RU2780160C2 - Recombinant non-pathogenic structures of marek's disease virus, encoding set of heterologous antigens - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: group of inventions is described, including a recombinant non-pathogenic Marek’s disease virus (hereinafter – rMDV_np) for protection of hens from a virus selected from a group consisting of Newcastle disease virus (hereinafter – NDV), infectious laryngotracheitis virus (hereinafter – ILTV), infectious bursitis virus (hereinafter – IBDV), Marek’s disease virus (hereinafter – MDV), and any combination thereof, an immunogenic composition containing rMDV_np, a vaccine containing the immunogenic composition, and a method for protection of chickens from a virus, including the administration of the vaccine. In one implementation, rMDV_np contains at least three heterologous nucleotide sequences located in an insignificant genome site of rMDV_np.

EFFECT: invention expands the arsenal of means for protection of chickens from a virus selected from a group consisting of NDV, ILTV, IBDV, MDV, and any combination thereof.

16 cl, 5 dwg, 9 tbl, 7 ex

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. § 119(e) of U.S. Provisional Applications Serial No. 62/571524, dated October 12, 2017, and Serial No. 62/729673, dated September 11, 2018, the contents of which are hereby incorporated by reference in their entirety.

FIELD OF TECHNOLOGY

The present invention relates to novel polyvalent recombinant nonpathogenic Marek's disease constructs encoding and expressing foreign antigens from three or more avian viruses, and methods of using these polyvalent recombinant nonpathogenic Marek's disease constructs in poultry vaccines.

BACKGROUND OF THE INVENTION

In addition to harming chickens, pathogenic poultry viruses are also costly for poultry farmers because most emerging diseases are contagious and the poultry industry relies heavily on limited, large-scale breeding centers. Vaccination of chickens is often the only viable means of controlling these viruses. Although attenuated or killed avian virus vaccines remain important in the marketplace, significant resources have been expended in recent years on the development of vaccines containing recombinant viral constructs that express pathogenic viral protein antigens. In addition, significant efforts have been made to develop stable and efficient polyvalent recombinant non-pathogenic Marek's disease viruses (abbreviated as rMDV _np ) that express foreign genes from a variety of viral pathogens. Such polyvalent vaccines are intended to serve to minimize the number of injections administered to chickens and thus reduce discomfort and stress for vaccinated chickens, as well as greatly reduce labor and material costs. Vaccination with such single multivalent constructs would also be preferable to alternative multivalent rMDV _np vaccines that contain multiple recombinant univalent rMDV _np constructs because these alternative vaccines, at least to date, result in protection against only one viral pathogen. The failure of such alternative vaccines appears to be due to the fact that one of the monovalent rMDV _np constructs outgrows the other monovalent rMDV _np constructs, thereby preventing the induction of a significant immune response by these monovalent rMDV _np constructs. In any case, despite significant past efforts to develop stable and efficient polyvalent rMDV _np vectors that actually express foreign genes from a variety of viral pathogens, in fact, such vaccines have been proposed for more than twenty years [see, for example, US 5965138] , but it has only recently been shown that a polyvalent vaccine that contains recombinant turkey herpes virus (abbreviated rHVT) encoding antigens from more than one other pathogen was stable and effective.

One viral disease in poultry that can be controlled by vaccination is Marek's disease. Marek's disease is a pathogenic disease that negatively affects chickens worldwide. Marek's disease occurs predominantly in chickens between the ages of 2 and 5 months. Clinical features include: progressive paralysis of one or more limbs, incoordination due to leg paralysis, limb droop due to wing involvement, and head droop due to neck muscle involvement. In acute cases, severe depression may occur. Bursal and thymic atrophy may also develop.

The etiological agent for Marek's disease is the Marek's disease virus serotype 1 (abbreviated as MDV1), a cellular virus that has a double-stranded DNA genome. MDV1 is a lymphotropic avian alpha herpesvirus that: (i) infects B cells, which can lead to cytolysis, and (ii) latently infects T cells, which can induce T cell lymphoma. Closely related to the virulent strain of MDV1, Marek's disease virus serotype 2 (abbreviated as MDV2), formerly known as chick herpesvirus 3, is a naturally attenuated strain of MDV that has been shown to be almost pathogenic in chickens [Petherbridge et al. J. J. Virological Methods 158: 11-17 (2009)]. SB-1 is a specific strain of MDV2 that has been shown to be useful in MDV1 vaccines [see, for example, Murthy and Calnek, Infection and Immunity 26 (2) 547-553 (1979)].

Another closely related alpha herpesvirus, Marek's disease virus serotype 3 (abbreviated as MDV3), more commonly known as turkey herpesvirus (abbreviated as HVT), is a non-pathogenic domestic turkey virus [see, for example, Kingham et al., J. of General Virology. 82: 1123-1135 (2001)]. Two commonly used HVT strains are the PB1 strain and the FC126 strain. Whereas HVT is also non-pathogenic in chickens, it elicits a long-lasting protective immune response in chickens against MDV1. Accordingly, HVT has been used in poultry vaccines against virulent MDV1 for many years, typically in combination with SB-1, which is more viremic than HVT but considered less safe. Alternatively, when flocks become infected with particularly virulent strains of MDV1, HVT can be combined with the Rispens vaccine. The Rispens vaccine is an isolate derived from a moderately virulent MDV1 strain that has been further attenuated by cell passaging. However, the Rispens strain retains some virulence against very susceptible chicken lines.

The sequence of the entire HVT genome has been disclosed [Afonso et al., J. Virology 75 (2): 971-978 (2001)], and like most alpha herpesviruses, HVT has a significant number of potential non-essential insertion sites [cf. for example, US 5187087; US 5830745; US 5834305; US 5853733; US 5928648; US 5961982; US 6121043; US 6299882 B1]. It has also been shown that HVT is amenable to genetic modification and, therefore, it has been used as a recombinant vector for many years [WO 87/04463]. Accordingly, recombinant HVT vectors have been reported to express foreign genes that encode antigens of, for example, Newcastle disease virus (NDV), [Sondermeijer et al., Vaccine, 11: 349-358 (1993); Reddy et al., Vaccine, 14: 469-477 (1996)], infectious bursitis virus (IBDV), [Darteil et al., Virology, 211: 481-490 (1995); Tsukamoto et al., J. of Virology 76 (11): 5637-5645 (2002)] and infectious laryngotracheitis virus (ILTV) [Johnson et al., Avian Disease, 54 (4): 1251-1259 (2010); W092/03554; US 6875856]. The complete genomic sequence of MDV2 is also known [see GenBank register number: AB049735.1 and Petherbridge et al., supra]. The genomic organization of MDV2 is very similar to HVT, and in the US region is particularly identical to the organization of HVT [see. Kingham et al., supra].

In addition, a recombinant chimeric virus known as a novel avian herpesvirus (NAHV) has been constructed in which certain regions of the HVT genome have been replaced by corresponding regions of the MDV1 genome. NAHV has also been used to express foreign genes that encode antigens from other poultry viruses [US 5965138; US 6913751].

Like MDV, infectious laryngotracheitis virus (abbreviated as ILTV or ILT) is an alpha herpesvirus that adversely affects chickens worldwide [Fuchs et al., Veterinary Research 38: 261-279 (2007)]. ILTV causes an acute respiratory disease in chickens characterized by respiratory depression, labored breathing, and bloody exudates. Virus replication is limited to cells in the respiratory tract, where in the trachea, infection causes tissue erosion and hemorrhage.

Newcastle disease is another highly contagious and debilitating disease in chickens. The etiological agent for Newcastle disease is the Newcastle disease virus (NDV). NDV belongs to the order Mononegavirales and belongs to the family Paramyxoviridae. Newcastle disease viruses have a non-segmented antisense single-stranded RNA genome. NDV has been grouped into three different pathotypes based on their virulence. Infection of poultry with non-pathogenic lentogenic strains of NDV is essentially asymptomatic. In contrast, mesogenic (moderately pathogenic) and velogenic (highly pathogenic) strains of NDV cause widespread disease that can be fatal. Most types of NDV infect the respiratory system and/or nervous system and can lead to suffocation and torticollis.

Infectious bursitis virus (abbreviated as IBDV or IBD), also called Gumboro disease virus, is the causative agent of infectious bursitis. IBDV causes an acute, highly contagious viral infection of chicken lymphoid tissue, the main target of which is the main immunological organ of the bird: the bursa of Fabricius. Disease rates in susceptible flocks are high with rapid weight loss and moderate to high mortality. Chickens that are recovering from the disease may be immunodeficient due to destruction of the bursa of Fabricius (or parts of it). This makes them particularly vulnerable to secondary infections.

IBDV is a member of the Birnaviridae family. Viruses of this family have a genome consisting of two segments (A and B) of double-stranded RNA. There are two IBDV serotypes, serotype 1 and 2, that can be differentiated using virus neutralization (VN) tests. Serotype 1 viruses have been shown to be pathogenic in chickens, while serotype 2 viruses have been shown to cause only subacute disease in turkeys. Historically, IBDV serotype 1 viruses consisted of only one type, which is now known as the "classic" IBD virus. More recently, so-called "variant" strains of IBDV have emerged. Classical and variant strains of IBDV can be identified and distinguished by a virus neutralization test using a panel of monoclonal antibodies or by RT-PCR [Wu et al., Avian Diseases, 51: 515-526 (2007)]. Well known classic IBDV strains include D78, Faragher 52/70 and STC while 89/03 is a well known variant strain. Many live or inactivated IBDV vaccines are commercially available, such as a live vaccine such as NOBILISR Gumboro D78 (MSD Animal Health). http://www.ncbi.nlm.nih.gov/pubmed/17626477

As stated above, since HVT can act both as an antigen that provides significant protection against Marek's disease and as a recombinant vector, it is currently used as a vector platform for polyvalent vaccines such as Innovax®-ILT (sold by Merck Animal Health) , which protects against ILTV; Innovax®-ND-SB (sold by Merck Animal Health) Vectormune® HVT-NDV (sold by Ceva), each of which protects against NDV; and Vaxxitek® HVT+IBD (Merial; formerly: Gallivac™ HVT-IBD) and Vectormune™ HVT-IBD (Ceva), both of which protect against IBDV. Notably, Innovax®-ILT contains two foreign genes, gD ILTV and gI ILTV, which have proven to be safe, effective and stable. However, these two foreign genes originate from the same pathogen, and in addition, they naturally overlap and must be co-expressed to ensure proper immunization against ILTV. More recently, a recombinant safe, effective and stable polyvalent vaccine containing HVT-ILTV-NDV has been disclosed [US 8932604 B2 and US 9409954 B2, the contents of which are incorporated herein by reference in their entirety). An early HVT-NDV-IBDV has also been discovered, although long-term testing during the development of the respective product found that one of the main constructs, HVP309, exhibited neither adequate genetic stability nor sustained expression of heterologous inserts [WO 2013/057235]. Subsequently, a more stable and efficient design was developed [WO 2016/102647]. Other recombinant HVT constructs have also been described [see, for example, US 9114108, US 9555016, US 9555096 and US 2018/0163230 A1].

However, despite the clear advantages of stable, polyvalent, recombinant MDV _np constructs that can efficiently express heterologous antigens from three or more different pathogens, and significant efforts to construct them, none have yet been obtained. Indeed, previous failures to construct such recombinant MDV _np constructs have led to the consensus in the art that insertion of foreign antigens from three or more different viral pathogens into the MDV _np construct overstresses the construct, resulting in an observed lack of stability. Accordingly, the suitability of any given polyvalent recombinant MDV _np as a vaccine remains unpredictable at best when the recombinant MDV _np contains a combination of heterologous antigens that are derived from a unique set of three or more poultry viruses. Therefore, there is a clear need to overcome collective industry shortcomings by developing new stable recombinant MDV _np vectors that can be used in multivalent vaccines as a single active agent to protect against three or more different non-MDV1 poultry virus pathogens.

The citation of any reference in this specification should not be construed as an admission that such reference is available as "prior art" of the present application.

SUMMARY OF THE INVENTION

Accordingly, the present invention relates to a novel polyvalent recombinant non-pathogenic Marek's disease virus (rMDV _np ) for use as a vector for the expression of foreign genes from a variety of viral pathogens. In specific embodiments, rMDV _np is recombinant turkey herpesvirus (rHVT). In alternative embodiments, rMDV _np is recombinant Marek's disease virus serotype 2 (rMDV2). rMDV _np , such as rHVT or rMDV2 of the present invention, can be used in safe and effective polyvalent vaccines against pathogenic poultry viruses.

Thus, the present invention relates to recombinant non-pathogenic Marek's disease virus (rMDV _np ) vectors (including HVT vectors) that encode and express antigens from three or more foreign avian viral pathogens. In specific embodiments, the rMDV _np encodes one or more laryngotracheitis virus (ILTV) antigens, one or more infectious bursitis virus (IBDV) antigens, and one or more Newcastle disease virus (NDV) antigens. In more specific embodiments, such rMDV _np vectors assist in protecting vaccinated chickens from clinical signs resulting from infection with pathogenic MDV, pathogenic IBDV, pathogenic NDV, and/or pathogenic ILTV. The vaccines are preferably effective for vaccinating healthy animals at 18-19 days of embryonic development, as well as for day-old chicks and older.

In specific embodiments, the recombinant non-pathogenic Marek's disease virus (rMDV _np ) comprises, at one or more non-essential sites in its genome, a first heterologous nucleotide sequence encoding one or more antigens from a first chicken pathogen, a second heterologous nucleotide sequence encoding one or more antigens from a second chicken pathogen. pathogen and a third heterologous nucleotide sequence encoding one or more antigens from the third chicken pathogen. In specific embodiments, the first avian pathogen, the second avian pathogen, and the third avian pathogen are all avian viruses. In more specific embodiments, the first avian pathogen, the second avian pathogen, and the third avian pathogen are different types of viruses, different from each other and different from the Marek's disease virus. In some embodiments of this type, the first chicken pathogen is infectious bursitis virus (IBDV), the second chicken pathogen is infectious laryngotracheitis virus (ILTV), and the third chicken pathogen is Newcastle disease virus (NDV).

In specific embodiments, the rMDV _np first heterologous nucleotide sequence encodes infectious bursitis virus viral protein 2 (IBDV VP2); and/or the second heterologous nucleotide sequence encodes laryngotracheitis virus glycoprotein D (gD ILTV), laryngotracheitis virus glycoprotein I (gI ILTV), or both gI ILTV and gD ILTV; and/or the third heterologous nucleotide sequence encodes a Newcastle disease virus (F NDV) fusion protein. In more specific embodiments of rMDV _np , the first heterologous nucleotide sequence encodes infectious bursitis virus viral protein 2 (IBDV VP2), the second heterologous nucleotide sequence encodes both infectious laryngotracheitis virus glycoprotein D (gD ILTV) and laryngotracheitis virus glycoprotein I (gI ILTV), and the third the heterologous nucleotide sequence encodes a Newcastle disease virus (F NDV) fusion protein.

Accordingly, in specific embodiments of the rMDV of the present invention, the first heterologous nucleotide sequence, the second heterologous nucleotide sequence, and the third heterologous nucleotide sequence are located at three different non-essential sites in the rMDV _np genome. In specific embodiments, the three different sites are individually selected from the group consisting of US2 site, UL54.5 site, UL7/8 site, UL40 site, UL43 site, UL45/46 site, UL55 site, US10 site, region between US10 and SORF3, the area between US2 and SORF3, the IG1 site, the IG2 site, and the IG3 site. In a particular embodiment of this type, the first non-essential site is a US2 site, the second non-essential site is a UL54.5 site, and the third non-essential site is a UL45/46 site.

In alternative embodiments, the first heterologous nucleotide sequence, the second heterologous nucleotide sequence, and the third heterologous nucleotide sequence are located at a first non-essential site in the rMDV _np genome or at a second non-essential site in the rMDV _np genome. In particular embodiments of this type, the first non-essential site and the second non-essential site are the same (ie, there is a single non-essential insertion site). In particular embodiments of this type, the only non-essential insertion site is the US2 site. In other embodiments of this type, the only non-essential insertion site is the UL54.5 site. In still other embodiments of this type, the only non-essential insertion site is the UL7/8 site. In other embodiments of this type, the only non-essential insertion site is the UL40 site. In yet other embodiments of this type, the only non-essential insertion site is the UL45/46 site. In yet other embodiments of this type, the only non-essential insertion site is the UL55 site. In yet other embodiments of this type, the only non-essential insertion site is the US10 site. Still in other embodiments of this type, the only non-essential insertion site is the region between US10 and SORF3. In yet other embodiments of this type, the only non-essential insertion site is the region between US2 and SORF3. In still other embodiments of this type, the only non-essential insertion site is the IG1 site. Still in other embodiments of this type, the only non-essential insertion site is the IG2 site. In yet other embodiments of this type, the only non-essential insertion site is the IG3 site. In still other embodiments of this type, the only non-essential insertion site is the UL43 site.

In other types of embodiments, the first non-essential site and the second non-essential site are different. The two different sites are individually selected from the group consisting of US2 site, UL54.5 site, UL7/8 site, UL40 site, UL43 site, UL45/46 site, UL55 site, US10 site, area between US10 and SORF3, area between US2 and SORF3, IG1 site, IG2 site, and IG3 site. In specific embodiments, the first heterologous nucleotide sequence and the second heterologous nucleotide sequence are located at the first non-essential site, and the third heterologous nucleotide sequence is located at the second non-essential site. In other embodiments, the first heterologous nucleotide sequence and the third heterologous nucleotide sequence are located at the first non-essential site, and the second heterologous nucleotide sequence is located at the second non-essential site. In yet other embodiments, the second heterologous nucleotide sequence and the third heterologous nucleotide sequence are located at the first non-essential site, and the first heterologous nucleotide sequence is located at the second non-essential site. In preferred embodiments of this type, the first heterologous nucleotide sequence encodes infectious bursitis virus viral protein 2 (IBDV VP2), the second heterologous nucleotide sequence encodes laryngotracheitis virus glycoprotein D (gD ILTV), infectious laryngotracheitis virus glycoprotein I (gI ILTV), or both, gI ILTV and gD ILTV, and a third heterologous nucleotide sequence encodes a Newcastle disease virus (F NDV) fusion protein.

In some embodiments of this type, an _np rMDV containing heterologous nucleotide sequences encoding ILTV gD protein, ILTV gI protein, and IBDV VP2 protein at a first non-essential site, and a heterologous nucleotide sequence that encodes an NDV F protein at a second non-essential site, is constructed such that that the heterologous nucleotide sequence encoding the IBDV VP2 protein is in the 5' region relative to the heterologous nucleotide sequence encoding the ILTV gD protein and the ILTV gI protein. In other embodiments of this type, the rMDV _np is designed such that the heterologous nucleotide sequence encoding the ILTV gD protein and the ILTV gI protein are in the 5' region relative to the heterologous nucleotide sequence encoding the IBDV VP2 protein.

In alternative embodiments, an rMDV _np comprising heterologous nucleotide sequences encoding an NDV F protein and an IBDV VP2 protein at a first non-essential site, and a heterologous nucleotide sequence that encodes an ILTV gD protein and an ILTV gI protein at a second non-essential site, are constructed such that that the heterologous nucleotide sequence encoding the NDV F protein is in the 5' region relative to the heterologous nucleotide sequence encoding the IBDV VP2 protein. In other embodiments of this type of rMDV _np is designed such that the heterologous nucleotide sequence encoding the IBDV VP2 protein is in the 5' region relative to the heterologous nucleotide sequence encoding the NDV F protein.

In other alternative embodiments, an rMDV _np comprising heterologous nucleotide sequences encoding NDV F protein, ILTV gD protein, and ILTV gI protein at a first non-essential site, and a heterologous nucleotide sequence encoding IBDV VP2 protein at a second non-essential site, is constructed such that the heterologous nucleotide sequence encoding the NDV F protein is in the 5' region relative to the heterologous nucleotide sequence encoding the ILTV gD protein and ILTV gI protein. In alternative embodiments of this type of rMDV _np is designed such that the heterologous nucleotide sequence encoding ILTV gD protein and ILTV gI protein is in the 5' region relative to the heterologous nucleotide sequence encoding NDV F protein.

In particular embodiments of this type, the first non-essential site is a US2 site and the second non-essential site is a UL54.5 site. In alternative embodiments, the first non-essential site is the UL54.5 site and the second non-essential site is the US2 site. In yet another embodiment of this type, the first non-essential site is a US2 site and the second non-essential site is a UL45/46 site. In yet another embodiment of this type, the first non-essential site is a UL45/46 site and the second non-essential site is a US2 site.

In yet another embodiment of this type, the first non-essential site is a UL54.5 site and the second non-essential site is a UL45/46 site. In another embodiment of this type, the first non-essential site is a UL45/46 site and the second non-essential site is a UL54.5 site. In yet another embodiment of this type, the first non-essential site is a US2 site and the second non-essential site is a UL55 site. In yet another embodiment of this type, the first non-essential site is a UL55 site and the second non-essential site is a US2 site.

Accordingly, the rMDV _np vectors of the present invention may contain heterologous nucleotide sequences that encode any combination of these foreign protein antigens. In specific embodiments, the ILTV gD protein comprises the amino acid sequence of SEQ ID NO: 2. In other embodiments, the ILTV gI protein comprises the amino acid sequence of SEQ ID NO: 4. In yet other embodiments, the IBDV VP2 protein comprises the amino acid sequence of SEQ ID NO: 6. Still in other embodiments, the NDV F protein comprises the amino acid sequence of SEQ ID NO: 8. In still other embodiments, the NDV F protein comprises the amino acid sequence of SEQ ID NO: 10. In addition, the present invention also provides rMDV _np vectors that contain any combination of sequences that encode one or more of these amino acid sequences, including specific embodiments that encode all of them.

In other embodiments, the rMDV _np ILTV gD protein comprises the amino acid sequence of SEQ ID NO: 2 and the ILTV gI protein comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments of this type, the ILTV gD protein comprises the amino acid sequence of SEQ ID NO: 2, protein ILTV gI contains the amino acid sequence of SEQ ID NO: 4, and the IBDV VP2 protein contains the amino acid sequence of SEQ ID NO: 6. In related embodiments, the ILTV gD protein contains the amino acid sequence of SEQ ID NO: 2, the ILTV gI protein contains the amino acid sequence of SEQ ID NO: 4 and the NDV F protein contains the amino acid sequence of SEQ ID NO: 8. In similar embodiments, the ILTV gD protein contains the amino acid sequence of SEQ ID NO: 2, the ILTV gI protein contains the amino acid sequence of SEQ ID NO: 4, and the NDV F protein contains the amino acid sequence SEQ ID NO: 10. In more specific embodiments, the gD protein ILTV contains the amino acid sequence of SEQ ID NO: 2, the ILTV gI protein contains the amino acid sequence of SEQ ID NO: 4, the IBDV VP2 protein contains the amino acid sequence of SEQ ID NO: 6, and the NDV F protein contains either the amino acid sequence of SEQ ID NO: 8 or the amino acid the sequence of SEQ ID NO: 10. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

In related embodiments, the ILTV gD protein is encoded by the nucleotide sequence of SEQ ID NO: 1. In other embodiments, the ILTV gI protein is encoded by the nucleotide sequence of SEQ ID NO: 3. In still other embodiments, the IBDV VP2 protein is encoded by the nucleotide sequence of SEQ ID NO: 5. Still in other embodiments, the NDV F protein is encoded by the nucleotide sequence of SEQ ID NO: 7. In still other embodiments, the NDV F protein is encoded by the nucleotide sequence of SEQ ID NO: 9.

Likewise, the rMDV _np vectors of the present invention may contain heterologous nucleic acids that contain any combination of such heterologous nucleotide sequences. In some embodiments, the rMDV _np comprises a first heterologous nucleic acid located at a first non-essential site in the rMDV _np genome and a second heterologous nucleic acid located at a second non-essential site in the rMDV _np genome, wherein the first heterologous nucleic acid contains both, the first heterologous nucleotide sequence and a second heterologous nucleotide sequence, while the second heterologous nucleic acid contains a third heterologous nucleotide sequence.

In some embodiments of this type, the first heterologous nucleic acid comprises heterologous nucleotide sequences encoding the infectious laryngotracheitis virus (ILTV) glycoprotein D (gD) protein, the infectious laryngotracheitis virus (ILTV) glycoprotein I (gI) protein, and the infectious bursitis virus protein 2 (VP2). (IBDV), while the second heterologous nucleic acid contains a heterologous nucleotide sequence that encodes the F protein of the Newcastle disease virus (NDV). In some embodiments of this type, the first heterologous nucleic acid is constructed and/or inserted into the rMDV _np genome such that the heterologous nucleotide sequence encoding the IBDV VP2 protein is in the 5' region relative to the heterologous nucleotide sequence encoding the ILTV gD protein and the gI protein. ILTV. In other embodiments of this type, the first heterologous nucleic acid is constructed and/or inserted into the rMDV _np genome such that the heterologous nucleotide sequence encoding the ILTV gD protein and the ILTV gI protein is in the 5' region relative to the heterologous nucleotide sequence encoding the VP2 protein. IBDV.

In alternative embodiments, the first heterologous nucleic acid comprises heterologous nucleotide sequences that encode NDV F protein and IBDV VP2 protein, while the second heterologous nucleic acid comprises heterologous nucleotide sequences that encode ILTV gD protein and ILTV gI protein. In some embodiments of this type, the first heterologous nucleic acid is constructed and/or inserted into the rMDV _np genome such that the heterologous nucleotide sequence encoding the NDV F protein is in the 5' region relative to the heterologous nucleotide sequence encoding the IBDV VP2 protein. In alternative embodiments of this type, the first heterologous nucleic acid is constructed and/or inserted into the rMDV _np genome such that the heterologous nucleotide sequence encoding the IBDV VP2 protein is in the 5' region relative to the heterologous nucleotide sequence encoding the NDV F protein.

In other alternative embodiments, the first heterologous nucleic acid comprises heterologous nucleotide sequences that encode NDV F protein, ILTV gD protein, and ILTV gI protein, while the second heterologous nucleic acid comprises a heterologous nucleotide sequence encoding IBDV VP2 protein. In some embodiments of this type, the first heterologous nucleic acid is constructed and/or inserted into the rMDV _np genome such that the heterologous nucleotide sequence encoding the NDV F protein is in the 5' region relative to the heterologous nucleotide sequence encoding the ILTV gD protein and the gI protein. ILTV. In alternative embodiments of this type, the first heterologous nucleic acid is constructed and/or inserted into the rMDV _np genome such that the heterologous nucleotide sequence encoding the ILTV gD protein and ILTV gI protein is in the 5' region relative to the heterologous nucleotide sequence encoding the NDV F protein. .

In addition, the present invention also relates to rMDV _np vectors that encode any combination of these nucleotide sequences, including specific embodiments in which the first heterologous nucleic acid encodes the nucleotide sequence of SEQ ID NOS: 1, 3 and 5, and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 7. In alternative embodiments, the first heterologous nucleic acid encodes the nucleotide sequence of SEQ ID NO: 1, 3, and 5, and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 9. In specific embodiments, rMDV _np is is rHVT. In alternative embodiments, rMDV _np is rMDV2.

In particular embodiments, two different non-essential rMDV _np sites are individually selected from the group consisting of US2 site, UL54.5 site, UL7/8 site, UL40 site, UL 43 site, UL45/46 site, UL55 site, US10 site, region between US10 and SORF3, regions between US2 and SORF3, site of intergenic region 1 (IG1), site of intergenic region 2 (IG2) and intergenic region (IG3).

In some embodiments, the first non-essential rMDV _np site is a US2 site, while the second non-essential rMDV _np site is a non-essential site other than the US2 site. In other embodiments, the first non-essential rMDV _np site is a UL 54.5 site, while the second non-essential rMDV _np site is a non-essential site other than the UL 54.5 site. In particular embodiments of this type, the first non-essential site is a US2 site and the second non-essential site is a UL54.5 site. In alternative embodiments, the first non-essential site is the UL54.5 site and the second non-essential site is the US2 site. In yet another embodiment of this type, the first non-essential site is a US2 site and the second non-essential site is a UL45/46 site. In yet another embodiment of this type, the first non-essential site is a UL45/46 site and the second non-essential site is a US2 site. In yet another embodiment of this type, the first non-essential site is a UL54.5 site and the second non-essential site is a UL45/46 site. In another embodiment of this type, the first non-essential site is a UL45/46 site and the second non-essential site is a UL54.5 site. In yet another embodiment of this type, the first non-essential site is a US2 site and the second non-essential site is a UL55 site. In yet another embodiment of this type, the first non-essential site is a UL55 site and the second non-essential site is a US2 site. In related embodiments, the first non-essential rMDV _np site is UL 54.5 and the second non-essential rMDV _np site is a UL7/8 site. In other embodiments, the first non-essential rMDV _np site is UL 54.5 and the second non-essential rMDV _np site is a US10 site. In related embodiments, the second non-essential rMDV _np site is a US2 site and the first non-essential rMDV _np site is a UL7/8 site. In other embodiments, the second non-essential rMDV _np site is a US2 site and the first non-essential rMDV _np site is a US10 site. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

In particular embodiments of this type, the first non-essential site and the second non-essential site are the same (ie, there is a single non-essential insertion site). In particular embodiments of this type, the only non-essential insertion site is the US2 site. In other embodiments of this type, the only non-essential insertion site is the UL54.5 site. In still other embodiments of this type, the only non-essential insertion site is the UL7/8 site. In other embodiments of this type, the only non-essential insertion site is the UL40 site. In yet other embodiments of this type, the only non-essential insertion site is the UL45/46 site. In yet other embodiments of this type, the only non-essential insertion site is the UL55 site. In yet other embodiments of this type, the only non-essential insertion site is the US10 site. Still in other embodiments of this type, the only non-essential insertion site is the region between US10 and SORF3. In other embodiments of this type, the only non-essential insertion site is the region between US2 and SORF3. In other embodiments of this type, the only non-essential insertion site is the IG1 site. In other embodiments of this type, the only non-essential insertion site is the IG2 site. In other embodiments of this type, the only non-essential insertion site is the IG3 site. In other embodiments of this type, the only non-essential insertion site is the UL43 site. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

Nucleotide sequences encoding ILTV gD protein, ILTV gI protein and IBDV VP2 protein may be under the functional control of exogenous promoters, ie promoters not found naturally in MDV _np . In some embodiments, these three nucleotide sequences are under the functional control of different promoters, i.e., the nucleotide sequence encoding the ILTV gD protein is under the functional control of the first promoter, and the nucleotide sequence encoding the ILTV gI protein is under the functional control of the second promoter, and the nucleotide sequence the IBDV VP2 protein coding sequence is under the functional control of a third promoter, the first promoter, the second promoter and the third promoter being different. In specific embodiments, the promoter for the nucleotide sequence encoding the ILTV gD protein is an endogenous ILTV gD promoter (ie, endogenous to ILTV). In some embodiments, the promoter for the nucleotide sequence encoding the ILTV gI protein is an endogenous ILTV gI promoter. In particular embodiments of this type, the promoter for the nucleotide sequence encoding the ILTV gD protein is the endogenous ILTV gD promoter and the promoter for the nucleotide sequence encoding the ILTV gI protein is the endogenous ILTV gI promoter. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

In some embodiments, at least one of the promoters operably linked to the nucleotide sequence encoding the NDV F protein, ILTV gD protein, ILTV gI protein, or IBDV VP2 protein is the mouse cytomegalovirus immediate early promoter (mCMV IE). In related embodiments, at least one of the promoters operably linked to the nucleotide sequence encoding the NDV F protein, ILTV gD protein, ILTV gI protein, or IBDV VP2 protein is a human cytomegalovirus (hCMV IE) immediate early promoter or a derivative thereof (e.g. , from strain AD169). In other embodiments, at least one of the promoters operably linked to the nucleotide sequence encoding the NDV F protein, ILTV gD protein, ILTV gI protein, or IBDV VP2 protein is a guinea-pig cytomegalovirus immediate early promoter. In other embodiments, at least one of the promoters operably linked to the nucleotide sequence encoding the NDV F protein, ILTV gD protein, ILTV gI protein, or IBDV VP2 protein is the chicken β-actin promoter. In other embodiments, at least one of the promoters operably linked to the nucleotide sequence encoding the NDV F protein, ILTV gD protein, ILTV gI protein, or IBDV VP2 protein is the pseudorabies virus (PRV) gpX promoter.

In specific embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the mCMV IE promoter. In related embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the immediate early promoter (hCMV IE) of human cytomegalovirus or a derivative thereof (eg, from strain AD169).

In other embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the guinea-pig cytomegalovirus immediate early promoter. In other embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the promoter of the chicken beta actin gene.

In some embodiments, the promoter operably linked to the nucleotide sequence encoding the NDV F protein is the human cytomegalovirus immediate early promoter (hCMV IE). In other embodiments, the promoter operably linked to the nucleotide sequence encoding the NDV F protein is the pseudorabies virus (PRV) gpX promoter. In related embodiments, the promoter operably linked to the nucleotide sequence encoding the NDV F protein is the chicken beta actin gene promoter. In other embodiments, the promoter operably linked to the nucleotide sequence encoding the NDV F protein is the simian virus 40 (SV40) promoter.

In more specific embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the mCMV IE promoter, the promoter for the nucleotide sequence encoding the ILTV gD protein is the endogenous gD ILTV promoter, the promoter for the nucleotide sequence encoding the ILTV gI protein is the endogenous ILTV gI promoter, and the promoter of the nucleotide sequence encoding the NDV F protein is the hCMV IE promoter. In other specific embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the hCMV IE promoter (or a derivative thereof), the promoter for the nucleotide sequence encoding the gD ILTV protein is the endogenous gD ILTV promoter, the promoter for the nucleotide sequence encoding the protein ILTV gI is the endogenous ILTV gI promoter, and the promoter of the nucleotide sequence encoding the NDV F protein is the hCMV IE promoter. In other specific embodiments, the promoter for the nucleotide sequence encoding the IBDV VP2 protein is the chicken β-actin promoter, the promoter for the nucleotide sequence encoding the ILTV gD protein is the endogenous ILTV gD promoter, the promoter for the nucleotide sequence encoding the ILTV gI protein, is the endogenous ILTV gI promoter, and the promoter of the nucleotide sequence encoding the NDV F protein is the hCMV IE promoter.

In some embodiments, the _np rMDV of the present invention, which includes inserts of nucleotide sequences encoding ILTV gD protein, ILTV gI protein, and IBDV VP2 protein, also includes one or more exogenous transcription terminator sequences. In particular embodiments of this type, the transcription terminator sequence is located in the 3' region relative to the nucleotide sequence encoding the IBDV VP2 protein. In specific embodiments, the nucleotide sequences encoding the ILTV gD protein and ILTV gI protein share one transcription terminator sequence, and the nucleotide sequence encoding the IBDV VP2 protein has a different one. In more specific embodiments, at least one of the transcription terminator sequences comprises a feline herpesvirus US-9 (FHV US-9) polyadenylation sequence. In still more specific embodiments, at least one of the transcription terminator sequences comprises a simian virus 40 (SV40) polyadenylation sequence.

In some embodiments, the NDV F protein also includes one or more exogenous transcription terminator sequences. In particular embodiments of this type, the transcription terminator sequence is located in the 3' region relative to the nucleotide sequence encoding the NDV F protein. In related embodiments, at least one of the transcription terminator sequences comprises a herpes simplex virus thymidine kinase (HSV TK) polyadenylation sequence. In alternative embodiments, at least one of the transcription terminator sequences comprises an immediate early polyadenylation sequence (hCMV IE) of human cytomegalovirus. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

The present invention also relates to rMDV _np which contains (i) mCMV IE promoter, chicken beta actin gene promoter or hCMV promoter, (ii) coding sequences for IBDV VP2 protein, (iii) transcription terminator sequence, (iv) ILTV gD promoter , (v) the coding sequence for ILTV gD protein, (vi) the ILTV gI promoter, and (vii) the coding sequence for ILTV gI protein at the UL54.5 site of the rMDV _np genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 21 is contained in the UL54.5 site of the rMDV _np genome. In specific embodiments, the rMDV _np further comprises (i) an hCMV IE promoter, (ii) a coding sequence for an NDV F protein, and (iii) an hCMV IE transcription terminator sequence at the US2 site of its genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 22 is contained in the US2 site of the rMDV _np genome.

The present invention also provides an _np rMDV comprising (i) an mCMV IE promoter, a chick beta actin gene promoter, or an hCMV promoter, (ii) a coding sequence for an IBDV VP2 protein, (iii) a transcription terminator sequence, (iv) an ILTV gD promoter, (v) the coding sequence for the ILTV gD protein, (vi) the ILTV gI promoter, and (vii) the coding sequence for the ILTV gI protein at the US2 site of the rMDV _np genome. In some embodiments of this type, the nucleotide sequence of SEQ ID NO: 24 is contained in the US2 site of the rMDV _np genome. In other embodiments of this type, the nucleotide sequence of SEQ ID NO: 25 is contained in the US2 site of the rMDV _np genome. In more specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 23 is contained in the US2 site of the rMDV _np genome. In specific embodiments, rMDV _np further comprises (i) an hCMV IE promoter, (ii) a coding sequence for an NDV F protein, and (iii) an hCMV IE transcription terminator sequence at the UL54.5 site of its genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 26 is contained in the UL54.5 site of the rMDV _np genome.

In addition, the present invention also relates to rMDV _np containing (i) ILTV gD promoter, (ii) coding sequence for ILTV gD protein, (iii) ILTV gI promoter, (iv) coding sequence for ILTV gI protein, (v) promoter hCMV IE, (vi) the coding sequence for the NDV F protein, and (vii) the hCMV IE transcription terminator sequence at the UL54.5 site of the _np rMDV genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 31 is contained in the UL54.5 site of the _np rMDV genome. In specific embodiments, the rMDV _np further comprises (i) an mCMV IE promoter, (ii) a coding sequence for the IBDV VP2 protein, and (iii) a transcription terminator sequence at the US2 site of the rMDV _np genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 32 is contained in the US2 site of the rMDV _np genome.

