATACCTCGCTCTGCTAA".'CCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTG 4500 TCTTACCGGGTTGGACTCAAC~ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAAC 4560 GGGGGGTTCGTGCACACAGCC:CAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCT 4620 ACAGCGTGAGCATTGAGAAAC~CGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCC 4680 GTATCTTTATAGTCCTGTCGc;GTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATG 4800 CTCGTCAGGGGGGCGGAGCC'PATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCT 4860 ATGCTTCCGGCTCGTATGTT(~TGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAAC 5220 AGCTATGACCATGATTACGC(: 5241 (2) INFORMATION FOR SES2 ID N0:24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '.1147 base pairs (B) TYPE: nucleic acid (C) STRANDEDLdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:24:
AAGCTTAATG TCGTAACAAC ~.'CCGCCCCGT TGACGCAAAT GGGCGGTAGG CGTGTACGGT 60 GGGAGGTCTA TATAAGCAGA GCTCGTT'.CAG TGAACCGTCT GCAGACTCTC TTCCGCATCG 120 CTGTCTGCGA GGGCCAGCTG ~.'TGGGCTCGC GGTTGAGGAC AAACTCTTCG CGGTCTTTCC 180 AGTACTCTTG GATCGGAAAC C:CGTCGGCCT CCGAACGGTA CTCCGCCACC GAGGGACCTG 240 TCGCAAGTCT AGAATGCACA AGGGAAT(:CC CAAAAGCTCC AAAACCCAAA CACATACCCA 360 ACAAGACCGC CCCCCACAAC C:CAGCACCGA ACTCGAAGAG ACCAGGACCT CCCGAGCACG 420 ACACAGCACA ACATCAGCTC ~~GCGATCCAC GCACTACGAT CCTCGAACAT CGGACAGACC 480 CGTCTCCTAC ACCATGAACA C~GACCAGGTC CCGCAAGCAA ACCAGCCACA GATTGAAGAA 540 CATCCCAGTT CACGGAAACC ~1CGAGGC(:AC CATCCAGCAC ATACCAGAGA GTGTCTCAAA 600 TATAGAGAATTGTACCAAAG(:AGAATTAGGTGAGTATGAGAAATTATTGAATTCAGTCCT900 CGAACCAATCAACCAAGCTT'PGACTCTAATGACCAAGAATGTGAAGCCCCTGCAGTCATT960 AGTGGCTACAGCTGCACAAA'CCACTGCAGGAATAGCTTTACATCAATCCAACCTCAATGC1080 TCAAGCAATCCAATCTCTTA(~AACCAGCCTTGAACAGTCTAACAAAGCTATAGAAGAAAT1140 TAGGGAGGCTACCCAAGAAA(:CGTCATTGCCGTTCAGGGAGTCCAGGACTACGTCAACAA1200 CGAACTCGTCCCTGCCATGC~~ACATATGTCATGTGAATTAGTTGGGCAGAGATTAGGGTT1260 AAGACTGCTTCGGTATTATA(:TGAGTTGTTGTCAATATTTGGCCCGAGTTTACGTGACCC1320 TATTTCAGCCGAGATATCAA'PTCAGGCACTGATTTATGCTCTTGGAGGAGAAATTCATAA1380 GATACTTGGGAAGTTGGGAT~~TTCTGGAAGTGATATGATTGCAATCTTGGAGAGTCGGGG1440 GATAAAAACAAAAATAACTC~~TGTTGA'rCTTCCCGGGAAATTCATCATCCTAAGTATCTC1500 ATACCCAACTTTATCAGAAG'CCAAGGGGGTTATAGTCCACAGACTGGAAGCGGTTTCTTA1560 CAACATAGGATCACAAGAGT(~GTACACCACTGTCCCGAGGTATATTGCAACTAATGGTTA1620 CCAGAACTCCCTGTATCCCA'.CGAGCCCACTCTTACAACAATGTATTAGGGGCGACACTTC1740 TAATATCGTCGCAAATTGTGC:TTCTATACTATGTAAGTGTTATAGCACAAGCACAATTAT1860 TAATCAGAGTCCTGATAAGT~"GCTGACATTCATTGCCTCCGATACCTGCCCACTGGTTGA1920 CAAAGTTGCCTTAGGCCCTG<:TATATCACTTGATAGGTTAGATGTAGGTACAAACTTAGG2040 GAACGCCCTTAAGAAACTGGATGATGC'.CAAGGTACTGATAGACTCCTCTAACCAGATCCT2100 TGAGACGGTTAGGCGCTCTTC:CTTCAATTTTGGCAGTCTCCTCAGCGTTCCTATATTAAG2160 TTGTACAGCCCTGGCTTTGT7:GTTGCTGATTTACTGTTGTAAAAGACGCTACCAACAGAC2220 ACTCAAGCAGCATACTAAGG7.'CGATCCGGCATTTAAACCTGATCTAACTGGAACTTCGAA2280 ATCCTATGTGAGATCACACTC~ACCGCGGAATTGTTGTTGTTAACTTGTTTATTGCAGCTT2340 TGCATTCTAGTTGTGGTTTG'PCCAAACTCATCAATGTATCTTATCATGTCTGGATCCCCC 2460 GGAATTCACTGGCCGTCGTT'rTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAAC 2520 CCGATCGCCCTTCCCAACAG'PTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATT 2640 TTCTCCTTACGCATCTGTGC(sGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCT 2700 GCTCTGATGCCGCATAGTTAi~GCCAGTACACTCCGCTATCGCTACGTGACTGGGTCATGG 2760 CTGCGCCCCGACACCCGCCAi~CACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGG 2820 CGTCATCACCGAAACGCGCGi~GGCAGTTCTTGAAGACGAAAGGGCCTCGTGATACGCCTA 2940 GGAAATGTGCGCGGAACCCC'CATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCG 3060 ATTCAACATTTCCGTGTCGC(:CTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTT 3180 GGTTACATCGAACTGGATCT(;AACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAA 3300 CGTTTTCCAATGATGAGCAC~~TTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATT 3360 GACGCCGGGCAAGAGCAACT(:GGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAG 3420 GCAATGGCAACAACGTTGCG(~AAACTA'.CTAACTGGCGAACTACTTACTCTAGCTTCCCGG 3720 CAACAATTAATAGACTGGAT(~GAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCC 3780 CTTCCGGCTGGCTGGTTTAT~'GCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGT 3840 GGGAGTCAGGCAACTATGGA".'GAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTG 3960 ATTAAGCATTGGTAACTGTC~~GACCAAGTTTACTCATATATACTTTAGATTGATTTAAAA 4020 . . 206 TCTTCTTGAGATCCTTTTTT'PCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCG4200 CACTTCAAGAACTCTGTAGCi~CCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTG4380 GCTGCTGCCAGTGGCGATAAc;TCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCG4440 ACGACCTACACCGAACTGAGe~TACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCC4560 GAAGGGAGAAAGGCGGACAG(~TATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACG4620 AGGGAGCTTCCAGGGGGAAA(~GCCTGG'rATCTTTATAGTCCTGTCGGGTTTCGCCACCTC4680 TGACTTGAGCGTCGATTTTT(~TGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCC4740 CCTGCGTTATCCCCTGATTC'.CGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACC4860 GCTCGCCGCAGCCGAACGAC(:GAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGC4920 CAATACGCAAACCGCCTCTC(~CCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACA4980 ATTAGGCACCCCAGGCTTTAC:ACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA5100 GCGGATAACAATTTCACACA(~GAAACAGCTATGACCATGATTACGCC 5147 (2) INFORMATION FOR SEQ ID N0:25:
( i ) SEQUENCE CHARACTERIS'CICS
(A) LENGTH: 8792 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE;: DNA (genomic) (xi) SEQUENCE
DESCJ~IPTION:
SEQ ID
N0:25:

AAAATAAGTTTAAAAGCTTTi~TTTTTCATACACGCGAGCGGTAAGGCTGCCGCCTTCAGG290 AAAAGTTACTCTGTAAACAG'PTCTTTCACAACAGCACAAAACATAGGTATTAGTTAACAG300 TTCATTTGGGCTATAATAAT~~TACATTTTCTTGGGTGGCAAAGCAAGGGTCGGTAATCTC360 AACAAAACCATCAACTGGAA'CGCAAGAATAGTCCAGCACGGTGGGTTCAATCTAAAAATG420 TCATAGCTTGTCTCAAAGCAGATTGTC'PTCTTTCCTCTGCCTTGGAAGTGGTTTGGTGAA540 GCACTACAGGTGTCTTTTCAACCTCTT'PCAGCACCCGCTCTATTACAGATCTCACCCACA600 CAGCACAGTTTTTAAGAGAA(:AATAGT'PTTGAAGGCTACAAGATTTACACTTAAGCACCA660 GCCAGTAATTATAAGTGCTT'CTAAGAACTACCCCTAGCTCAGGGTTAATGCACCTTTTAA720 GAGTGTTTCTAAATATAATA(:TCCCCACATAGTTAATTTCATCAGGCCTGCTAGAATTTA840 CAAACTCTCGGTACCACATA'.CACTTTT'CATTCATAGCCCCACCCTTAATAAAGTCCTCAA900 TCACTTTCTGAACCACATGC'.CTGCTAGCCATGCATTGTAAAGACAAGCTGTTAGAGCAGT960 CATCCCACAAAATAGGAATT~.'CATAGCATAAAGCAAAGCAATTACAATATTTAGGAACTC1140 TCACCACAGCAGTCACGTGAC:ATGTTGTCTCAGCAGTGCAGTTGCCTTCCATCCTACAAT1200 TATGAACAAAAACTAAACAC':.'TCTAACAAAGATACAGTGACAATCTCCCTTCCTCTAAAA1260 GCATTGTTTACATTAGGGTG~~TTATTAACAACGTCAGAAATTTCTTTAATTAAAGTGCCT1320 TTAAAATGTGCAAGAGCATC~1TCATACTCAAAACCAAGCTGAGAGTAAAAGACCACCTTA1380 AAAGTAATCCCAGGCTTGTT':.'TTATCAACAGCCTTAAACATGCTTTCACAAAATATAGAA1440 GCAGTAACATCATCAATGGTC~TCGAAGAGAAACTCCATAGGAGACTCCAGCATTGATCCA1500 AGCTCTCTAACAAAATCTTC<~TCAAAATGAATAATGCCCTTTACACAAACGCGGGGCAGA1560 AGTGGTAAAGTCATGCCCATC~GGTGAGGCCAAAATCCTTAAAAAAGCTATCTAAGTAGTT1680 GGTCATCCCCTCAGTTi~AAA.~1GTTTTGCAGCTGGGTGGTGCATACCACATAGTGCCAGCT 1740 TATAGCTACAAAGACCTGCA'rCCCCTCCTTAGCAGACAGCTCTTGCACACACGCAGTAAC 1800 CAAGGAGAGCi~AAATTTCAGCAAGCGCAGGCTCAACAGTAATAGTGAAGCAGAGGCATTT 1920 CAGACGAGGCTCACTAGCTGCAGTCGCCATTTATGAGGTCTGCAATi~AAAAACAACTCAT 1980 CAGCAGCTGAAAAAGTGCAC'PTTGACCTCATTAAGCCACTGCATATGCi~AGTCCTCATCT 2040 AGGGGCCCGCTGTGGCTGGAi~AGCCTGCGCACAGCCCGAAGGTTAAAAATGGACTGTAAC 2220 AGCATTGAAACCCCGCGACAc:AGGTCAGTCTCGCGGTCTTGATCTCTTATTATAGCGACC 2280 AAATGGTCCTTCAGAGTGATt~TTGCAC'rCATAGAAGTAGGCAGCTCCGGCAGCCATTCTG 2340 CAAAATAACAAAACACCACTi~.AGCATAGCACCATCACCAAGCATGAAAACAGGTAAAAAC 2400 AAAAGCAACACTTACTTATT(:AGCAGTCACAAGAATGTTGGGCTCCCAAGTGACAGACi~A2460 GCCTAATGCAAGGTGGGCACi~GTCTCCGGAATAAGTTGACAAAAGTCACGCCGCAAAGCT 2520 TCCTGAAGAGAAACGGCGGTi~GCCTGGATATCTGCAACGGACCCAAAACCTTCAGTGTCA 2580 CTTCCAATAAACAGATi~AAA(:TCTAAATAGTCCCCACTTAi~AACCGAAACAGCCGCGGCA 2640 TCAGAAGGCAAAAAGTCTGTAP.GCTCTAGCTGAGCACACACACTCTCCACTAGACACTTG 2760 GACGGCTGGCTGTCAGAGGA(~AGCTATGAGGATGAAATGCCAAGCACAGCGTTTATATAG 2880 TCCTCAAAGTAGGGCGTGTG(~AAAACGi~AAAGGAATATAACGGGGCGTTTGAGGAAGTGG 2940 TGCCAAGTACAGTCATAAAA'.CGTGGGCGCGTGGTAAATGTTAAGTGCAGTTTCCCTTTGG 3000 CGGTTGGCCCGGAAAGTTCA(:AAAAAG'CACAGCACGTCCTTGTCACCGTGTCAACCACAA 3060 i~ACCACAAATAGGCACAACGC:CCAAAAACCCGGGTCGACACGCGTGAATTCACCGGTTCG 3120 AGCTTAATGTCGTAACAACTC:CGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTG 3180 GGAGGTCTATATAAGCAGAGC:TCGTTTAGTGAACCGTCTGCAGACTCTCTTCCGCATCGC 3240 TGTCTGCGAGGGCCAGCTGT"..'GGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCA 3300 GTACTCTTGGATCGGAAACCC:GTCGGC(:TCCGAACGGTACTCCGCCACCGAGGGACCTGA 3360 GCGAGTCCGCATCGACCGGA7.'CGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGT 3420 ATCATGACTAGGCCCAGTCACCAGTAC'rTGGTCATAAAACTGATGCCTAATGTTTCACTT 3960 AGGGAGGCTACCCAAGAAAC(:GTCATTGCCGTTCAGGGAGTCCAGGACTACGTCAACAAC 4320 AGACTGCTTCGGTATTATAC'CGAGTTG'rTGTCAATATTTGGCCCGAGTTTACGTGACCCT 4440 