The present invention also relates to rMDV _np containing (i) ILTV gD promoter, (ii) coding sequence for ILTV gD protein, (iii) ILTV gI promoter, (iv) coding sequence for ILTV gI protein, (v) hCMV IE promoter, (vi) the coding sequence for the NDV F protein; and (vii) the hCMV IE transcription terminator sequence at the US2 site of the rMDV _np genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 28 is contained in the US2 site of the rMDV _np genome. In specific embodiments, the rMDV _np further comprises (i) an mCMV IE promoter, (ii) a coding sequence for the IBDV VP2 protein, and (iii) a transcription terminator sequence at the UL54.5 site of the rMDV _np genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 27 is contained in the UL54.5 site of the _np rMDV genome.

The present invention also provides an rMDV _np comprising (i) an mCMV IE promoter, (ii) a coding sequence for an IBDV VP2 protein, (iii) a transcription terminator sequence, (iv) an hCMV IE promoter, (v) a coding sequence for an NDV F protein, and (vi) the hCMV IE transcription terminator sequence at the US2 site of the rMDV _np genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 30 is contained in the US2 site of the rMDV _np genome. In specific embodiments, rMDV _np further comprises (i) an ILTV gD promoter, (ii) a coding sequence for ILTV gD protein, (iii) an ILTV gI promoter, and (iv) a coding sequence for ILTV gI protein at the UL54.5 site of its genome. In specific embodiments of this type, the nucleotide sequence of SEQ ID NO: 29 is contained in the UL54.5 site of the rMDV _np genome.

The present invention also provides an rMDV _np comprising (i) an mCMV IE promoter, (ii) a coding sequence for an IBDV VP2 protein, (iii) a transcription terminator sequence, (iv) an hCMV IE promoter, (v) a coding sequence for an NDV F protein, and (vi) the hCMV IE transcription terminator sequence at the UL54.5 site of the rMDV _np genome. In specific embodiments, the _np rMDV further comprises (i) an ILTV gD promoter, (ii) a coding sequence for ILTV gD protein, (iii) an ILTV gI promoter, and (iv) a coding sequence for ILTV gI protein within the US2 site of its genome.

The present invention also provides an rMDV _np comprising (i) an mCMV IE promoter, (ii) a coding sequence for an IBDV VP2 protein, (iii) a transcription terminator sequence, (iv) an hCMV IE promoter, (v) a coding sequence for an NDV F protein, and (vi) hCMV IE transcription terminator sequence at US2 site of rMDV _np genome and (i) ILTV gD promoter, (ii) coding sequence for ILTV gD protein, (iii) ILTV gI promoter, and (iv) coding sequence for ILTV gI protein, contained in the UL7/8 site of its genome.

In more specific embodiments, the implementation of the present invention relates to rHVT, which contains a first heterologous nucleic acid and a second heterologous nucleic acid. First heterologous nucleic acid containing (i) cytomegalovirus immediate early promoter (IE mCMV IE), (ii) coding sequence for IBDV VP2 protein, (iii) transcription terminator sequence, (iv) ILTV gD promoter, (v) coding sequence for protein gD ILTV, (vi) promoter gI ILTV and (vii) coding sequence for protein gI ILTV. In specific embodiments of this type, the specific order from 5' to 3' of the nucleotide sequences of this recombinant nucleic acid is (i) - (vii). In a more specific embodiment, the transcription terminator sequence contains an SV40 polyadenylation sequence. In an even more specific embodiment of this type, the first heterologous nucleic acid is located at the US2 site of the rHVT genome. The second heterologous nucleic acid comprises a recombinant nucleic acid which contains, in the 5' to 3' direction in the following order, (i) the cytomegalovirus immediate early promoter (hCMV IE), (ii) the coding sequence for the NDV F protein, and (iii) the transcription terminator sequence . In a more specific embodiment, the transcription terminator sequence comprises an immediate early polyadenylation sequence (hCMV IE) of human cytomegalovirus. In more specific embodiments of this type, the second heterologous nucleic acid is located at the UL54.5 site of the rHVT genome.

Accordingly, the present invention includes recombinant HVTs (rHVTs) that contain two heterologous nucleic acids each inserted into separate non-essential sites in the HVT genome. In some embodiments, the first heterologous nucleic acid comprises (i) mouse cytomegalovirus immediate early promoter (mCMV IE), (ii) a coding sequence for IBDV VP2 protein, (iii) a transcription terminator sequence, (iv) an ILTV gD promoter, (v) a coding sequence for ILTV gD protein, (vi) ILTV gI promoter and (vii) coding sequence for ILTV gI protein. In specific embodiments of this type, the specific order from 5' to 3' of the nucleotide sequences of this recombinant nucleic acid is (i) - (vii). The second heterologous nucleic acid contains in the 5' to 3' direction in the following order (i) an immediate early promoter (hCMV IE) of cytomegalovirus, (ii) a coding sequence for an NDV F protein, and (iii) a transcription terminator sequence. In specific embodiments, the first heterologous nucleic acid is inserted into the US2 site and the second heterologous nucleic acid is inserted into the UL54.5 site. In specific embodiments of this type, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 23 and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 26. In alternative embodiments, the second heterologous nucleic acid is inserted into the US2 site and the first heterologous nucleic acid the acid is inserted into the UL54.5 site. In some embodiments of this type, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 21 and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 22.

In other embodiments, the first heterologous nucleic acid comprises (i) mouse cytomegalovirus immediate early promoter (mCMV IE), (ii) a coding sequence for IBDV VP2 protein, (iii) a transcription terminator sequence, (iv) human cytomegalovirus immediate early promoter (hCMV IE). ), (v) a coding sequence for the NDV F protein, and (vi) a transcription terminator sequence. In specific embodiments of this type, the specific order from 5' to 3' for the nucleotide sequences of this recombinant nucleic acid is (i) - (vi). The second heterologous nucleic acid contains in the 5' to 3' direction in the following order (i) the ILTV gD promoter, (ii) the ILTV gD protein coding sequence, (iii) the ILTV gI promoter, and (iv) the ILTV gI protein coding sequence. In specific embodiments, the first heterologous nucleic acid is inserted into the US2 site and the second heterologous nucleic acid is inserted into the UL54.5 site. In specific embodiments, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 30 and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 29. In alternative embodiments, the second heterologous nucleic acid is inserted at the US2 site and the first heterologous nucleic acid is inserted to UL54.5 site.

In other embodiments, the first heterologous nucleic acid comprises (i) an ILTV gD promoter, (ii) a coding sequence for ILTV gD protein, (iii) an ILTV gI promoter, (iv) a coding sequence for ILTV gI protein, (v) an immediate early promoter ( hCMV IE) of human cytomegalovirus, (vi) a coding sequence for the NDV F protein, and (vii) a transcription terminator sequence. In specific embodiments of this type, the specific order from 5' to 3' of the nucleotide sequences of this recombinant nucleic acid is (i) - (vii). The second heterologous nucleic acid contains in the 5' to 3' direction in the following order (i) an immediate early promoter (mCMV IE) of mouse cytomegalovirus, (ii) a coding sequence for the IBDV VP2 protein, (iii) a transcription terminator sequence. In specific embodiments, the first heterologous nucleic acid is inserted into the US2 site and the second heterologous nucleic acid is inserted into the UL54.5 site. In specific embodiments of this type, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 28 and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 27. In alternative embodiments, the second heterologous nucleic acid is inserted at the US2 site and the first heterologous nucleic acid the acid is inserted into the UL54.5 site. In particular embodiments of this type, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 31 and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 32.

The present invention also provides methods for producing any rMDV _np of the present invention (eg, rHVT). In some embodiments, the first heterologous nucleic acid is designed to contain a nucleotide sequence that encodes an ILTV gD protein, a nucleotide sequence that encodes an ILTV gI protein, and a nucleotide sequence that encodes an IBDV VP2 protein. In specific embodiments of this type, promoters for the nucleotide sequences that encode ILTV gD protein and ILTV gI protein, respectively, are their respective endogenous promoters. In appropriate embodiments, the promoter for the nucleotide sequence encoding the IB2 VPD protein is the mCMV IE promoter, the chicken beta actin gene promoter, or the hCMV promoter.

The first heterologous nucleic acid is then inserted into a non-essential rMDV _np site of the present invention. In some embodiments, the first heterologous nucleic acid is an expression cassette. In specific embodiments of this type, the expression cassette contains the nucleotide sequence of SEQ ID NO: 21.

The method may further include generating a second heterologous nucleic acid that also inserts into a non-essential rMDV _np site. In specific embodiments, the second heterologous nucleic acid comprises a human cytomegalovirus immediate early promoter (hCMV IE), a coding sequence for an NDV F protein, and a transcription terminator sequence. In some embodiments, the second heterologous nucleic acid is an expression cassette. In specific embodiments of this type, the expression cassette comprises the nucleotide sequence of SEQ ID NO: 22. In specific embodiments, the first heterologous nucleic acid is inserted into the first non-essential rMDV _np site and the second heterologous nucleic acid is inserted into the second non-essential rMDV _np site. In some embodiments, the first non-essential rMDV _np site is a UL 54.5 site. In related embodiments, the second non-essential rMDV _np site is the US2 site. In alternative embodiments, the first non-essential rMDV _np site is a US2 site and the second non-essential rMDV _np site is a UL 54.5 site. In specific embodiments of this type, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 23 and the second heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 26. In other embodiments of this type, the first heterologous nucleic acid comprises the nucleotide sequence of SEQ ID NO: 24, and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 26. In other embodiments of this type, the first heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 25, and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 26.

In some embodiments, the method for producing rMDV _np is a method for producing rHVT. In alternative embodiments, the method for producing rMDV _np is a method for producing rMDV2.

Accordingly, in one aspect, the present invention relates to immunogenic compositions and/or vaccines that contain the rMDV _np of the present invention (eg, rHVT). In specific embodiments, these immunogenic compositions and/or vaccines are stable, safe, and have relatively strong antigen expression and/or potency. Alternatively or in addition, the immunogenic compositions and/or vaccines that contain the rMDV _np of the present invention assist in protecting chickens from disease caused by ILTV and/or IBDV and/or NDV and/or MDV1 after administration of the immunogenic compositions and/or MDV1 to chickens. /or vaccines.

The present invention also relates to immunogenic compositions and/or vaccines that contain any rMDV _np of the present invention (e.g. rHVT) which are further combined with additional IBDV, ILTV, NDV and/or MDV antigen(s) to improve and extend the immunogenicity provided . In a particular embodiment of this type, the antigen is an attenuated or moderately live IBDV variant (eg, IBDV 89/03). In another specific embodiment of this type, the antigen is attenuated (or moderately live) Newcastle disease virus (NDV), such as NDV C2. In yet another specific embodiment of this type, the antigen is attenuated Marek's disease virus, eg SB1. In addition, the present invention also relates to immunogenic compositions and/or vaccines that contain any rMDV _np of the present invention, which additionally encodes an antigen for a pathogen other than MDV, ILTV or NDV.

The present invention also relates to methods for protecting poultry against disease caused by ILTV and/or IBDV and/or NDV and/or MDV1 by administering such vaccines and/or immunogenic compositions to birds (eg chickens). In specific embodiments of this type, the vaccine of the present invention is administered subcutaneously. In other embodiments, the implementation of the vaccine of the present invention is administered in ovo.

These and other aspects of the present invention will be better appreciated with reference to the following figures and detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is a schematic drawing of fragments of inserts for creating HVT/IBDV/ILTV/NDV structures as described in Example 2 below. Briefly, this is a schematic representation of the HVT genome, consisting of two unique regions, each flanked by repeat regions, and cloned fragments required for the reconstruction of the HVT/IBDV/ILTV/NDV 67014 virus. Orientation of the inserted gene cassettes (mIE-IBDV-vp2 and ILTV -gD/gI or hIE-NDV-F) relative to interrupted genes (UL54.5 or US2), with flanking genes shown enlarged. Legend: TRL: long terminal repeat region, UL: unique long region; IRL: long internal repeat region, IRS: short internal repeat region; US: unique short area; TRS: region of short terminal repeats.

Figure 2 is a schematic drawing of fragments of the insert to create the design # 2 HVT/IBDV/ILT/NDV. The two HVT insertion sites are UL54.5 and US2. [Cm. See also the description for Figure 1 above].

Figure 3 is a schematic drawing of fragments of the insert to create the design # 3 HVT/IBDV/ILT/NDV. The two HVT insertion sites are UL54.5 and US2. [Cm. See also the description for Figure 1 above].

Figure 4 is a schematic drawing of fragments of inserts to create the design No. 4 HVT/IBDV/ILT/NDV. The two HVT insertion sites are UL54.5 and US2. [Cm. See also the description for Figure 1 above].

Figure 5 is a schematic drawing of fragments of inserts to create the design No. 5 HVT/IBDV/ILT/NDV. The two HVT insertion sites are UL54.5 and US2. [Cm. See also the description for Figure 1 above].

DETAILED DESCRIPTION OF THE INVENTION

The present invention overcomes the existing problem in creating one rMDV _np vector that encodes and expresses antigens from three or more foreign pathogenic chicken viruses. In specific embodiments, the rMDV _np of the present invention encodes and expresses foreign antigenic proteins from three or more avian viruses. In specific embodiments, the avian viruses are Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), and infectious bursitis virus (IBDV). Such rMDV _np vectors may be used in vaccines and/or immunogenic compositions that aid in protection against Marek's disease, infectious bursitis (Gumboro's disease), infectious laryngotracheitis virus and/or Newcastle disease virus. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2. The present invention also relates to immunogenic compositions and/or vaccines which contain any rMDV _np of the present invention in combination with an additional IBDV, ILTV, NDV and/or MDV antigen and/or one or more antigens from a number of chicken pathogens other than MDV , ILTV, NDV, or IBDV. In a completely different aspect, a recombinant vector that encodes and expresses foreign antigens from the NDV, ILTV, IBDV series is not rMDV _np but is a chimeric Marek's disease virus that contains certain genomic sequences from MDV1, replacing their counterparts in the HVT vector, for example novel avian herpes virus (NAHV) [see, for example, US 6913751].

Prior to the present invention, an HVT vector has already been constructed containing the NDV gene inserted in the US10 region. This HVT-NDV vector has been shown to be stable and to express sufficient levels of the corresponding NDV gene product, NDV F protein, to protect vaccinated chickens from virulent NDV challenge. In addition, an HVT vector has already been constructed containing a pair of ILTV genes inserted into the UL54.5 HVT region. This HVT-ILTV vector has been shown to be stable and to express sufficient levels of the respective ILTV gene products, ILTV gI and gD proteins, to protect vaccinated chickens from virulent ILTV infecting virus. More recently, other polyvalent constructs have also been described.

More specifically, a polyvalent HVT construct for protection against both NDV and ILTV was developed based on successful constructs involving insertion of the NDV-F gene at the US10 site and insertion of the ILTV gD and gI genes at the UL54.5 site in separate constructs [cf. , US 8932604 B2]. However, unexpectedly, upon passage of this polyvalent construct in tissue cell culture, the recombinant virus lost the ability to express the ILTVgD, ILTVgI, and F NDV proteins. This is supported by a number of duplicate recombinant HVT constructs. Indeed, these recombinant viruses were unstable and unsuitable for further development as vaccines. These results indicate that the creation of a single multivalent rHVT vector that can stably express both the NDV F protein and the ILTVgD and ILTVgI proteins was not a simple process that can be extrapolated from existing information. Indeed, if such stable and efficient multivalent rHVT vectors were even possible, their design would have to be based on an unpredictable set of complex interactions that minimally affect the relationship between the insertion sites used and the foreign nucleotide sequences to be inserted. Accordingly, the design of rHVT constructs remains unpredictable from the prior art. This may appear to be an even greater problem for rMDV _np , which encodes heterologous antigens from three or more avian pathogens.

Despite the apparent difficulties outlined above and the general consensus in the field that the insertion of foreign antigens from three or more different viral pathogens into an MDV _np construct results in construct overload resulting in a lack of stability, the present invention surprisingly provides stable recombinant MDV _np vectors. in which two genes from ILTV, one gene from IBDV and one gene from NDV were inserted into one MDV _np . Accordingly, such a single rMDV _np construct can be used as the sole active ingredient in a vaccine that provides protection against the four major pathogenic poultry viruses.

In specific embodiments of the present invention, the nucleotide sequences encoding the four foreign antigens are inserted into one or more non-essential regions of the genome of a single HVT. Accordingly, such a recombinant HVT vector should be capable of being used to provide protection against MDV, NDV, IBDV and ILTV infections. Previously, several different rHVT vectors were needed to protect against these four viruses, which can influence each other's antigenicity.

Therefore, the present invention is advantageous over existing methods in that it should provide simultaneous protection against MDV, NDV, IBDV and ILTV infections by inoculating poultry and/or poultry eggs with only one recombinant MDV _np . In particular, this allows additional vaccines to be administered in ovo, since there is a limitation on the volume of injection into the egg, and this further saves manufacturing costs, since only one rather than two vectors are required.

In addition, the present invention includes embodiments that include different rMDV _np constructs in the same vaccine and/or immunogenic compositions. In certain embodiments of this type, the vaccine and/or immunogenic composition contains both rMDV2 and rHVT, each of which encodes one or more foreign antigens. Indeed, in contrast to the combination of two rHVTs, which inevitably results in significant outgrowth of one construct relative to the other, it is reported that combining rHVT with rMDV2 does not result in significant outgrowth. Therefore, in specific embodiments, the vaccine of the present invention contains rHVT which encodes ILTV gD protein, ILTV gI protein, IBDV VP2 protein and NDV F protein, with rMDV2 which encodes another avian viral antigen. So far, it has not been shown that rMDV _np , which encodes and expresses foreign antigens from three different poultry viruses and is still stable, is also capable of expressing sufficient levels of the respective antigens to protect vaccinated chickens from virulent challenge by the respective three viruses, as well as against virulent MDV.

Accordingly, the present invention relates to immunogenic compositions and/or vaccines that contain any rMDV _np of the present invention. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2. In addition, the present invention relates to methods of assisting in protecting (and in some embodiments protecting) poultry against disease caused by ILTV and/or IBDV and/or NDV and/or MDV1 by administering such a vaccine and/or immunogenic compositions of the present invention. In specific embodiments, the poultry is a chicken. In specific embodiments of this type, the vaccine of the present invention is administered subcutaneously. In other embodiments, the implementation of the vaccine of the present invention is administered in ovo. In preferred embodiments, the rMDV _np vaccine of the present invention is both safe, stable and effective.

To more fully appreciate the present invention, the following definitions are provided.

The use of specific terms for convenience of description is not meant to be limiting in any way. Thus, for example, reference to a composition containing a "polypeptide" includes a reference to one or more such polypeptides.

As used herein, the term "non-pathogenic Marek's disease virus" or "MDV _np " or "npMDV" is a virus of the MDV family that has little or no pathogenicity in poultry. The term "MDV _np " includes naturally occurring MDVs that have been passaged or otherwise similarly manipulated, but does not include viral constructs in which a specific region of the genome of one MDV serotype is replaced with a corresponding region of another MDV serotype to form a chimeric virus, such as a novel avian herpes virus (NAHV). In some embodiments, MDV _np is HVT. In other embodiments, MDV _np is MDV2. In particular embodiments of this type, MDV2 is SB1.

As used herein, an MDV _np that has been genetically modified to encode a heterologous nucleotide sequence (eg, a foreign gene) is defined as "recombinant MDV _np " or "rMDV _np ". The term "rMDV _np " includes naturally occurring MDV _np that have been genetically modified to encode a heterologous nucleotide sequence, but does not include viral constructs in which a specific region of the MDV genome of one serotype is replaced by a corresponding region of the MDV serotype of another serotype to form a chimeric virus such as the new avian herpesvirus (NAHV).

As used herein, the term "novel avian herpesvirus" ("NAHV") is a recombinant chimeric virus containing a unique long viral genomic region that naturally occurs in turkey herpesvirus (HVT) and a unique short viral genomic region that naturally occurs in Marek's disease virus 1 (MDV1) [see, US 5965138, US 6183753, US 6913751 B2]. In a preferred embodiment, NAHV includes a unique long viral genomic region that occurs naturally in turkey herpesvirus (HVT), a unique short viral genomic region that naturally occurs in Marek's disease virus 1 (MDV1), and repeat regions of HVT viruses [ cm. US 6913751 B2].

As used herein, the term "optional site" is a site in the MDV _np genome (or alternatively the NAVH genome) at which insertion of a heterologous nucleotide sequence at that site does not prevent MDV _np (or NAVH) from replicating in the host cell. Non-essential sites are typically identified by the open reading frame in which they reside, such as the US2 site, or the region between two open reading frames, such as the UL7/8 site. The use of the term "non-essential site" is in no way even intended to suggest that there is only one unique nucleotide position in the nucleotide sequence of a given open reading frame (or in the region between two open reading frames) where a heterologous nucleic acid insertion must be made in order to so that MDV _np (or NAVH) retains the ability to replicate in the host cell.

As used herein, when rMDV _np (or NAHV) is said to have a given nucleic acid "inserted" at a non-essential site in the rMDV _np (or NAHV genome), this means that the given nucleic acid is a heterologous nucleic acid, which resides in this non-essential MDV _np (or NAHV) site. Accordingly, an rMDV _np comprising a first nucleic acid inserted at a first non-essential site in the rMDV _np genome and a second nucleic acid inserted at a second non-essential site in the rMDV _np genome is equivalent to an rMDV _np containing a first heterologous nucleic acid located at the first non-essential site in the rMDV _np genome, and a second heterologous nucleic acid located at a second non-essential site in the rMDV _np genome, and vice versa.

As used herein, the term "poultry" may include chickens, turkeys, ducks, geese, quails, and pheasants.

As used herein, the term "vaccine" is a composition that is suitable for use in an animal (including, in some embodiments, a human, while in other embodiments specifically not a human) containing one or more antigens, usually in combination with a pharmaceutically acceptable carrier, such as a liquid containing water, and which, when administered to an animal, elicits an immune response strong enough to minimally assist in protection against disease resulting from infection with a wild-type microorganism, i.e. strong enough to aid in prevention disease and/or preventing, ameliorating or curing the disease.

As used herein, the term "helps protect" does not require complete protection against any signs of infection. For example, "helps protect" may mean that protection is sufficient so that after infection, the symptoms of the underlying infection are at least reduced, and/or that one or more of the underlying cellular, physiological, or biochemical causes or mechanisms causing the symptoms reduced and/or eliminated. It is understood that "attenuated" as used herein means the relative state of the infection, including the molecular state of the infection, and not just the physiological state of the infection.

The vaccines of the present invention contain at least one stable rMDV _np of the present invention. rMDV _np is considered phenotypically stable when at least 90% of viral plaques examined are positive for the expression of the introduced foreign antigen, as demonstrated by binding of antibodies specific for the expressed protein in an immunofluorescence assay after at least 10 tissue cell passages. cultures after initial seeding or after re-isolation of the virus from vaccinated birds.

The vaccines of the present invention are also effective and preferably provide at least at least 70% protection against NDV and/or at least 70% protection against IBDV and/or at least 70% protection against ILTV and/or at least 60% protection against MDV from clinical signs or lesions associated with the disease. More preferably, the vaccine minimally provides at least 80% protection against NDV, at least 80% protection against IBDV, at least 80% protection against ILTV, and at least 70% protection against MDV against clinical signs or lesions associated with the disease. . Even more preferably, the vaccines comply with the guidelines established by the USDA and codified in the Code of Federal Regulations, Title 9, Part 113 (9CFR 113) "Standard Requirements for Animal Products" for live virus vaccines, which must provide at least at least 90% protection in the case of NDV, IBDV and ILTV, and at least 80% in the case of MDV, against clinical signs or lesions associated with the disease, in order to obtain a license.

Used in the present description, the term "polyvalent vaccine" is a vaccine that contains two or more different antigens. In a specific embodiment of this type, a multivalent vaccine stimulates the recipient's immune system against two or more different pathogens.

As used herein, the term "adjuvant" is a substance that is capable of facilitating or enhancing a cascade of immunological events, which ultimately leads to a better immunological response, i.e. integrated systemic response to an antigen. An adjuvant is not normally required for an immunological response to occur, but promotes or enhances that response.

Used in the present description, the term "pharmaceutically acceptable" is used as an adjective to indicate that the modified noun is suitable for use in a pharmaceutical product. When used, for example, to describe an excipient in a pharmaceutical vaccine, it characterizes the excipient as being compatible with the other ingredients of the composition and not adversely harmful to the intended recipient.

As used herein, the term "systemic administration" is administration to the body's circulatory system (including the cardiovascular and lymphatic system), thereby affecting the body as a whole, and not at a specific locus, such as the gastrointestinal tract (for example, through oral or rectal administration) and into the respiratory system (eg via intranasal administration). Systemic administration can be performed, for example, by injection into the muscle tissue (intramuscularly), into the dermis (intradermal or transdermal), under the skin (subcutaneously), under the mucous membrane (submucosal), into the veins (intravenous), etc.

As used herein, the term "parenteral administration" includes subcutaneous injections, submucosal injections, intravenous injections, intramuscular injections, intradermal injections, and infusion.

The term "about" is used interchangeably with the term "about" and means that the value is within twenty-five percent of the specified value, that is, a peptide containing "about" 100 amino acid residues may contain from 75 to 125 amino acid residues.

Used in the present description, the term "polypeptide" is used interchangeably with the terms "protein" and "peptide" and refers to a polymer containing two or more amino acids linked by peptide bonds. As used herein, the term "polypeptide" includes a substantial fragment or segment and encompasses a stretch of amino acid residues of at least about 8 amino acids, typically at least about 12 amino acids, typically at least about 16 amino acids, preferably , at least about 20 amino acids, and in particularly preferred embodiments, at least about 30 or more amino acids, such as 35, 40, 45, 50, etc. Such fragments may have ends that begin and/or end in almost all positions, for example, starting with residues 1, 2, 3, etc. and ending, for example, 155, 154, 153, etc. in all practical combinations.

Optionally, the polypeptide may lack some of the amino acid residues that are encoded by the gene or mRNA. For example, a gene or mRNA molecule may encode a sequence of amino acid residues at the N-terminus of a polypeptide (ie, a signal sequence) that is cleaved from the protein and therefore may not be part of the final protein.

As used herein, the term "antigenic fragment" in relation to a particular protein (for example, a protein antigen) is a fragment of that protein (including large fragments that lack only one amino acid from a full-length protein) that is antigenic, that is, capable of interacting specifically with an antigen-recognizing molecule of the immune system, such as an immunoglobulin (antibody) or a T-cell antigen receptor. For example, an antigenic fragment of an IBDV VP2 protein is an antigenic fragment of a VP2 protein. Preferably, the antigenic fragment of the present invention is immunodominant for antibody and/or T-cell receptor recognition. In specific embodiments, an antigenic fragment with respect to a given protein antigen is a fragment of that protein that retains at least 25% of the antigenicity of the full-length protein. In preferred embodiments, the antigenic fragment retains at least 50% of the antigenicity of the full length protein. In more preferred embodiments, it retains at least 75% of the antigenicity of the full length protein. Antigenic fragments may be as small as 5-10 amino acids or, on the other hand, may be large fragments lacking only one amino acid from a full-length protein. In specific embodiments, the implementation of the antigenic fragment contains from 25 to 100 amino acid residues.

In this context, an amino acid sequence is 100% "homologous" to a second amino acid sequence if the two amino acid sequences are identical and/or differ only by neutral or conservative substitutions, as defined below. Accordingly, an amino acid sequence is about 80% "homologous" to a second amino acid sequence if about 80% of the two amino acid sequences are identical and/or differ only by neutral or conservative substitutions.

Functionally equivalent amino acid residues can often be replaced by residues in the sequence resulting in a conservative amino acid substitution. Such changes are defined by the term "conservative substitution" as used herein. For example, one or more amino acid residues in a sequence can be replaced with another amino acid of similar polarity that acts as a functional equivalent, resulting in a silent change. Amino acid substitutions in a sequence may be selected from other members of the class to which the amino acid belongs. For example, non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. Amino acids containing aromatic ring structures are phenylalanine, tryptophan and tyrosine. Polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. Positively charged (with properties of a base) amino acids include arginine, lysine, and histidine. Negatively charged (acidic) amino acids include aspartic acid and glutamic acid. It is expected that such changes will not affect the apparent molecular weight determined by polyacrylamide gel electrophoresis or isoelectric point.

Particularly preferred conservative substitutions are: Lys for Arg and vice versa so that a positive charge can be retained; Glu for Asp and vice versa, so that a negative charge can be retained; Ser for Thr so that free -OH can be retained; and Gln for Asn so that free NH2 can be retained. Amino acids can also be assigned to the following similarity groups: (1) proline, alanine, glycine, serine, and threonine; (2) glutamine, asparagine, glutamic acid and aspartic acid; (3) histidine, lysine and arginine; (4) cysteine; (5) valine, leucine, isoleucine, methionine; and (6) phenylalanine, tyrosine, and tryptophan.

In a related embodiment, two highly homologous DNA sequences can be identified by their own homology or by the homology of the amino acids they encode. Such sequence comparisons can be performed using standard software available from sequence data banks. In a particular embodiment, the two highly homologous DNA sequences encode amino acid sequences having about 80% identity, more preferably about 90% identity, and even more preferably about 95% identity. More specifically, the two highly homologous amino acid sequences share about 80% identity, even more preferably about 90% identity, and even more preferably about 95% identity.

As used herein, percent protein and DNA sequence identity can be determined using software such as MacVector v9 commercially available from Accelrys (Burlington, Massachusetts) and the Clustal W algorithm with default alignment parameters and default parameters for identity. . See, for example, Thompson et al. 1994 Nucleic Acids Res. 22:4673-4680. ClustalW is available as a free download for Dos, Macintosh and Unix platforms, for example from EMBLI, the European Bioinformatics Institute. The current download link is at http://www.ebi.ac.uk/clustalw/. These and other available programs can also be used to determine sequence similarity using the same or similar default settings.

As used herein, the terms "polynucleotide" or "nucleic acid" or "nucleic acid molecule" are used interchangeably and refer to a molecule containing nucleotides, including, but not limited to, RNA, cDNA, genomic DNA, and even synthetic DNA sequences. The terms are also intended to encompass nucleic acid molecules that include any of the art-known analogues of DNA and RNA bases.

A "coding sequence" for a nucleic acid, or a "sequence encoding" a particular protein or peptide, is a nucleotide sequence that is transcribed and translated into a polypeptide in vitro or in vivo when under the control of appropriate regulatory elements.

The boundaries of the coding sequence are defined by a start codon at the 5' end and a translation stop codon at the 3' end. A coding sequence may include, without limitation, prokaryotic sequences, cDNA from eukaryotic mRNA, genomic DNA sequences from eukaryotic (eg, avian) DNA, and even synthetic DNA sequences. The transcription termination sequence may be located 3' from the coding sequence.

"Operably linked" refers to an arrangement of elements in which the described components are configured to perform their normal function. Thus, control elements operably linked to a coding sequence are able to influence the expression of the coding sequence. Control elements need not be contiguous with a coding sequence if they function to direct its expression. Thus, for example, intermediate non-translated but transcribed sequences may be present between the promoter and the coding sequence, and the promoter may still be considered "operably linked" to the coding sequence.

As used herein, the term "transcription terminator sequence" is used interchangeably with the term "polyadenylation regulatory element" and is a sequence that is typically located 3' from a coding region of DNA and that may be required to terminate transcription of that coding region. DNA sequences. A transcription terminator is a DNA regulatory element involved in terminating the transcription of a coding region in RNA. Typically, such an element encodes a section, such as a hairpin structure, that has a secondary structure that can cause the RNA polymerase complex to stop transcription. Thus, the transcription terminator is always located 3' from the stop codon from the region to be translated, the 3' untranslated region.

As used herein, the term "expression cassette" is a recombinant nucleic acid that minimally contains a promoter and a heterologous coding sequence operably linked to that promoter. In many such embodiments, the expression cassette further comprises a transcription terminator sequence. Accordingly, insertion of an expression cassette at a non-essential site in the rMDV _np genome can result in the expression of a heterologous rMDV _np coding sequence. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

As used herein, a "heterologous nucleotide sequence" is a nucleotide sequence that is added to the nucleotide sequence of the present invention by recombinant methods to form a nucleic acid that does not occur naturally. In specific embodiments, the "heterologous nucleotide sequence" of the present invention may encode a protein antigen (e.g., encoded by a "foreign gene" relative to the rMDV _np vectors of the present invention), such as IBDV VP2 protein, ILTV gI protein, ILTV gD protein, and/or FNDV. In this case, such protein antigens can be called "foreign antigens" or, more specifically, "foreign protein antigens".

Heterologous nucleotide sequences may also encode fusion (eg, chimeric) proteins. In addition, the heterologous nucleotide sequence may encode peptides and/or proteins that have regulatory and/or structural properties. In other such embodiments, the heterologous nucleotide sequence may encode a protein or peptide that functions as a means of detecting the protein or peptide encoded by the nucleotide sequence of the present invention upon expression of the recombinant nucleic acid. In yet another embodiment, the heterologous nucleotide sequence can function as a means of detecting the nucleotide sequence of the present invention. A heterologous nucleotide sequence may contain non-coding sequences, including restriction sites, regulatory sites, promoters, and the like. "Heterologous nucleic acid" includes a heterologous nucleotide sequence.

Insertion of a nucleic acid encoding an antigen of the present invention into an rMDV _np vector is easily achieved when the ends of both the nucleic acid and the vector contain compatible restriction sites. If this is not possible, it may be necessary to modify the ends of the nucleotide sequence and/or the vector by hydrolyzing single-stranded nucleic acid overhangs (e.g., DNA overhangs) generated by restriction endonuclease digestion to produce blunt ends, or to achieve the same result by filling in the single-stranded ends with an appropriate polymerase. Alternatively, the desired sites can be obtained, for example, by ligating nucleotide sequences (linkers) at the ends. Such linkers may contain specific oligonucleotide sequences that define the desired restriction sites. Restriction sites can also be created using the polymerase chain reaction (PCR). [See, for example, Saiki et al., Science 239: 487-491 (1988)]. The digested vector and DNA fragments can also be modified, if necessary, by homopolymer tailing. Alternatively, the recombinant nucleotide sequence may be synthesized de novo.

Protein antigens and nucleic acids encoding protein antigens

The ILTV gD gene appears to encode a 434 amino acid long glycoprotein with a molecular weight of 48,477 daltons, although others have suggested that a start codon located in the 3' region that results in an ILTV gD protein containing only 377 amino acid residues is the actual start codon [Wild et al., Virus Genes 12: 104-116 (1996)]. The gI ILTV gene encodes a 362 amino acid long glycoprotein with a molecular weight of 39,753 daltons [US 6,875,856, hereby incorporated by reference]. Nucleic acids encoding natural and/or laboratory variants of gD ILTV and gI ILTV can be replaced by those given as examples.

In specific embodiments, the rMDV _np comprises a recombinant nucleic acid (eg, an expression cassette) that encodes an ILTV gD protein comprising the amino acid sequence of SEQ ID NO: 2, or an antigenic fragment thereof. In related embodiments, rMDV _np comprises a recombinant nucleic acid that encodes an ILTV gD protein containing an amino acid sequence that is greater than 90% and/or greater than 95% and/or greater than 98% and/or greater than 99 % identity with the amino acid sequence of SEQ ID NO: 2. In specific embodiments, the ILTV gD protein is encoded by the nucleotide sequence of SEQ ID NO: 1. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

In some embodiments, the rMDV _np comprises a recombinant nucleic acid (eg, an expression cassette) that encodes an ILTV gI protein comprising the amino acid sequence of SEQ ID NO: 4, or an antigenic fragment thereof. In related embodiments, the rMDV _np comprises a recombinant nucleic acid that encodes an ILTV gI protein containing an amino acid sequence that is greater than 90% and/or greater than 95% and/or greater than 98% and/or greater than 99 % identity with the amino acid sequence of SEQ ID NO: 4. In specific embodiments, the ILTV gI protein is encoded by the nucleotide sequence of SEQ ID NO: 3. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

As mentioned earlier, IBDV is a member of the Birnaviridae family, whose genome consists of two segments (A and B) of double-stranded RNA. The larger A segment encodes a 110 kDa polyprotein, which is subsequently cleaved by autoproteolysis to form the mature viral proteins VP2, VP3, and VP4. Of these, VP2 and VP3 are the structural capsid proteins for the virion, with the VP2 protein being the main protective immunogen. In the case of IBDV, there are two serotypes, serotype 1 and 2, which can be distinguished using virus neutralization (VN) tests. Serotype 1 viruses have been shown to be pathogenic in chickens, while IBDV serotype 2 only causes subacute disease in turkeys. Historically, IBDV serotype 1 viruses consisted of only one type, which is known as the "classic" IBD virus, but so-called "variant" strains of IBDV have subsequently emerged. In specific embodiments, the IBDV VP2 gene encodes a VP2 protein from classical type IBDV. Such genes are well known and sequence information is readily available [see, for example, GenBank registry. number: D00869 (F52/70), D00499 (STC) or AF499929 (D78)]. Alternatively, this gene can be obtained from a classical IBDV genome isolated from a natural source using routine Birnavirus manipulation techniques. The classic type of IBDV can be easily identified using serology or molecular biology.

In specific embodiments, the rMDV _np comprises a recombinant nucleic acid (eg, an expression cassette) that encodes an IBDV VP2 protein comprising the amino acid sequence of SEQ ID NO: 6, or an antigenic fragment thereof. In related embodiments, the rMDV _np comprises a recombinant nucleic acid that encodes an IBDV VP2 protein containing an amino acid sequence that is greater than 90% and/or greater than 95% and/or greater than 98% and/or greater than 99% identity with the amino acid sequence of SEQ ID NO: 6. In specific embodiments, the IBDV VP2 protein is encoded by the nucleotide sequence of SEQ ID NO: 5. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

Routine IBDV vaccinations are given as early as possible in the life of the poultry using attenuated IBDV strains, but should only be given when the anti-IBDV MDA level has been sufficiently reduced, which is usually sometime between 15 and 20 days post-hatch. Many live or inactivated IBDV vaccines are commercially available, such as a live vaccine such as Nobilis™ Gumboro D78 (Merck Animal Health).

NDV has a non-segmented, negative sense, single-stranded RNA genome of approximately 15 kb. and contains six genes, including the NDV F protein gene, which encodes the so-called "fusion" glycoprotein (F protein). The F protein is involved in the attachment and entry of NDV into host cells, and as an immunodominant protein, it may be the basis of an effective immune response against NDV. The NDV F protein is expressed as a native F0 protein, which is activated upon cleavage by extracellular peptidases.

For example, the NDV F protein gene can be derived from NDV clone 30, a common lentogenic NDV vaccine strain. In some embodiments, the rMDV _np comprises a recombinant nucleic acid (eg, an expression cassette) that encodes an NDV F protein comprising the amino acid sequence of SEQ ID NO: 8, or an antigenic fragment thereof. In related embodiments, the rMDV _np comprises a recombinant nucleic acid that encodes an NDV F protein containing an amino acid sequence that is greater than 90% and/or greater than 95% and/or greater than 98% and/or greater than 99% identity with the amino acid sequence of SEQ ID NO: 8. In specific embodiments, the NDV F protein is encoded by the nucleotide sequence of SEQ ID NO: 7. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

In related embodiments, the rMDV _np comprises a recombinant nucleic acid (eg, an expression cassette) that encodes an NDV F protein comprising the amino acid sequence of SEQ ID NO: 10, or an antigenic fragment thereof. In other embodiments, the rMDV _np comprises a recombinant nucleic acid that encodes an NDV F protein containing an amino acid sequence that is greater than 90% and/or greater than 95% and/or greater than 98% and/or greater than 99 % identity with the amino acid sequence of SEQ ID NO: 10. In specific embodiments, the NDV F protein is encoded by the nucleotide sequence of SEQ ID NO: 9. In specific embodiments, rMDV _np is rHVT. In alternative embodiments, rMDV _np is rMDV2.

Nucleic acids encoding natural and/or laboratory-derived variants of the F protein gene are equally applicable from either lentogenic, mesogenic, or velogenic NDV because the F protein gene sequence itself is highly conserved across these various NDV pathotypes.

Nucleotide and/or protein sequences for the chick pathogen protein antigens encoded by the rMDV _np of the present invention can also be found in public databases such as GenBank or the Protein Information Resource.

Promoters and regulatory elements of polyadenylation

A promoter is a functional region in the genome of an organism that directs transcription of a coding region located 3' from it. Therefore, the promoter is located 5' from the coding region of the gene. Promoter-driven mRNA synthesis begins at a "transcriptional start site" (TSS). The resulting mRNA, in turn, is translated into protein, starting at the start codon of the gene, which is the first ATG sequence in the open reading frame (the first AUG in mRNA). Typically, the TSS is located 30-40 nucleotides in the 5' region from the start codon. TSS can be determined by sequencing the 5' end of the mRNA of the gene, for example, by the RACE method. Typically, promoters are within about 1000 nucleotides in the 5' region of the start codon's A position, which is commonly referred to as A+1, and most promoters are located between nucleotides -500 and A+1.

Promoter nomenclature is usually based on the name of the gene that controls expression. For example, the mouse cytomegalovirus "immediate early gene 1" (mCMVIE1) promoter "mCMV-IE1 gene promoter" refers to a promoter that naturally drives the expression of mCMV immediate early gene 1 (IE1 gene) and, accordingly, is located directly at the 5'- regions from this gene. Because the IE1 gene is such a well-described and well-recognised gene, and since the genomes of several mCMVs have been sequenced (in whole or in part), such a promoter can easily be identified by conventional methods. For example, in a basic protocol, a promoter can be obtained by subcloning the approximate region between two consecutive genes, for example, from the poly A signal of the gene in the 5' region to the TSS gene in the 3' region. The promoter can then be identified by standard tests, for example by expression of the marker gene by progressively smaller regions of the putative promoter.

Generally, promoters contain a number of recognizable regulatory regions, such as an enhancer region, which is involved in the binding of regulatory factors that affect the timing, duration, conditions, and rate of transcription. While the enhancer region is usually located in the 5' region, the promoter also contains another 3' region, which is involved in the binding of transcription factors and the control of the RNA polymerase itself. This 3' region typically contains a number of conserved promoter sequence elements such as the TATA block, the CAAT block, and the GC block.

A promoter containing both an enhancer and a 3' region is referred to as a "full" promoter, while a promoter containing only a 3' region is referred to as a "central" promoter.

A promoter for the expression of a (heterologous) gene in a (viral) vector must be able to efficiently drive the transcription of this 3' coding sequence. This is commonly referred to as a promoter "operably linked" to a coding sequence such that the gene is "under control" of the promoter or "driven" by the promoter. Typically, this means that in an expression cassette, the promoter and the coding sequence of the gene are in the same nucleic acid, in effective proximity, and without signals or sequences between them that could interfere with efficient transcription of the coding sequence.

The mCMV-IE1 gene promoter is well known in the art and can be readily obtained from a variety of commercial sources such as commercial cloning and expression plasmid suppliers. The IE1 gene is also referred to as the "IE core gene". The mCMV-IE1 protein is also referred to as pp89. Dörsch-Häsler et al. [Proc. Nat. Acad. Sci., 82: 8325-8329 (1985)] described the mCMV IE1 gene promoter in 1985, and the use of this promoter in heterologous expression is also described in WO 87/03.905 and EP 728842. The nucleotide sequence of the complete mCMV IE locus is available from GenBank under Registry . number L06816.1 (since March 2004). The mCMV itself can be obtained from ATCC: Source Register. number VR-194, and recently this information is available under the register. number VR-1399.

In one embodiment, the mCMV-IE1 gene promoter is a complete promoter including both the promoter core region and the enhancer region for the mCMV-IE1 gene. The complete mCMV-IE1 gene promoter is approximately 1.4 kb in size. However, the present invention also allows for some variance in the length of not only the mCMV IE1 gene promoter, but also other elements that make up the recombinant DNA expression cassette used in the present invention. This may be the result of differences in the precise conditions used for cloning and construction. For example, this variance may arise from the use of different restriction enzyme sites, primers for PCR cloning, or different conditions for tailoring the ends of the cloning molecules used. Therefore, there may be some variation in the length - smaller or larger - of the constituent elements without affecting the stability and relatively strong expression of the antigen and/or the efficiency of the entire expression cassette. In this case, these differences in length are not significant and are within the scope of the invention. Therefore, the mCMV-IE1 gene promoter is "about 1.4 kb" is: 1.4 kb. ± approx. 25%. In specific embodiments, the promoter is 1.4 kb. ± approx. 20%. In other embodiments, the implementation of the dispersion may be 1.4 kb. ± about 15%, 1.4 kb. ± about 12%, 1.4 kb. ± about 10%, 1.4 kb. ± about 8%, 1.4 kb. ± about 6%, 1.4 kb. ± about 5%, 1.4 kb. ± 4%, 1.4 kbp ± 3%, 1.4 kbp ± 2% or even 1.4 kb. ± 1%.

Similarly, promoter element homologues or variants that are equally effective and stable can be used. Therefore, in some embodiments, the mCMV-IE1 gene promoter of the present invention may be an approximately 1.4 kb DNA molecule that contains a nucleotide sequence having at least 95%, 96%, 97%, 98 % or even 99% nucleotide sequence identity with the nucleotide sequence of SEQ ID NO: 13. In a specific embodiment, the mCMV-IE1 gene promoter consists of the nucleotide sequence of SEQ ID NO: 13.

Many alternative promoters can be used to drive the expression of a heterologous gene encoding a protein antigen or antigenic fragment thereof in rMDV _np of the present invention. Examples include the pseudorabies virus (PRV) gpX promoter [see, WO 87/04463], the Rous sarcoma virus LTR promoter, the SV40 early gene promoter, the chicken beta actin gene promoter containing the nucleotide sequence of SEQ ID NO: 17, the hCMV strain IE promoter Towne SEQ ID NO: 16, hCMV promoter IE derivative (from strain AD169) containing the nucleotide sequence of SEQ ID NO: 14, ILTV gD promoter containing the nucleotide sequence of SEQ ID NO: 11, and ILTV gI promoter containing the nucleotide sequence of SEQ ID NO: : 12, [see, for example, US 6183753 B1], human cytomegalovirus immediate early gene 1 (IE hCMV1) promoter [US 5830745; US 5980906] and the chicken beta-actin gene promoter [EP 1 298 139 B1]. A particular heterologous promoter for the IBDV VP2 gene is the mouse cytomegalovirus (mCMV IE1) promoter. In a specific embodiment of this type of IE1 mCMV contains the nucleotide sequence of SEQ ID NO: 13 [see, for example, EP 728,842; PCT/EP2015/081121].

The inclusion of a polyadenylation regulatory element in the 3' region of the DNA coding region is often required to terminate transcription of the DNA coding sequence. Accordingly, many genes contain a polyadenylation regulatory element at the 3' end of their coding sequence. Many such regulatory elements have been identified and can be used in the rMDV _np of the present invention. Specific examples of polyadenylation regulatory elements exemplified herein include the feline herpesvirus (FHV) US-9 polyadenylation signal containing the nucleotide sequence of SEQ ID NO: 18 and the human herpes simplex virus (HSV) thymidine kinase polyadenylation signal containing the nucleotide sequence . SEQ ID NO: 19. The terminator and polyadenylation sequence can also be derived from the feline herpesvirus (FHV) glycoprotein B (gB) gene, from the immediate early (IE) gene of human cytomegalovirus (hCMV), strain AD 169, or from simian virus 40 (SV40). ),

Vaccines and immunogenic compositions

The present invention relates to the use of recombinant MDV _np , nucleic acid molecules used to construct MDV _np , or host cells for growing them, or any combination thereof, in accordance with the present invention, to obtain a vaccine for poultry. Accordingly, the present invention relates to vaccines and/or immunogenic compositions that include the polyvalent recombinant MDV _np of the present invention. Such vaccines can be used to help prevent and/or prevent infectious bursitis (Gumboro's disease) and/or Marek's disease and/or diseases associated with ILTV infections and/or diseases associated with NDV infections. The vaccine according to the present invention can be used for prophylactic and/or therapeutic treatment and thus can prevent the onset and/or progression of infection and/or the clinical signs of these diseases.

The recombinant MDV _np of the present invention can be grown by any number of methods currently used in the art. For example, the recombinant MDV _np of the present invention can be grown using in vitro cultures of primary chick cells, see for example the examples below where chick embryonic fibroblast (CEF) cells were used. CEF can be prepared by trypsinization of chick embryos. CEFs can also be placed in monolayers and then infected with MDV _np . This particular process can easily be scaled up to industrial production.

Therefore, another aspect of the invention relates to a method for preparing a vaccine of the invention, comprising the steps of infecting host cells with the recombinant MDV _np of the present invention, harvesting the infected host cells, and then mixing the harvested infected host cells with a pharmaceutically acceptable carrier. Suitable methods of infection, culture and harvest are well known in the art and are described and exemplified herein.

Typically, infected host cells are harvested when they are intact to obtain recombinant MDV _np in its cell-associated form. These cells can be placed in a suitable carrier composition to provide storage and freezing stability. Infected cells can be filled into glass ampoules, which are sealed, frozen and stored in liquid nitrogen. Accordingly, in some embodiments of the present invention, the vaccines and/or immunogenic compositions of the present invention are stored frozen and, accordingly, contain a cryopreservative such as dimethyl sulfoxide (DMSO) to keep infected cells frozen.

Alternatively, when the recombinant MDV _np is a recombinant HVT, it can be isolated from the host cell, such as by sonication at the end of the culture, and then transferred to a stabilizer and lyophilized for stable storage, or otherwise reduced in storage liquid volume, and then reconstituted in liquid diluent before or during administration. Such recovery can be achieved using, for example, vaccine grade water. In some embodiments, the lyophilized portion of a multivalent vaccine may contain one or more antigens, and the diluent may contain one or more other antigens.

In specific embodiments, the implementation of the vaccine of the present invention (or part thereof) may be in lyophilized form, for example, in the form of tablets and/or spheres, which are obtained by the method described in WO 2010/125084, hereby incorporated by reference in its entirety. In particular, reference is made to the examples, from page 15, line 28 to page 27, line 9 of WO 2010/125084, describing a method for preparing such rapidly disintegrating tablets/spheres. Such lyophilized forms can be readily dissolved in a diluent to allow systemic administration of the vaccine.

Vaccines and immunogenic compositions may, but need not, include physiologically compatible buffers and saline and the like, as well as pharmaceutically acceptable adjuvants. Adjuvants may be useful in improving the immune response and/or improving the stability of vaccine preparations. Adjuvants are usually described as non-specific stimulants of the immune system, but may also be useful for targeting specific arms of the immune system. One or more compounds that have this activity may be added to the vaccine. Therefore, specific vaccines of the present invention may further contain an adjuvant. Examples of chemical compounds that can be used as adjuvants include, but are not limited to, aluminum compounds (e.g., aluminum hydroxide), metabolizable and non-metabolizable oils, mineral oils, including mannide oleate derivatives in mineral oil solution (e.g., MONTANIDE ISA 70 from Seppic SA, France) and light mineral oils such as DRAKEOL 6VR, block polymers, ISCOMs (Immune Boosting Complexes), vitamins and minerals (including but not limited to: vitamin E, vitamin A, selenium and vitamin B12) and CARBOPOL®.

Other suitable adjuvants sometimes referred to as immunostimulants include, but are not limited to: cytokines, growth factors, chemokines, cell culture supernatants of lymphocytes, monocytes, cells from lymphoid organs, cell preparations and/or extracts from plants, bacteria or parasites (Staphylococcus aureus or lipopolysaccharide preparations) or mitogens. Typically, the adjuvant is administered simultaneously with the antigen of the present invention. However, adjuvants may also be administered or alternatively may be administered during the two week period prior to vaccination and/or during the period following vaccination, i. as long as the antigen, such as the recombinant MDV _np of the present invention, is retained in the tissue.

The vaccines and/or immunogenic compositions of the present invention can be administered by any route, for example, in ovo, parenteral administration, including intramuscular injection, subcutaneous injection, intravenous injection, intradermal injection, scarification, oral administration, or any combination thereof.

In addition, the polyvalent recombinant MDV _np of the present invention can be used and/or combined with additional IBDV, ILTV, NDV and/or MDV antigens to improve and extend the provided immunogenicity and/or antigens to other pathogens to provide immune protection against such other pathogens. These additional antigens may be live or killed whole microorganisms, other recombinant vectors, cell homogenates, extracts, proteins, or any other such derivative, provided that they do not adversely affect the safety and stability of the relatively strong expression of the antigen and/or the efficacy of the vaccine. according to the present invention.

The combination of the polyvalent recombinant MDV _np of the present invention with an additional MDV, IBDV, NDV and/or ILTV antigen may be useful in cases where polyvirulent field strains of MDV, IBDV, NDV or ILTV predominate, such as in a particular geographic region. In this regard, the combination of the polyvalent recombinant MDV _np of the present invention with MDV1, MDV2 or HVT includes the Rispens strain (MDV1), the SB1 strain (MDV2), the FC126 strain (HVT) and/or the PB1 strain (HVT). To improve the response against IBDV, the polyvalent recombinant MDV _np can be combined with an IBDV vaccine strain such as a soft live IBDV vaccine strain, e.g. D78 (cloned intermediate), PBG98, Cu-1, ST-12 (intermediate) or 89/ 03 (live variant of the Delaware strain) in a polyvalent vaccine.

Examples of other microorganisms that can be used as antigens with the polyvalent recombinant MDV _np of the present invention include: (i) viruses such as infectious bronchitis virus, avian influenza virus, adenovirus, egg drop syndrome virus, infectious bursitis virus, chicken anemia, avian encephalomyelitis virus, avian pox virus, turkey rhinotracheitis virus, duck distemper virus (duck viral enteritis), pigeon pox virus, avian leukemia virus, avian pneumovirus and reovirus, (ii) bacteria such as Escherichia coli, Salmonella spec ., Ornitobacterium rhinotracheale, Haemophilis paragallinarum, Pasteurella multocida, Erysipelothrix rhusiopathiae, Erysipelas spec., Erysipelas spec.,. (iii) parasites such as Eimeria spec., and (iv) fungi such as Aspergillus spec. In specific embodiments of the present invention, the recombinant MDV _np of the present invention may be combined with a mild live NDV vaccine strain, such as the C2 vaccine strain. Many of these strains are used in commercial vaccines.

The combination vaccine can be prepared in a variety of ways, including combining the recombinant MDV _np of the present invention with virus or bacterial or fungal or parasite or host cell preparations, or a mixture of any and/or all of them. In specific embodiments, the components for such a combination vaccine are conveniently manufactured separately and then combined and loaded into the same vaccine container.

As described above, the vaccine according to the invention can advantageously be used to provide safe and effective immune protection to the chicken, for example against one or more poultry diseases, by a single inoculation at a very young age or in ovo. Alternatively, as should be apparent to any person skilled in the art of poultry vaccines, the combinations described above may also include vaccination schedules in which the polyvalent recombinant MDV _np of the present invention and the additional antigen are not administered simultaneously; for example, recombinant MDV _np can be applied in ovo and NDV strain C2 and/or IBDV (eg 89/03) can be applied at a subsequent time/date.

Accordingly, the vaccines of the present invention may be administered to an avian subject in a single dose or in multiple doses. For example, the vaccine of the present invention can be applied on the day of hatching and/or in ovo on day 16-18 (day of embryonic development) ED. When multiple doses are administered, they may be administered either simultaneously or sequentially, in a manner and at a time consistent with the formulation of the vaccine, and in such amount as will be immunologically effective. Therefore, the vaccine of the present invention can effectively serve as a primary vaccination, which can subsequently be followed and amplified by a booster vaccination of an identical vaccine, or by using another vaccine preparation, such as a classic inactivated adjuvanted whole virus vaccine. Alternatively, the vaccine of the present invention may be administered to the bird solely as a booster.

The volume per dose of the vaccine of the present invention can be optimized according to the intended mode of administration: in ovo inoculation is typically used with a volume of 0.05 to 0.5 ml/egg, and parenteral injection is typically performed with a volume between 0.1 and 1 ml/bird. In any event, optimization of the vaccine dose volume is within the ability of the skilled artisan.

SEQUENCE TABLE

SEQ ID
NO: Description Type of one gD ILTV Glycoprotein nucleic acid 2 gD ILTV Glycoprotein Amino acid 3 gI ILTV Glycoprotein nucleic acid four gI ILTV Glycoprotein Amino acid 5 VP2 IBDV nucleic acid 6 VP2 IBDV Amino acid 7 F NDV Protein (Clone 30) nucleic acid eight F NDV Protein (Clone 30) Amino acid 9 F NDV Protein (B1 Hitchner) nucleic acid ten F NDV Protein (B1 Hitchner) amino acid eleven gD ILTV promoter nucleic acid 12 gI ILTV promoter nucleic acid 13 mCMV IE promoter nucleic acid fourteen hCMV IE promoter (from strain AD169) nucleic acid fifteen hCMV IE promoter (truncated) nucleic acid 16 hCMV IE promoter (Towne strain) nucleic acid 17 Chicken β-actin promoter nucleic acid eighteen FHV US-9 polyadenylation signal nucleic acid 19 HSV TK polyadenylation signal nucleic acid twenty SV40 polyadenylation signal nucleic acid 21 484-1050-2641-10859 (HVT/IBDV/ILTV/NDV 670-14)
mCMV IEpro-VP2-SV40pA/ILTV/HVT UL54.5 nucleic acid 22 (HVT/IBDV/ILTV/NDV 670-14)
hCMV IEpro-F-IE(term)/HVT US2 1322-48.1 nucleic acid 23 (HVT/IBDV/ILTV/NDV #2)
228509-ILT-435Vec6 (mCMV IEpro-VP2-SV40pA/ILTV/HVT) nucleic acid 24 1333-85.B6 (ILTV/chicken β-actin pro-VP2-FHV US9pA/HVT) nucleic acid 25 1386-04.4#1 (ILTV/hCMV IEpro-VP2-HSV TKpA/HVT) nucleic acid 26 654-45:325341_IE-F/1C1 (HVT/IBDV/ILT/NDV #2)
hCMV IEpro-F-IE(therm)/HVT UL54.5 nucleic acid 27 VP2/1C1#8
(HVT/IBDV/ILT/NDV #3)
mCMV IEpro-VP2-SV40pA/HVT UL54.5 nucleic acid 28 1332-47.A2
(HVT/IBDV/ILT/NDV #3)
ILT/hCMV IEpro-F-IE(therm)/HVT US2 nucleic acid 29 1332-23.7
(HVT/IBDV/ILT/NDV #4)
ILT/HVT UL54.5 nucleic acid thirty 435Vec60
(HVT/IBDV/ILT/NDV #4)
mCMV IEpro-VP2-SV40pA/
hCMV IEpro-F-IE(term)/HVT US2 nucleic acid 31 1332-29.4
(HVT/IBDV/ILT/NDV #5)
ILT/hCMV IEpro-F-IE(therm)/HVT UL54.5 nucleic acid 32 435Vec6
(HVT/IBDV/ILT/NDV #5)
mCMV IEpro-VP2-SV40pA/HVT US2 nucleic acid

The present invention can be better understood by reference to the following non-limiting examples, which are presented as illustrations of the invention. The following examples are presented to more fully illustrate embodiments of the invention and should in no way be construed as limiting the broad scope of the invention.

EXAMPLES

Example 1

CONSTRUCTION OF RECOMBINANT HVT/ILTV/IBDV/NDV VIRAL VECTORS

Recombinant polyvalent non-pathogenic Marek's disease constructs have been generated that encode and express (i) two infectious laryngotracheitis virus protein antigens, (ii) infectious bursitis virus protein antigen, and (iii) Newcastle disease virus protein antigen. The present invention overcomes the problem of vaccine interference when two recombinant HVT vaccines such as Innovax®-ILT (sold by Merck Animal Health) and Vaxxitek® (sold by Merial) are administered to the same animal. In addition, the present invention uniquely provides a first recombinant non-pathogenic Marek's disease virus (rMDV _np ) that encodes antigens from three different non-MDV viral pathogens.

Recombinant turkey herpesvirus (HVT) constructs were created in which antigenic donor material from three poultry pathogens, infectious laryngotracheitis virus (ILTV), Newcastle disease virus (NDV), and infectious bursitis virus (IBDV), was inserted into the HVT vector [see also, US 8932604 B2, WO 2013/057,235, WO 2016/102647 and US application serial number 62/351471, filed June 17, 2016, the contents of which are incorporated herein by reference in their entirety]. Donor materials include a 3.563 kb SalI-HindIII fragment. from ILTV, strain for NVSL infection, Lot No. 83-2 [nucleotide positions 10532-14094; Wild et al., Virus Genes 12: 104-116 (1996): Register. # U28832], encoding the full length glycoprotein D (gD) and glycoprotein I (gI) genes, as well as partial coding regions from glycoprotein E (amino acids 1-101) and ORF5 (amino acids 734-985); an expression cassette consisting of the coding region for NDV, strain Clone 30, fusion protein gene (F) (nucleotide positions 4544-6205; Romer-Oberdorfer et al., (1999); Registry # Y18898) driven by a viral promoter and followed by terminator sequence follows; and an expression cassette consisting of a coding region for an IBDV strain, Faragher, type F52/70, a viral protein 2 (vp2) gene driven by a viral promoter and followed by a terminator sequence. In an exemplary embodiment, the promoter driving IBDV VP2 expression is derived from the immediate early (IE) gene of murine cytomegalovirus (mCMV) strain ATCC VR-194, while the promoter for NDV F expression is derived from the immediate early (IE) gene of human cytomegalovirus (hCMV) , strain AD 169. The terminator and polyadenylation sequence for IBDV VP2 is derived from simian virus 40 (SV40), while the terminator and polyadenylation sequence for F NDV is derived from the human cytomegalovirus (hCMV) immediate early (IE) gene. The donor material for the first heterologous nucleic acid was inserted at the UL54.5 site (position 111240/111241, Afonso et al., J. Virology 75 (2): 971-978 (2001); registry # AF291866, between amino acid residues 21 and 22), while the donor material for the second heterologous nucleic acid was inserted at the US2 site (position 140540/140541, Afonso et al., (2001) supra; registry # AF291866, between amino acid residues 124 and 125) [cf. Figure 1].

Genetic and phenotypic stability is a major component of the safety and relatively strong antigen expression and/or efficacy profile of any new recombinant viral vaccine candidate.

IBDV/ILTV and NDV expression cassettes inserted into the HVT backbone are not essential for viral replication and therefore may be lost due to mutation during amplification of viral material during tissue cell culture passages. A satisfactory vaccine candidate should not easily mutate to lose expression of the foreign gene insert. A candidate vaccine is considered stable if at least 90% of viral plaques can be shown to express the introduced foreign antigenic protein after 10 or more passages in tissue culture.

The ability to generate herpesviruses by cosmid reconstruction has previously been demonstrated for the pseudorabies virus [van Zijl et al., J. Virology 62: 2191-2195 (1988)]. Subsequently, this procedure was used to construct recombinant HVT vectors [see US 5853733, herein incorporated by reference in relation to the methodology disclosed in relation to the construction of recombinant HVT vectors] and used to construct the recombinant HVT/IBDV/ILTV/NDV vectors of the present invention. In this method, the entire HVT genome is cloned into bacterial vectors as several large overlapping subgenomic fragments constructed using standard recombinant DNA techniques [Maniatis et al., Molecular Cloning, Cold Spring Harbor Laboratory press, Cold Spring Harbor, New York (1982); and Sambrook et al., Molecular Cloning, Cold Spring Harbor Laboratory press, Cold Spring Harbor, New York (1989)]. The HVT FC126 strain cosmid library was derived from fragmented viral DNA cloned into the pWE15 cosmid vector (Stratagene, currently Agilent Technologies, Santa Clara, CA). In addition, several large genomic DNA fragments were isolated by restriction digestion with BamHI and cloned into either pWE15 or the pSP64 plasmid vector (Promega, Madison Wis.). As described in US Pat. No. 5,853,733, co-transfection of these fragments into chick embryonic fibroblast (CEF) cells results in regeneration of the HVT genome mediated by homologous recombination across overlapping regions of the fragments. If the insert is inserted directly into one or more subgenomic fragments prior to co-transfection, this procedure results in a high frequency of viruses containing the insert. Five overlapping subgenomic clones are required to generate the complete HVT FC126 genome and serve as the basis for the creation of all recombinant HVT/IBDV/ILTV/NDV viruses.

Construction HVT/IBDV/ILTV/NDV 670-14.1-1A1 or A2

The triple recombinant HVT viral vector, HVT/IBDV/ILTV/NDV 670-14.1-1, contains an IBDV/ILTV expression cassette inserted into the HVT UL54.5 site and an NDV expression cassette inserted into the HVT US2 site. Cosmid regeneration of HVT/IBDV/ILTV/NDV 670-14.1-1 was performed essentially as described in US 5,853,733 [eg, FIG. 8 of US 5853733]. To enable integration into the UL54.5 region of the FC126 HVT genome, the region overlapped by cosmids nr. 407-31.1C1 in US 5853733 was now generated from three smaller plasmids: 672-01.A40 and 672-07.C40 and one transfer plasmid (484-1050-2641-10859) overlapping these two and containing IBDV/ILTV expressing cassettes at the UL54.5 gene locus. To enable integration into the US region of the FC126 HVT genome, the region overlapped by cosmid nr. 378-50 in US 5853733 was now derived from three smaller plasmids: pSY640 and 556-60.6 and one transfer plasmid (1322-48.1) overlapping the two and containing the NDV expression cassettes at the US2 gene locus.

A set of nine linearized constructs: 2 cosmids and 7 plasmids were transfected together into chicken embryonic fibroblasts (CEF) using the standard CaCl ₂ transfection protocol and the resulting viral material was twice purified from plaques.

Description of subgenomic fragments for generating FC126 HVT

Subgenomic clone 407-32.2C3

Cosmid 407-32.2C3 contains a region of HVT genomic DNA approximately 40170 bases long [Left end - position 39754; Afonso et al. (2001), supra; register. #AF291866]. This region includes the HVT BamHI fragments F', L, P, N1, E, D and the 2092 bp fragment B.

Subgenomic clone 172-07.BA2

Plasmid 172-07.BA2 contains the 25931 bp region of HVT genomic DNA. It was created by cloning HVT BamHI fragment B [position 37663 to 63593; Afonso et al., 2001, supra; Register. # AF291866], into plasmid pSP64 (Promega, Madison Wis.).

Subgenomic clone 407-32.5G6

Cosmid 407-32.5G6 contains a 39404 bp region of HVT genomic DNA [position 61852-101255; Afonso et al. (2001), supra; Register. #AF291866]. This region includes the HVT BamHI fragments H, C, Q, K1, M, K2, plus the 1742 base pairs of the B fragment and the 3880 base pairs of the J fragment.

Subgenomic clone 407-31.1C1

Cosmid 407-31.1C1 contains a 37444 bp region of HVT genomic DNA [position 96.095-133.538; Afonso et al. (2001), supra; register. #AF291866]. This region includes the HVT BamHI fragments J, G, I, F, O, plus the 1281 base pairs of the K2 fragment and the 6691 base pairs of the A fragment.

Subgenomic clone 378-50

Cosmid 378-50 contains a 28897 bp region of HVT genomic DNA [see Fig. 8 of US 5853733]. This region includes HVT BamHI fragment A. It was constructed by cloning HVT BamHI fragment A [position 126848-155744; Afonso et al. (2001), supra; register. #AF291866] to the pWE15 cosmid.

Additional fragments of inserts for generation

HVT/IBDV/ILTV/NDV 670-14.1-1A1 or A2 (see Figure 1)

Subgenomic clone 484-1050-2641-10859

Insert plasmid 484-1050-2641-10859 contains an 8636 bp region of the HVT unique long genomic region [positions 109489-118124; Afonso et al., 2001, supra; register. # AF291866], cloned into a derivative of plasmid pNEB193 (removed AatII-PvuII). It is flanked by AscI sites and includes HVT BamHI fragments I, S, as well as 1337 base pairs of the G fragment and 1177 base pairs of the F fragment. Inserted into the XhoI site in the UL54.5 HVT open reading frame [position 111240/111241; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 21 and 22] 2 elements: expression cassette consisting of MCMV promoter IE, IBDV classic type F52/70, strain Faragher, viral protein gene 2 (VP2) and SV40 polyadenylation signal, then SalI-HindIII fragment of 351 bp from ILTV, NVSL challenge strain, lot #83-2 [positions 10532-14094; Wild et al., Virus Genes 12: 104-116 (1996); register. # U28832] encoding full length genes for glycoprotein D (gD) and glycoprotein I (gI), plus partial coding regions from glycoprotein E (amino acids 1-101) and ORF5 (amino acids 734-985). The VP2 IBDV, gD ILTV, and gI ILTV genes are transcribed in the opposite direction to the HVT UL54.5 gene.

Subgenomic clone 672-01.A40

Plasmid 672-01.A40 contains a 14731 bp region of HVT genomic DNA derived from a unique long region [positions 96095-110825; Afonso et al., 2001, supra; Register. # AF291866], cloned into a derivative of plasmid pNEB193. This region includes the HVT BamHI fragments G, J and 1281 base pairs of K2.

Subgenomic clone 672-07.C40

Plasmid 672-07.C40 contains 12520 base pairs of HVT genomic DNA derived from a unique long region [positions 116948-129467; Afonso et al., 2001, supra; register. # AF291866], cloned into a derivative of plasmid pNEB193. This region includes HVT BamHI fragments F, O and 2620 bp A.

Subgenomic clone 1322-48.1

Insert plasmid 1322-48.1 contains an EcoRI fragment of 7311 base pairs of unique short HVT regions [positions 126880-144190; Afonso et al., 2001, supra; register. # AF291866], cloned into plasmid pSP64 (Promega, Madison, WI.). Inserted into a unique StuI site in the US2 HVT gene [positions 140540/140541; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 124 and 125] - an expression cassette consisting of the IE HCMV promoter, strain NDV, clone 30, fusion gene (F) and transcription terminator from the IE HCMV gene. The NDV F gene is transcribed in the opposite direction to the HVT US2 gene.

Subgenomic clone pSY640

Plasmid pSY640 contains approximately 13600 base pairs of HVT genomic DNA (positions 126848-140540; Afonso et al., 2001, supra; registry # AF291866) derived from BamHI fragment A. To create this plasmid, the DNA region located in the 5'-region from the US2 gene, starting from the StuI site located in the US2 gene and continuing to the end of the BamHI fragment of A. (Promega, Madison WI.).

Subgenomic clone 556-60.6

Plasmid 556-60.6 contains approximately 12,500 base pairs of HVT genomic DNA derived from BamHI fragment A (approx. positions 143300-155744; Afonso et al., 2001, supra; registry # AF291866]. The 3' regions from the US2 gene (beginning at the StuI site located in the US2 gene and continuing to the end of the BamHI fragment A) were cloned into pSP64 (Promega, Madison WI) and then exonuclease excised to excise ~150 bp from the StuI site .n and re-cloned into the plasmid vector pBR322.

Standard protocol for CaCl ₂ transfection

Secondary CEFs are plated in 6-well culture plates and incubated at 38°C with 5% CO ₂ for 24 hours and form a confluent monolayer. For each well, a total of 0.5 μg cosmid and plasmid DNA was mixed in Hepes buffer and 125 mM CaCl ₂ was added dropwise until the inevitable precipitation occurred. This mixture was added to a monolayer of CEF cells and incubated for 2-3 hours. The supernatant was removed and a top layer of 15% glycerol was added and kept on the cells for one minute. It was then removed, washed with phosphate buffered saline (PBS), fresh culture medium was added and the cells were incubated for two days. The infection was then expanded twice by harvesting cells by trypsinization and seeding onto larger plates, first on 6 cm plates and then on 3 cm plates 3 days later, until 50-90% CPE was reached. The amplified transfected cells were then harvested by trypsinization and dilutions from 10 ^-3 to 10 ^-4 were plated on 6 cm CEF monolayer dishes and incubated. The next day, the plates were covered with agar and a number of individual HVT/IBDV/ILTV/NDV plaques were isolated and amplified on CEF. Each viral material was purified from the plaques a second time by infecting fused CEF monolayers on 6 cm plates with 1 round purified material diluted to 10 ^-4 -10 ^-5 and by incubating the cells. The next day, the plates were covered with agar and a number of individual HVT/IBDV/ILTV/NDV plaques were isolated and amplified on CEF.

EXAMPLE 2

RECOMBINANT HVT/IBDV/ILTV/NDV VIRAL MATERIAL IS PHENOTYPICALLY STABLE FOR THE EXPRESSION OF IBDV, ILTV AND NDV PROTEINS

AFTER SEQUENTIAL PASSAGES IN TISSUE CELL CULTURE

Two plaque-purified HVT/IBDV/ILTV/NDV isolates, each from a separate co-transfection material, were serially passaged 15 times on secondary CEF cells and evaluated for expression of the inserted ILTV, NDV and IBDV genes in an immunofluorescence assay.

GENERATION OF MATERIAL AFTER PASSAGES OF TISSUE CELL CULTURE

For each tissue cell culture passage, confluent monolayers of secondary CEFs were inoculated with 50-100 µl of viral material and incubated at 38° C., 5% CO ₂ for 2-5 days until CPE became evident. Cells were then harvested by trypsinization, passage 1 (P1). The process was repeated to prepare further material for passages (P2 - P15).

PHENOTYPICAL STABILITY ANALYSIS

Monolayers of secondary CEFs were plated on six well plates. Cells were inoculated with viral material collected after passages of various levels: P0 - P15 or diluent alone. The plates were inoculated at several dilutions to achieve a countable number of plaques per well and incubated at 38°C, 5% CO ₂ . After a five day incubation, the supernatant was decanted and the CEF monolayers were fixed with 70% acetone for approximately 20 minutes at 15-30°C. The acetone solution was decanted and cells were allowed to air dry before staining with gD ILTV (polyclonal rabbit anti-gD ILTV), gI ILTV (polyclonal rabbit anti-gI ILTV), F NDV (Mab #57) and VP2 IBDV (MCA GDV-R63) primary antibodies. After an approximately 1.5 hour blocking step, 5% goat serum in PBS+0.5% Triton-X 100, 2 ml per well, was added to the plates and then incubated at 36-39° C. in a humidified incubator. Primary antibodies were diluted appropriately, added at 2 ml per well and then incubated at 36-39ºC for 1.3 hours in a humidified incubator.

After antibody incubation, the plates were washed three times with PBS+0.5% Triton-X 100. A FITC-labeled secondary antibody solution (rabbit anti-mouse or goat anti-rabbit) was prepared at a 1:50 dilution and 2 ml was added to each well. After incubation, the plates were washed three times with PBS+0.5% Triton-X 100 and examined for fluorescence, and plaques were scored positive or negative for fluorescence staining. The plates were then examined under a light microscope and the plaques were counted. The percentage of fluorescent plaques at each passage level is described in Table 1 below. Both isolates maintain acceptable levels of expression for all four antigens (over 90%) at passage 15.