ATTTCAGCCGAGATATCAAT'CCAGGCACTGATTTATGCTCTTGGAGGAGAAATTCATAAG 4500 ATACTTGGGAAGTTGGGATA'CTCTGGAAGTGATATGATTGCAATCTTGGAGAGTCGGGGG 4560 ATAAAAACAAAAATAACTCA'CGTTGATCTTCCCGGGAAATTCATCATCCTAAGTATCTCA 4620 TACCCAACTTTATCAGAAGT(:AAGGGGGTTATAGTCCACAGACTGGAAGCGGTTTCTTAC 4680 AACATAGGATCACAAGAGTG(~TACACCACTGTCCCGAGGTATATTGCAACTAATGGTTAC 4740 TTAATATCTAATTTTGATGAGTCATCT'PGTGTATTCGTCTCAGAGTCAGCCATTTGTAGC 4800 AATCAGAGTCCTGATAAGTT(~CTGACATTCATTGCCTCCGATACCTGCCCACTGGTTGAA 5040 AAAGTTGCCTTAGGCCCTGC~.'ATATCACTTGATAGGTTAGATGTAGGTACAAACTTAGGG 5160 AACGCCCTTAAGAAACTGGA'PGATGCTAAGGTACTGATAGACTCCTCTAACCAGATCCTT5220 CAATTGTTGTTGTTAACTTG'PTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCA5580 AAAGTCCGCGGAACTCGCCC'PGTCGTAAAACCACGCCTTTGACGTCACTGGACATTCCCG5760 TGGGAACACCCTGACCAGGGCGTGACC'PGAACCTGACCGTCCCATGACCCCGCCCCTTGC5820 CTAGTTCTAGAGCGGCCGCCi~CCGCGGTGGAGCTCCAGCTTTTGTTCCCTTTAGTGAGGG5940 TTAATTCCGAGCTTGGCGTAATCATGG'PCATAGCTGTTTCCTGTGTGAAATTGTTATCCG6000 TGAGTGAGCTAACTCACATTf~ATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAAC6120 GCGGTATCAGCTCACTCAAA(~GCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCA6300 CTGGCGTTTTTCCATAGGCT(:CGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGT6420 CTCGTGCGCTCTCCTGTTCC(~ACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCT6540 TCGGGAAGCGTGGCGCTTTC'.PCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTC6600 TCCGGTAACTATCGTCTTGAC~TCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCA6720 GCCACTGGTAACAGGATTAG(:AGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAG6780 TGGTGGCCTAACTACGGCTA(:ACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAG6840 ATTTTGGTCATGAGATTATC;~AAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGA7080 AGTTTTAAATCAATCTAAAG'rATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTA7140 AGGGCCGAGCGCAGAAGTGG'rCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGT7380 TGCCGGGAAGCTAGAGTAAG'L'AGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATT7440 GCTACAGGCATCGTGGTGTCi~CGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCC7500 CAACGATCAAGGCGAGTTACi~TGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTC7560 GGTCCTCCGATCGTTGTCAG~S.AGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCA7620 GCACTGCATAATTCTCTTAC'CGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAG7680 GCAAAAACAGGAAGGCAAAA'CGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGA7980 ATACTCATACTCTTCCTTTT'.CCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATG8040 AGCGGATACATATTTGAATG'.CATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTT8100 CCCCGAAAAGTGCCACCTGGGAAATTG'CAAACGTTAATATTTTGTTAAAATTCGCGTTAA8160 ATTTTTGTTAAATCAGCTCA'.~'TTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATA8220 AATCAAAAGAATAGACCGAGATAGGGT'CGAGTGTTGTTCCAGTTTGGAACAAGAGTCCAC8280 CACTACGTGAACCATCACCC'"AATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAA8400 ATCGGAACCCTAAAGGGAGCC:CCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGG8460 GCCATTCGCCATTCAGGCTGC:GCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGC8640 , 212 GGTTTTCCCAGTCACGACGT'rGTAAAACGACGGCCAGTGA ATTGTAATAC GACTCACTAT8760 (2) INFORMATION FOR SEQ ID N0:26:
(i) SEQUENCE CHAR~CTERIS'rICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:26:

(2) INFORMATION FOR SES~ ID N0:27:
( i ) SEQUENCE CHAR~~CTERIS'PICS
(A) LENGTH: :?1 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDNESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:27:
GTGCTGGCTG GCACGGGCAT '.C 21 (2) INFORMATION FOR SEQ ID N0:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDED2dESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:28:
ATGTCCACCA AAGTCCCCTC 'I' 21 (2) INFORMATION FOR SEc~ ID N0:29:
( i ) SEQUENCE CHARi~.CTERIS'rICS
(A) LENGTH: :?1 base pairs (B) TYPE: nucleic acid (C) STRANDEDt4ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:29:
CCCGGGGCGT CGTATGGATA 'C 21 (2) INFORMATION FOR SEQ ID N0:30:
( i ) SEQUENCE CHARACTERIS'CICS
(A) LENGTH: :?4 base pairs (B) TYPE: nucleic acid (C) STRANDEDLdESS: single (D) TOPOLOGY:; linear (ii) MOLECULE TYPE.; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:30:

(2) INFORMATION FOR SEQ ID N0:31:
(i) SEQUENCE CHAR~~CTERISTICS:
(A) LENGTH: ~?4 base pairs (B) TYPE: nu<:leic acid (C) STRANDED1VESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC1~IPTION: SEQ ID N0:31:

(2) INFORMATION FOR SECT ID N0:32:
( i ) SEQUENCE CHARe~CTERIS'PICS
(A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDtQESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:32:

(2) INFORMATION FOR SEQ ID N0:33:
( i ) SEQUENCE CHARACTERIS'CICS
(A) LENGTH: :?4 base pairs (B) TYPE: nucleic acid (C) STRANDEDtdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:33:
GGTTGTGTGG AAGACCCGGG C~GCG 24 (2) INFORMATION FOR SEQ ID N0:34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~t5 base pairs . 215 (B) TYPE: nu~~leic acid (C) STRANDED1VESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:34:
AGATCTGCTA ATGGAACGCG 'PATCGCTGCC CCCACAGTAC AGCAA 45 (2) INFORMATION FOR SEQ ID N0:35:
(i) SEQUENCE CHARi~CTERISTICS:
(A) LENGTH: 58 base pairs (B) TYPE: nucleic acid (C) STRANDEDI4ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:35:

(2) INFORMATION FOR SEQ ID N0:36:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 58 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: s:i.ngle (D) TOPOLOGY:; linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:36:

(2) INFORMATION FOR SEQ ID N0:37:
(i) SEQUENCE CHAR1~CTERISTICS:

(A) LENGTH: 38 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:37:
CTAGTCATCT TAACGCGTGT CCTCAACATC ACCCGCGA 3g (2) INFORMATION FOR SE(~ ID N0:38:
(i) SEQUENCE CHARi~CTERIS'rICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDI4ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:38:
CTTGCTTGTT ATTAAAAAAA (~ 21 (2) INFORMATION FOR SEQ ID N0:39:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :?1 base pairs (B) TYPE: nucleic arid (C) STRANDEDIdESS: single (D) TOPOLOGY:; linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:39:

TCACGCCCTG GTCAGGGTGT 't 21 (2) INFORMATION FOR SEQ ID N0:40:
(i) SEQUENCE CHAR:3CTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDIVESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:40:
GCCATCGCGT CAACCTGA lg (2) INFORMATION FOR SEQ ID N0:41:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :L8 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:41:

(2) INFORMATION FOR SEQ ID N0:42:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~?1 base pairs (B) TYPE: nucleic acid (C) STRANDEDtdESS: single (D) TOPOLOGYr. linear , , 218 (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC:ftIPTION: SEQ ID N0:42:
TCACGCCCTG GTCAGGGTGT 'I 21 (2) INFORMATION FOR SEQ ID N0:93:
(i) SEQUENCE CHARR~CTERISTICS:
(A) LENGTH: :18 base pairs (B) TYPE: nucleic acid (C) STRANDEDtJESS: single ( D) TOPOLOGY : linea:r (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:43:
ACCACGCGCC CACATTTT lg (2) INFORMATION FOR SEQ ID N0:44:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) STRANDEDtdESS: single (D) TOPOLOGY:; linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:44:

(2) INFORMATION FOR SE(2 ID N0:45:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 43 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDP~ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC1~IPTION: SEQ ID N0:45:
CGATATCCGT TAAGTTTGTA 'I'CGTAATGCT CCCCTACCAA GAC 43 (2) INFORMATION FOR SEQ ID N0:46:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 40 base pairs (B) TYPE: nucleic acid (C) STRANDEDtQESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:46:
GGGATAAAAA TTAACGGTTA (:ATGAGAATC TTATACGGAC 40 (2) INFORMATION FOR SES2 ID N0:47:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:47:
GGGCTGAAGC TTGCTGGCCG (:TCATTAGAC AAGCGAATGA GGGAC 45 (2) INFORMATION FOR SE(2 ID N0:48:
(i) SEQUENCE CHARACTERIS'~ICS:
(A) LENGTH: Ei2 base pairs (B) TYPE: nucleic acid (C) STRANDED1VESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:48:

(2) INFORMATION FOR SEc~ ID N0:49:
(i) SEQUENCE CHARi~CTERIS'rICS:
(A) LENGTH: 64 base pairs (B) TYPE: nucleic acid (C) STRANDEDI~ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:49:

(2) INFORMATION FOR SEQ ID N0:50:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~15 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:50:

TATCTCGAAT TCCCGCGGCT 'rTAAATGGAC GGAACTCTTT TCCCC 45 (2) INFORMATION FOR SEQ ID N0:51:
( i ) SEQUENCE CHARi~CTERI S'rICS
(A) LENGTH: 62 base pairs (B) TYPE: nucleic acid ( C ) STRANDEDIJESS : s.ingle (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:51:
GATCTTTTGT TAACAAAAAC 'CAATCAGCTA TCGCGAATCG ATTCCCGGGG GATCCGGTAC 60 (2) INFORMATION FOR SES~ ID N0:52:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 62 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:52:

(2) INFORMATION FOR SEQ ID N0:53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 43 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:53:
CGATATCCGT TAAGTTTGTA 'PCGTAATCTG CAGCCCGGGG GGG 43 (2) INFORMATION FOR SE(~ ID N0:54:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 44 base pairs (B) TYPE: nucleic acid (C) STRANDEDI4ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:54:

(2) INFORMATION FOR SEQ ID N0:55:
( i ) SEQUENCE CHARACTERIS')?ICS
(A) LENGTH: :?9 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:55:

(2) INFORMATION FOR SE(2 ID N0:56:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :36 base pairs (B) TYPE: nucleic acid , 223 (C) STRANDEDI~IESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:56:

(2) INFORMATION FOR SEC2 ID N0:57:
(i) SEQUENCE CHARi~.CTERIS'PICS:
(A) LENGTH: :l8 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:57:

(2) INFORMATION FOR SES2 ID N0:58:
( i ) SEQUENCE CHARACTERIS'CICS
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDtdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:58:
GTAATTAGTA AAATTCACCT "G 22 (2) INFORMATION FOR SEQ ID N0:59:
(i) SEQUENCE CHAR~~CTERISTICS:
(A) LENGTH: :_8 base pairs , 224 (B) TYPE: nucleic acid (C) STRANDED1VESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:59:
TGGAGATCGC GGAAGTCG lg (2) INFORMATION FOR SE(2 ID N0:60:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :L8 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:60:

(2) INFORMATION FOR SEQ ID N0:61:
( i ) SEQUENCE CHARACTERI S'C ICS
(A) LENGTH: .L7 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:61:

(2) INFORMATION FOR SE()_ ID N0:62:
(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 33 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:62:
ATGCTCCCGC GGTTAACGGT 'PACATGAGAA TCT 33 (2) INFORMATION FOR SEQ ID N0:63:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ci9 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:63:
CATAAATTAT TTCATTATCG <:GATATCCGT TAAGTTTGTA TCGTAATGCA CAAGGGAATC 60 (2) INFORMATION FOR SEQ ID N0:64:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~l8 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESC1~IPTION: SEQ ID N0:64:

(2) INFORMATION FOR SEc~ ID N0:65:
(i) SEQUENCE CHARi~CTERISTICS:
(A) LENGTH: :35 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:65:

(2) INFORMATION FOR SEQ ID N0:66:
(i) SEQUENCE CHARACTERIS'~ICS:
(A) LENGTH: 28 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE.. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:66:
GGGGGTACCT TTGAGAGTAC (:ACTTCAG 28 (2) INFORMATION FOR SE()_ ID N0:67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 49 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:67:
GGGTCTAGAG CGGCCGCTTA 'CAAAGATCTA AAATGCATAA TTTC 44 (2) INFORMATION FOR SEQ ID N0:68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :35 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY; linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:68:

(2) INFORMATION FOR SEQ ID N0:69:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 82 base pairs (B) TYPE: nucleic acid (C) STRANDEDLdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:69:

(2) INFORMATION FOR SEQ ID N0:70:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 82 base pairs (B) TYPE: nucleic acid (C) STRANDEDI4ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:70:

(2) INFORMATION FOR SES2 ID N0:71:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '10 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY; linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:71:
AGCTTCCCGG GTTAATTAAT ~.~AGTCATCAG GCAGGGCGAG AACGAGACTA TCTGCTCGTT 60 (2) INFORMATION FOR SE() ID N0:72:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~~0 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION; SEQ ID N0:72:

AGCTCTAATT AATTAACGAG (:AGATAG'PCT CGTTCTCGCC CTGCCTGATG ACTAATTAAT 60 (2) INFORMATION FOR SE(2 ID N0:73:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~~2 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:73:
ATCATCGAGC TCGCGGCCGC (:TATCAAAAG TCTTAATGAG TT 42 (2) INFORMATION FOR SEQ ID N0:74:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '13 base pairs (B) TYPE: nucleic acid (C) STRANDEDt~ESS: single (D) TOPOLOGY:; linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:74:
GAATTCCTCG AGCTGCAGCC <:GGGTTTTTA TAGCTAATTA GTCATTTTTT CGTAAGTAAG 60 (2) INFORMATION FOR SEQ ID N0:75:
( i ) SEQUENCE CHARACTERIS'CICS
(A) LENGTH: ~~2 base pairs (B) TYPE: nucleic acid . , 230 (C) STRANDED2dESS: single (D) TOPOLOGY:. linear (ii.) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:75:

(2) INFORMATION FOR SES2 ID N0:76:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: <l5 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:76:

(2) INFORMATION FOR SE()_ ID N0:77:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~t2 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE.: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:77:
ACTGTACTCG AGTCTAGAAT C~CACAAGGGA ATCCCCAAAA GC 42 (2) INFORMATION FOR SEQ ID N0:78:
(i) SEQUENCE CHARACTERISTICS:

. 231 (A) LENGTH: :L8 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:78:

(2) INFORMATION FOR SEi2 ID N0:79:
(i) SEQUENCE CHAR~~CTERISTICS:
(A) LENGTH: 39 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY;. linear (ii) MOLECULE TYPE;. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:79:

(2) INFORMATION FOR SEQ ID N0:80:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: .7 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY;: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:80:

. 232 (2) INFORMATION FOR SEc~ ID N0:81:
(i) SEQUENCE CHARe~CTERISTICS:
(A) LENGTH: :31 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:81:

(2) INFORMATION FOR SE(2 ID N0:82:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 32 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2dESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:82:
TCTACTGCAG CCGGTGTCTT C:TATGGAGGT CA 32 (2) INFORMATION FOR SEQ ID N0:83:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~?4 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:83:
TCTTCGCCCC CGTTTTCACC i~TGG 24 (2) INFORMATION FOR SEt2 ID N0:84:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:84:

(2) INFORMATION FOR SEQ ID N0:85:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '71 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2dESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:85:
AATTCGGTAC CAAGCTTCTT ~.'ATTCTATAC TTAAAAAGTG AAAATAAATA CAAAGGTTCT 60 (2) INFORMATION FOR SEQ ID N0:86:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~~0 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:86:
CGCATCGCTG TCTGCGAGGG c:CAGCTGTTG GGCTCGCGGT TGAGGACAAA CTCTTCGCGG 60 TCTTTCCAGT 7p (2) INFORMATION FOR SEC2 ID N0:87:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '70 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:87:
ACTCTTGGAT CGGAAACCCG '.L'CGGCCTCCG AACGTACTCC GCCACCGAGG GACCTGAGCG 60 (2) INFORMATION FOR SEQ ID N0:88:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: Ei0 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2JESS: single (D) TOPOLOGY;. linear (ii) MOLECULE TYPE;. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:88:
GACCGGATCG GAAAACCTCT C:GAGAAAGGC GTCTAACCAG TCACAGTCGC AAGCCCGGGT 60 (2) INFORMATION FOR SE~)_ ID N0:89:
(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs (B) TYPE: nucleic acid (C) STRANDEDI~ESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:89:
CTTTGTATTT ATTTTCACTT 'CTTAAGTATA GAATAAAGAA GCTTGGTACC G 51 (2) INFORMATION FOR SES2 ID N0:90:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '12 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDNESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:90:
GAAGAGTTTG TCCTCAACCG C:GAGCCCAAC AGCTGGCCCT CGCAGACAGC GATGCGGAAG 60 (2) INFORMATION FOR SEQ ID N0:91:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: '~3 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:91:

(2) INFORMATION FOR SEQ ID N0:92:
(i) SEQUENCE CHARi~CTERISTICS:
(A) LENGTH: '75 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:92:

(2) INFORMATION FOR SES2 ID N0:93:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 36 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2dESS: single (D) TOPOLOGY.. linear (ii) MOLECULE TYPE.. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:93:
ATCGTCCTGC AGACTCTCTT (:CGCATCGCT GTCTGC 36 (2) INFORMATION FOR SEQ ID N0;94:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~?9 base pairs (B) TYPE: nucleic acid (C) STRANDEDIQESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:94:
GCTCTAGACT TGCGACTGTG i~CTGGTTAG 2g (2) INFORMATION FOR SEC2 ID N0:95:
(i) SEQUENCE CHARACTERIS'PICS:
(A) LENGTH: :?0 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE.: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:95:

(2) INFORMATION FOR SEQ ID N0:96:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE.. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:96:

(2) INFORMATION FOR SE()_ ID N0:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ..7 base pairs (B) TYPE: nu~~leic acid (C) STRANDED1VESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC12IPTION: SEQ ID N0:97:

(2) INFORMATION FOR SEQ ID N0:98:
( i ) SEQUENCE CHARi~CTERIS'rICS
(A) LENGTH: :33 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:98:

(2) INFORMATION FOR SEQ ID N0:99:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: .L2 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2dESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:99:

(2) INFORMATION FOR SE(2 ID N0:100:

(1) SEQUENCE CHARi~CTERISTICS:
(A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:100:

(2) INFORMATION FOR SE(2 ID N0:101:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:101:

(2) INFORMATION FOR SE(2 ID N0:102:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 48 base pairs (B) TYPE: nucleic acid (C) STRANDED2dESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:102:

(2) INFORMATION FOR SEc~ ID N0:103:
(i) SEQUENCE CHARi~CTERISTICS:
(A) LENGTH: 48 base pairs (B) TYPE: nucleic acid (C) STRANDEDIQESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:103:
CCCAAAAACC CGCAAAGCGC 'CTGGCCACTT AAGTGCGCAC AGCTGGGG 48 (2) INFORMATION FOR SEQ ID N0:104:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 33 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY:; linear (ii) MOLECULE TYPE:; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:104:

(2) INFORMATION FOR SE(2 ID N0:105:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: .39 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single , , 241 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESC1~IPTION: SEQ ID N0:105:
ATCGTAGATA TCCTCGAGTT i~CGCCCCGCC CTGCCACTC 39 (2) INFORMATION FOR SE(z ID N0:106:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :33 base pairs (B) TYPE: nucleic acid (C) STRANDEDIQESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:106:

(2) INFORMATION FOR SEQ ID N0:107:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :?1 base pairs (B) TYPE: nucleic acid (C) STRANDEDIdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:107:

(2) INFORMATION FOR SEQ ID N0:108:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 24 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:108:

(2) INFORMATION FOR SE(~ ID N0:109:
(i) SEQUENCE CHARACTERIS'PICS:
(A) LENGTH: :?1 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:109:
CGCGCACAAA CTGGTAGGTG (: 21 (2) INFORMATION FOR SEQ ID N0:110:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~l2 base pairs (B) TYPE: nucleic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:110:
AGATCTGCTA ATGGAACGCG ~.'ATCAAGTTT AATAATATTA TC 42 (2) INFORMATION FOR SEQ ID NO:111:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :39 base pairs _, 243 (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:

(2) INFORMATION FOR SE(2 ID N0:112:
(i) SEQUENCE CHARI~CTERISTICS:
(A) LENGTH: .L8 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:112:

(2) INFORMATION FOR SES2 ID N0:113:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 30 base pairs (B) TYPE: nu<:leic acid (C) STRANDED2dESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:. DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:113:
ATCAGTACGC GTATGGGCCA C:ACACGGAGG 30 (2) INFORMATION FOR SE()_ ID N0:114:
(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:114:
ATCAGTAGAT CTGTTATTAG 'CGATATCAAA 30 (2) INFORMATION FOR SE(,2 ID N0:115:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~12 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY. linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:115:

(2) INFORMATION FOR SEQ ID N0:116:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 42 base pairs (B) TYPE: nu<:leic acid (C) STRANDEDPdESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:116:

, 245 (2) INFORMATION FOR SE(.2 ID N0:117:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 36 base pairs (B) TYPE: nuc=leic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE; DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:117:

(2) INFORMATION FOR SEQ ID N0:118:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY:: linear (ii) MOLECULE TYPE:: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:118:
ATCAGTACGC GTGTTATTAG '.'GATATCAAA 30 (2) INFORMATION FOR SES2 ID N0:119:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 39 base pairs (B) TYPE: nucleic acid (C) STRANDEDIJESS: single (D) TOPOLOGY:. linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:119:
ATCCGTACGC GTTAGAGGGC e~AAGCCCGTG CAGCAGCGC 39 (2) INFORMATION FOR SEC, ID N0:120:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: :39 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:120:

Claims

CLAIMS:

1. A recombinant canine adenovirus type 2 (CAV2) containing a deletion in the E3 region of the CAV2 genome and an insertion of heterologous DNA in the E3 region or in the region located between the E4 region and the right ITR
region of the CAV2 genome, wherein the CAV2 replicates in a host.

2. The CAV2 of claim 1 which is packaged as an infectious CAV2.

3. The CAV2 of claim 1 or 2, wherein the heterologous DNA encodes an expression product selected from the group consisting of an epitope of interest, a biological response modulator, a growth factor, a recognition sequence, a therapeutic gene product, and a fusion protein.

4. The CAV2 of claim 3, wherein the heterologous DNA
encodes an antigenic epitope of interest.

5. The CAV2 of claim 4, wherein the antigenic epitope of interest is an antigen of a veterinary pathogen or a veterinary toxin.

6. The CAV2 of claim 5, wherein the antigenic epitope of interest is selected from the group consisting of a Morbillivirus antigen, a rabies glycoprotein, an avian influenza antigen, a bovine leukemia virus antigen, a Newcastle Disease Virus (NDV) antigen, Feline Leukemia virus (FeLV) envelope protein, Rous associated virus type 1 (RAV-1) env, matrix and/or preplomer of infectious bronchitis virus, a herpesvirus glycoprotein, a flavivirus antigen, an immunodeficiency virus antigen, a parvovirus antigen, an equine influenza antigen, a Marek's Disease virus antigen, a poxvirus antigen, and an infectious bursal disease virus antigen.

7. The CAV2 of claim 6, wherein the Morbillivirus antigen comprises canine distemper virus hemagglutinin (HA) or fusion (F) proteins.

8. The CAV2 of claim 4, wherein the antigenic epitope of interest is an antigen of a human pathogen or toxin.

9. The CAV2 of claim 8, wherein the antigenic epitope of interest is selected from the group consisting of a Morbillivirus antigen, a rabies glycoprotein, an influenza antigen, a herpesvirus antigen, a flavivirus antigen, a hepatitis virus antigen, an immunodeficiency virus antigen, a Hantaan virus antigen, a C. tetani antigen, a mumps antigen, a pneumococcal antigen, a Borrelia antigen, a Plasmodium antigen, and a chicken pox antigen.

10. The CAV2 of any one of claims 1 to 9, wherein the heterologous DNA includes a promoter.

11. The CAV2 of claim 10, wherein the promoter is a herpesvirus promoter.

12. The CAV2 of claim 10, wherein the promoter is a cytomegalovirus (CMV) promoter.

13. The CAV2 of claim 12, wherein the promoter is a murine CMV-IE promoter.

14. The CAV2 of claim 12, wherein the promoter is a human CMV-IE promoter.

15. The CAV2 of claim 12, wherein the promoter is a truncated transcriptionally active human CMV-IE promoter, the nucleotide sequence therefor being set forth at nucleotides 2-93 of SEQ ID NO: 10.

16. An immunogenic or vaccine composition containing the CAV2 of any one of claims 1 to 15, and a pharmaceutically acceptable carrier or diluent.