TABLE 1

EXPRESSION STABILITY AFTER PASSAGE IN TISSUE CELL CULTURE

Expression percentage Virus number Passage level ILT-gD ILT-gI IBDV-VP2 NDV-F 640-14.1-1A1 P0 100% 100% 100% 100% P5 100% 100% 100% 100% P10 100% 100% 100% 100% P15 100% 100% 93% 99% 640-14.1-1A2 P0 100% 100% 100% 100% P5 100% 100% 100% 100% P10 100% 100% 100% 100% P15 100% 100% 96% 100%

EXAMPLE 3

RECOMBINANT HVT/IBDV/ILTV/NDV VIRAL MATERIAL IS PHENOTYPICALLY STABLE FOR EXPRESSION OF ILTV, NDV AND IBDV PROTEINS AFTER VACCINATION AND EXTRACTION FROM BIRDS

Vaccines were prepared from two isolates of HVT/IBDV/ILTV/NDV 670-14.1-1, isolate A1 and isolate A2, and were used to inoculate two groups of chicks at 21 days of age (21) subcutaneously. A third group of birds were vaccinated with one diluent to serve as a negative control group. Pooled spleen samples from three birds were collected twice a week for four weeks post-vaccination and processed for virus isolation on chick embryonic fibroblast (CEF) cells. When the cytopathic effect was clearly visible, the monolayers were fixed and the plaques were analyzed for the expression of IBDV VP2, ILTV gD and ILTV gI and F NDV proteins by immunofluorescence assay (IFA) with antibodies specific for each of these proteins.

PHENOTYPICAL STABILITY ANALYSIS

Monolayers of secondary CEFs were plated on six-well plates. Cells were inoculated with 5x10 ⁶ spleen cells and incubated at 38°C, 5% CO ₂ . After five days of incubation, the supernatant was decanted and the CEF monolayers were fixed with 70% acetone for approximately 20 minutes at 15-30°C. The acetone solution was decanted and cells were allowed to air dry before staining with primary antibodies gD ILTV (polyclonal rabbit anti-gD ILTV), gI ILTV (polyclonal rabbit anti-gI ILTV), F NDV (Mab #57) and VP2 IBDV (MCA GDV-R63 ). After a blocking step of approximately 0.5 hour, 5% goat serum in PBS+0.5% Triton-X 100, 2 ml per well, was added to the plates and then incubated at 36-39° C. in a humidified incubator. Primary antibodies were diluted appropriately, added at 2 ml per well, and then incubated at 36-39 ºC for 1 hour in a humidified incubator. After antibody incubation, the plates were washed three times with PBS+0.5% Triton-X 100. A FITC-labeled secondary antibody solution (rabbit anti-mouse or goat anti-rabbit) was prepared at a 1:50 dilution and 2 ml was added to each well. The plates were incubated for 1 hour at 36º-39ºC in a humidified incubator. After incubation, the plates were washed three times with PBS+0.5% Triton-X 100 and examined in the fluorescent range, and the plaques were scored as positive (+) or negative (-) for fluorescent staining. The plates were then examined under a light microscope and the plaques were counted. The percentage of fluorescent plaques at each passage level is presented in Table 2 below.

TABLE 2

EXPRESSION STABILITY AFTER PASSAGE IN BIRDS (D18)

Vaccine Vaccine dose (PFU) Expression percentage NDV
F ILTVgI gD ILTV VP2 IBDV HVT/IBDV/ILTV/NDV
670-14.1-1A1 (p10) 1660 100% 100% 100% 100% HVT/IBDV/ILTV/NDV
670-14.1-1A2 (p10) 4800 100% 100% 100% 100% Diluent ND 0 0 0 0

EXAMPLE 4

HVT/IBDV/ILTV/NDV

PERFORMANCE DATA FOR TWO SPECIAL DESIGN ISOLATES

The following four studies were conducted to demonstrate the efficacy of a single HVT/IBDV/ILTV/NDV 670-14 construct as a vaccine candidate for protection against challenge with virulent infectious laryngotracheitis virus (ILTV) or virulent infectious bursitis virus (IBDV), or virulent Marek's disease virus ( MDV).

In the first study, specific pathogen-free (SPF) day-old chicks were vaccinated with candidate vaccine HVT/IBDV/ILTV/NDV 670-14.1-1A2. The controls included the second group, which remained unvaccinated. 28 days post-vaccination, vaccinated chickens and unvaccinated control chickens were challenged with virulent ILTV/USDA Lot LT 963 via the intratracheal (IT) route of administration. The birds were then observed for clinical signs of disease for 10 days. In Table 3, the results show that the 670-14.1-1A2 vaccine provides partial protection against challenge. A second study was conducted using a second vaccine clone, 670-14.1-1A1. In Table 4, the results show a marked improvement in protection. Accordingly, the following two studies were performed with candidate vaccine 670-14.1-1A1. These results suggest that HVT/IBDV/ILTV/NDV may be both stable and effective.

TABLE 3

Immunization with ILTV after vaccination with isolate 670-14.1-1A2

Group Vaccine identification Vaccination infection at 4 weeks Age actual dose Path #Infected/Total %Infected %Protected 5 Infected controls 1 day NA SC 15/15 100% 0% 6 Non-infected controls 1 day NA SC 0/10 0% ND 7 HVT/IBDV/ILTV/NDV 1 day 2394 SC 14/22 64% 36%

TABLE 4

ILTV IMMUNIZATION AFTER VACCINATION WITH ISOLATE 670-14.1-1A1

Group Vaccine identification Vaccination infection at 4 weeks Age actual dose Path Affected/Total Age actual dose 5 HVT/IBDV/ILTV/NDV 1 day 1359 SC 9/20 45% 55% 6 Infected controls 1 day ND SC 10/10 100% ND 7 Non-infected controls 1 day ND SC 0/10 0% ND

In the third study, specific pathogen-free (SPF) day-old chicks were vaccinated with the HVT/IBDV/ILTV/NDV 670-14.1-1A1 vaccine. Controls included a second unvaccinated group. 28 days after vaccination, vaccinated chickens and unvaccinated control chickens were challenged with a virulent strain of IBDV/CS89 via the intraocular (IO) route of administration. The birds were then observed for clinical signs of disease for 10 days, and the bursa was collected and histologically examined for total lesions consistent with IBDV and scored according to the European Pharmacopoeia 9.0 (04/2013:0587). The results presented in Table 5 show that the 670-14.1-1A1 vaccine provides 100% protection against challenge.

TABLE 5

IBDV CONTAMINATION AFTER VACCINATION WITH ISOLATE 670-14.1-1A1#

Group Vaccine identification Vaccination Week 4 challenge with virulent CS89 IBDV Age Path Dose
(0.2 ml) # Hit/Total % Protection 3 placebo
(Marek Thinner) 1 day SC N/A 0/15 0 four Negative controls
(not infected) 1 day N/A N/A 10/10 ND 5 HVT/IBD/ILTV/NDV 1 day SC 1.810 21/21 100

#P10

In the fourth study, specific pathogen-free (SPF) day-old chicks were vaccinated with the HVT/IBD/ILTV/NDV 670-14.1-1A1 vaccine. Controls included a second group of unvaccinated chickens. Five days after vaccination, vaccinated chickens and unvaccinated control chickens were challenged with a virulent strain of MDV/GA by the intra-abdominal (IA) route. The birds were then observed for clinical signs of disease for 50 days and examined for cumulative lesions consistent with MDV after death or euthanasia. The results presented in Table 6 show that the 670-14.1-1A1 vaccine provided 95% protection against challenge. Overall, these results indicate that the HVT/IBD/ILTV/NDV vaccine can be both stable and effective. This also results in the fact that the upper limit of foreign antigens encoded in a multivalent HVT construct has not been reached for stable and effective multivalent HVT vaccines.

TABLE 6

MDV CONTAMINATION AFTER VACCINATION WITH 670-14.1-1A1 ISOLATE

Group Vaccine identification Vaccination
SC Infection
MDV-GA5 Necropsy results on day 50 Age Dose Age at infection # Infected/Total %
Infected %
Protection 3 HVT/IBDV/ILTV/NDV 1 day 3328 Day 5 1/22 5% 95% four HVT 1 day 2372 Day 5 0/22 0% 100% 5 Diluent 1 day ND Day 5 15/22 68% ND 6 Diluent 1 day ND - 0/12 0% ND

EXAMPLE 5

OPTIONAL CONSTRUCTIONS HVT/IBDV/ILTV/NDV

HVT/IBDV/ILTV/NDV #2 construction

The triple recombinant HVT viral vector, HVT/IBDV/ILTV/NDV #2, contains an NDV expression cassette inserted into the HLT UL54.5 site and an IBDV/ILTV expression cassette inserted into the HVT US2 site. The regeneration of the HVT/IBDV/ILTV/NDV #2 cosmid was carried out essentially as described in US 5,853,733 [eg, FIG. 8 of US 5853733]. To enable integration into the UL54.5 region of the FC126 HVT genome, the region overlapped by cosmids nr. 407-32.1C1 in US 5853733 was now provided from three smaller plasmids: 672-01.A40 and 672-07.C40 and one transfer plasmid (654-45: 325341_IE-F/1C1) overlapping the two and containing the expression cassette NDV at the UL54.5 gene locus. To enable integration into the US region of the FC126 HVT genome, the region overlapped by cosmid nr. 378-50 in US 5853733 was now provided from three smaller plasmids: pSY640 and 556-60.6 and one transfer plasmid (228509-ILT-435Vec6) overlapping these two and containing the IBDV/ILTV expression cassettes at the US2 gene locus,

A set of nine linearized constructs: 2 cosmids and 7 plasmids were transfected together into chicken embryonic fibroblasts (CEF) using the standard CaCl ₂ transfection protocol and the resulting viral material was twice purified from plaques.

HVT/IBDV/ILTV/NDV #3 construction

The triple recombinant HVT viral vector, HVT/IBDV/ILTV/NDV #3, contains an IBDV expression cassette inserted into the HVT UL54.5 site and an ILTV/NDV expression cassette inserted into the HVT US2 site. Cosmid regeneration of HVT/IBDV/ILTV/NDV #3 was performed essentially as described in US 5,853,733 [eg, FIG. 8 of US 5853733]. To enable integration into the UL54.5 region of the FC126 HVT genome, the region overlapped by cosmids nr. 407-32.1C1 in US 5853733 was now provided from three smaller plasmids: 672-01.A40 and 672-07.C40 and one transfer plasmid (VP2/1C1 #8) overlapping the two and containing the IBDV expression cassette at the gene locus UL54.5. To enable integration into the US region of the FC126 HVT genome, the region overlapped by cosmid nr. 378-50 in US 5853733 was now provided from three smaller plasmids: pSY640 and 556-60.6 and one transfer plasmid (1332-47.A2) overlapping the two and containing the ILTV/NDV expression cassettes at the US2 gene locus.

A set of nine linearized constructs: 2 cosmids and 7 plasmids were transfected together into chicken embryonic fibroblasts (CEF) using the standard CaCl ₂ transfection protocol and the resulting viral material was twice purified from plaques.

HVT/IBDV/ILTV/NDV #4 construction

The triple recombinant HVT viral vector, HVT/IBDV/ILTV/NDV #4, contains an ILTV expression cassette inserted into the HLT UL54.5 site and an IBDV/NDV expression cassette inserted into the HVT US2 site. Cosmid regeneration of HVT/IBDV/ILTV/NDV #4 was performed essentially as described in US 5,853,733 [eg, FIG. 8 of US 5853733]. To enable integration into the UL54.5 region of the FC126 HVT genome, the region overlapped by cosmids nr. 407-32.1C1 in US 5853733 was now provided from three smaller plasmids: 672-01.A40 and 672-07.C40 and one transfer plasmid (1332-23.7) overlapping the two and containing the ILTV expression cassette at the UL54 gene locus. 5. To enable integration into the US region of the FC126 HVT genome, the region overlapped by cosmid nr. 378-50 in US 5,853,733 was now provided from three smaller plasmids: pSY640 and 556-60.6 and one transfer plasmid (435Vec60) overlapping the two and containing the IBDV/NDV expression cassettes at the US2 gene locus.

A set of nine linearized constructs: 2 cosmids and 7 plasmids were transfected together into chicken embryonic fibroblasts (CEF) using the standard CaCl ₂ transfection protocol and the resulting viral material was twice purified from plaques.

Design HVT/IBDV/ILTV/NDV #5

A triple recombinant HVT viral vector, HVT/IBDV/ILTV/NDV #5, contains ILTV/NDV expression cassettes inserted into the HVT UL54.5 site and an IBDV expression cassette inserted into the HVT US2 site. Cosmid regeneration of HVT/IBDV/ILTV/NDV # 5 was carried out essentially as described in US 5853733 [for example, FIG. 8 of US 5853733]. To enable integration into the UL54.5 region of the FC126 HVT genome, the region overlapped by cosmids nr. 407-32.1C1 in US 5853733 was now provided from three smaller plasmids: 672-01.A40 and 672-07.C40 and one transfer plasmid (1332-29.4) overlapping the two and containing the ILTV/NDV expression cassettes at the gene locus UL54.5. To enable integration into the US region of the FC126 HVT genome, the region overlapped by cosmid nr. 378-50 in US 5,853,733 was now provided from three smaller plasmids: pSY640 and 556-60.6 and one transfer plasmid (435Vec6) overlapping the two and containing the IBDV expression cassette at the US2 gene locus.

A set of nine linearized constructs: 2 cosmids and 7 plasmids were transfected together into chicken embryonic fibroblasts (CEF) using the standard CaCl ₂ transfection protocol and the resulting viral material was twice purified from plaques.

Additional fragments of inserts for generating HVT/IBDV/ILTV/NDV #2 (see Figure 2)

Subgenomic clone 654-45: 325341_IE-F/1C1

Insert plasmid 654-45: 325341_IE-F/1C1 contains 8636 base pairs of the HVT unique long genomic region [positions 109489-118124; Afonso et al., 2001, supra; register. # AF291866], cloned into a derivative of plasmid pNEB193 (removed AatII-PvuII). It is flanked by AscI sites and includes HVT BamHI fragments I, S, as well as 1337 base pairs of the G fragment and 1177 base pairs of the F fragment. Inserted into the XhoI site in the UL54.5 HVT open reading frame [position 111240/111241; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 21 and 22] - an expression cassette consisting of the IE HCMV promoter, strain NDV, clone 30, fusion gene (F) and transcription terminator from the IE HCMV gene. The NDV F gene is transcribed in the opposite direction to the HVT UL54.5 gene.

Subgenomic clone 228509-ILT-435Vec6 [see International Application PCT/EP2017/064662]

Insert plasmid 228509-ILT-435Vec6 contains an EcoRI fragment of 7311 base pairs of unique short HVT regions [positions 126880-144190; Afonso et al., 2001, supra; register. # AF291866], cloned into plasmid pSP64 (Promega, Madison, WI.). Inserted into a unique StuI site in the US2 HVT gene [positions 140540/140541; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 124 and 125] two elements: an expression cassette consisting of the MCMV IE promoter, classical type IBDV F52/70, Faragher strain, viral protein 2 (VP2) gene, and SV40 polyadenylation signal; then a SalI-HindIII fragment of 351 bp from ILTV, strain for NVSL infection, Lot No. 83-2 [positions 10532-14094; Wild et al., Virus Genes 12: 104-116 (1996); register. # U28832] encoding full length genes for glycoprotein D (gD) and glycoprotein I (gI), plus partial coding regions from glycoprotein E (amino acids 1-101) and ORF5 (amino acids 734-985). The VP2 IBDV, gD ILTV, and gI ILTV genes are transcribed in the opposite direction to the US2 HVT gene.

Additional fragments of inserts for generating HVT/IBDV/ILTV/NDV #3 (see Figure 3)

Subgenomic clone VP2/1C1 #8

The VP2/1C1 #8 insert plasmid contains an 8636 bp region of the HVT unique long genomic region [positions 109489-118124; Afonso et al., 2001, supra; register. # AF291866], cloned into a derivative of plasmid pNEB193 (removed AatII-PvuII). It is flanked by AscI sites and includes HVT BamHI fragments I, S, as well as 1337 base pairs of the G fragment and 1177 base pairs of the F fragment. Inserted into the XhoI site in the UL54.5 HVT open reading frame [position 111240/111241; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 21 and 22] - an expression cassette consisting of the MCMV IE promoter, the classic type IBDV F52/70, the Faragher strain, the viral protein 2 gene (VP2) and the SV40 polyadenylation signal. The IBDV VP2 gene is transcribed in the opposite direction to the HVT UL54.5 gene.

Subgenomic clone 1332-47.A2

Insert plasmid 1332-47.A2 contains an EcoRI fragment of 7311 base pairs of unique short HVT regions [positions 126880-144190; Afonso et al., 2001, supra; register. # AF291866], cloned into plasmid pSP64 (Promega, Madison, WI.). Inserted into a unique StuI site in the US2 HVT gene [positions 140540/140541; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 124 and 125] two elements: SalI-HindIII fragment of 3563 base pairs from ILTV, NVSL infection strain, lot # 83-2 [positions 10532-14094; Wild et al., Virus Genes 12: 104-116 (1996); Register. # U28832], encoding the full-length genes for glycoprotein D (gD) and glycoprotein I (gI), as well as partial coding regions from glycoprotein E (amino acids 1-101) and ORF5 (amino acids 734-985); followed by an expression cassette consisting of the HCMV IE promoter, NDV strain, clone 30, fusion gene (F), and a transcription terminator from the HCMV IE gene. The gD ILTV, gI ILTV, and F NDV genes are transcribed in the opposite direction to the US2 HVT gene.

Additional fragments of inserts for generating HVT/IBDV/ILTV/NDV #4 (see Figure 4)

Subgenomic clone 1332-23.7

Insert plasmid 1332-23.7 contains an 8636 bp region of the HVT unique long genomic region [positions 109489-118124; Afonso et al., 2001, supra; register. # AF291866], cloned into a derivative of plasmid pNEB193 (removed AatII-PvuII). It is flanked by AscI sites and includes HVT BamHI fragments I, S, as well as 1337 base pairs of the G fragment and 1177 base pairs of the F fragment. Inserted into the XhoI site in the UL54.5 HVT open reading frame [position 111240/111241; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 21 and 22] - SalI-HindIII fragment of 3563 base pairs from ILTV, NVSL challenge strain, lot # 83-2 [positions 10532-14094; Wild et al., Virus Genes 12: 104-116 (1996); register. # U28832] encoding full length genes for glycoprotein D (gD) and glycoprotein I (gI), plus partial coding regions from glycoprotein E (amino acids 1-101) and ORF5 (amino acids 734-985). The gD ILTV and gI ILTV genes are transcribed in the opposite direction to the UL54.5 HVT gene.

Subgenomic clone 435Vec60

Insert plasmid 435Vec60 contains an EcoRI fragment of 7311 base pairs of unique short HVT regions [positions 126880-144190; Afonso et al., 2001, supra; register. # AF291866], cloned into plasmid pSP64 (Promega, Madison, WI.). Inserted into a unique StuI site in the US2 HVT gene [positions 140540/140541; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 124 and 125] two elements: an expression cassette consisting of the MCMV IE promoter, classical type IBDV F52/70, Faragher strain, viral protein 2 (VP2) gene, and SV40 polyadenylation signal; followed by an expression cassette consisting of the HCMV IE promoter, NDV strain, clone 30, fusion gene (F), and a transcription terminator from the HCMV IE gene. Both the VP2 IBDV and F NDV genes are transcribed in the opposite direction to the US2 HVT gene.

Additional fragments of inserts for generating HVT/IBDV/ILTV/NDV # 5 (see Figure 5)

Subgenomic clone 1332-29.4 [see, US 9409954 B2]

Insert plasmid 1332-29.4 contains an 8636 bp region of the HVT unique long genomic region [positions 109489-118124; Afonso et al., 2001, supra; register. # AF291866], cloned into a derivative of plasmid pNEB193 (removed AatII-PvuII). It is flanked by AscI sites and includes HVT BamHI fragments I, S, as well as 1337 base pairs of the G fragment and 1177 base pairs of the F fragment. Inserted into the XhoI site in the UL54.5 HVT open reading frame [position 111240/111241; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 21 and 22] two elements: SalI-HindIII fragment of 3563 base pairs from ILTV, NVSL challenge strain, lot # 83-2 [positions 10532-14094; Wild et al., Virus Genes 12: 104-116 (1996); Register. # U28832], encoding the full-length genes for glycoprotein D (gD) and glycoprotein I (gI), as well as partial coding regions from glycoprotein E (amino acids 1-101) and ORF5 (amino acids 734-985); followed by an expression cassette consisting of the HCMV IE promoter, NDV strain, clone 30, fusion gene (F), and a transcription terminator from the HCMV IE gene. The gD ILTV, gI ILTV, and F NDV genes are transcribed in the opposite direction to the UL54.5 HVT gene.

Subgenomic clone 435Vec6

Insert plasmid 435Vec6 contains an EcoRI fragment of 7311 base pairs of unique short HVT regions [positions 126880-144190; Afonso et al., 2001, supra; register. # AF291866], cloned into plasmid pSP64 (Promega, Madison, WI.). Inserted into a unique StuI site in the US2 HVT gene [positions 140540/140541; Afonso et al., 2001, supra; register. # AF291866, between amino acid residues 124 and 125] - an expression cassette consisting of the MCMV IE promoter, classic type IBDV F52/70, Faragher strain, viral protein 2 (VP2) gene, and SV40 polyadenylation signal. The IBDV VP2 gene is transcribed in the opposite direction to the HVT US2 gene.

The sequences used in the HVT/ILTV/IBDV/NDV viral vectors disclosed in this example are shown in Example 7 below as SEQ ID NOS: 23 and 26-32.

EXAMPLE 6

FAIL DESIGNS

Recombinant viral vaccine vectors are by definition designed to carry and express foreign genes. If the transcription and expression of these foreign genes provide a disadvantage in the growth of the recombinant virus compared to the parent virus, it is possible that these genes will be lost during vaccine production. For this reason, candidate vaccines must be tested for genetic and phenotypic stability.

In addition, the USDA and codified in the Code of Federal Regulations Title 9 Part 113 (9CFR 113) Standard Requirements for Animal Origin Products is used as protection criteria. Live virus vaccines must provide at least 90% protection, in the case of NDV, IBDV, and ILTV, and at least 80%, in the case of MDV, against disease-associated clinical signs or lesions in order to be licensed.

The genetic stability of the viral constructs was determined by Southern blot analysis after a certain number of passages in tissue cell culture, the highest expected level of vaccine production, and compared with DNA from the original isolate. DNA isolated from viral material will be digested with restriction enzymes, transferred to a membrane and hybridized with probes designed to detect the presence of inserted foreign genes. Genetic stability can also be determined using PCR analysis. PCR primers designed to anneal DNA within or flanking foreign DNA can be used to amplify fragments of known size from viral DNA templates, both before and after passage in tissue culture.

The phenotypic stability of viral constructs was determined by immunological staining of individual viral plaques with antibodies directed against the protein products of these inserted foreign genes. The protection provided by these recombinant vaccines is based on the expression of these protein products to stimulate the animal's immune system. In most cases, if the percentage of viruses staining positive for foreign protein expression fell below 90%, the ability of the viruses to grow in tissue cell culture was likely to be adversely affected and therefore they were not suitable vaccine candidates.

As can be easily seen from Tables 7A and 7B below, most of the rMDV _np constructs do not meet these two criteria, namely stability with relatively high antigen expression and/or potency. Table 7A shows a number of recombinant HVT constructs with multiple heterologous inserts, in which one of the heterologous inserts encodes an IBDV antigen. As the results show, all of the constructs in Table 7A failed to satisfy stability under relatively strong criteria for antigen expression and/or potency.

TABLE 7A

DOUBLE RECOMBINANT HVT AND IBDV VIRAL CONSTRUCTS:

Name/
Designation Insert site Insert IBDV promoter IBDV
Expression Stability HVT003 UL43 [IBDV] polyprotein
[Ecoli] Bgal PRV gX Weak stable HVT 016 UL43 [IBDV]VP2
[Ecoli] Bgal hCMVIE strong not stable HVT 056 US2 [MDV] gA, gB
[IBDV]VP2 hCMVIE strong not stable HVT 060 US2 [MDV] gA, gB
[IBDV]VP2,
16kDa ORF IE-VP2,
gX-16dk ORF strong not stable HVT 137 US2
UL54.5 [MDV] gA, gB, gC
[IBDV]VP2 [BHV]VP8
(shell) Weak stable HVT 143 US2
US2
UL54.5 [MDV] gA, gB, gD
[NDV] HN, F
[IBDV]VP2 [BHV]VP8
(shell) Weak not stable HVT/NDV/IBDV
1312-92 US2
UL7/UL8 [IBDV]VP2
[NDV] F hCMVIE strong not stable HVT/NDV/IBDV
1312-94 US2
UL7/UL8 [IBDV]VP2
[NDV] F hCMVIE strong not stable HVT/NDV/IBDV
1312-95 US2
UL7/UL8 [IBDV]VP2
[NDV] F hCMVIE strong not stable HVT/NDV/IBDV
1329-54 US2 [IBDV]VP2
[NDV] F FHV gB strong not stable

Table 7B below shows a series of eleven recombinant HVT constructs and one single NAHV construct, each containing a plurality of heterologous inserts, in which at least one of the heterologous inserts encodes an NDV or ILTV antigen. As the results show, all of the constructs shown in Table 7B are not stable under relatively strong criteria for antigen expression and/or potency.

TABLE 7B

DOUBLE RECOMBINANT HVT AND NAHV VIRAL CONSTRUCTS:

Name Insert site Insert Stability NDV Protection MDV
Protection ILTV
Protection HVT 048 US2 [MDV] gA, gB
[NDV] F stable Good *Protective -- HVT 049 US2 [MDV] gA, gB
[NDV]HN stable Weak
(< 20%) Not tested -- HVT 050 US2 [MDV] gA, gB
[NDV] F, HN stable Good *Protective -- HVT 053 US2 [MDV] gA, gB
[ILTV] gB, gD not stable -- Not tested Not HVT 078 US2 [MDV] gA, gB, gD
[NDV]HN, F not stable Not tested Not tested -- HVT 079 US2 [MDV] gA, gB, gD
[ILTV] gB, gD not stable -- Not tested (71-100%) HVT 106 US2 [MDV]gA, gB, gD
[NDV]HN, F stable **Unknown Not tested -- HVT 123 UL54.5+US2 [ILTV] gD, gB/UL54.5
[MDV] gA, gD, gB/US2 not stable -- Not tested Not tested HVT 125 UL54.5+US2 [ILTV] gDgI, gB/UL54.5
[MDV] gA, gD, gB/US2 not stable -- Not tested Not tested HVT 128 UL54.5+US2 [NDV] HN, F /UL54.5
[MDV] gA, gD, gB/US2 not stable Not tested Not tested -- HVT 139 UL54.5+US2 [ILTV] gDgI /UL54.5
[MDV] gA, gD, gB/US2 not stable -- Not tested Not tested HVY-198 (NAHV) US2* (MDV) [NDV]F+
[ILTV] gD, gI not stable

*Defensive, but subsequently unsuccessful in field trials. ** Only 75% of birds are seroconverted to F NDV

EXAMPLE 7

SEQUENCES

The following sequences were used in exemplary rHVT constructs. The coding sequences below include single stop codons that can be easily replaced with alternative stop codons without changing the properties of the protein antigens that encode the coding sequences.

SEQ ID NO 1: gD ILTV Glycoprotein (1134 bp)

atggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaatacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagcttaactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccg ccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaa

SEQ ID NO 2: gD ILTV Glycoprotein (377 amino acids)

MDRHLFLRNAFWTIVLLSSFASQSTAAVTYDYILGRRALDALTIPAVGPYNRYLTRVSRGCDVVELNPISNVDDMISAAKEKEKGGPFEASVVWFYVIKGDDGEDKYCPIYRKEYRECGDVQLLSECAVQSAQMWAVDYVPSTLVSRNGAGLTIFSPTAALSGQYLLTLKIGRFAQTALVTLEVNDRCLKIGSQLNFLPSKCWTTEQYQTGFQGEHLYPIADTNTRHADDVYRGYEDILQRWNNLLRKKNPSAPDPRPDSVPQEIPAVTKKAEGRTPDAESSEKKAPPEDSEDDMQAEASGENPAALPEDDEVPEDTEHDDPNSDPDYYNDMPAVIPVEETTKSSNAVSMPIFAAFVACAVALVGLLVWSIVKCARS

SEQ ID NO 3: gI ILTV Glycoprotein (1089 bp)

atggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctaccaggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctcgaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaaca agggggctgacggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtga

SEQ ID NO 4: gI ILTV Glycoprotein (362 amino acids)

MASLLGTLALLAATLAPFGAMGIVITGNHVSARIDDDHIVIVAPRPEATIQLQLFFMPGQRPHKPYSGTVRVAFRSDITNQCYQELSEERFENCTHRSSSVFVGCKVTEYTFSASNRLTGPPHPFKLTIRNPRPNDSGMFYVIVRLDDTKEPIDVFAIQLSVYQFANTAATRGLYSKASCRTFGLPTVQLEAYLRTEESWRNWQAYVATEATTTSAEATTPTPVTATSASELEAEHFTFPWLENGVDHYEPTPANENSNVTVRLGTMSPTLIGVTVAAVVSATIGLVIVISIVTRNMCTPHRKLDTVSQDDEERSQTRRESRKFGPMVACEINKGADQDSELVELVAIVNPSALSSPDSIKM

SEQ ID NO 5: VP2 IBDV (1362 bp)

atgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaaattgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactgttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctttgatgggactgcggtaatcaccagagctgtggccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccaatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctag cagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgattccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtaa

SEQ ID NO 6: VP2 IBDV (453 amino acids)

MTNLQDQTQQIVPFIRSLLMPTTGPASIPDDTLEKHTLRSETSTYNLTVGDTGSGLIVFFPGFPGSIVGAHYTLQSNGNYKFDQMLLTAQNLPASYNYCRLVSRSLTVRSSTLPGGVYALNGTINAVTFQGSLSELTDVSYNGLMSATANINDKIGNVLVGEGVTVLSLPTSYDLGYVRLGDPIPAIGLDPKMVATCDSSDRPRVYTITAADDYQFSSQYQPGGVTITLFSANIDAITSLSIGGELVFQTSVQGLVLGATIYLIGFDGTAVITRAVAADNGLTAGTDNLMPFNLVIPTNEITQPITSIKLEIVTSKSGGQAGDQMSWSASGSLAVTIHGGNYPGALRPVTLVAYERVATGSVVTVAGVSNFELIPNPELAKNLVTEYGRFDPGAMNYTKLILSERDRLGIKTVWPTREYTDFREYFMEVADLNSPLKIAGAFGFKDIIRAIRR

SEQ ID NO: 7: F NDV Protein coding sequence (Clone 30; 1662 bp)

atgggccccagaccttctaccaagaacccagtacctatgatgctgactgtccgagtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcggctgcaggaattgtggttacaggagacaaagccgtcaacatatacacctcatcccagacaggatcaatcatagttaagctcctcccgaatctgcccaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcaccccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcggtgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagtttgttaatgaccaatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaattgactacagtattcggaccacaaatcacttcacctgctttaaacaagctgactattcaggcactttacaatctagctggtggaaatatggattacttattgactaagttaggtgtagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctatacgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaagctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaagtggtgacacaggtcggttctgtgatagaagaacttg acacctcatactgtatagaaactgacttacatttatattgtacaagaatagtaacgttccctatgtcccctggtatttattcctgcttgagcggcaatacgtcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatacatgactatcaaaggttcagtcatcgccaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttttatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataataacaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgctttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgactagcacatctgctctcattacctatatcgtgttgactatcatatctcttgtttttggtatacttagcctgattctagcatgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataatactctagatcagatgagagccactacaaaaatgtga

SEQ ID NO: 8: F NDV Protein (Clone 30; 553 amino acids)

MGPRPSTKNPVPMMLTVRVALVLSCICPANSIDGRPLAAAGIVVTGDKAVNIYTSSQTGSIIVKLLPNLPKDKEACAKAPLDAYNRTLTTLLTPLGDSIRRIQESVTTSGGGRQGRLIGAIIGGVALGVATAAQITAAAALIQAKQNAANILRLKESIAATNEAVHEVTDGLSQLAVAVGKMQQFVNDQFNKTAQELDCIKIAQQVGVELNLYLTELTTVFGPQITSPALNKLTIQALYNLAGGNMDYLLTKLGVGNNQLSSLIGSGLITGNPILYDSQTQLLGIQVTLPSVGKLNNMRATYLETLSVSTTRGFASALVPKVVTQVGSVIEELDTSYCIETDLHLYCTRIVTFPMSPGIYSCLSGNTSACMYSKTEGALTTPYMTIKGSVIANCKMTTCRCVNPPGIISQNYGEAVSLIDKQSCNVLSLGGITLRLSGEFDVTYQKNISIQDSQVIITGNLDISTELGNVNNSISNALNKLEESNRKLDKVNVKLTSTSALITYIVLTIISLVFGILSLILACYLMYKQKAQQKTLLWLGNNTLDQMRATTKM

SEQ ID NO: 9: F NDV Protein coding sequence: (B1 Hitchner; 1698 bp)

atggatcgatcccggttggcgccctccaggtgcaggatgggctccagaccttctaccaagaacccagcacctatgatgctgactatccgggtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcagctgcaggaattgtggttacaggagacaaagcagtcaacatatacacctcatcccagacaggatcaatcatagttaagctcctcccgaatctgccaaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcaccccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcggtgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagttcgttaatgaccaatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaatcgactacagtattcggaccacaaatcacttcacctgccttaaacaagctgactattcaggcactttacaatctagctggtgggaatatggattacttattgactaagttaggtatagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctatacgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaacctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaag tggtgacacgggtcggttctgtgatagaagaacttgacacctcatactgtatagaaactgacttagatttatattgtacaagaatagtaacgttccctatgtcccctggtatttactcctgcttgagcggcaatacatcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatatatgactatcaaaggctcagtcatcgctaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttttatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataataacaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgccttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgaccagcacatctgctctcattacctatatcgttttgactatcatatctcttgtttttggtatacttagcctgattctagcatgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataataccctagatcagatgagagccactacaaaaatgtga

SEQ ID NO: 10: F NDV Protein (B1 Hitchner; 565 amino acids)

MDRSRLAPSRCRMGSRPSTKNPAPMMLTIRVALVLSCICPANSIDGRPLAAAGIVVTGDKAVNIYTSSQTGSIIVKLLPNLPKDKEACAKAPLDAYNRTLTTLLTPLGDSIRRIQESVTTSGGGRQGRLIGAIIGGVALGVATAAQITAAAALIQAKQNAANILRLKESIAATNEAVHEVTDGLSQLAVAVGKMQQFVNDQFNKTAQELDCIKIAQQVGVELNLYLTESTTVFGPQITSPALNKLTIQALYNLAGGNMDYLLTKLGIGNNQLSSLIGSGLITGNPILYDSQTQLLGIQVTLPSVGNLNNMRATYLETLSVSTTRGFASALVPKVVTRVGSVIEELDTSYCIETDLDLYCTRIVTFPMSPGIYSCLSGNTSACMYSKTEGALTTPYMTIKGSVIANCKMTTCRCVNPPGIISQNYGEAVSLIDKQSCNVLSLGGITLRLSGEFDVTYQKNISIQDSQVIITGNLDISTELGNVNNSISNALNKLEESNRKLDKVNVKLTSTSALITYIVLTIISLVFGILSLILACYLMYKQKAQQKTLLWLGNNTLDQMRATTKM

SEQ ID NO 11: gD ILTV promoter (527 bp)

aaacagctgtactacagagtaaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatagcgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgct

SEQ ID NO 12: gI ILTV promoter (264 bp)

tgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagc

SEQ ID NO 13: mCMV IE promoter (1391 bp)

aactccgcccgttttatgactagaaccaatagtttttaatgccaaatgcactgaaatcccctaatttgcaaagccaaacgccccctatgtgagtaatacggggactttttacccaatttcccacgcggaaagccccctaatacactcatatggcatatgaatcagcacggtcatgcactctaatggcggcccatagggactttccacatagggggcgttcaccatttcccagcataggggtggtgactcaatggcctttacccaagtacattgggtcaatgggaggtaagccaatgggtttttcccattactggcaagcacactgagtcaaatgggactttccactgggttttgcccaagtacattgggtcaatgggaggtgagccaatgggaaaaacccattgctgccaagtacactgactcaatagggactttccaatgggtttttccattgttggcaagcatataaggtcaatgtgggtgagtcaatagggactttccattgtattctgcccagtacataaggtcaatagggggtgaatcaacaggaaagtcccattggagccaagtacactgcgtcaatagggactttccattgggttttgcccagtacataaggtcaataggggatgagtcaatgggaaaaacccattggagccaagtacactgactcaatagggactttccattgggttttgcccagtacataaggtcaatagggggtgagtcaacaggaaagttccattggagccaagtacattgagtcaatagggactttccaatgggttttgcccagtacataaggtcaatgggaggtaagccaatgggtttttcccattactggcacgtatactgagtcattagggactttccaatgggttttgcccagtacataaggtcaataggggtgaatcaacaggaaagtcccattggagccaagtacactgagtcaatagggactttccattgggttttgcccagtacaaaaggtcaatagggg gtgagtcaatgggtttttcccattattggcacgtacataaggtcaataggggtgagtcattgggtttttccagccaatttaattaaaacgccatgtactttcccaccattgacgtcaatgggctattgaaactaatgcaacgtgacctttaaacggtactttcccatagctgattaatgggaaagtaccgttctcgagccaatacacgtcaatgggaagtgaaagggcagccaaaacgtaacaccgccccggttttcccctggaaattccatattggcacgcattctattggctgagctgcgttctacgtgggtataagaggcgcgaccagcgtcggtaccgtcgcagtcttcggtctgaccaccgtagaacgcagagctcctcgctgcag

SEQ ID NO 14: hCMV IE promoter, from strain AD169 (301 bp)

ggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatag

SEQ ID NO: 15: hCMV IE Promoter (Short; 360 bp)

cgcgccaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtagcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccata

SEQ ID NO: 16: hCMV IE Promoter (Towne strain; 1191 bp)

gtgaataataaaatgtgtgtttgtccgaaatacgcgtttgagatttctgtcccgactaaattcatgtcgcgcgatagtggtgtttatcgccgatagagatggcgatattggaaaaatcgatatttgaaaatatggcatattgaaaatgtcgccgatgtgagtttctgtgtaactgatatcgccatttttccaaaagttgatttttgggcatacgcgatatctggcgatacgcttatatcgtttacgggggatggcgatagacgcctttggtgacttgggcgattctgtgtgtcgcaaatatcgcagtttcgatataggtgacagacgatatgaggctatatcgccgatagaggcgacatcaagctggcacatggccaatgcatatcgatctatacattgaatcaatattggccattagccatattattcattggttatatagcataaatcaatattggctattggccattgcatacgttgtatccatatcataatatgtacatttatattggctcatgtccaacattaccgccatgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacg tcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagaacgccatccacgctgttttgacctccatagaagacaccgg

SEQ ID NO 17: chicken β-actin promoter (692 bp)

(Note: "nnn" denotes an undefined sequence in a highly GC-rich region. Maybe 3-5 "g")

nnn

SEQ ID NO 18: FHV US-9 polyadenylation signal (55 bp)

caataaacatagcatacgttatgacatggtctaccgcgtcttatatggggacgac

SEQ ID NO 19: HSV TK polyadenylation signal (370 bp)

gatccataattgattgacgggagatgggggaggctaactgaaacacggaaggagacaataccggaaggaacccgcgctatgacggcaataaaaagacagaataaaacgcacgggtgttgggtcgtttgttcataaacgcggggttcggtcccagggctggcactctgtcgataccccaccgagaccccattggggccaatacgcccgcgtttcttccttttccccaccccaccccccaagttcgggtgaaggcccagggctcgcagccaacgtcggggcggcaggccctgccatagccactggccccgtgggttagggacggggtcccccatggggaatggtttatggttcgtgggggttattattttga

SEQ ID NO 20: SV40 polyadenylation signal (199 bp)

agcttcagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttcg

SEQ ID NO 21: 484-1050-2641-10859(mCMV IEpro-VP2-SV40pA/ILTV/HVT UL54.5 region (15252 bp) (HVT/IBDV/ILTV/NDV 670-14 Virus)

ggcgcgccactggagaacggcatgaccgcaaaaggcgttgtagagatcgatcccacgaactctcaggcgatcgtgtcagtcgccataaacagcgacgatcgtctccaggatctgaacggttttcttctcaacgatcatcagtatatgaggaactgaacctgatatttagccgagggaaacgcaggttaaaaaccctatcaagcgattgcgattttcgcgtatctagtaaaaatagatgggcttcggtactagccttcgccgccaactctgaatatgcccttcgtggacctcatataacatggcattgtttgttggatgcggggccggaattaagaagaacattcgaaatacgagcaaaaatttcggccctggcatgtgctgcgcgagaatcggtacttcggggagaaagttttatcggagctttgggtagtgcagaggaaactctatcttggttgaaaatgcatgcgaccctgcacttgattctggttaaccacgatccaatttttaagacggctggcgcggtcctagataacctccgcttaaaactagccccaatattgatgtgcagatataacacagaaaaacgatcaatggaagacatgctacggcggtcatctcccgaagacatcaccgattccctaacaatgtgcctgattatgttatcgcgcattcgtcgtaccatgcgcaccgcaggaaataaatatagctatatgatagatccaatgaatcgtatgtctaattacactccaggcgaatgtatgacaggtatattgcgatatattgacgaacatgctagaaggtgtcctgatcacatatgtaatttgtatatcacatgtacacttatgccgatgtatgtgcacgggcgatatttctattgtaattcatttttttgttagtaaactaccacaggctgtccggaaatctaagttaatgaataaagtagatggttaatactcattgcttagaattggactacttttaattctctttaatgttcgta ttaaataaaaacatctttaataaacttcagcctcttcgcttattgtagaaattgagtattcaaaatcatgttcaaagccgtcttcggagagtgtactcgccacggtggttggaacatcactatgtctacacgtcaaatttaagcacgtcaggtctgtcgaggacaagaaatggttaactagtgtttcaattattcttataaacgttaagcattgtaagccccccggccgtccgcagcaacaatttactagtatgccgtgggctccgggactatcacggatgtccaattcgcacatgcatataatttttctagggtctctcatttcgagaaatcttcggggatccatcagcaatgcgggctgtagtcccgattcccgtttcaaatgaaggtgctccaacacggtcttcaaagcaaccggcataccagcaaacacagactgcaactccccgctgcaatgattggttataaacagtaatctgtcttctggaagtatatttcgcccgacaatccacggcgcccccaaagttaaaaaccatccatgtgtatttgcgtcttctctgttaaaagaatattgactggcattttcccgttgaccgccagatatccaaagtacagcacgatgttgcacggacgactttgcagtcaccagccttcctttccacccccccaccaacaaaatgtttatcgtaggacccatatccgtaataaggatgggtctggcagcaaccccataggcgcctcggcgtggtagttctcgaggccttaagcttaaggatcccccaactccgcccgttttatgactagaaccaatagtttttaatgccaaatgcactgaaatcccctaatttgcaaagccaaacgccccctatgtgagtaatacggggactttttacccaatttcccacgcggaaagccccctaatacactcatatggcatatgaatcagcacggtcatgcactctaatggcggcccatagggactttccacatagggggcgtt caccatttcccagcataggggtggtgactcaatggcctttacccaagtacattgggtcaatgggaggtaagccaatgggtttttcccattactggcaagcacactgagtcaaatgggactttccactgggttttgcccaagtacattgggtcaatgggaggtgagccaatgggaaaaacccattgctgccaagtacactgactcaatagggactttccaatgggtttttccattgttggcaagcatataaggtcaatgtgggtgagtcaatagggactttccattgtattctgcccagtacataaggtcaatagggggtgaatcaacaggaaagtcccattggagccaagtacactgcgtcaatagggactttccattgggttttgcccagtacataaggtcaataggggatgagtcaatgggaaaaacccattggagccaagtacactgactcaatagggactttccattgggttttgcccagtacataaggtcaatagggggtgagtcaacaggaaagttccattggagccaagtacattgagtcaatagggactttccaatgggttttgcccagtacataaggtcaatgggaggtaagccaatgggtttttcccattactggcacgtatactgagtcattagggactttccaatgggttttgcccagtacataaggtcaataggggtgaatcaacaggaaagtcccattggagccaagtacactgagtcaatagggactttccattgggttttgcccagtacaaaaggtcaatagggggtgagtcaatgggtttttcccattattggcacgtacataaggtcaataggggtgagtcattgggtttttccagccaatttaattaaaacgccatgtactttcccaccattgacgtcaatgggctattgaaactaatgcaacgtgacctttaaacggtactttcccatagctgattaatgggaaagtaccgttctcgagccaatacacgtcaatgggaag tgaaagggcagccaaaacgtaacaccgccccggttttcccctggaaattccatattggcacgcattctattggctgagctgcgttctacgtgggtataagaggcgcgaccagcgtcggtaccgtcgcagtcttcggtctgaccaccgtagaacgcagagctcctcgctgcaggcggccgctctagaactcgtcgatcgcagcgatgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaaattgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactgttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctt tgatgggactgcggtaatcaccagagctgtggccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccaatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgattccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtagatccagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttcggatcctctagagtcgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagggtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactaca gagtaaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatagcgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggc caatacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagcttaactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgcta ccaggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctcgaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctcccaatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagag cccctgttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcctaacttttgcgacgcgacacgtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgaaagcttcccgggttaattaaggccctcgaggatacatccaaagaggttgagtattctctctacacttcttgttaaatggaaagtgcatttgcttgttcttacaatcggcccgagtctcgttcacagcgcctcgttcacacttaaaccacaaatagtctacaggctatatgggagccagactgaaactcacatatgactaatattcgggggtgttagtcacgtgtagcccattgtgtgcatataacgatgttggacgcgtccttattcgcggtgtacttgatactatggcagcgagcatgggatattcatcctcgtcatcgttaacatctctacgggttcagaatgtttggcatgtcgtcgatcctttgcccatcgttgcaaattacaagtccgatcgccatgaccgcgataagcctgtaccatgtggcattagggtgacatctcgatcatacattataagaccaacgtgcgagtcttccaaagacctgcacgccttcttcttcggattgtcaacgggttcttcagaatctatgcccatatctggcgttgagaccattgtgcgtttaatgaacaataaagcggcatgccatggaaaggagggctgcagatctccattttctcacgccactatcctggacgctgtagacgataattataccatgaa tatagagggggtatgtttccactgccactgtgatgataagttttctccagattgttggatatctgcattttctgctgccgaacaaacttcatcgctatgcaaagagatgcgtgtgtacacgcgccggtggagtatacgggaaactaaatgttcatagaggtctttgggctatatgttattaaataaaataattgaccagtgaacaatttgtttaatgttagtttattcaatgcattggttgcaaatattcattacttctccaatcccaggtcattctttagcgagatgatgttatgacattgctgtgaaaattactacaggatatatttttaagatgcaggagtaacaatgtgcatagtaggcgtagttatcgcagacgtgcaacgcttcgcatttgagttaccgaagtgcccaacagtgctgcggttatggtttatgcgcacagaatccatgcatgtcctaattgaaccatccgatttttcttttaatcgcgatcgatgtttgggcaactgcgttatttcagatctaaaaaatttaccctttatgaccatcacatctctctggctcataccccgcttggataagatatcatgtagattccgccctaagaaatgcaaactaacattattgtcggttccatatacacttccatcttgtccttcgaaaataacaaactcgcgcaatagaccgtccgtacatgcatggccgatgtgtgtcaacatcattggtctgctagatcccgatgggacgaatcgtacagtcgtcgctccagcattggcaaaaatccccagataccctccatgcggcaaatctaaattgcgaccccgaagagactgcaccaaagtcttatcgacgcacgctgatttttttgaacagcgggagcccattatcttcagtggagcgtagacgggcgaggctaattatgtgacatagcaacactgcatgtatgtttttataaatcaataagagtacataatttattacgtatcatttccgtttg taatatactgtatacatcatccacactattagtcagcactagcgcgcgggcgcacgttacaatagcagcgtgcccgttatctatattgtccgatatttacacataacatttcatcgacatgattaaatacctaagtactgcacacagatgtttaatgtatatcgtcatataaattatatcgctaggacagacccaaacgacctttatcccaaacagtcagatcctcttctcaagtgtcgatttctgttatggaatatgcataccctggcccagaaattgcacgcacgagcgtagtgaatgcgtcattggttttacatttaaaggctaaatgcacaaattctttagacgacagcacatcgttaaatagcatctctagcgttcttatgaatgctaagcattggagtcctcctggtcggccacaataacagctgagtatcataccctgagctccggggttgtcgcacatagcggattcgtataaacataggattttccgcgaatccatcagttgcaaaaatctgttaggctccatcaacaacgctggatttacttcagatccacgcgtaaagtaatggtgctcgaataccgtttttagagttgtcggcatttcaaggaacaaagaattcatttcttcattgcaacgacgcgccagaaatcccaagacctctttgggtagtatgttcttgcctataaaacacggcgttccaagtgccaggaaccacgcatgtgttactgttggggcgtattcagaaataaagcggggtttatgcggcttttgaagctcggatatccaaagtatcgcttgctgatgaacgagcgatgtagctgttacaaaacctcctttccatcctccagtcaacataatatttatcggcctacctatgtccgtaataagtattggtcgggcaattattccgtatgaggtcttgcaggaataagctcttagggacagccagcttggatatggtgcgaaacagaccttctcggcttcagaatgtcg ctccgcagtctcttcgtgtcggtgcatcttagatccaccatcaatgtgtgcagcattgactcccgcccgtcgaatattccttttgttacgatgcagtaatgagcacgatcatgggcggggcgatgacgttctatttgcatgtctgcgaacaatttgcgtcagtcatacagctatggagtgggccatttctggccgtcaacttaaaaacgcgaaccgcagacatatgtatttgcatgcaaagacgtatcttcgtatttctgggcatcttcaaatgctctggccaatatggcaatgaatttggattcgtttgacgccgatggtatgcagtgcaaatgtgccaatagcccacatccgaaaaagttatttgtcatacaagcaggtgttaagtagcaatcacataaaggcaccagacgcctcatggcatcataatgaatagctccttctccccactggaaccactgacaaaatctgcgagtatattccgcaaaccacattttatttctcatagaaactaccctaaatccttttaacgggaagaagaatcctagatagtgcttgaagtcatgactgttactgctgcaataacactgtatattatttataaattccgtttgtctaggtatctgatgtaggcattccgatccctttactattgcgtcttcacgaccaaatgggaatgcgccaaaatccccacacctcatcaccctggaggcagattgtgtattattaatatccgccgattgaagcacaaaacggtacggtactgttcctaattctggtatagattctatggtcaaaagtctgcatatccccgacattgccatgagatcacacagtccaagtagcatgtttattgagtcactcagactgtcaacgtccctcgccgcaccaccaatcgaaaataaagtatctacgcaagttatagctccgcattttctatcgctagcagcaatcgcgacgcaaaacataaaggccatgttgggatttgaactctctgggg ggcttgttatcttctgcaccgtcgcagtcgcagttttccgaaatttatgtctaatatattttccggccgtgctccaatcggccgaaaagaatctgcgtattaccagactcattgacgggccgataaagaccataaaacaaaattcctgtgcactccctcctccagttttgccatcgtccaagtcccgtaactttttttgcgtttcgaggagcaagcgttcgttatccctacccacacttgttttccaccgttttcttattataagcggttgtatcgccaacgcgtcaccgcaggttgtcacatacagtgatggcatacttgaacgtgcaacaacgcgctcgctttgcaaatctaagtcattgaccatcaaatcgcgttgagaggatagccaggcatcttttttcctagtatggtgacggtgcagccaccccaactcagttcttgtaaaaaaagctattggcgggaatttatgttctgaggtgcattctatatttatgagtccatcaaatgccattaaccagattcgtattttttcgctcgacccggcatcactatggatacaatacctttctatggcccatttcagctctcgaaccaaccacacggacaattgactaacataagtatgatctttatcacagtcgcacccatctgagttatatttatggcatccgagcgctcttactgtacggtcggatacacccatggtttttcctttatatagtcgggttatagtctgtcgggtttggcggtagcacggagtagtttgatttttaagaatcgaaaaccggcttggagagaccactgtcgaatatttgtccgtatactctacacgtgagtgttgtccattcctaggtatattcatctgttcggataccttcaattgctgttcaggcataaccttaaagcatatgttatgttgtacatcaaaacttggtgagttatgttcgattgccgcgcataaagaatcgtacatgagcgtttctgctaacatactatc tatattctcacacgcccctgcatatactgttcctattccaaattcacgttttgccccatcggctatctgctcccaaaaagttgtaatataggtgccgctgggtgcgaaattttcatcagttgtattcctgataaactgaatcactttacataatttttgccacatatctgcgtgcagccatagtatcgaacccgtgggctcggagacgacagtgcgtacaatgggtattttacctttccccaacaaaataatggtatacaagttaggtccgtacctagaccttaatgtttccaattcttctgaatcactgcactctcgtaggggagtaacggtaataatttcgtctctgagccccgttttgcgttgaaaactaatcacattagataatgtgcaatcggtttcttttatccggatacatctaagtattatgacatcggtggtcattgtttccatcaacgaccatcttttacgatcgcccatactactcatggacgttgtcggtgttgaaaaatcaccagaattgcaacggatctctgggtaccatgctgctgatggaattggcggttttaattgttgtttcagtctattattgctatctttggcggggttgaataatgtggggggagagtgattgcaggaatccgaatgggtcaataaaacgaccgtgctccgttctgccggcgccgatccgattgaagctatatacttcgcttctctccccacttttccaatttgatccggaaataaaacggccccggacaacagtatcgtacgatccggatccggatcctgcttgcctacagaagaatcaacatctcgccccaatattctggtcaaaactggctcgctcatggcaacgcggacgtttcccccggtggccagtcttaatggttaatgttcttttcggcaatcttatacatcagcgggttgcgtgaatactggtcacagttcagtcatttactacacaccagcaatacgacgacggacagtaccgtcccga cgaacgcgacgcccaaaattgctatcgcgaccgcgtccgaggcgatgtcgtacgggcggtgcggggttggatcctcggcaaagagatcctcgtaattcggcggtgggagcggagggtaaagacgcgggtggggatctccctccggaccgcgcgccgggcgcggttcgaaaatgctttccgcctcgctcagtgtcaacgccaagtattcgggcgggctgggggccggaatatctcccgcgacttcttctatcggcgcggaattggagtcgcggtcgtggcgcgcttctagcgtcgtcaacggaagtccattttcggggtctcccggtgggcgttcagcgtccatcgtcgtatatgctctaacacacgtctcgctatattaaaaaaaagaagagtatcggtcagtgtcgagtgtcgccgacaatgtcgcgagttctcggcgatttaatttttggaactgctccctatgaatcccgtaactgtagcgcccgcgcagaaagccgccatcagaccaactacgtgtctgttcgatgtttgcccgccgatcgctttaccgattaaggttccggcgagaaatgacatgctcgatccaagaacaaagtttttcgcggtaaacaacaacatagttaccgtgcgagatggagaaaccacatctcccgaattagtagaggaaagcccgcgctgtcggtttggggacatatcgatcttttttgtgtttttcctaggacccttttgccagatcgtacaaagtcgcgtcttatgagcggacgttcttactgcagctcggtaggagtggggcagggttagatttcgtcggcgtttcggcccccgtatgcgccgcgccaccctcttcgccgagctctttatgcgcggtgggggtgagcgcttccggagttgcgatctccgatctcgagccgcagcccggcggtgtctctttcagtggagcgttagcgccatcatgtggttcgtggcggtggaaaggctattatgtgttaggggagag accacgtgatcggcatgcaaatgagcaaggcgaacgcgtcagcgttcgcactgcgaaccaataatatatatattatactattggctttaggtgcgaacgtccggctagtccaatagcggggtcgcgtttcgtaccacgtgttatagaccgccctaaactcgcactcgggggtccggccgcgcccagacagggcggagacgtgccacaggggctttaaaacaccgcttcgggcaccgttcatctcggcgcgcc

SEQ ID NO 22: 1322-48.1 hCMV IEpro-F-IE(term)/HVT US2 region (12692 bp) (HVT/IBDV/ILTV/NDV 670-14 Virus)

ctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggctggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatcgacgctctcccttatgcgactcctgcattaggaagcagcccagtagtaggttgaggccgttgagcaccgccgccgcaaggaatggtgcatgcaaggagatggcgcccaacagtcccccggccacggggcctgccaccatacccacgccgaaacaagcgctcatgagcccgaagtggcgagcccgatcttccccatcggtgatgtcggcgatataggcgccagcaaccgcacctgtggcgccggtgatgccggccacgatgcgtccggcgtagaggatctggctagcgatgaccctgctgattggttcgctgaccatttccggggtgcggaacggcgttaccagaaactcagaaggttcgtccaaccaaaccgactctgacggcagtttacgagagagatgatagggtctgcttcagtaagccagatgctacacaattaggcttgtacatattgtcgttagaacgcggctacaattaatacataaccttatgtatcatacacatacgatttaggtgacactatagaatacaagctagcttgggctgcaggtcgactctagaggatcgttaattaacgatccccgggcgagctcgaattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcggg taccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaaatgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatgg ctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttactgtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctg aatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaactgtagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgata cacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagcttcgcgccaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtagcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccggttgcgccgccaccatgggccccagaccttctaccaagaacccagtacctatgatgctgactgtccgagtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcggctgcaggaattgtggttacaggagacaaagccgtcaacatatacacctcatc ccagacaggatcaatcatagttaagctcctcccgaatctgcccaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcaccccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcggtgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagtttgttaatgaccaatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaattgactacagtattcggaccacaaatcacttcacctgctttaaacaagctgactattcaggcactttacaatctagctggtggaaatatggattacttattgactaagttaggtgtagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctatacgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaagctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaagtggtgacacaggtcggttctgtgatagaagaacttgacacctcatactgtatagaaactgacttacatttatattgtacaagaatagtaacgttccctatgtcccctggtatttattcctgcttgagcggcaatacgtcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatacatgactatcaaaggttca gtcatcgccaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttttatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataataacaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgctttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgactagcacatctgctctcattacctatatcgtgttgactatcatatctcttgtttttggtatacttagcctgattctagcatgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataatactctagatcagatgagagccactacaaaaatgtgaggatctctcgaggaattctagatcccacgtcactattgtatactctatattatactctatgttatactctgtaatcctactcaataaacgtgtcacgcctgtgaaaccgtactaagtctcccgtgtcttcttatcaccatcaggtgacatcctcgcccaggctgtcaatcatgccggtatcgattccagtagcaccggccccacgctgacaacccactcttgcagcgttagcagcgcccctcttaacaagccgacccccaccagcgtcgcggttactaacactcctctccccgacctgcaactagtaagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacaataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatt tggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactggagctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtccgaaattacatacgtggcggcgatggaagtagatgttgagtcttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaaatatcgaaagcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaacaatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattataaaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgc agttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggtgaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcgagttataagaaatggggggatgccaatggatgttgaatatgtcatttctaaaatgcttacgtttgaccaggagttcagaccttctgctaaggaaatattgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtccgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattag aatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatatatttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccgagaattagatggtttatatacgcgcgtagtagttgtcaacggggactttactacggccgatataatgtttaatgttaaagtggcatgtgccttttcaaagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgacatttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtatacc aggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcactacggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccgctgctattctattgcgcatgcaacatattacgccgactgtcctataatcagttctacggtattcagaggatgccgggacgccgttgtttatactaggccccacagcagaattcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcct gttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcaatgctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccg gttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgt

SEQ ID NO 23: 228509-ILT-435Vec6 (mCMV IEpro-VP2-SV40pA/ILTV/HVT US2 region) (14113 bp) (HVT/IBDV/ILTV/NDV #2 Virus)

gaattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcgggtaccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaa atgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatggctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttac tgtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctgaatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaact gtagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgatacacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagctgcggccgctctagaactagtggatcccccgggctgcagcccaatgtggaattcgcccttgcacattgttactcctgcatcttaaaaatatatcctgtagtaattttcacagcaatgtcataacatcatctcgctaaagaatgacctgggattggagaagtaatgaatatttgcaaccaatgcattgaataaactaacattaaacgaattcactagtggatcccccaactccgcccgttttatgactagaaccaatagtttttaatgccaaatgcactgaaatcccctaatttgcaaagccaaacgccccctatgtgagtaatacggggacttt ttacccaatttcccacgcggaaagccccctaatacactcatatggcatatgaatcagcacggtcatgcactctaatggcggcccatagggactttccacatagggggcgttcaccatttcccagcataggggtggtgactcaatggcctttacccaagtacattgggtcaatgggaggtaagccaatgggtttttcccattactggcaagcacactgagtcaaatgggactttccactgggttttgcccaagtacattgggtcaatgggaggtgagccaatgggaaaaacccattgctgccaagtacactgactcaatagggactttccaatgggtttttccattgttggcaagcatataaggtcaatgtgggtgagtcaatagggactttccattgtattctgcccagtacataaggtcaatagggggtgaatcaacaggaaagtcccattggagccaagtacactgcgtcaatagggactttccattgggttttgcccagtacataaggtcaataggggatgagtcaatgggaaaaacccattggagccaagtacactgactcaatagggactttccattgggttttgcccagtacataaggtcaatagggggtgagtcaacaggaaagttccattggagccaagtacattgagtcaatagggactttccaatgggttttgcccagtacataaggtcaatgggaggtaagccaatgggtttttcccattactggcacgtatactgagtcattagggactttccaatgggttttgcccagtacataaggtcaataggggtgaatcaacaggaaagtcccattggagccaagtacactgagtcaatagggactttccattgggttttgcccagtacaaaaggtcaatagggggtgagtcaatgggtttttcccattattggcacgtacataaggtcaataggggtgagtcattgggtttttccagccaatttaattaaaacgccatgtactttcccacca ttgacgtcaatgggctattgaaactaatgcaacgtgacctttaaacggtactttcccatagctgattaatgggaaagtaccgttctcgagccaatacacgtcaatgggaagtgaaagggcagccaaaacgtaacaccgccccggttttcccctggaaattccatattggcacgcattctattggctgagctgcgttctacgtgggtataagaggcgcgaccagcgtcggtaccgtcgcagtcttcggtctgaccaccgtagaacgcagagctcctcgctgcaggcggccgctctagaactcgtcgatcgcagcgatgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaaattgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacact gttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctttgatgggactgcggtaatcaccagagctgtggccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccaatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgattccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtaagcttcagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttcggatcctctagagtcgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagg gtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactacagagtaaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatagcgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctga atgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaatacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagcttaactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgcc ccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctaccaggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctcgaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctccc aatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagagcccctgttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcctaacttttgcgacgcgacacgtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgaagcttagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacgataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatttggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactggagctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtccgaaattacatacgtggcggcgatggaagtagatgttgagtc ttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaaatatcgaaagcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaacaatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattataaaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgcagttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggtgaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcga gttataagaaatggggggatgccaatggatgttgaatatgtcatttctaaaatgcttacgtttgaccaggagttcagaccttctgctaaggaaatattgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtccgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattagaatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatatatttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccga gaattagatggtttatatacgcgcgtagtagttgtcaacggggactttactacggccgatataatgtttaatgttaaagtggcatgtgccttttcaaagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgacatttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtataccaggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcactacggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccg ctgctattctattgcgcatgcaacatattacgccgactgtcctataatcagttctacggtattcagaggatgccgggacgccgttgtttatactaggccccacagcagaattc

SEQ ID NO 24: 1333-85.B6 (ILTV/chicken β-actin pro-VP2-FHV US9pA/HVT US2 region) (13064 bp)

gaattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcgggtaccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaa atgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatggctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttac tgtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctgaatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaact gtagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgatacacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagcttaattaagtaccgagctcgaattggcgcgcccgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagggtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactacagagtaaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtctt ttcgacacgtctcttatagcgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaatacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgttt aaagatcgggtcgcagcttaactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctaccaggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccg agtacacgttctccgcctcgaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctcccaatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagagcccctgttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcc taacttttgcgacgcgacacgtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgggcgcgccggatcagatctccatggtcgaggtgagccccacgttctgcttcactctccccatctcccccccctccccacccccaattttgtatttatttattttttaattattttgtgcagcgatgggggcgggggggggggnnncgcgcgccaggcggggcggggcggggcgaggggcggggcggggcgaggcggagaggtgcggcggcagccaatcagagcggcgcgctccgaaagtttccttttatggcgaggcggcggcggcggcggccctataaaaagcgaagcgcgcggcgggcgggagtcgctgcgcgctgccttcgccccgtgccccgctccgccgccgcctcgcgccgcccgccccggctctgactgaccgcgttactcccacaggtgagcgggcgggacggcccttctcctccgggctgtaattagcggcaggaaggaaatgggcggggagggccttcgtgcgtcgccgcgccgccgtccccttctccctctccagcctcggggctgtccgcggggggacggctgccttcgggggggacggggcagggcggggttcggcttctggcgtgtgaccggcggctctagagcctctgctaaccatgttcatgccttcttctttttcctacagctcctgggcaacgtgctggttattgtgctgtctcatcattttggcaaagaattgcagatctggatctatgacaaacctgcaagatcaaacccaacagattgttc cgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaagttgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactgttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctttgatgggactgcggtaatcaccagagctgtggccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccgatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccct ccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgatcccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtaagatccgatctctcgattaattaacaataaacatagcatacgttatgacatggtctaccgcgtcttatatggggacgacaagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacgataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatttggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactggagctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtccgaaattacatacgtggcggcgatggaagtagatgttgagtcttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaaatatcgaa agcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaacaatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattataaaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgcagttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggtgaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcgagttataagaaatggggggatgccaatggatgttgaatatgtcatttcta aaatgcttacgtttgaccaggagttcagaccttctgctaaggaaatattgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtccgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattagaatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatatatttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccgagaattagatggtttatatacgcgcgtagtagttgtcaacggggacttta ctacggccgatataatgtttaatgttaaagtggcatgtgccttttcaaagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgacatttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtataccaggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcactacggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccgctgctattctattgcgcatgcaacatattacgccgactgtcctataatc agttctacggtattcagaggatgccgggacgccgttgtttatactaggccccacagcagaattc

SEQ ID NO 25: 1386-04.4#1 (ILTV/hCMV IEpro-VP2-HSV TKpA/HVT US2 region) (13017 bp)

gaattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcgggtaccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaa atgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatggctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttac tgtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctgaatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaact gtagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgatacacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagcttgttaattaagtcgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagggtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactacagagtaaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatag cgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaatacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagctt aactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctaccaggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctc gaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctcccaatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagagcccctgttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcctaacttttgcgacgcgaca cgtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgaaagcttaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggcgcgccggatctatgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaac gccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaagttgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactgttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctttgatgggactgcggtaatcaccagagctgtggccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccgatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgatcccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtaagatccataattga ttgacgggagatgggggaggctaactgaaacacggaaggagacaataccggaaggaacccgcgctatgacggcaataaaaagacagaataaaacgcacgggtgttgggtcgtttgttcataaacgcggggttcggtcccagggctggcactctgtcgataccccaccgagaccccattggggccaatacgcccgcgtttcttccttttccccaccccaccccccaagttcgggtgaaggcccagggctcgcagccaacgtcggggcggcaggccctgccatagccactggccccgtgggttagggacggggtcccccatggggaatggtttatggttcgtgggggttattattttgaagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacaataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatttggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactggagctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtccgaaattacatacgtggcggcgatggaagtagatgttgagtcttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaaatatcgaaagcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaa caatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattataaaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgcagttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggtgaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcgagttataagaaatggggggatgccaatggatgttgaatatgtcatttctaaaatgcttacgtttgaccaggagttcagaccttctgctaaggaaata ttgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtccgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattagaatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatatatttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccgagaattagatggtttatatacgcgcgtagtagttgtcaacggggactttactacggccgatataatgtttaatgttaaagtggcatgtgccttttca aagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgacatttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtataccaggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcactacggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccgctgctattctattgcgcatgcaacatattacgccgactgtcctataatcagttctacggtattcagaggatgccgggacgccgttgtttatactag gccccacagcagaattc

SEQ ID NO 26: 654-45:325341_IE-F/1C1 (HVT/IBDV/ILT/NDV #2 Virus)

HCMV IEpro-F-IEpA/HVT UL54.5 region (11017 bp)

ggcgcgccactggagaacggcatgaccgcaaaaggcgttgtagagatcgatcccacgaactctcaggcgatcgtgtcagtcgccataaacagcgacgatcgtctccaggatctgaacggttttcttctcaacgatcatcagtatatgaggaactgaacctgatatttagccgagggaaacgcaggttaaaaaccctatcaagcgattgcgattttcgcgtatctagtaaaaatagatgggcttcggtactagccttcgccgccaactctgaatatgcccttcgtggacctcatataacatggcattgtttgttggatgcggggccggaattaagaagaacattcgaaatacgagcaaaaatttcggccctggcatgtgctgcgcgagaatcggtacttcggggagaaagttttatcggagctttgggtagtgcagaggaaactctatcttggttgaaaatgcatgcgaccctgcacttgattctggttaaccacgatccaatttttaagacggctggcgcggtcctagataacctccgcttaaaactagccccaatattgatgtgcagatataacacagaaaaacgatcaatggaagacatgctacggcggtcatctcccgaagacatcaccgattccctaacaatgtgcctgattatgttatcgcgcattcgtcgtaccatgcgcaccgcaggaaataaatatagctatatgatagatccaatgaatcgtatgtctaattacactccaggcgaatgtatgacaggtatattgcgatatattgacgaacatgctagaaggtgtcctgatcacatatgtaatttgtatatcacatgtacacttatgccgatgtatgtgcacgggcgatatttctattgtaattcatttttttgttagtaaactaccacaggctgtccggaaatctaagttaatgaataaagtagatggttaatactcattgcttagaattggactacttttaattctctttaatgttcgta ttaaataaaaacatctttaataaacttcagcctcttcgcttattgtagaaattgagtattcaaaatcatgttcaaagccgtcttcggagagtgtactcgccacggtggttggaacatcactatgtctacacgtcaaatttaagcacgtcaggtctgtcgaggacaagaaatggttaactagtgtttcaattattcttataaacgttaagcattgtaagccccccggccgtccgcagcaacaatttactagtatgccgtgggctccgggactatcacggatgtccaattcgcacatgcatataatttttctagggtctctcatttcgagaaatcttcggggatccatcagcaatgcgggctgtagtcccgattcccgtttcaaatgaaggtgctccaacacggtcttcaaagcaaccggcataccagcaaacacagactgcaactccccgctgcaatgattggttataaacagtaatctgtcttctggaagtatatttcgcccgacaatccacggcgcccccaaagttaaaaaccatccatgtgtatttgcgtcttctctgttaaaagaatattgactggcattttcccgttgaccgccagatatccaaagtacagcacgatgttgcacggacgactttgcagtcaccagccttcctttccacccccccaccaacaaaatgtttatcgtaggacccatatccgtaataaggatgggtctggcagcaaccccataggcgcctcggcgtggtagttctcgaggccttaattaagtaccgagctcgaattggcgcgccaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatca acgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggcgcgccggatccatgggccccagaccttctaccaagaacccagtacctatgatgctgactgtccgagtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcggctgcaggaattgtggttacaggagacaaagccgtcaacatatacacctcatcccagacaggatcaatcatagttaagctcctcccgaatctgcccaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcaccccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcggtgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagtttgttaatgaccaatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaattgactacagtattcggaccacaaatcacttcacctgctttaaacaagctgactattcaggcactttacaatctagctggtggaaatatggattacttattgactaagttaggtgtagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctata cgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaacctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaagtggtgacacaggtcggttctgtgatagaagaacttgacacctcatactgtatagaaactgacttagatttatattgtacaagaatagtaacgttccctatgtcccctggtatttattcctgcttgagcggcaatacgtcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatacatgactatcaaaggttcagtcatcgccaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttttatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataataacaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgctttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgactagcacatctgctctcattacctatatcgttttgactatcatatctcttgtttttggtatacttagcccgattctagcatgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataatactctagatcagatgagagccactacaaaaatgtgaggatctctcgaggaattctagatcccacgtcactattgtatactctatattatactctatgttatactctgtaatcctactcaataaacgtgtcacgcctgtgaaaccgtactaagtctcccgtgtcttcttatcaccatcaggtgacatcctcgcccaggctgt caatcatgccggtatcgattccagtagcaccggccccacgctgacaacccactcttgcagcgttagcagcgcccctcttaacaagccgacccccaccagcgtcgcggttactaacactcctctcccctcgaggatacatccaaagaggttgagtattctctctacacttcttgttaaatggaaagtgcatttgcttgttcttacaatcggcccgagtctcgttcacagcgcctcgttcacacttaaaccacaaatagtctacaggctatatgggagccagactgaaactcacatatgactaatattcgggggtgttagtcacgtgtagcccattgtgtgcatataacgatgttggacgcgtccttattcgcggtgtacttgatactatggcagcgagcatgggatattcatcctcgtcatcgttaacatctctacgggttcagaatgtttggcatgtcgtcgatcctttgcccatcgttgcaaattacaagtccgatcgccatgaccgcgataagcctgtaccatgtggcattagggtgacatctcgatcatacattataagaccaacgtgcgagtcttccaaagacctgcacgccttcttcttcggattgtcaacgggttcttcagaatctatgcccatatctggcgttgagaccattgtgcgtttaatgaacaataaagcggcatgccatggaaaggagggctgcagatctccattttctcacgccactatcctggacgctgtagacgataattataccatgaatatagagggggtatgtttccactgccactgtgatgataagttttctccagattgttggatatctgcattttctgctgccgaacaaacttcatcgctatgcaaagagatgcgtgtgtacacgcgccgttgagtatacgggaaactaaatgttcatagaggtctttgggctatatgttattaaataaaataattgaccagtgaacaatttgtttaatgttagtttattcaatgcat tggttgcaaatattcattacttctccaatcccaggtcattctttagcgagatgatgttatgacattgctgtgaaaattactacaggatatatttttaagatgcaggagtaacaatgtgcatagtaggcgtagttatcgcagacgtgcaacgcttcgcatttgagttaccgaagtgcccaacagtgctgcggttatggtttatgcgcacagaatccatgcatgtcctaattgaaccatccgatttttcttttaatcgcgatcgttgtttgggcaactgcgttatttcagatctaaaaaatttaccctttatgaccatcacatctctctggctcataccccgcttggataagatatcatgtagattccgccctaagaaatgcaaactaacattattgtcggttccatatacacttccatcttgtccttcgaaaataacaaactcgcgcaatagaccgtccgtacatgcatggccgatgtgtgtcaacatcattggtctgctagatcccgatgggacgaatcgtacagtcgtcgctccagcattggcaaaaatccccagataccctccatgcggcaaatctaaattgcgaccccgaagagactgcaccaaagtcttatcgacgcacgctgatttttttgaacagcgggagcccattatcttcagtggagcgtagacgggcgaggctaattatgtgacatagcaacactgcatgtatgtttttataaatcaataagagtacataatttattacgtatcatttccgtttgtaatatactgtatacatcatccacactattagtcagcactagcgcgcgggcgcacgttacaatagcagcgtgcccgttatctatattgtccgatatttacacataacatttcatcgacatgattaaatacctaagtactgcacacagatgtttaatgtatatcgtcatataaattatatcgctaggacagacccaaacgacctttatcccaaacagtcagatcctcttctcaagt gtcgatttctgttatggaatatgcataccctggcccagaaattgcacgcacgagcgtagtgaatgcgtcattggttttacatttaaaggctaaatgcacaaattctttagacgacagcacatcgttaaatagcatctctagcgttcttatgaatgctaagcattggagtcctcctggtcggccacaataacagctgagtatcataccctgagctccggggttgtcgcacatagcggattcgtataaacataggattttccgcgaatccatcagttgcaaaaatctgttaggctccatcaacaacgctggatttacttcagatccacgcgtaaagtaatggtgctcgaataccgtttttagagttgtcggcatttcaaggaacaaagaattcatttcttcattgcaacgacgcgccagaaatcccaagacctctttgggtagtatgttcttgcctataaaacacggcgttccaagtgccaggaaccacgcatgtgttactgttggggcgtattcagaaataaagcggggtttatgcggcttttgaagctcggatatccaaagtatcgcttgctgatgaacgagcgatgtagctgttacaaaacctcctttccatcctccagtcaacataatatttatcggcctacctatgtccgtaataagtattggtcgggcaattattccgtatgaggtcttgcaggaataagctcttagggacagccagcttggatatggtgcgaaacagaccttctcggcttcagaatgtcgctccgcagtctcttcgtgtcggtgcatcttagatccaccatcaatgtgtgcagcattgactcccgcccgtcgaatattccttttgttacgatgcagtaatgagcacgatcatgggcggggcgatgacgttctatttgcatgtctgcgaacaatttgcgtcagtcatacagctatggagtgggccatttctggccgtcaacttaaaaacgcgaaccgcagacatatgtatttgcat gcaaagacgtatcttcgtatttctgggcatcttcaaatgctctggccaatatggcaatgaatttggattcgtttgacgccgatggtatgcagtgcaaatgtgccaatagcccacatccgaaaaagttatttgtcatacaagcaggtgttaagtagcaatcacataaaggcaccagacgcctcatggcatcataatgaatagctccttctccccactggaaccactgacaaaatctgcgagtatattccgcaaaccacattttatttctcatagaaactaccctaaatccttttaacgggaagaagaatcctagatagtgcttgaagtcatgactgttactgctgcaataacactgtatattatttataaattccgtttgtctaggtatctgatgtaggcattccgatccctttactattgcgtcttcacgaccaaatgggaatgcgccaaaatccccacacctcatcaccctggaggcagattgtgtattattaatatccgccgattgaagcacaaaacggtacggtactgttcctaattctggtatagattctatggtcaaaagtctgcatatccccgacattgccatgagatcacacagtccaagtagcatgtttattgagtcactcagactgtcaacgtccctcgccgcaccaccaatcgaaaataaagtatctacgcaagttatagctccgcattttctatcgctagcagcaatcgcgacgcaaaacataaaggccatgttgggatttgaactctctggggggcttgttatcttctgcaccgtcgcagtcgcagttttccgaaatttatgtctaatatattttccggccgtgctccaatcggccgaaaagaatctgcgtattaccagactcattgacgggccgataaagaccataaaacaaaattcctgtgcactccctcctccagttttgccatcgtccaagtcccgtaactttttttgcgtttcgaggagcaagcgttcgttatccctacccac acttgttttccaccgttttcttattataagcggttgtatcgccaacgcgtcaccgcaggttgtcacatacagtgatggcatacttgaacgtgcaacaacgcgctcgctttgcaaatctaagtcattgaccatcaaatcgcgttgagaggatagccaggcatcttttttcctagtatggtgacggtgcagccaccccaactcagttcttgtaaaaaaagctattggcgggaatttatgttctgaggtgcattctatatttatgagtccatcaaatgccattaaccagattcgtattttttcgctcgacccggcatcactatggatacaatacctttctatggcccatttcagctctcgaaccaaccacacggacaattgactaacataagtatgatctttatcacagtcgcacccatctgagttatatttatggcatccgagcgctcttactgtacggtcggatacacccatggtttttcctttatatagtcgggttatagtctgtcgggtttggcggtagcacggagtagtttgatttttaagaatcgaaaaccggcttggagagaccactgtcgaatatttgtccgtatactctacacgtgagtgttgtccattcctaggtatattcatctgttcggataccttcaattgctgttcaggcataaccttaaagcatatgttatgttgtacatcaaaacttggtgagttatgttcgattgccgcgcataaagaatcgtacatgagcgtttctgctaacatactatctatattctcacacgcccctgcatatactgttcctattccaaattcacgttttgccccatcggctatctgctcccaaaaagttgtaatataggtgccgctgggtgcgaaattttcatcagttgtattcctgataaactgaatcactttacataatttttgccacatatctgcgtgcagccatagtatcgaacccgtgggctcggagacgacagtgcgtacaatgggtattttacct ttccccaacaaaataatggtatacaagttaggtccgtacctagaccttaatgtttccaattcttctgaatcactgcactctcgtaggggagtaacggtaataatttcgtctctgagccccgttttgcgttgaaaactaatcacattagataatgtgcaatcggtttcttttatccggatacatctaagtattatgacatcggtggtcattgtttccatcaacgaccatcttttacgatcgcccatactactcatggacgttgtcggtgttgaaaaatcaccagaattgcaacggatctctgggtaccatgctgctgatggaattggcggttttaattgttgtttcagtctattattgctatctttggcggggttgaataatgtggggggagagtgattgcaggaatccgaatgggtcaataaaacgaccgtgctccgttctgccggcgccgatccgattgaagctatatacttcgcttctctccccacttttccaatttgatccggaaataaaacggccccggacaacagtatcgtacgatccggatccggatcctgcttgcctacagaagaatcaacatctcgccccaatattctggtcaaaactggctcgctcatggcaacgcggacgtttcccccggtggccagtcttaatggttaatgttcttttcggcaatcttatacatcagcgggttgcgtgaatactggtcacagttcagtcatttactacacaccagcaatacgacgacggacagtaccgtcccgacgaacgcgacgcccaaaattgctatcgcgaccgcgtccgaggcgatgtcgtacgggcggtgcggggttggatcctcggcaaagagatcctcgtaattcggcggtgggagcggagggtaaagacgcgggtggggatctccctccggaccgcgcgccgggcgcggttcgaaaatgctttccgcctcgctcagtgtcaacgccaagtattcgggcgggctgggggccggaatatctcc cgcgacttcttctatcggcgcggaattggagtcgcggtcgtggcgcgcttctagcgtcgtcaacggaagtccattttcggggtctcccggtgggcgttcagcgtccatcgtcgtatatgctctaacacacgtctcgctatattaaaaaaaagaagagtatcggtcagtgtcgagtgtcgccgacaatgtcgcgagttctcggcgatttaatttttggaactgctccctatgaatcccgtaactgtagcgcccgcgcagaaagccgccatcagaccaactacgtgtctgttcgatgtttgcccgccgatcgctttaccgattaaggttccggcgagaaatgacatgctcgatccaagaacaaagtttttcgcggtaaacaacaacatagttaccgtgcgagatggagaaaccacatctcccgaattagtagaggaaagcccgcgctgtcggtttggggacatatcgatcttttttgtgtttttcctaggacccttttgccagatcgtacaaagtcgcgtcttatgagcggacgttcttactgcagctcggtaggagtggggcagggttagatttcgtcggcgtttcggcccccgtatgcgccgcgccaccctcttcgccgagctctttatgcgcggtgggggtgagcgcttccggagttgcgatctccgatctcgagccgcagcccggcggtgtctctttcagtggagcgttagcgccatcatgtggttcgtggcggtggaaaggctattatgtgttaggggagagaccacgtgatcggcatgcaaatgagcaaggcgaacgcgtcagcgttcgcactgcgaaccaataatatatatattatactattggctttaggtgcgaacgtccggctagtccaatagcggggtcgcgtttcgtaccacgtgttatagaccgccctaaactcgcactcgggggtccggccgcgcccagacagggcggagacgtgccacaggggctttaaaacaccgcttcgggcacc gttcatctcggcgcgcc

SEQ ID NO 27: VP2/1C1#8 (HVT/IBDV/ILT/NDV #3 Virus)

MCMV IEpro-VP2-SV40pA/HVT UL54.5 region (11665 bp)

ggcgcgccactggagaacggcatgaccgcaaaaggcgttgtagagatcgatcccacgaactctcaggcgatcgtgtcagtcgccataaacagcgacgatcgtctccaggatctgaacggttttcttctcaacgatcatcagtatatgaggaactgaacctgatatttagccgagggaaacgcaggttaaaaaccctatcaagcgattgcgattttcgcgtatctagtaaaaatagatgggcttcggtactagccttcgccgccaactctgaatatgcccttcgtggacctcatataacatggcattgtttgttggatgcggggccggaattaagaagaacattcgaaatacgagcaaaaatttcggccctggcatgtgctgcgcgagaatcggtacttcggggagaaagttttatcggagctttgggtagtgcagaggaaactctatcttggttgaaaatgcatgcgaccctgcacttgattctggttaaccacgatccaatttttaagacggctggcgcggtcctagataacctccgcttaaaactagccccaatattgatgtgcagatataacacagaaaaacgatcaatggaagacatgctacggcggtcatctcccgaagacatcaccgattccctaacaatgtgcctgattatgttatcgcgcattcgtcgtaccatgcgcaccgcaggaaataaatatagctatatgatagatccaatgaatcgtatgtctaattacactccaggcgaatgtatgacaggtatattgcgatatattgacgaacatgctagaaggtgtcctgatcacatatgtaatttgtatatcacatgtacacttatgccgatgtatgtgcacgggcgatatttctattgtaattcatttttttgttagtaaactaccacaggctgtccggaaatctaagttaatgaataaagtagatggttaatactcattgcttagaattggactacttttaattctctttaatgttcgta ttaaataaaaacatctttaataaacttcagcctcttcgcttattgtagaaattgagtattcaaaatcatgttcaaagccgtcttcggagagtgtactcgccacggtggttggaacatcactatgtctacacgtcaaatttaagcacgtcaggtctgtcgaggacaagaaatggttaactagtgtttcaattattcttataaacgttaagcattgtaagccccccggccgtccgcagcaacaatttactagtatgccgtgggctccgggactatcacggatgtccaattcgcacatgcatataatttttctagggtctctcatttcgagaaatcttcggggatccatcagcaatgcgggctgtagtcccgattcccgtttcaaatgaaggtgctccaacacggtcttcaaagcaaccggcataccagcaaacacagactgcaactccccgctgcaatgattggttataaacagtaatctgtcttctggaagtatatttcgcccgacaatccacggcgcccccaaagttaaaaaccatccatgtgtatttgcgtcttctctgttaaaagaatattgactggcattttcccgttgaccgccagatatccaaagtacagcacgatgttgcacggacgactttgcagtcaccagccttcctttccacccccccaccaacaaaatgtttatcgtaggacccatatccgtaataaggatgggtctggcagcaaccccataggcgcctcggcgtggtagttctcgaggccttaagcttaaggatcccccaactccgcccgttttatgactagaaccaatagtttttaatgccaaatgcactgaaatcccctaatttgcaaagccaaacgccccctatgtgagtaatacggggactttttacccaatttcccacgcggaaagccccctaatacactcatatggcatatgaatcagcacggtcatgcactctaatggcggcccatagggactttccacatagggggcgtt caccatttcccagcataggggtggtgactcaatggcctttacccaagtacattgggtcaatgggaggtaagccaatgggtttttcccattactggcaagcacactgagtcaaatgggactttccactgggttttgcccaagtacattgggtcaatgggaggtgagccaatgggaaaaacccattgctgccaagtacactgactcaatagggactttccaatgggtttttccattgttggcaagcatataaggtcaatgtgggtgagtcaatagggactttccattgtattctgcccagtacataaggtcaatagggggtgaatcaacaggaaagtcccattggagccaagtacactgcgtcaatagggactttccattgggttttgcccagtacataaggtcaataggggatgagtcaatgggaaaaacccattggagccaagtacactgactcaatagggactttccattgggttttgcccagtacataaggtcaatagggggtgagtcaacaggaaagttccattggagccaagtacattgagtcaatagggactttccaatgggttttgcccagtacataaggtcaatgggaggtaagccaatgggtttttcccattactggcacgtatactgagtcattagggactttccaatgggttttgcccagtacataaggtcaataggggtgaatcaacaggaaagtcccattggagccaagtacactgagtcaatagggactttccattgggttttgcccagtacaaaaggtcaatagggggtgagtcaatgggtttttcccattattggcacgtacataaggtcaataggggtgagtcattgggtttttccagccaatttaattaaaacgccatgtactttcccaccattgacgtcaatgggctattgaaactaatgcaacgtgacctttaaacggtactttcccatagctgattaatgggaaagtaccgttctcgagccaatacacgtcaatgggaag tgaaagggcagccaaaacgtaacaccgccccggttttcccctggaaattccatattggcacgcattctattggctgagctgcgttctacgtgggtataagaggcgcgaccagcgtcggtaccgtcgcagtcttcggtctgaccaccgtagaacgcagagctcctcgctgcaggcggccgctctagaactcgtcgatcgcagcgatgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaaattgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactgttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctt tgatgggactgcggtaatcaccagagctgtggccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccaatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgattccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtagatccagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttcggatcctctagagtcgaggatacatccaaagaggttgagtattctctctacacttcttgttaaatggaaagtgcatttgcttgttcttacaatcggcccgagtctcgttcacagcgcctcgttcacacttaaaccacaaatagtctacaggctatatgggagccagactgaaactcacatatgactaatattcgggggtgttagtcacgtgtagcccattgtgtgcatataacgatgtt ggacgcgtccttattcgcggtgtacttgatactatggcagcgagcatgggatattcatcctcgtcatcgttaacatctctacgggttcagaatgtttggcatgtcgtcgatcctttgcccatcgttgcaaattacaagtccgatcgccatgaccgcgataagcctgtaccatgtggcattagggtgacatctcgatcatacattataagaccaacgtgcgagtcttccaaagacctgcacgccttcttcttcggattgtcaacgggttcttcagaatctatgcccatatctggcgttgagaccattgtgcgtttaatgaacaataaagcggcatgccatggaaaggagggctgcagatctccattttctcacgccactatcctggacgctgtagacgataattataccatgaatatagagggggtatgtttccactgccactgtgatgataagttttctccagattgttggatatctgcattttctgctgccgaacaaacttcatcgctatgcaaagagatgcgtgtgtacacgcgccggtggagtatacgggaaactaaatgttcatagaggtctttgggctatatgttattaaataaaataattgaccagtgaacaatttgtttaatgttagtttattcaatgcattggttgcaaatattcattacttctccaatcccaggtcattctttagcgagatgatgttatgacattgctgtgaaaattactacaggatatatttttaagatgcaggagtaacaatgtgcatagtaggcgtagttatcgcagacgtgcaacgcttcgcatttgagttaccgaagtgcccaacagtgctgcggttatggtttatgcgcacagaatccatgcatgtcctaattgaaccatccgatttttcttttaatcgcgatcgatgtttgggcaactgcgttatttcagatctaaaaaatttaccctttatgaccatcacatctctctggctcataccccgcttggataagat atcatgtagattccgccctaagaaatgcaaactaacattattgtcggttccatatacacttccatcttgtccttcgaaaataacaaactcgcgcaatagaccgtccgtacatgcatggccgatgtgtgtcaacatcattggtctgctagatcccgatgggacgaatcgtacagtcgtcgctccagcattggcaaaaatccccagataccctccatgcggcaaatctaaattgcgaccccgaagagactgcaccaaagtcttatcgacgcacgctgatttttttgaacagcgggagcccattatcttcagtggagcgtagacgggcgaggctaattatgtgacatagcaacactgcatgtatgtttttataaatcaataagagtacataatttattacgtatcatttccgtttgtaatatactgtatacatcatccacactattagtcagcactagcgcgcgggcgcacgttacaatagcagcgtgcccgttatctatattgtccgatatttacacataacatttcatcgacatgattaaatacctaagtactgcacacagatgtttaatgtatatcgtcatataaattatatcgctaggacagacccaaacgacctttatcccaaacagtcagatcctcttctcaagtgtcgatttctgttatggaatatgcataccctggcccagaaattgcacgcacgagcgtagtgaatgcgtcattggttttacatttaaaggctaaatgcacaaattctttagacgacagcacatcgttaaatagcatctctagcgttcttatgaatgctaagcattggagtcctcctggtcggccacaataacagctgagtatcataccctgagctccggggttgtcgcacatagcggattcgtataaacataggattttccgcgaatccatcagttgcaaaaatctgttaggctccatcaacaacgctggatttacttcagatccacgcgtaaagtaatggtgctcgaatacc gtttttagagttgtcggcatttcaaggaacaaagaattcatttcttcattgcaacgacgcgccagaaatcccaagacctctttgggtagtatgttcttgcctataaaacacggcgttccaagtgccaggaaccacgcatgtgttactgttggggcgtattcagaaataaagcggggtttatgcggcttttgaagctcggatatccaaagtatcgcttgctgatgaacgagcgatgtagctgttacaaaacctcctttccatcctccagtcaacataatatttatcggcctacctatgtccgtaataagtattggtcgggcaattattccgtatgaggtcttgcaggaataagctcttagggacagccagcttggatatggtgcgaaacagaccttctcggcttcagaatgtcgctccgcagtctcttcgtgtcggtgcatcttagatccaccatcaatgtgtgcagcattgactcccgcccgtcgaatattccttttgttacgatgcagtaatgagcacgatcatgggcggggcgatgacgttctatttgcatgtctgcgaacaatttgcgtcagtcatacagctatggagtgggccatttctggccgtcaacttaaaaacgcgaaccgcagacatatgtatttgcatgcaaagacgtatcttcgtatttctgggcatcttcaaatgctctggccaatatggcaatgaatttggattcgtttgacgccgatggtatgcagtgcaaatgtgccaatagcccacatccgaaaaagttatttgtcatacaagcaggtgttaagtagcaatcacataaaggcaccagacgcctcatggcatcataatgaatagctccttctccccactggaaccactgacaaaatctgcgagtatattccgcaaaccacattttatttctcatagaaactaccctaaatccttttaacgggaagaagaatcctagatagtgcttgaagtcatgactgttactgctgcaataaca ctgtatattatttataaattccgtttgtctaggtatctgatgtaggcattccgatccctttactattgcgtcttcacgaccaaatgggaatgcgccaaaatccccacacctcatcaccctggaggcagattgtgtattattaatatccgccgattgaagcacaaaacggtacggtactgttcctaattctggtatagattctatggtcaaaagtctgcatatccccgacattgccatgagatcacacagtccaagtagcatgtttattgagtcactcagactgtcaacgtccctcgccgcaccaccaatcgaaaataaagtatctacgcaagttatagctccgcattttctatcgctagcagcaatcgcgacgcaaaacataaaggccatgttgggatttgaactctctggggggcttgttatcttctgcaccgtcgcagtcgcagttttccgaaatttatgtctaatatattttccggccgtgctccaatcggccgaaaagaatctgcgtattaccagactcattgacgggccgataaagaccataaaacaaaattcctgtgcactccctcctccagttttgccatcgtccaagtcccgtaactttttttgcgtttcgaggagcaagcgttcgttatccctacccacacttgttttccaccgttttcttattataagcggttgtatcgccaacgcgtcaccgcaggttgtcacatacagtgatggcatacttgaacgtgcaacaacgcgctcgctttgcaaatctaagtcattgaccatcaaatcgcgttgagaggatagccaggcatcttttttcctagtatggtgacggtgcagccaccccaactcagttcttgtaaaaaaagctattggcgggaatttatgttctgaggtgcattctatatttatgagtccatcaaatgccattaaccagattcgtattttttcgctcgacccggcatcactatggatacaatacctttctatggcccatttcagc tctcgaaccaaccacacggacaattgactaacataagtatgatctttatcacagtcgcacccatctgagttatatttatggcatccgagcgctcttactgtacggtcggatacacccatggtttttcctttatatagtcgggttatagtctgtcgggtttggcggtagcacggagtagtttgatttttaagaatcgaaaaccggcttggagagaccactgtcgaatatttgtccgtatactctacacgtgagtgttgtccattcctaggtatattcatctgttcggataccttcaattgctgttcaggcataaccttaaagcatatgttatgttgtacatcaaaacttggtgagttatgttcgattgccgcgcataaagaatcgtacatgagcgtttctgctaacatactatctatattctcacacgcccctgcatatactgttcctattccaaattcacgttttgccccatcggctatctgctcccaaaaagttgtaatataggtgccgctgggtgcgaaattttcatcagttgtattcctgataaactgaatcactttacataatttttgccacatatctgcgtgcagccatagtatcgaacccgtgggctcggagacgacagtgcgtacaatgggtattttacctttccccaacaaaataatggtatacaagttaggtccgtacctagaccttaatgtttccaattcttctgaatcactgcactctcgtaggggagtaacggtaataatttcgtctctgagccccgttttgcgttgaaaactaatcacattagataatgtgcaatcggtttcttttatccggatacatctaagtattatgacatcggtggtcattgtttccatcaacgaccatcttttacgatcgcccatactactcatggacgttgtcggtgttgaaaaatcaccagaattgcaacggatctctgggtaccatgctgctgatggaattggcggttttaattgttgtttcagtctat tattgctatctttggcggggttgaataatgtggggggagagtgattgcaggaatccgaatgggtcaataaaacgaccgtgctccgttctgccggcgccgatccgattgaagctatatacttcgcttctctccccacttttccaatttgatccggaaataaaacggccccggacaacagtatcgtacgatccggatccggatcctgcttgcctacagaagaatcaacatctcgccccaatattctggtcaaaactggctcgctcatggcaacgcggacgtttcccccggtggccagtcttaatggttaatgttcttttcggcaatcttatacatcagcgggttgcgtgaatactggtcacagttcagtcatttactacacaccagcaatacgacgacggacagtaccgtcccgacgaacgcgacgcccaaaattgctatcgcgaccgcgtccgaggcgatgtcgtacgggcggtgcggggttggatcctcggcaaagagatcctcgtaattcggcggtgggagcggagggtaaagacgcgggtggggatctccctccggaccgcgcgccgggcgcggttcgaaaatgctttccgcctcgctcagtgtcaacgccaagtattcgggcgggctgggggccggaatatctcccgcgacttcttctatcggcgcggaattggagtcgcggtcgtggcgcgcttctagcgtcgtcaacggaagtccattttcggggtctcccggtgggcgttcagcgtccatcgtcgtatatgctctaacacacgtctcgctatattaaaaaaaagaagagtatcggtcagtgtcgagtgtcgccgacaatgtcgcgagttctcggcgatttaatttttggaactgctccctatgaatcccgtaactgtagcgcccgcgcagaaagccgccatcagaccaactacgtgtctgttcgatgtttgcccgccgatcgctttaccgattaaggttccggcgagaaatgacatgctcgatc caagaacaaagtttttcgcggtaaacaacaacatagttaccgtgcgagatggagaaaccacatctcccgaattagtagaggaaagcccgcgctgtcggtttggggacatatcgatcttttttgtgtttttcctaggacccttttgccagatcgtacaaagtcgcgtcttatgagcggacgttcttactgcagctcggtaggagtggggcagggttagatttcgtcggcgtttcggcccccgtatgcgccgcgccaccctcttcgccgagctctttatgcgcggtgggggtgagcgcttccggagttgcgatctccgatctcgagccgcagcccggcggtgtctctttcagtggagcgttagcgccatcatgtggttcgtggcggtggaaaggctattatgtgttaggggagagaccacgtgatcggcatgcaaatgagcaaggcgaacgcgtcagcgttcgcactgcgaaccaataatatatatattatactattggctttaggtgcgaacgtccggctagtccaatagcggggtcgcgtttcgtaccacgtgttatagaccgccctaaactcgcactcgggggtccggccgcgcccagacagggcggagacgtgccacaggggctttaaaacaccgcttcgggcaccgttcatctcggcgcgcc

SEQ ID NO 28: 1332-47.A2 (HVT/IBDV/ILT/NDV # 3 Virus)

ILT/hCMV IEpro-F-IE(therm)/HVT US2 region (13253 bp)

aattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcgggtaccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaaa tgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatggctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttact gtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctgaatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaactg tagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgatacacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagcttgttaattaagtcgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagggtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactacagagtaaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatagc gagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaatacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagctta actttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctaccaggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctcg aacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctcccaatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagagcccctgttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcctaacttttgcgacgcgacac gtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgaaagcttcgcgccaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtagcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccggttgcgccgccaccatgggccccagaccttctaccaagaacccagtacctatgatgctgactgtccgagtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcggctgcaggaattgtggttacaggagacaaagccgtcaacatatacacctcatcccagacaggatcaatcatagttaagctcctcccgaatctgcccaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcaccccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcgg tgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagtttgttaatgaccaatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaattgactacagtattcggaccacaaatcacttcacctgctttaaacaagctgactattcaggcactttacaatctagctggtggaaatatggattacttattgactaagttaggtgtagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctatacgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaagctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaagtggtgacacaggtcggttctgtgatagaagaacttgacacctcatactgtatagaaactgacttacatttatattgtacaagaatagtaacgttccctatgtcccctggtatttattcctgcttgagcggcaatacgtcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatacatgactatcaaaggttcagtcatcgccaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttttatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataata acaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgctttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgactagcacatctgctctcattacctatatcgtgttgactatcatatctcttgtttttggtatacttagcctgattctagcatgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataatactctagatcagatgagagccactacaaaaatgtgaggatctctcgaggaattctagatcccacgtcactattgtatactctatattatactctatgttatactctgtaatcctactcaataaacgtgtcacgcctgtgaaaccgtactaagtctcccgtgtcttcttatcaccatcaggtgacatcctcgcccaggctgtcaatcatgccggtatcgattccagtagcaccggccccacgctgacaacccactcttgcagcgttagcagcgcccctcttaacaagccgacccccaccagcgtcgcggttactaacactcctctccccgacctgcaactagtaagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacaataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatttggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactgg agctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtccgaaattacatacgtggcggcgatggaagtagatgttgagtcttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaaatatcgaaagcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaacaatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattataaaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgcagttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggt gaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcgagttataagaaatggggggatgccaatggatgttgaatatgtcatttctaaaatgcttacgtttgaccaggagttcagaccttctgctaaggaaatattgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtccgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattagaatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatata tttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccgagaattagatggtttatatacgcgcgtagtagttgtcaacggggactttactacggccgatataatgtttaatgttaaagtggcatgtgccttttcaaagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgacatttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtataccaggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcacta cggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccgctgctattctattgcgcatgcaacatattacgccgactgtcctataatcagttctacggtattcagaggatgccgggact

SEQ ID NO 29: 1332-23.7 (HVT/IBDV/ILT/NDV # 4 Virus)

ILT/HVT UL54.5 region (12248 bp)

ggcgcgccactggagaacggcatgaccgcaaaaggcgttgtagagatcgatcccacgaactctcaggcgatcgtgtcagtcgccataaacagcgacgatcgtctccaggatctgaacggttttcttctcaacgatcatcagtatatgaggaactgaacctgatatttagccgagggaaacgcaggttaaaaaccctatcaagcgattgcgattttcgcgtatctagtaaaaatagatgggcttcggtactagccttcgccgccaactctgaatatgcccttcgtggacctcatataacatggcattgtttgttggatgcggggccggaattaagaagaacattcgaaatacgagcaaaaatttcggccctggcatgtgctgcgcgagaatcggtacttcggggagaaagttttatcggagctttgggtagtgcagaggaaactctatcttggttgaaaatgcatgcgaccctgcacttgattctggttaaccacgatccaatttttaagacggctggcgcggtcctagataacctccgcttaaaactagccccaatattgatgtgcagatataacacagaaaaacgatcaatggaagacatgctacggcggtcatctcccgaagacatcaccgattccctaacaatgtgcctgattatgttatcgcgcattcgtcgtaccatgcgcaccgcaggaaataaatatagctatatgatagatccaatgaatcgtatgtctaattacactccaggcgaatgtatgacaggtatattgcgatatattgacgaacatgctagaaggtgtcctgatcacatatgtaatttgtatatcacatgtacacttatgccgatgtatgtgcacgggcgatatttctattgtaattcatttttttgttagtaaactaccacaggctgtccggaaatctaagttaatgaataaagtagatggttaatactcattgcttagaattggactacttttaattctctttaatgttcgta ttaaataaaaacatctttaataaacttcagcctcttcgcttattgtagaaattgagtattcaaaatcatgttcaaagccgtcttcggagagtgtactcgccacggtggttggaacatcactatgtctacacgtcaaatttaagcacgtcaggtctgtcgaggacaagaaatggttaactagtgtttcaattattcttataaacgttaagcattgtaagccccccggccgtccgcagcaacaatttactagtatgccgtgggctccgggactatcacggatgtccaattcgcacatgcatataatttttctagggtctctcatttcgagaaatcttcggggatccatcagcaatgcgggctgtagtcccgattcccgtttcaaatgaaggtgctccaacacggtcttcaaagcaaccggcataccagcaaacacagactgcaactccccgctgcaatgattggttataaacagtaatctgtcttctggaagtatatttcgcccgacaatccacggcgcccccaaagttaaaaaccatccatgtgtatttgcgtcttctctgttaaaagaatattgactggcattttcccgttgaccgccagatatccaaagtacagcacgatgttgcacggacgactttgcagtcaccagccttcctttccacccccccaccaacaaaatgtttatcgtaggacccatatccgtaataaggatgggtctggcagcaaccccataggcgcctcggcgtggtagttctcgaggccttaattaagtcgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagggtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactacagag taaccgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatagcgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaa tacttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagcttaactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctacca ggaacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctcgaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctcccaatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagagccc ctgttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcctaacttttgcgacgcgacacgtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgaaagcttcccgggttaattaaggccctcgaggatacatccaaagaggttgagtattctctctacacttcttgttaaatggaaagtgcatttgcttgttcttacaatcggcccgagtctcgttcacagcgcctcgttcacacttaaaccacaaatagtctacaggctatatgggagccagactgaaactcacatatgactaatattcgggggtgttagtcacgtgtagcccattgtgtgcatataacgatgttggacgcgtccttattcgcggtgtacttgatactatggcagcgagcatgggatattcatcctcgtcatcgttaacatctctacgggttcagaatgtttggcatgtcgtcgatcctttgcccatcgttgcaaattacaagtccgatcgccatgaccgcgataagcctgtaccatgtggcattagggtgacatctcgatcatacattataagaccaacgtgcgagtcttccaaagacctgcacgccttcttcttcggattgtcaacgggttcttcagaatctatgcccatatctggcgttgagaccattgtgcgtttaatgaacaataaagcggcatgccatggaaaggagggctgcagatctccattttctcacgccactatcctggacgctgtagacgataattataccatgaatat agagggggtatgtttccactgccactgtgatgataagttttctccagattgttggatatctgcattttctgctgccgaacaaacttcatcgctatgcaaagagatgcgtgtgtacacgcgccgttgagtatacgggaaactaaatgttcatagaggtctttgggctatatgttattaaataaaataattgaccagtgaacaatttgtttaatgttagtttattcaatgcattggttgcaaatattcattacttctccaatcccaggtcattctttagcgagatgatgttatgacattgctgtgaaaattactacaggatatatttttaagatgcaggagtaacaatgtgcatagtaggcgtagttatcgcagacgtgcaacgcttcgcatttgagttaccgaagtgcccaacagtgctgcggttatggtttatgcgcacagaatccatgcatgtcctaattgaaccatccgatttttcttttaatcgcgatcgttgtttgggcaactgcgttatttcagatctaaaaaatttaccctttatgaccatcacatctctctggctcataccccgcttggataagatatcatgtagattccgccctaagaaatgcaaactaacattattgtcggttccatatacacttccatcttgtccttcgaaaataacaaactcgcgcaatagaccgtccgtacatgcatggccgatgtgtgtcaacatcattggtctgctagatcccgatgggacgaatcgtacagtcgtcgctccagcattggcaaaaatccccagataccctccatgcggcaaatctaaattgcgaccccgaagagactgcaccaaagtcttatcgacgcacgctgatttttttgaacagcgggagcccattatcttcagtggagcgtagacgggcgaggctaattatgtgacatagcaacactgcatgtatgtttttataaatcaataagagtacataatttattacgtatcatttccgtttgtaat atactgtatacatcatccacactattagtcagcactagcgcgcgggcgcacgttacaatagcagcgtgcccgttatctatattgtccgatatttacacataacatttcatcgacatgattaaatacctaagtactgcacacagatgtttaatgtatatcgtcatataaattatatcgctaggacagacccaaacgacctttatcccaaacagtcagatcctcttctcaagtgtcgatttctgttatggaatatgcataccctggcccagaaattgcacgcacgagcgtagtgaatgcgtcattggttttacatttaaaggctaaatgcacaaattctttagacgacagcacatcgttaaatagcatctctagcgttcttatgaatgctaagcattggagtcctcctggtcggccacaataacagctgagtatcataccctgagctccggggttgtcgcacatagcggattcgtataaacataggattttccgcgaatccatcagttgcaaaaatctgttaggctccatcaacaacgctggatttacttcagatccacgcgtaaagtaatggtgctcgaataccgtttttagagttgtcggcatttcaaggaacaaagaattcatttcttcattgcaacgacgcgccagaaatcccaagacctctttgggtagtatgttcttgcctataaaacacggcgttccaagtgccaggaaccacgcatgtgttactgttggggcgtattcagaaataaagcggggtttatgcggcttttgaagctcggatatccaaagtatcgcttgctgatgaacgagcgatgtagctgttacaaaacctcctttccatcctccagtcaacataatatttatcggcctacctatgtccgtaataagtattggtcgggcaattattccgtatgaggtcttgcaggaataagctcttagggacagccagcttggatatggtgcgaaacagaccttctcggcttcagaatgtcgctcc gcagtctcttcgtgtcggtgcatcttagatccaccatcaatgtgtgcagcattgactcccgcccgtcgaatattccttttgttacgatgcagtaatgagcacgatcatgggcggggcgatgacgttctatttgcatgtctgcgaacaatttgcgtcagtcatacagctatggagtgggccatttctggccgtcaacttaaaaacgcgaaccgcagacatatgtatttgcatgcaaagacgtatcttcgtatttctgggcatcttcaaatgctctggccaatatggcaatgaatttggattcgtttgacgccgatggtatgcagtgcaaatgtgccaatagcccacatccgaaaaagttatttgtcatacaagcaggtgttaagtagcaatcacataaaggcaccagacgcctcatggcatcataatgaatagctccttctccccactggaaccactgacaaaatctgcgagtatattccgcaaaccacattttatttctcatagaaactaccctaaatccttttaacgggaagaagaatcctagatagtgcttgaagtcatgactgttactgctgcaataacactgtatattatttataaattccgtttgtctaggtatctgatgtaggcattccgatccctttactattgcgtcttcacgaccaaatgggaatgcgccaaaatccccacacctcatcaccctggaggcagattgtgtattattaatatccgccgattgaagcacaaaacggtacggtactgttcctaattctggtatagattctatggtcaaaagtctgcatatccccgacattgccatgagatcacacagtccaagtagcatgtttattgagtcactcagactgtcaacgtccctcgccgcaccaccaatcgaaaataaagtatctacgcaagttatagctccgcattttctatcgctagcagcaatcgcgacgcaaaacataaaggccatgttgggatttgaactctctggggggct tgttatcttctgcaccgtcgcagtcgcagttttccgaaatttatgtctaatatattttccggccgtgctccaatcggccgaaaagaatctgcgtattaccagactcattgacgggccgataaagaccataaaacaaaattcctgtgcactccctcctccagttttgccatcgtccaagtcccgtaactttttttgcgtttcgaggagcaagcgttcgttatccctacccacacttgttttccaccgttttcttattataagcggttgtatcgccaacgcgtcaccgcaggttgtcacatacagtgatggcatacttgaacgtgcaacaacgcgctcgctttgcaaatctaagtcattgaccatcaaatcgcgttgagaggatagccaggcatcttttttcctagtatggtgacggtgcagccaccccaactcagttcttgtaaaaaaagctattggcgggaatttatgttctgaggtgcattctatatttatgagtccatcaaatgccattaaccagattcgtattttttcgctcgacccggcatcactatggatacaatacctttctatggcccatttcagctctcgaaccaaccacacggacaattgactaacataagtatgatctttatcacagtcgcacccatctgagttatatttatggcatccgagcgctcttactgtacggtcggatacacccatggtttttcctttatatagtcgggttatagtctgtcgggtttggcggtagcacggagtagtttgatttttaagaatcgaaaaccggcttggagagaccactgtcgaatatttgtccgtatactctacacgtgagtgttgtccattcctaggtatattcatctgttcggataccttcaattgctgttcaggcataaccttaaagcatatgttatgttgtacatcaaaacttggtgagttatgttcgattgccgcgcataaagaatcgtacatgagcgtttctgctaacatactatctata ttctcacacgcccctgcatatactgttcctattccaaattcacgttttgccccatcggctatctgctcccaaaaagttgtaatataggtgccgctgggtgcgaaattttcatcagttgtattcctgataaactgaatcactttacataatttttgccacatatctgcgtgcagccatagtatcgaacccgtgggctcggagacgacagtgcgtacaatgggtattttacctttccccaacaaaataatggtatacaagttaggtccgtacctagaccttaatgtttccaattcttctgaatcactgcactctcgtaggggagtaacggtaataatttcgtctctgagccccgttttgcgttgaaaactaatcacattagataatgtgcaatcggtttcttttatccggatacatctaagtattatgacatcggtggtcattgtttccatcaacgaccatcttttacgatcgcccatactactcatggacgttgtcggtgttgaaaaatcaccagaattgcaacggatctctgggtaccatgctgctgatggaattggcggttttaattgttgtttcagtctattattgctatctttggcggggttgaataatgtggggggagagtgattgcaggaatccgaatgggtcaataaaacgaccgtgctccgttctgccggcgccgatccgattgaagctatatacttcgcttctctccccacttttccaatttgatccggaaataaaacggccccggacaacagtatcgtacgatccggatccggatcctgcttgcctacagaagaatcaacatctcgccccaatattctggtcaaaactggctcgctcatggcaacgcggacgtttcccccggtggccagtcttaatggttaatgttcttttcggcaatcttatacatcagcgggttgcgtgaatactggtcacagttcagtcatttactacacaccagcaatacgacgacggacagtaccgtcccgacgaa cgcgacgcccaaaattgctatcgcgaccgcgtccgaggcgatgtcgtacgggcggtgcggggttggatcctcggcaaagagatcctcgtaattcggcggtgggagcggagggtaaagacgcgggtggggatctccctccggaccgcgcgccgggcgcggttcgaaaatgctttccgcctcgctcagtgtcaacgccaagtattcgggcgggctgggggccggaatatctcccgcgacttcttctatcggcgcggaattggagtcgcggtcgtggcgcgcttctagcgtcgtcaacggaagtccattttcggggtctcccggtgggcgttcagcgtccatcgtcgtatatgctctaacacacgtctcgctatattaaaaaaaagaagagtatcggtcagtgtcgagtgtcgccgacaatgtcgcgagttctcggcgatttaatttttggaactgctccctatgaatcccgtaactgtagcgcccgcgcagaaagccgccatcagaccaactacgtgtctgttcgatgtttgcccgccgatcgctttaccgattaaggttccggcgagaaatgacatgctcgatccaagaacaaagtttttcgcggtaaacaacaacatagttaccgtgcgagatggagaaaccacatctcccgaattagtagaggaaagcccgcgctgtcggtttggggacatatcgatcttttttgtgtttttcctaggacccttttgccagatcgtacaaagtcgcgtcttatgagcggacgttcttactgcagctcggtaggagtggggcagggttagatttcgtcggcgtttcggcccccgtatgcgccgcgccaccctcttcgccgagctctttatgcgcggtgggggtgagcgcttccggagttgcgatctccgatctcgagccgcagcccggcggtgtctctttcagtggagcgttagcgccatcatgtggttcgtggcggtggaaaggctattatgtgttaggggagagacca cgtgatcggcatgcaaatgagcaaggcgaacgcgtcagcgttcgcactgcgaaccaataatatatatattatactattggctttaggtgcgaacgtccggctagtccaatagcggggtcgcgtttcgtaccacgtgttatagaccgccctaaactcgcactcgggggtccggccgcgcccagacagggcggagacgtgccacaggggctttaaaacaccgcttcgggcaccgttcatctcggcgcgcc

SEQ ID NO 30: 435Vec60 (HVT/IBDV/ILT/NDV #4 Virus)

mCMV IEpro-VP2-SV40pA/hCMV IEpro-F-IE(therm)/HVT US2 region (13068 bp)

aattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcgggtaccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaaa tgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatggctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttact gtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctgaatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaactg tagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgatacacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagctgcggccgctctagaactagtggatcccccgggctgcagcccaatgtggaattcgcccttgcacattgttactcctgcatcttaaaaatatatcctgtagtaattttcacagcaatgtcataacatcatctcgctaaagaatgacctgggattggagaagtaatgaatatttgcaaccaatgcattgaataaactaacattaaacgaattcactagtggatcccccaactccgcccgttttatgactagaaccaatagtttttaatgccaaatgcactgaaatcccctaatttgcaaagccaaacgccccctatgtgagtaatacggggactttt tacccaatttcccaagcggaaagccccctaatacactcatatggcatatgaatcagcacggtcatgcactctaatggcggcccatagggactttccacatagggggcgttcaccatttcccagcataggggtggtgactcaatggcctttacccaagtacattgggtcaatgggaggtaagccaatgggtttttcccattactggcaagcacactgagtcaaatgggactttccactgggttttgcccaagtacattgggtcaatgggaggtgagccaatgggaaaaacccattgctgccaagtacactgactcaatagggactttccaatgggtttttccattgttggcaagcatataaggtcaatgtgggtgagtcaatagggactttccattgtattctgcccagtacataaggtcaatagggggtgaatcaacaggaaagtcccattggagccaagtacactgcgtcaatagggactttccattgggttttgcccagtacataaggtcaataggggatgagtcaatgggaaaaacccattggagccaagtacactgactcaatagggactttccattgggttttgcccagtacataaggtcaatagggggtgagtcaacaggaaagtcccattggagccaagtacattgagtcaatagggactttccaatgggttttgcccagtacataaggtcaatgggaggtaagccaatgggtttttcccattactggcacgtatactgagtcattagggactttccaatgggttttgcccagtacataaggtcaataggggtgaatcaacaggaaagtcccattggagccaagtacactgagtcaatagggactttccattgggttttgcccagtacaaaaggtcaatagggggtgagtcaatgggtttttcccattattggcacgtacataaggtcaataggggtgagtcattgggtttttccagccaatttaattaaaacgccatgtactttcccaccat tgacgtcaatgggctattgaaactaatgcaacgtgacctttaaacggtactttcccatagctgattaatgggaaagtaccgttctcgagccaatacacgtcaatgggaagtgaaagggcagccaaaacgtaacaccgccccggttttcccctggaaattccatattggcacgcattctattggctgagctgcgttctacgtgggtataagaggcgcgaccagcgtcggtaccgtcgcagtcttcggtctgaccaccgtagaacgcagagctcctcgctgcaggcggccgctctagaactcgtcgatcgcagcgatgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaaattgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactg ttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctttgatgggactgcggtaatcaccagagctgtagccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccaatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgattccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtaagcttgatctagagcggccgcggggatccagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttcggatcctctagagtcgacaattatttcatttaataacatatagcccaaagacctctatgaacatttagtttcccgtatactcaacggcgcgtgtacacacaaggg cgaattccacagtggatatcaagcttaattaagtaccgagctcgaattggcgcgccaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggcgcgccggatccatgggccccagaccttctaccaagaacccagtacctatgatgctgactgtccgagtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcggctgcaggaattgtggttacaggagacaaagccgtcaacatatacacctcatcccagacaggatcaatcatagttaagctcctcccgaatctgcccaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcaccccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcggtgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagtttgttaatgacc aatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaattgactacagtattcggaccacaaatcacttcacctgctttaaacaagctgactattcaggcactttacaatctagctggtggaaatatggattacttattgactaagttaggtgtagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctatacgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaacctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaagtggtgacacaggtcggttctgtgatagaagaacttgacacctcatactgtatagaaactgacttagatttatattgtacaagaatagtaacgttccctatgtcccctggtatttattcctgcttgagcggcaatacgtcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatacatgactatcaaaggttcagtcatcgccaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttttatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataataacaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgctttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgactagcacatctgctctcattacctatatcgttttgactatcatatctcttgtttttggtatacttagcccgattctagc atgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataatactctagatcagatgagagccactacaaaaatgtgaggatctctcgaggaattctagatcccacgtcactattgtatactctatattatactctatgttatactctgtaatcctactcaataaacgtgtcacgcctgtgaaaccgtactaagtctcccgtgtcttcttatcaccatcaggtgacatcctcgcccaggctgtcaatcatgccggtatcgattccagtagcaccggccccacgctgacaacccactcttgcagcgttagcagcgcccctcttaacaagccgacccccaccagcgtcgcggttactaacactcctctccccgacctgcaactagtgcggccgcagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacgataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatttggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactggagctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtcccgaaattacatacgtggcggcgatggaagtagatgttgagtcttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaa atatcgaaagcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaacaatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattataaaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgcagttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggtgaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcgagttataagaaatggggggatgccaatggatgttgaatatgt catttctaaaatgcttacgtttgaccaggagttcagaccttctgctaaggaaatattgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtccgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattagaatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatatatttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccgagaattagatggtttatatacgcgcgtagtagttgtcaacgg ggactttactacggccgatataatgtttaatgttaaagtggcatgtgccttttcaaagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgacatttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtataccaggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcactacggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccgctgctattctattgcgcatgcaacatattacgccgactgtc ctataatcagttctacggtattcagaggatgccgggacgccgttgtttatactaggccccacagcaga

SEQ ID NO 31: 1332-29.4 (HVT/IBDV/ILT/NDV #5 Virus)

ILT/hCMV IEpro-F-IEpA(therm)/HVT UL54.5 area (14598 bp)

cgcgccactggagaacggcatgaccgcaaaaggcgttgtagagatcgatcccacgaactctcaggcgatcgtgtcagtcgccataaacagcgacgatcgtctccaggatctgaacggttttcttctcaacgatcatcagtatatgaggaactgaacctgatatttagccgagggaaacgcaggttaaaaaccctatcaagcgattgcgattttcgcgtatctagtaaaaatagatgggcttcggtactagccttcgccgccaactctgaatatgcccttcgtggacctcatataacatggcattgtttgttggatgcggggccggaattaagaagaacattcgaaatacgagcaaaaatttcggccctggcatgtgctgcgcgagaatcggtacttcggggagaaagttttatcggagctttgggtagtgcagaggaaactctatcttggttgaaaatgcatgcgaccctgcacttgattctggttaaccacgatccaatttttaagacggctggcgcggtcctagataacctccgcttaaaactagccccaatattgatgtgcagatataacacagaaaaacgatcaatggaagacatgctacggcggtcatctcccgaagacatcaccgattccctaacaatgtgcctgattatgttatcgcgcattcgtcgtaccatgcgcaccgcaggaaataaatatagctatatgatagatccaatgaatcgtatgtctaattacactccaggcgaatgtatgacaggtatattgcgatatattgacgaacatgctagaaggtgtcctgatcacatatgtaatttgtatatcacatgtacacttatgccgatgtatgtgcacgggcgatatttctattgtaattcatttttttgttagtaaactaccacaggctgtccggaaatctaagttaatgaataaagtagatggttaatactcattgcttagaattggactacttttaattctctttaatgttcgtatt aaataaaaacatctttaataaacttcagcctcttcgcttattgtagaaattgagtattcaaaatcatgttcaaagccgtcttcggagagtgtactcgccacggtggttggaacatcactatgtctacacgtcaaatttaagcacgtcaggtctgtcgaggacaagaaatggttaactagtgtttcaattattcttataaacgttaagcattgtaagccccccggccgtccgcagcaacaatttactagtatgccgtgggctccgggactatcacggatgtccaattcgcacatgcatataatttttctagggtctctcatttcgagaaatcttcggggatccatcagcaatgcgggctgtagtcccgattcccgtttcaaatgaaggtgctccaacacggtcttcaaagcaaccggcataccagcaaacacagactgcaactccccgctgcaatgattggttataaacagtaatctgtcttctggaagtatatttcgcccgacaatccacggcgcccccaaagttaaaaaccatccatgtgtatttgcgtcttctctgttaaaagaatattgactggcattttcccgttgaccgccagatatccaaagtacagcacgatgttgcacggacgactttgcagtcaccagccttcctttccacccccccaccaacaaaatgtttatcgtaggacccatatccgtaataaggatgggtctggcagcaaccccataggcgcctcggcgtggtagttctcgaggccttaattaagtcgacggcagagtcgcagacgcccctattggacgtcaaaattgtagaggtgaagttttcaaacgatggcgaagtaacggcgacttgcgtttccaccgtcaaatctccctatagggtagaaactaattggaaagtagacctcgtagatgtaatggatgaaatttctgggaacagtcccgccggggtttttaacagtaatgagaaatggcagaaacagctgtactacagagta accgatggaagaacatcggtccagctaatgtgcctgtcgtgcacgagccattctccggaaccttactgtcttttcgacacgtctcttatagcgagggaaaaagatatcgcgccagagttatactttacctctgatccgcaaacggcatactgcacaataactctgccgtccggcgttgttccgagattcgaatggagccttaataatgtttcactgccggaatatttgacggccacgaccgttgtttcgcataccgctggccaaagtacagtgtggaagagcagcgcgagagcaggcgaggcgtggatttctggccggggaggcaatatatacgaatgcaccgtcctcatctcagacggcactcgcgttactacgcgaaaggagaggtgcttaacaaacacatggattgcggtggaaaacggtgctgctcaggcgcagctgtattcactcttttctggacttgtgtcaggattatgcgggagcatatctgctttgtacgcaacgctatggaccgccatttatttttgaggaatgctttttggactatcgtactgctttcttccttcgctagccagagcaccgccgccgtcacgtacgactacattttaggccgtcgcgcgctcgacgcgctaaccataccggcggttggcccgtataacagatacctcactagggtatcaagaggctgcgacgttgtcgagctcaacccgatttctaacgtggacgacatgatatcggcggccaaagaaaaagagaaggggggccctttcgaggcctccgtcgtctggttctacgtgattaagggcgacgacggcgaggacaagtactgtccaatctatagaaaagagtacagggaatgtggcgacgtacaactgctatctgaatgcgccgttcaatctgcacagatgtgggcagtggactatgttcctagcacccttgtatcgcgaaatggcgcgggactgactatattctcccccactgctgcgctctctggccaata cttgctgaccctgaaaatcgggagatttgcgcaaacagctctcgtaactctagaagttaacgatcgctgtttaaagatcgggtcgcagcttaactttttaccgtcgaaatgctggacaacagaacagtatcagactggatttcaaggcgaacacctttatccgatcgcagacaccaatacacgacacgcggacgacgtatatcggggatacgaagatattctgcagcgctggaataatttgctgaggaaaaagaatcctagcgcgccagaccctcgtccagatagcgtcccgcaagaaattcccgctgtaaccaagaaagcggaagggcgcaccccggacgcagaaagcagcgaaaagaaggcccctccagaagactcggaggacgacatgcaggcagaggcttctggagaaaatcctgccgccctccccgaagacgacgaagtccccgaggacaccgagcacgatgatccaaactcggatcctgactattacaatgacatgcccgccgtgatcccggtggaggagactactaaaagttctaatgccgtctccatgcccatattcgcggcgttcgtagcctgcgcggtcgcgctcgtggggctactggtttggagcatcgtaaaatgcgcgcgtagctaatcgagcctagaataggtggtttcttcctacatgccacgcctcacgctcataatataaatcacatggaatagcataccaatgcctattcattgggacgttcgaaaagcatggcatcgctacttggaactctggctctccttgccgcgacgctcgcacccttcggcgcgatgggaatcgtgatcactggaaatcacgtctccgccaggattgacgacgatcacatcgtgatcgtcgcgcctcgccccgaagctacaattcaactgcagctatttttcatgcctggccagagaccccacaaaccctactcaggaaccgtccgcgtcgcgtttcggtctgatataacaaaccagtgctaccagg aacttagcgaggagcgctttgaaaattgcactcatcgatcgtcttctgtttttgtcggctgtaaagtgaccgagtacacgttctccgcctcgaacagactaaccggacctccacacccgtttaagctcactatacgaaatcctcgtccgaacgacagcgggatgttctacgtaattgttcggctagacgacaccaaagaacccattgacgtcttcgcgatccaactatcggtgtatcaattcgcgaacaccgccgcgactcgcggactctattccaaggcttcgtgtcgcaccttcggattacctaccgtccaacttgaggcctatctcaggaccgaggaaagttggcgcaactggcaagcgtacgttgccacggaggccacgacgaccagcgccgaggcgacaaccccgacgcccgtcactgcaaccagcgcctccgaacttgaagcggaacactttacctttccctggctagaaaatggcgtggatcattacgaaccgacacccgcaaacgaaaattcaaacgttactgtccgtctcgggacaatgagccctacgctaattggggtaaccgtggctgccgtcgtgagcgcaacgatcggcctcgtcattgtaatttccatcgtcaccagaaacatgtgcaccccgcaccgaaaattagacacggtctcgcaagacgacgaagaacgttcccaaactagaagggaatcgcgaaaatttggacccatggttgcgtgcgaaataaacaagggggctgaccaggatagtgaacttgtggaactggttgcgattgttaacccgtctgcgctaagctcgcccgactcaataaaaatgtgattaagtctgaatgtggctctccaatcatttcgattctctaatctcccaatcctctcaaaaggggcagtatcggacacggactgggaggggcgtacacgatagttatatggtacagcagaggcctctgaacacttaggaggagaattcagccggggagagcccct gttgagtaggcttgggagcatattgcaggatgaacatgttagtgatagttctcgcctcttgtcttgcgcgcctaacttttgcgacgcgacacgtcctctttttggaaggcactcaggctgtcctcggggaagatgatcccagaaacgttccggaagggactgtaatcaaatggacaaaagtcctgcggaacgcgtgcaagatgaaggcggccgatgtctgctcttcgcctaactattgctttcatgatttaatttacgacggaggaaagaaagactgcccgcccgcgggacccctgtctgcaaacctggtaattttactaaagcgcggcgaaagcttcgcgccaggtcaattccctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtagcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccggttgcgccgccaccatgggccccagaccttctaccaagaacccagtacctatgatgctgactgtccgagtcgcgctggtactgagttgcatctgtccggcaaactccattgatggcaggcctcttgcggctgcaggaattgtggttacaggagacaaagccgtcaacatatacacctcatcccagacaggatcaatcatagttaagctcctcccgaatctgcccaaggataaggaggcatgtgcgaaagcccccttggatgcatacaacaggacattgaccactttgctcacc ccccttggtgactctatccgtaggatacaagagtctgtgactacatctggaggggggagacaggggcgccttataggcgccattattggcggtgtggctcttggggttgcaactgccgcacaaataacagcggccgcagctctgatacaagccaaacaaaatgctgccaacatcctccgacttaaagagagcattgccgcaaccaatgaggctgtgcatgaggtcactgacggattatcgcaactagcagtggcagttgggaagatgcagcagtttgttaatgaccaatttaataaaacagctcaggaattagactgcatcaaaattgcacagcaagttggtgtagagctcaacctgtacctaaccgaattgactacagtattcggaccacaaatcacttcacctgctttaaacaagctgactattcaggcactttacaatctagctggtggaaatatggattacttattgactaagttaggtgtagggaacaatcaactcagctcattaatcggtagcggcttaatcaccggtaaccctattctatacgactcacagactcaactcttgggtatacaggtaactctaccttcagtcgggaagctaaataatatgcgtgccacctacttggaaaccttatccgtaagcacaaccaggggatttgcctcggcacttgtcccaaaagtggtgacacaggtcggttctgtgatagaagaacttgacacctcatactgtatagaaactgacttacatttatattgtacaagaatagtaacgttccctatgtcccctggtatttattcctgcttgagcggcaatacgtcggcctgtatgtactcaaagaccgaaggcgcacttactacaccatacatgactatcaaaggttcagtcatcgccaactgcaagatgacaacatgtagatgtgtaaaccccccgggtatcatatcgcaaaactatggagaagccgtgtctctaatagataaacaatcatgcaatgttt tatccttaggcgggataactttaaggctcagtggggaattcgatgtaacttatcagaagaatatctcaatacaagattctcaagtaataataacaggcaatcttgatatctcaactgagcttgggaatgtcaacaactcgatcagtaatgctttgaataagttagaggaaagcaacagaaaactagacaaagtcaatgtcaaactgactagcacatctgctctcattacctatatcgtgttgactatcatatctcttgtttttggtatacttagcctgattctagcatgctacctaatgtacaagcaaaaggcgcaacaaaagaccttattatggcttgggaataatactctagatcagatgagagccactacaaaaatgtgaggatctctcgaggaattctagatcccacgtcactattgtatactctatattatactctatgttatactctgtaatcctactcaataaacgtgtcacgcctgtgaaaccgtactaagtctcccgtgtcttcttatcaccatcaggtgacatcctcgcccaggctgtcaatcatgccggtatcgattccagtagcaccggccccacgctgacaacccactcttgcagcgttagcagcgcccctcttaacaagccgacccccaccagcgtcgcggttactaacactcctctccccgacctgcaactagtaagcttcccgggttaattaaggccctcgaggatacatccaaagaggttgagtattctctctacacttcttgttaaatggaaagtgcatttgcttgttcttacaatcggcccgagtctcgttcacagcgcctcgttcacacttaaaccacaaatagtctacaggctatatgggagccagactgaaactcacatatgactaatattcgggggtgttagtcacgtgtagcccattgtgtgcatataacgatgttggacgcgtccttattcgcggtgtacttgatactatggcagcgagcatgggatattcatcc tcgtcatcgttaacatctctacgggttcagaatgtttggcatgtcgtcgatcctttgcccatcgttgcaaattacaagtccgatcgccatgaccgcgataagcctgtaccatgtggcattagggtgacatctcgatcatacattataagaccaacgtgcgagtcttccaaagacctgcacgccttcttcttcggattgtcaacgggttcttcagaatctatgcccatatctggcgttgagaccattgtgcgtttaatgaacaataaagcggcatgccatggaaaggagggctgcagatctccattttctcacgccactatcctggacgctgtagacgataattataccatgaatatagagggggtatgtttccactgccactgtgatgataagttttctccagattgttggatatctgcattttctgctgccgaacaaacttcatcgctatgcaaagagatgcgtgtgtacacgcgccgttgagtatacgggaaactaaatgttcatagaggtctttgggctatatgttattaaataaaataattgaccagtgaacaatttgtttaatgttagtttattcaatgcattggttgcaaatattcattacttctccaatcccaggtcattctttagcgagatgatgttatgacattgctgtgaaaattactacaggatatatttttaagatgcaggagtaacaatgtgcatagtaggcgtagttatcgcagacgtgcaacgcttcgcatttgagttaccgaagtgcccaacagtgctgcggttatggtttatgcgcacagaatccatgcatgtcctaattgaaccatccgatttttcttttaatcgcgatcgttgtttgggcaactgcgttatttcagatctaaaaaatttaccctttatgaccatcacatctctctggctcataccccgcttggataagatatcatgtagattccgccctaagaaatgcaaactaacattattgtcggttccatatacactt ccatcttgtccttcgaaaataacaaactcgcgcaatagaccgtccgtacatgcatggccgatgtgtgtcaacatcattggtctgctagatcccgatgggacgaatcgtacagtcgtcgctccagcattggcaaaaatccccagataccctccatgcggcaaatctaaattgcgaccccgaagagactgcaccaaagtcttatcgacgcacgctgatttttttgaacagcgggagcccattatcttcagtggagcgtagacgggcgaggctaattatgtgacatagcaacactgcatgtatgtttttataaatcaataagagtacataatttattacgtatcatttccgtttgtaatatactgtatacatcatccacactattagtcagcactagcgcgcgggcgcacgttacaatagcagcgtgcccgttatctatattgtccgatatttacacataacatttcatcgacatgattaaatacctaagtactgcacacagatgtttaatgtatatcgtcatataaattatatcgctaggacagacccaaacgacctttatcccaaacagtcagatcctcttctcaagtgtcgatttctgttatggaatatgcataccctggcccagaaattgcacgcacgagcgtagtgaatgcgtcattggttttacatttaaaggctaaatgcacaaattctttagacgacagcacatcgttaaatagcatctctagcgttcttatgaatgctaagcattggagtcctcctggtcggccacaataacagctgagtatcataccctgagctccggggttgtcgcacatagcggattcgtataaacataggattttccgcgaatccatcagttgcaaaaatctgttaggctccatcaacaacgctggatttacttcagatccacgcgtaaagtaatggtgctcgaataccgtttttagagttgtcggcatttcaaggaacaaagaattcatttcttcattgcaacgacgcg ccagaaatcccaagacctctttgggtagtatgttcttgcctataaaacacggcgttccaagtgccaggaaccacgcatgtgttactgttggggcgtattcagaaataaagcggggtttatgcggcttttgaagctcggatatccaaagtatcgcttgctgatgaacgagcgatgtagctgttacaaaacctcctttccatcctccagtcaacataatatttatcggcctacctatgtccgtaataagtattggtcgggcaattattccgtatgaggtcttgcaggaataagctcttagggacagccagcttggatatggtgcgaaacagaccttctcggcttcagaatgtcgctccgcagtctcttcgtgtcggtgcatcttagatccaccatcaatgtgtgcagcattgactcccgcccgtcgaatattccttttgttacgatgcagtaatgagcacgatcatgggcggggcgatgacgttctatttgcatgtctgcgaacaatttgcgtcagtcatacagctatggagtgggccatttctggccgtcaacttaaaaacgcgaaccgcagacatatgtatttgcatgcaaagacgtatcttcgtatttctgggcatcttcaaatgctctggccaatatggcaatgaatttggattcgtttgacgccgatggtatgcagtgcaaatgtgccaatagcccacatccgaaaaagttatttgtcatacaagcaggtgttaagtagcaatcacataaaggcaccagacgcctcatggcatcataatgaatagctccttctccccactggaaccactgacaaaatctgcgagtatattccgcaaaccacattttatttctcatagaaactaccctaaatccttttaacgggaagaagaatcctagatagtgcttgaagtcatgactgttactgctgcaataacactgtatattatttataaattccgtttgtctaggtatctgatgtaggcattccgatcccttt actattgcgtcttcacgaccaaatgggaatgcgccaaaatccccacacctcatcaccctggaggcagattgtgtattattaatatccgccgattgaagcacaaaacggtacggtactgttcctaattctggtatagattctatggtcaaaagtctgcatatccccgacattgccatgagatcacacagtccaagtagcatgtttattgagtcactcagactgtcaacgtccctcgccgcaccaccaatcgaaaataaagtatctacgcaagttatagctccgcattttctatcgctagcagcaatcgcgacgcaaaacataaaggccatgttgggatttgaactctctggggggcttgttatcttctgcaccgtcgcagtcgcagttttccgaaatttatgtctaatatattttccggccgtgctccaatcggccgaaaagaatctgcgtattaccagactcattgacgggccgataaagaccataaaacaaaattcctgtgcactccctcctccagttttgccatcgtccaagtcccgtaactttttttgcgtttcgaggagcaagcgttcgttatccctacccacacttgttttccaccgttttcttattataagcggttgtatcgccaacgcgtcaccgcaggttgtcacatacagtgatggcatacttgaacgtgcaacaacgcgctcgctttgcaaatctaagtcattgaccatcaaatcgcgttgagaggatagccaggcatcttttttcctagtatggtgacggtgcagccaccccaactcagttcttgtaaaaaaagctattggcgggaatttatgttctgaggtgcattctatatttatgagtccatcaaatgccattaaccagattcgtattttttcgctcgacccggcatcactatggatacaatacctttctatggcccatttcagctctcgaaccaaccacacggacaattgactaacataagtatgatctttatcacagtcgcacc catctgagttatatttatggcatccgagcgctcttactgtacggtcggatacacccatggtttttcctttatatagtcgggttatagtctgtcgggtttggcggtagcacggagtagtttgatttttaagaatcgaaaaccggcttggagagaccactgtcgaatatttgtccgtatactctacacgtgagtgttgtccattcctaggtatattcatctgttcggataccttcaattgctgttcaggcataaccttaaagcatatgttatgttgtacatcaaaacttggtgagttatgttcgattgccgcgcataaagaatcgtacatgagcgtttctgctaacatactatctatattctcacacgcccctgcatatactgttcctattccaaattcacgttttgccccatcggctatctgctcccaaaaagttgtaatataggtgccgctgggtgcgaaattttcatcagttgtattcctgataaactgaatcactttacataatttttgccacatatctgcgtgcagccatagtatcgaacccgtgggctcggagacgacagtgcgtacaatgggtattttacctttccccaacaaaataatggtatacaagttaggtccgtacctagaccttaatgtttccaattcttctgaatcactgcactctcgtaggggagtaacggtaataatttcgtctctgagccccgttttgcgttgaaaactaatcacattagataatgtgcaatcggtttcttttatccggatacatctaagtattatgacatcggtggtcattgtttccatcaacgaccatcttttacgatcgcccatactactcatggacgttgtcggtgttgaaaaatcaccagaattgcaacggatctctgggtaccatgctgctgatggaattggcggttttaattgttgtttcagtctattattgctatctttggcggggttgaataatgtggggggagagtgattgcaggaatccgaatg ggtcaataaaacgaccgtgctccgttctgccggcgccgatccgattgaagctatatacttcgcttctctccccacttttccaatttgatccggaaataaaacggccccggacaacagtatcgtacgatccggatccggatcctgcttgcctacagaagaatcaacatctcgccccaatattctggtcaaaactggctcgctcatggcaacgcggacgtttcccccggtggccagtcttaatggttaatgttcttttcggcaatcttatacatcagcgggttgcgtgaatactggtcacagttcagtcatttactacacaccagcaatacgacgacggacagtaccgtcccgacgaacgcgacgcccaaaattgctatcgcgaccgcgtccgaggcgatgtcgtacgggcggtgcggggttggatcctcggcaaagagatcctcgtaattcggcggtgggagcggagggtaaagacgcgggtggggatctccctccggaccgcgcgccgggcgcggttcgaaaatgctttccgcctcgctcagtgtcaacgccaagtattcgggcgggctgggggccggaatatctcccgcgacttcttctatcggcgcggaattggagtcgcggtcgtggcgcgcttctagcgtcgtcaacggaagtccattttcggggtctcccggtgggcgttcagcgtccatcgtcgtatatgctctaacacacgtctcgctatattaaaaaaaagaagagtatcggtcagtgtcgagtgtcgccgacaatgtcgcgagttctcggcgatttaatttttggaactgctccctatgaatcccgtaactgtagcgcccgcgcagaaagccgccatcagaccaactacgtgtctgttcgatgtttgcccgccgatcgctttaccgattaaggttccggcgagaaatgacatgctcgatccaagaacaaagtttttcgcggtaaacaacaacatagttaccgtgcgagatggagaaaccac atctcccgaattagtagaggaaagcccgcgctgtcggtttggggacatatcgatcttttttgtgtttttcctaggacccttttgccagatcgtacaaagtcgcgtcttatgagcggacgttcttactgcagctcggtaggagtggggcagggttagatttcgtcggcgtttcggcccccgtatgcgccgcgccaccctcttcgccgagctctttatgcgcggtgggggtgagcgcttccggagttgcgatctccgatctcgagccgcagcccggcggtgtctctttcagtggagcgttagcgccatcatgtggttcgtggcggtggaaaggctattatgtgttaggggagagaccacgtgatcggcatgcaaatgagcaaggcgaacgcgtcagcgttcgcactgcgaaccaataatatatatattatactattggctttaggtgcgaacgtccggctagtccaatagcggggtcgcgtttcgtaccacgtgttatagaccgccctaaactcgcactcgggggtccggccgcgcccagacagggcggagacgtgccacaggggctttaaaacaccgcttcgggcaccgttcatctcgg

SEQ ID NO 32: 435Vec6 (HVT/IBDV/ILT/NDV #5)

mCMV IEpro-VP2-SV40pA/HVT US2 region (10681 bp)

aattccagactaaatgccccggcccaatttgtcaagtgtgcagtcacggaggcgtcgaccgtgtccccggcattaaacaggaaagcgttaaagtttttgaatgttaggtcacaggtacaaacataaatgtttgtacaaacaggtaacaggtacaaacataaatgccccggcataaatgtcccttacggcggatcgaaacgacattaggcatactcgggtaccattttgcattccgatcagcacggatgaaattaggcaggaatgcggtttatattatgcggcattggacaaacgatatggcattgattggcagtttatgaatgtcttcatgttgggcgtaaacggattcctattggttcagaagacaacgacgatatatttagagagaaaaagctacccagcataggataaacacacattgagcattgagagacataggtatcggtatggatgggaaaactacacacgtgaacaccaaacgacttatatactcgagcggtgatactactgagcaagaatgcactgcatctgagccactgaatgaagactgtgatgaaaatgtgaccatcgatggaattggagaagaatatgcgcagttcttcatgtccccgcaatgggtcccaaatctacatcgcttgagcgaggataccaaaaaggtataccgatgtatggtttccaacagactcaattattttccctattatgaggcgttcaggcggtctttgtttgatatgtatatgctaggtcggttggggcgtcgacttaagcgatctgactgggagactattatgcatctgtcaccaacgcaaagtcggcgtctacatagaactttaagatttgtggagcgtagaattatcccatctaacagttatatacgcacatcgggccacgttccgccttcgagggcacttccgacagatacgaatttaaagatggatgaataattaaattggaaagagtaactacattaatcgagcgtcatgacggcgtcccgtgaaaa tgggaattttctactcgaaacaccgtgacatttgacagacctggaattgttattctgatatatagtgggtgtgtctggccggcaacatacataatgtgcatgcgaaaccactttttcagtgtacgctgacattgtgcaacacggaggggtagcatctacatacaatatatgttgattaatgattggagaaaaaactatgcagctcgccgatcatatggctaactcgccttcgtctatatggcggaccccgcgggaaaaatcgacgtaccatctgatttacaacaccagtaatgaacatgtcgcatccctgcccagatctgtgcgcccattggcgcggatcgttgtgaatgccgccgaaacacttcaggtcggtatgagagccgggaggccgccatcagcaggagtttggcgagaggtgtttgatagaatgatgacagccttccgtgaccacgagcctactgcgacatttaatgctgcaaatcccattagaaaaatggtcgagacagttctacagaataatgaagagcccccgcggacgcatgctgaaatgggtaatcgccttatgaacattatgtactggtgttgcttgggacacgcaggacaatgctcgatatggcagttgtacgagacgaatcaggccattttaagtttattagatgaagtggttatcggcacaacaaatcccttttgcaccctcgagcaatactggaagccattatgcaccgcaatcgccaacaaggggacctcatcgcttgttgaggatgccaaagtggccgagtacctggttagcatgcgcaaattgatataacataggcacgctctgatgttacagaccacaataccgcatacatttattgtaaggttgttaataaaggtttattctatgtaagactacaatactttcgacattgcttgtatacatattaaatactttctcaagttcctattacataaaatgggatctatcattacattcgttaagagtctggataattttact gtttgccagcttcgatcttggaacgtactgtggatagtgccttacttggaatcgtgaaaatttgaaacgtccattatttggatatcttccggttgtcccatatcccgccctggtaccgctcggataccttgcccgtatggattcgtattgacagtcgcgcaatcggggaccaacaacgcgtgggtccacactcattcggaaattttccgatgattctgaatatttattgccgctcgttacgagtcgttggacatatctgtaatacatttcttcttctgaaggatcgctgcacatttgatctatacattggccaggatgttcaagtctcagatgttgcattctggcacagcacaactttatggcatttccgatgtaatcgtccggcagccctgggggagttctatattcgcatattgggatggtaaggacaatagcagatctcgcaacctccagggaggctataataacgtttttaaaggatggatttctcataaaaatctgtcgcaaattacactgagaatatcctttactagcgccgattgagagcatcgtcgtccaattttctaaatggaaagaaaacaaggcgggcaagagtgttccaaacattttcattttcggcgaatctctcaaatcccatggcgtgcaattgattgcaaaattggcacttccgttcacgtttgtatctccaaactctaagacacttttaattgaaaaactacgttctagtgtggaaagaaacctataggcagaccatagaactatttgacaccacatatctttttgtatgtcaaactgaccatgatcgtatgttgctgaatgcactagggcaattcgctcgcgcgactccatacattgaataattccacacgtcagctcatcggttagcaaggtccagtagttgaagtcatttatttttccccgcggctggccaaatctacctctgggaatatccaagttgtcgaatatgatcgcaccggctctggtcatggtgaaggaactg tagcataaagacgcaggtatcataggggtaatatttttttattcactcacatactaaaagtaacgcatattagcaccatgtatgggctatcaattgacatttgcgtagcactacatcacgattatgtacaacataatgggacaacatatggcaagtagatgcaatttcctcacactagttgggtttatctactattgaattttcccctatctgtgatacacttgggagcctctacaagcatattgccatcatgtacgtttttatctactgtcttaacgcccatgggaacggaggcgtcgtcgtcatgtattggacggcaacataggcagcaacacaaattgcgtttaggtggggtgcatgtggactcgataccaagcccctgcagctggggaacgtctggtggagagccgataatttgatatacgcacgccatattactgtcgttgaagtacgccttatcttctatgttttcaaatttaggttcccaagtggacgtgagaagtgtttgtatctcacatggaatggcccaaggcattccagcccaggtgcctggtactttaatggcaaacaaacgttttggtagaggtattgattctattgcagttctgcagatatctgcagccccgagtatccacaggctatacgatacgttatcggaggcaagctgcggccgctctagaactagtggatcccccgggctgcagcccaatgtggaattcgcccttgcacattgttactcctgcatcttaaaaatatatcctgtagtaattttcacagcaatgtcataacatcatctcgctaaagaatgacctgggattggagaagtaatgaatatttgcaaccaatgcattgaataaactaacattaaacgaattcactagtggatcccccaactccgcccgttttatgactagaaccaatagtttttaatgccaaatgcactgaaatcccctaatttgcaaagccaaacgccccctatgtgagtaatacggggactttt tacccaatttcccaagcggaaagccccctaatacactcatatggcatatgaatcagcacggtcatgcactctaatggcggcccatagggactttccacatagggggcgttcaccatttcccagcataggggtggtgactcaatggcctttacccaagtacattgggtcaatgggaggtaagccaatgggtttttcccattactggcaagcacactgagtcaaatgggactttccactgggttttgcccaagtacattgggtcaatgggaggtgagccaatgggaaaaacccattgctgccaagtacactgactcaatagggactttccaatgggtttttccattgttggcaagcatataaggtcaatgtgggtgagtcaatagggactttccattgtattctgcccagtacataaggtcaatagggggtgaatcaacaggaaagtcccattggagccaagtacactgcgtcaatagggactttccattgggttttgcccagtacataaggtcaataggggatgagtcaatgggaaaaacccattggagccaagtacactgactcaatagggactttccattgggttttgcccagtacataaggtcaatagggggtgagtcaacaggaaagtcccattggagccaagtacattgagtcaatagggactttccaatgggttttgcccagtacataaggtcaatgggaggtaagccaatgggtttttcccattactggcacgtatactgagtcattagggactttccaatgggttttgcccagtacataaggtcaataggggtgaatcaacaggaaagtcccattggagccaagtacactgagtcaatagggactttccattgggttttgcccagtacaaaaggtcaatagggggtgagtcaatgggtttttcccattattggcacgtacataaggtcaataggggtgagtcattgggtttttccagccaatttaattaaaacgccatgtactttcccaccat tgacgtcaatgggctattgaaactaatgcaacgtgacctttaaacggtactttcccatagctgattaatgggaaagtaccgttctcgagccaatacacgtcaatgggaagtgaaagggcagccaaaacgtaacaccgccccggttttcccctggaaattccatattggcacgcattctattggctgagctgcgttctacgtgggtataagaggcgcgaccagcgtcggtaccgtcgcagtcttcggtctgaccaccgtagaacgcagagctcctcgctgcaggcggccgctctagaactcgtcgatcgcagcgatgacaaacctgcaagatcaaacccaacagattgttccgttcatacggagccttctgatgccaacaaccggaccggcgtccattccggacgacaccctggagaagcacactctcaggtcagagacctcgacctacaatttgactgtgggggacacagggtcagggctaattgtctttttccctggattccctggctcaattgtgggtgctcactacacactgcagagcaatgggaactacaagttcgatcagatgctcctgactgcccagaacctaccggccagctacaactactgcagactagtgagtcggagtctcacagtgaggtcaagcacactccctggtggcgtttatgcactaaacggcaccataaacgccgtgaccttccaaggaagcctgagtgaactgacagatgttagctacaatgggttgatgtctgcaacagccaacatcaacgacaaaattgggaatgtcctggtaggggaaggggtcactgtcctcagcctacccacatcatatgatcttgggtatgtgaggcttggtgaccccattcccgctatagggcttgacccaaaaatggtagctacatgcgacagcagtgacaggcccagagtctacaccataactgcagccgatgattaccaattctcatcacagtaccaaccaggtggggtaacaatcacactg ttctcagccaacattgatgctatcacaagcctcagcattgggggagagctcgtgtttcaaacaagcgtccaaggccttgtactgggcgccaccatctaccttataggctttgatgggactgcggtaatcaccagagctgtagccgcagataatgggctgacggccggcaccgacaatcttatgccattcaatcttgtcattccaaccaatgagataacccagccaatcacatccatcaaactggagatagtgacctccaaaagtggtggtcaggcaggggatcagatgtcatggtcggcaagtgggagcctagcagtgacgatccatggtggcaactatccaggggccctccgtcccgtcacactagtagcctacgaaagagtggcaacaggatccgtcgttacggtcgctggggtgagtaacttcgagctgattccaaatcctgaactagcaaagaacctggttacagaatacggccgatttgacccaggagccatgaactacacaaaattgatactgagtgagagggaccgtcttggcatcaagaccgtctggccaacaagggagtacactgattttcgtgagtacttcatggaggtggccgacctcaactctcccctgaagattgcaggagcatttggcttcaaagacataatccgggctataaggaggtaagcttgatctagagcggccgcggggatccagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttcggatcctctagagtcgacaattattttatttaataacatatagcccaaagacctctatgaacatttagtttcccgtatactcaacggcgcgtgtacacacaaggg cgaattccacagtggatatcaagcttagcttgcctccgattctagcattacatagccggtcagtagatcctgccattcggtagcgcaaccggctacatcttcaaacagtctcacgataaatgcatctctcgttcctgccaatccggaaccgggcataccactcccgcctgccgatttaattctcacaattgggcgatgccggcggggcaaaacgaatgtggatttggcaaaccgacacaggtctgctgtacggactaatatgggcacacccacatcattcttcagatgctccatgcattgttctatgagaaagatccatagggtggaggcagcgtcacgagatcgcccaggcaatcgatcgcattcgtctagtaaagtgacgagagttatcatgcacacacccatgcccacgccttccgaataactggagctgtggaagatcggaaacgtctttttgactgccggtctcgtactactttcgcacaggtgtatacccggacgcgtactatatattttatatcatccaacgtccgaaattacatacgtggcggcgatggaagtagatgttgagtcttcgaaagtaagtgcctcgaatatgggtattgtctgtgaaaatatcgaaagcggtacgacggttgcagaaccgtcgatgtcgccagatactagtaacaatagcttcgataacgaagacttccgtgggcctgaatacgatgtggagataaataccagaaaatctgctaatcttgatcgtatggaatcttcgtgccgtgaacaacgagcggcgtgcgaacttcgaaagtgttcgtgtcctacgtctgccgtgcgcatgcaatacagtattctttcatctctcgctccgggttcagagggtcatgtatatatatgtactagatacggggacgcggaccaaaaaaaatgcatagtgaaggcagtcgttggaggaaagaatcccgggagggaagtggatattttaaaaaccatctcacataaatcaattata aaattaatccatgcctataaatggaaaaatgttgtgtgtatggcaatgcgtgtatatcgttatgatcttttcacatatattgacggagtcggccctatgccccttcaacagatgatctatattcaacgtggactactagaggcgctagcatacatacatgaaaggggcatcattcaccgagacgtaaagacggagaatatattcttggataatcacgaaaatgcagttttgggtgacttcggtgctgcatgccaactaggagattgtatagatacgccccaatgttacggttggagcggaactgtggaaacaaattcgccggaattatctgcacttgatccgtattgcacaaaaacagatatttggagtgccggattggttctatatgagatggcaattaaaaatgtaccattgtttagtaagcaggtgaaaagttcgggatctcagctgagatccataatacggtgcatgcaagtgcatgaactggagtttccccgcaacgattctaccaacctctgtaaacatttcaaacaatatgcggttcgtgtacgaccgccttataccattcctcgagttataagaaatggggggatgccaatggatgttgaatatgtcatttctaaaatgcttacgtttgaccaggagttcagaccttctgctaaggaaatattgaatatgcccctatttactaaggcgccgattaacctgcttaatatcacaccctctgacagtgtctaacggtatacaggcgggagcgggtcgtggcgtcatcatcaccacttgagaatttatattttgaattgttgattgataaattaacctgattcattgagaactgaaacgccatattggtttcttggatatgtctacaacaattagttaaattgctatgttctactgcgagtaacatttgataagttgtaagagacgggcgactcatgtcgaagttgacgaatataaagtacataacgtgtttagaatacccagaatccgaatagtc cgcgggggcgtcttctcgcgtgagtaccaaatactgagttgaacttgaaaatgctaaatctgtgacactctttgtgtgatgattattgtcaccacttcgaagatggcttcgacattcatgatgttctggtgtttgtttggaatcgtaatagcgcttgtttcgtccaagtctgacaacaaagaaaatctgaagaattatatcacggataagtcaaccaatattagaatacccacgccattatttgtatcaacggaaaactcttatcccacaaaacatgtaatctacgatgaaaactgtggcttcgctgtactcaatcctataagtgaccccaaatatgtccttttgagccagcttctaatgggaaggcgcaaatatgatgcgacggtcgcgtggtttgttctcggtaaaatgtgtgccagattaatatatttgcgcgaattttataactgctcgacaaatgagccttttggcacatgttctatgagctctcctggatggtgggacaggcgctacgtctcaaccagtttcatttctcgcgacgaattacagctggtttttgcagcgccgtcccgagaattagatggtttatatacgcgcgtagtagttgtcaacggggactttactacggccgatataatgtttaatgttaaagtggcatgtgccttttcaaagactggaatagaagatgatacattatgcaaaccctttcatttctttgccaatgcaacattgcacaatttaaccatgattagatcggtaactcttcgagcgcacgaaagccatttaaaggaatgggtggcacggagaggtggtaacgtccctgcagtgctacttgagtctaccatgtatcatgcatccaatctgcctagaaatttcagggatttctacataaagtctccagatgattataagtataatcacctagatgggccatctgtaatgctcatcactgacagacctagtgaagatttggatgggaggctcgttcaccaaagtgaca tttttactactacaagtcctataaaacaggtccggtatgaagagcatcagtcacatacaaagcagtatcctgtaaacaaaatacaagctataatttttttgatagggttaggctcgttcattggaagcatattcgtagttttggtagtatggattatacgcagatattgcaatggagcgcggagtgggggaacgccccccagtcctcgccggtatgtgtataccaggctatgatcacgtgtgaaacttgggcggacctgtatcatatgtacaccgtccctattcgtttatagccagtacgtgttatctgcacatagaggaacatgtgtcatactgggatcgcatgcatggtatgtgtgactctaatattattctgtatcataataaaaacacagtgcatggtatatagaggatcgctggtaagcactacggtagaccaatcggctcagattgcattctttggcatcgataccgttgttaatttatatggcaaagtcttgttcatgggagatcagtatttggaggaaatatactctggaacgatggaaatactcaaatggaatcaagctaaccgctgctattctattgcgcatgcaacatattacgccgactgtcctataatcagttctacggtattcagaggatgccgggacgccgttgtttatactaggccccacagcag

The present invention should not be limited in scope to the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

In addition, it should be understood that all base sizes or amino acid sizes and all molecular weights or molecular weight values given for nucleic acids or polypeptides are approximate and are provided for description.

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SEQUENCE LIST

<110> Intervet, Inc.

Intervet International BV

Cook, Stephanie M

Morsey, Mohamad

Tarpey, Ian

<120> RECOMBINANT NON-PATHOGENIC MARK DISEASE VIRUS CONSTRUCTS,

ENCODING MULTIPLE HETEROLOGUE ANTIGENS

<130> 24522

<150> US 62/571,524

<151> 2017-10-12

<150> US 62/729,673

<151> 2018-09-11

<160> 32

<170>PatentIn version 3.5

<210> 1

<211> 1134

<212> DNA

<213> Infectious laryngotracheitis virus

<400> 1

atggaccgcc atttattttt gaggaatgct ttttggacta tcgtactgct ttcttccttc 60

gctagccaga gcaccgccgc cgtcacgtac gactacattt taggccgtcg cgcgctcgac 120

gcgctaacca taccggcggt tggcccgtat aacagatacc tcactagggt atcaagaggc 180

tgcgacgttg tcgagctcaa cccgatttct aacgtggacg acatgatatc ggcggccaaa 240

gaaaaagaga aggggggccc tttcgaggcc tccgtcgtct ggttctacgt gattaagggc 300

gacgacggcg aggacaagta ctgtccaatc tatagaaaag agtacaggga atgtggcgac 360

gtacaactgc tatctgaatg cgccgttcaa tctgcacaga tgtgggcagt ggactatgtt 420

cctagcaccc ttgtatcgcg aaatggcgcg ggactgacta tattctcccc cactgctgcg 480

ctctctggcc aatacttgct gaccctgaaa atcgggagat ttgcgcaaac agctctcgta 540

actctagaag ttaacgatcg ctgtttaaag atcgggtcgc agcttaactt tttaccgtcg 600

aaatgctgga caacagaaca gtatcagact ggatttcaag gcgaacacct ttatccgatc 660

gcagacacca atacacgaca cgcggacgac gtatatcggg gatacgaaga tattctgcag 720

cgctggaata atttgctgag gaaaaagaat cctagcgcgc cagaccctcg tccagatagc 780

gtcccgcaag aaattcccgc tgtaaccaag aaagcggaag ggcgcacccc ggacgcagaa 840

agcagcgaaa agaaggcccc tccagaagac tcggaggacg acatgcaggc agaggcttct 900

ggagaaaatc ctgccgccct ccccgaagac gacgaagtcc ccgaggacac cgagcacgat 960

gatccaaact cggatcctga ctattacaat gacatgcccg ccgtgatccc ggtggaggag 1020

actactaaaa gttctaatgc cgtctccatg cccatattcg cggcgttcgt agcctgcgcg 1080

gtcgcgctcg tggggctact ggtttggagc atcgtaaaat gcgcgcgtag ctaa 1134

<210> 2

<211> 377

<212> Protein

<213> Infectious laryngotracheitis virus

<400> 2

Met Asp Arg His Leu Phe Leu Arg Asn Ala Phe Trp Thr Ile Val Leu

1 5 10 15

Leu Ser Ser Phe Ala Ser Gln Ser Thr Ala Ala Val Thr Tyr Asp Tyr

20 25 30

Ile Leu Gly Arg Arg Ala Leu Asp Ala Leu Thr Ile Pro Ala Val Gly

35 40 45

Pro Tyr Asn Arg Tyr Leu Thr Arg Val Ser Arg Gly Cys Asp Val Val

50 55 60

Glu Leu Asn Pro Ile Ser Asn Val Asp Asp Met Ile Ser Ala Ala Lys

65 70 75 80

Glu Lys Glu Lys Gly Gly Pro Phe Glu Ala Ser Val Val Trp Phe Tyr

85 90 95

Val Ile Lys Gly Asp Asp Gly Glu Asp Lys Tyr Cys Pro Ile Tyr Arg

100 105 110

Lys Glu Tyr Arg Glu Cys Gly Asp Val Gln Leu Leu Ser Glu Cys Ala

115 120 125

Val Gln Ser Ala Gln Met Trp Ala Val Asp Tyr Val Pro Ser Thr Leu

130 135 140

Val Ser Arg Asn Gly Ala Gly Leu Thr Ile Phe Ser Pro Thr Ala Ala

145 150 155 160

Leu Ser Gly Gln Tyr Leu Leu Thr Leu Lys Ile Gly Arg Phe Ala Gln

165 170 175

Thr Ala Leu Val Thr Leu Glu Val Asn Asp Arg Cys Leu Lys Ile Gly

180 185 190

Ser Gln Leu Asn Phe Leu Pro Ser Lys Cys Trp Thr Thr Glu Gln Tyr

195 200 205

Gln Thr Gly Phe Gln Gly Glu His Leu Tyr Pro Ile Ala Asp Thr Asn

210 215 220

Thr Arg His Ala Asp Asp Val Tyr Arg Gly Tyr Glu Asp Ile Leu Gln

225 230 235 240

Arg Trp Asn Asn Leu Leu Arg Lys Lys Asn Pro Ser Ala Pro Asp Pro

245 250 255

Arg Pro Asp Ser Val Pro Gln Glu Ile Pro Ala Val Thr Lys Lys Ala

260 265 270

Glu Gly Arg Thr Pro Asp Ala Glu Ser Ser Glu Lys Lys Ala Pro Pro

275 280 285

Glu Asp Ser Glu Asp Asp Met Gln Ala Glu Ala Ser Gly Glu Asn Pro

290 295 300

Ala Ala Leu Pro Glu Asp Asp Glu Val Pro Glu Asp Thr Glu His Asp

305 310 315 320

Asp Pro Asn Ser Asp Pro Asp Tyr Tyr Asn Asp Met Pro Ala Val Ile

325 330 335

Pro Val Glu Glu Thr Thr Lys Ser Ser Asn Ala Val Ser Met Pro Ile

340 345 350

Phe Ala Ala Phe Val Ala Cys Ala Val Ala Leu Val Gly Leu Leu Val

355 360 365

Trp Ser Ile Val Lys Cys Ala Arg Ser

370 375

<210> 3

<211> 1089

<212> DNA

<213> Infectious laryngotracheitis virus

<400> 3

atggcatcgc tacttggaac tctggctctc cttgccgcga cgctcgcacc cttcggcgcg 60

atgggaatcg tgatcactgg aaatcacgtc tccgccagga ttgacgacga tcacatcgtg 120

atcgtcgcgc ctcgccccga agctacaatt caactgcagc tatttttcat gcctggccag 180

agaccccaca aaccctactc aggaaccgtc cgcgtcgcgt ttcggtctga tataacaaac 240

cagtgctacc aggaacttag cgaggagcgc tttgaaaatt gcactcatcg atcgtcttct 300

gtttttgtcg gctgtaaagt gaccgagtac acgttctccg cctcgaacag actaaccgga 360

cctccacacc cgtttaagct cactatacga aatcctcgtc cgaacgacag cgggatgttc 420

tacgtaattg ttcggctaga cgacaccaaa gaacccattg acgtcttcgc gatccaacta 480

tcggtgtatc aattcgcgaa caccgccgcg actcgcggac tctattccaa ggcttcgtgt 540

cgcaccttcg gattacctac cgtccaactt gaggcctatc tcaggaccga ggaaagttgg 600

cgcaactggc aagcgtacgt tgccacggag gccacgacga ccagcgccga ggcgacaacc 660

ccgacgcccg tcactgcaac cagcgcctcc gaacttgaag cggaacactt tacctttccc 720

tggctagaaa atggcgtgga tcattacgaa ccgacacccg caaacgaaaa ttcaaacgtt 780

actgtccgtc tcgggacaat gagccctacg ctaattgggg taaccgtggc tgccgtcgtg 840

agcgcaacga tcggcctcgt cattgtaatt tccatcgtca ccagaaacat gtgcaccccg 900

caccgaaaat tagacacggt ctcgcaagac gacgaagaac gttcccaaac tagaagggaa 960

tcgcgaaaat ttggacccat ggttgcgtgc gaaataaaca agggggctga cggatagt 1020

gaacttgtgg aactggttgc gattgttaac ccgtctgcgc taagctcgcc cgactcaata 1080

aaaatgtga 1089

<210> 4

<211> 362

<212> Protein

<213> Infectious laryngotracheitis virus

<400> 4

Met Ala Ser Leu Leu Gly Thr Leu Ala Leu Leu Ala Ala Thr Leu Ala

1 5 10 15

Pro Phe Gly Ala Met Gly Ile Val Ile Thr Gly Asn His Val Ser Ala

20 25 30

Arg Ile Asp Asp Asp His Ile Val Ile Val Ala Pro Arg Pro Glu Ala

35 40 45

Thr Ile Gln Leu Gln Leu Phe Phe Met Pro Gly Gln Arg Pro His Lys

50 55 60

Pro Tyr Ser Gly Thr Val Arg Val Ala Phe Arg Ser Asp Ile Thr Asn

65 70 75 80

Gln Cys Tyr Gln Glu Leu Ser Glu Glu Arg Phe Glu Asn Cys Thr His

85 90 95

Arg Ser Ser Ser Val Phe Val Gly Cys Lys Val Thr Glu Tyr Thr Phe

100 105 110

Ser Ala Ser Asn Arg Leu Thr Gly Pro Pro His Pro Phe Lys Leu Thr

115 120 125

Ile Arg Asn Pro Arg Pro Asn Asp Ser Gly Met Phe Tyr Val Ile Val

130 135 140

Arg Leu Asp Asp Thr Lys Glu Pro Ile Asp Val Phe Ala Ile Gln Leu

145 150 155 160

Ser Val Tyr Gln Phe Ala Asn Thr Ala Ala Thr Arg Gly Leu Tyr Ser

165 170 175

Lys Ala Ser Cys Arg Thr Phe Gly Leu Pro Thr Val Gln Leu Glu Ala

180 185 190

Tyr Leu Arg Thr Glu Glu Ser Trp Arg Asn Trp Gln Ala Tyr Val Ala

195 200 205

Thr Glu Ala Thr Thr Thr Ser Ala Glu Ala Thr Thr Pro Thr Pro Val

210 215 220

Thr Ala Thr Ser Ala Ser Glu Leu Glu Ala Glu His Phe Thr Phe Pro

225 230 235 240

Trp Leu Glu Asn Gly Val Asp His Tyr Glu Pro Thr Pro Ala Asn Glu

245 250 255

Asn Ser Asn Val Thr Val Arg Leu Gly Thr Met Ser Pro Thr Leu Ile

260 265 270

Gly Val Thr Val Ala Ala Val Val Ser Ala Thr Ile Gly Leu Val Ile

275 280 285

Val Ile Ser Ile Val Thr Arg Asn Met Cys Thr Pro His Arg Lys Leu

290 295 300

Asp Thr Val Ser Gln Asp Asp Glu Glu Arg Ser Gln Thr Arg Arg Glu

305 310 315 320

Ser Arg Lys Phe Gly Pro Met Val Ala Cys Glu Ile Asn Lys Gly Ala

325 330 335

Asp Gln Asp Ser Glu Leu Val Glu Leu Val Ala Ile Val Asn Pro Ser

340 345 350

Ala Leu Ser Ser Pro Asp Ser Ile Lys Met

355 360

<210> 5

<211> 1362

<212> DNA

<213> Infectious bursitis virus

<400> 5

atgacaaacc tgcaagatca aacccaacag attgttccgt tcatacggag ccttctgatg 60

ccaacaaccg gaccggcgtc cattccggac gacaccctgg agaagcacac tctcaggtca 120

gagacctcga cctacaattt gactgtgggg gacacagggt cagggctaat tgtctttttc 180

cctggattcc ctggctcaat tgtgggtgct cactacacac tgcagagcaa tgggaactac 240

aagttcgatc agatgctcct gactgcccag aacctaccgg ccagctacaa ctactgcaga 300

ctagtgagtc ggagtctcac agtgaggtca agcacactcc ctggtggcgt ttatgcacta 360

aacggcacca taaacgccgt gaccttccaa ggaagcctga gtgaactgac agatgttagc 420

tacaatgggt tgatgtctgc aacagccaac atcaacgaca aaattgggaa tgtcctggta 480

ggggaagggg tcactgtcct cagcctaccc acatcatatg atcttgggta tgtgaggctt 540

ggtgacccca ttcccgctat agggcttgac ccaaaaatgg tagctacatg cgacagcagt 600

gacaggccca gagtctacac cataactgca gccgatgatt accaattctc atcacagtac 660

caaccaggtggggtaacaat cacactgttc tcagccaaca ttgatgctat cacaagcctc 720

agcattgggg gagagctcgt gtttcaaaca agcgtccaag gccttgtact gggcgccacc 780

atctacctta taggctttga tgggactgcg gtaatcacca gagctgtggc cgcagataat 840

gggctgacgg ccggcaccga caatcttatg ccattcaatc ttgtcattcc aaccaatgag 900

ataacccagc caatcacatc catcaaactg gagatagtga cctccaaaag tggtggtcag 960

gcaggggatc agatgtcatg gtcggcaagt gggagcctag cagtgacgat ccatggtggc 1020

aactatccag gggccctccg tcccgtcaca ctagtagcct acgaaagagt ggcaacagga 1080

tccgtcgtta cggtcgctgg ggtgagtaac ttcgagctga ttccaaatcc tgaactagca 1140

aagaacctgg ttacagaata cggccgattt gacccaggag ccatgaacta cacaaaattg 1200

atactgagtg agagggaccg tcttggcatc aagaccgtct ggccaacaag ggagtacact 1260

gattttcgtg agtacttcat ggaggtggcc gacctcaact ctcccctgaa gattgcagga 1320

gcatttggct tcaaagacat aatccggggct ataaggaggt aa 1362

<210> 6

<211> 453

<212> Protein

<213> Infectious bursitis virus

<400> 6

Met Thr Asn Leu Gln Asp Gln Thr Gln Gln Ile Val Pro Phe Ile Arg

1 5 10 15

Ser Leu Leu Met Pro Thr Thr Gly Pro Ala Ser Ile Pro Asp Asp Thr

20 25 30

Leu Glu Lys His Thr Leu Arg Ser Glu Thr Ser Thr Tyr Asn Leu Thr

35 40 45

Val Gly Asp Thr Gly Ser Gly Leu Ile Val Phe Phe Pro Gly Phe Pro

50 55 60

Gly Ser Ile Val Gly Ala His Tyr Thr Leu Gln Ser Asn Gly Asn Tyr

65 70 75 80

Lys Phe Asp Gln Met Leu Leu Thr Ala Gln Asn Leu Pro Ala Ser Tyr

85 90 95

Asn Tyr Cys Arg Leu Val Ser Arg Ser Leu Thr Val Arg Ser Ser Thr

100 105 110

Leu Pro Gly Gly Val Tyr Ala Leu Asn Gly Thr Ile Asn Ala Val Thr

115 120 125

Phe Gln Gly Ser Leu Ser Glu Leu Thr Asp Val Ser Tyr Asn Gly Leu

130 135 140

Met Ser Ala Thr Ala Asn Ile Asn Asp Lys Ile Gly Asn Val Leu Val

145 150 155 160

Gly Glu Gly Val Thr Val Leu Ser Leu Pro Thr Ser Tyr Asp Leu Gly

165 170 175

Tyr Val Arg Leu Gly Asp Pro Ile Pro Ala Ile Gly Leu Asp Pro Lys

180 185 190

Met Val Ala Thr Cys Asp Ser Ser Asp Arg Pro Arg Val Tyr Thr Ile

195 200 205

Thr Ala Ala Asp Asp Tyr Gln Phe Ser Ser Gln Tyr Gln Pro Gly Gly

210 215 220

Val Thr Ile Thr Leu Phe Ser Ala Asn Ile Asp Ala Ile Thr Ser Leu

225 230 235 240

Ser Ile Gly Gly Glu Leu Val Phe Gln Thr Ser Val Gln Gly Leu Val

245 250 255

Leu Gly Ala Thr Ile Tyr Leu Ile Gly Phe Asp Gly Thr Ala Val Ile

260 265 270

Thr Arg Ala Val Ala Ala Asp Asn Gly Leu Thr Ala Gly Thr Asp Asn

275 280 285

Leu Met Pro Phe Asn Leu Val Ile Pro Thr Asn Glu Ile Thr Gln Pro

290 295 300

Ile Thr Ser Ile Lys Leu Glu Ile Val Thr Ser Lys Ser Gly Gly Gln

305 310 315 320

Ala Gly Asp Gln Met Ser Trp Ser Ala Ser Gly Ser Leu Ala Val Thr

325 330 335

Ile His Gly Gly Asn Tyr Pro Gly Ala Leu Arg Pro Val Thr Leu Val

340 345 350

Ala Tyr Glu Arg Val Ala Thr Gly Ser Val Val Thr Val Ala Gly Val

355 360 365

Ser Asn Phe Glu Leu Ile Pro Asn Pro Glu Leu Ala Lys Asn Leu Val

370 375 380

Thr Glu Tyr Gly Arg Phe Asp Pro Gly Ala Met Asn Tyr Thr Lys Leu

385 390 395 400

Ile Leu Ser Glu Arg Asp Arg Leu Gly Ile Lys Thr Val Trp Pro Thr

405 410 415

Arg Glu Tyr Thr Asp Phe Arg Glu Tyr Phe Met Glu Val Ala Asp Leu

420 425 430

Asn Ser Pro Leu Lys Ile Ala Gly Ala Phe Gly Phe Lys Asp Ile Ile

435 440 445

Arg Ala Ile Arg Arg

450

<210> 7

<211> 1662

<212> DNA

<213> Newcastle disease virus

<400> 7

atgggcccca gaccttctac caagaaccca gtacctatga tgctgactgt ccgagtcgcg 60

ctggtactga gttgcatctg tccggcaaac tccattgatg gcaggcctct tgcggctgca 120

ggaattgtgg ttacaggaga caaagccgtc aacatataca cctcatccca gacaggatca 180

atcatagtta agctcctccc gaatctgccc aaggataagg aggcatgtgc gaaagccccc 240

ttggatgcat acaacaggac attgaccact ttgctcaccc cccttggtga ctctatccgt 300

aggatacaag agtctgtgac tacatctgga ggggggagac aggggcgcct tataggcgcc 360

attattggcg gtgtggctct tggggttgca actgccgcac aaataacagc ggccgcagct 420

ctgatacaag ccaaacaaaa tgctgccaac atcctccgac ttaaagagag cattgccgca 480

accaatgagg ctgtgcatga ggtcactgac ggattatcgc aactagcagt ggcagttggg 540

aagatgcagc agtttgttaa tgaccaattt aataaaacag ctcaggaatt agactgcatc 600

aaaattgcac agcaagttgg tgtagagctc aacctgtacc taaccgaatt gactacagta 660

ttcggaccac aaatcacttc acctgcttta aacaagctga ctattcaggc actttacaat 720

ctagctggtg gaaatatgga ttacttattg actaagttag gtgtaggggaa caatcaactc 780

agctcattaa tcggtagcgg cttaatcacc ggtaacccta ttctatacga ctcacagact 840

caactcttgg gtatacaggt aactctacct tcagtcggga agctaaataa tatgcgtgcc 900

acctacttgg aaaccttatc cgtaagcaca accaggggat ttgcctcggc acttgtccca 960

aaagtggtga cacaggtcgg ttctgtgata gaagaacttg acacctcata ctgtatagaa 1020

actgacttac atttatattg tacaagaata gtaacgttcc ctatgtcccc tggtatttat 1080

tcctgcttga gcggcaatac gtcggcctgt atgtactcaa agaccgaagg cgcacttact 1140

acaccataca tgactatcaa aggttcagtc atcgccaact gcaagatgac aacatgtaga 1200

tgtgtaaacc ccccgggtat catatcgcaa aactatggag aagccgtgtc tctaatagat 1260

aaacaatcat gcaatgtttt atccttaggc gggataactt taaggctcag tggggaattc 1320

gatgtaactt atcagaagaa tatctcaata caagattctc aagtaataat aacaggcaat 1380

cttgatatct caactgagct tgggaatgtc aacaactcga tcagtaatgc tttgaataag 1440

ttagaggaaa gcaacagaaa actagacaaa gtcaatgtca aactgactag cacatctgct 1500

ctcattacct atatcgtgtt gactatcata tctcttgttt ttggtatact tagcctgatt 1560

ctagcatgct acctaatgta caagcaaaag gcgcaacaaa agaccttatt atggcttggg 1620

aataatactc tagatcagat gagagccact acaaaaatgt ga 1662

<210> 8

<211> 553

<212> Protein

<213> Newcastle disease virus

<400> 8

Met Gly Pro Arg Pro Ser Thr Lys Asn Pro Val Pro Met Met Leu Thr

1 5 10 15

Val Arg Val Ala Leu Val Leu Ser Cys Ile Cys Pro Ala Asn Ser Ile

20 25 30

Asp Gly Arg Pro Leu Ala Ala Ala Gly Ile Val Val Thr Gly Asp Lys

35 40 45

Ala Val Asn Ile Tyr Thr Ser Ser Gln Thr Gly Ser Ile Ile Val Lys

50 55 60

Leu Leu Pro Asn Leu Pro Lys Asp Lys Glu Ala Cys Ala Lys Ala Pro

65 70 75 80

Leu Asp Ala Tyr Asn Arg Thr Leu Thr Thr Leu Leu Thr Pro Leu Gly

85 90 95

Asp Ser Ile Arg Arg Ile Gln Glu Ser Val Thr Thr Ser Gly Gly Gly

100 105 110

Arg Gln Gly Arg Leu Ile Gly Ala Ile Ile Gly Gly Val Ala Leu Gly

115 120 125

Val Ala Thr Ala Ala Gln Ile Thr Ala Ala Ala Ala Leu Ile Gln Ala

130 135 140

Lys Gln Asn Ala Ala Asn Ile Leu Arg Leu Lys Glu Ser Ile Ala Ala

145 150 155 160

Thr Asn Glu Ala Val His Glu Val Thr Asp Gly Leu Ser Gln Leu Ala

165 170 175

Val Ala Val Gly Lys Met Gln Gln Phe Val Asn Asp Gln Phe Asn Lys

180 185 190

Thr Ala Gln Glu Leu Asp Cys Ile Lys Ile Ala Gln Gln Val Gly Val

195 200 205

Glu Leu Asn Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gly Pro Gln

210 215 220

Ile Thr Ser Pro Ala Leu Asn Lys Leu Thr Ile Gln Ala Leu Tyr Asn

225 230 235 240

Leu Ala Gly Gly Asn Met Asp Tyr Leu Leu Thr Lys Leu Gly Val Gly

245 250 255

Asn Asn Gln Leu Ser Ser Leu Ile Gly Ser Gly Leu Ile Thr Gly Asn

260 265 270

Pro Ile Leu Tyr Asp Ser Gln Thr Gln Leu Leu Gly Ile Gln Val Thr

275 280 285

Leu Pro Ser Val Gly Lys Leu Asn Asn Met Arg Ala Thr Tyr Leu Glu

290 295 300

Thr Leu Ser Val Ser Thr Thr Arg Gly Phe Ala Ser Ala Leu Val Pro

305 310 315 320

Lys Val Val Thr Gln Val Gly Ser Val Ile Glu Glu Leu Asp Thr Ser

325 330 335

Tyr Cys Ile Glu Thr Asp Leu His Leu Tyr Cys Thr Arg Ile Val Thr

340 345 350

Phe Pro Met Ser Pro Gly Ile Tyr Ser Cys Leu Ser Gly Asn Thr Ser

355 360 365

Ala Cys Met Tyr Ser Lys Thr Glu Gly Ala Leu Thr Thr Pro Tyr Met

370 375 380

Thr Ile Lys Gly Ser Val Ile Ala Asn Cys Lys Met Thr Thr Cys Arg

385 390 395 400

Cys Val Asn Pro Pro Gly Ile Ile Ser Gln Asn Tyr Gly Glu Ala Val

405 410 415

Ser Leu Ile Asp Lys Gln Ser Cys Asn Val Leu Ser Leu Gly Gly Ile

420 425 430

Thr Leu Arg Leu Ser Gly Glu Phe Asp Val Thr Tyr Gln Lys Asn Ile

435 440 445

Ser Ile Gln Asp Ser Gln Val Ile Ile Thr Gly Asn Leu Asp Ile Ser

450 455 460

Thr Glu Leu Gly Asn Val Asn Asn Ser Ile Ser Asn Ala Leu Asn Lys

465 470 475 480

Leu Glu Glu Ser Asn Arg Lys Leu Asp Lys Val Asn Val Lys Leu Thr

485 490 495

Ser Thr Ser Ala Leu Ile Thr Tyr Ile Val Leu Thr Ile Ile Ser Leu

500 505 510

Val Phe Gly Ile Leu Ser Leu Ile Leu Ala Cys Tyr Leu Met Tyr Lys

515 520 525

Gln Lys Ala Gln Gln Lys Thr Leu Leu Trp Leu Gly Asn Asn Thr Leu

530 535 540

Asp Gln Met Arg Ala Thr Thr Lys Met

545 550

<210> 9

<211> 1698

<212> DNA

<213> Newcastle disease virus

<400> 9

atggatcgat cccggttggc gccctccagg tgcaggatgg gctccagacc ttctaccaag 60

aacccagcac ctatgatgct gactatccgg gtcgcgctgg tactgagttg catctgtccg 120

gcaaactcca ttgatggcag gcctcttgca gctgcaggaa ttgtggttac aggagacaaa 180

gcagtcaaca tatacacctc atcccagaca ggatcaatca tagttaagct cctcccgaat 240

ctgccaaagg ataaggaggc atgtgcgaaa gcccccttgg atgcatacaa caggacattg 300

accactttgc tcacccccct tggtgactct atccgtagga tacaagagtc tgtgactaca 360

tctggagggg ggagacaggg gcgccttata ggcgccatta ttggcggtgt ggctcttggg 420

gttgcaactg ccgcacaaat aacagcggcc gcagctctga tacaagccaa acaaaatgct 480

gccaacatcc tccgacttaa agagagcatt gccgcaacca atgaggctgt gcatgaggtc 540

actgacggat tatcgcaact agcagtggca gttgggaaga tgcagcagtt cgttaatgac 600

caatttaata aaacagctca ggaattagac tgcatcaaaa ttgcacagca agttggtgta 660

gagctcaacc tgtacctaac cgaatcgact acagtattcg gaccacaaat cacttcacct 720

gccttaaaca agctgactat tcaggcactt tacaatctag ctggtgggaa tatggattac 780

ttattgacta agttaggtat agggaacaat caactcagct cattaatcgg tagcggctta 840

atcaccggta accctattct atacgactca cagactcaac tcttgggtat acaggtaact 900

ctaccttcag tcgggaacct aaataatatg cgtgccacct acttggaaac cttatccgta 960

agcacaacca ggggatttgc ctcggcactt gtcccaaaag tggtgacacg ggtcggttct 1020

gtgatagaag aacttgacac ctcatactgt atagaaactg acttagattt atattgtaca 1080

agaatagtaa cgttccctat gtcccctggt atttactcct gcttgagcgg caatacatcg 1140

gcctgtatgt actcaaagac cgaaggcgca cttactacac catatatgac tatcaaaggc 1200

tcagtcatcg ctaactgcaa gatgacaaca tgtagatgtg taaacccccc gggtatcata 1260

tcgcaaaact atggagaagc cgtgtctcta atagataaac aatcatgcaa tgttttatcc 1320

ttaggcggga taactttaag gctcagtggg gaattcgatg taacttatca gaagaatatc 1380

tcaatacaag attctcaagt aataataaca ggcaatcttg atatctcaac tgagcttggg 1440

aatgtcaaca actcgatcag taatgccttg aataagttag aggaaagcaa cagaaaacta 1500

gacaaagtca atgtcaaact gaccagcaca tctgctctca ttacctatat cgttttgact 1560

atcatatctc ttgtttttgg tatacttagc ctgattctag catgctacct aatgtacaag 1620

caaaaggcgc aacaaaagac cttattatgg cttgggaata ataccctaga tcagatgaga 1680

gccactacaa aaatgtga 1698

<210> 10

<211> 565

<212> Protein

<213> Newcastle disease virus

<400> 10

Met Asp Arg Ser Arg Leu Ala Pro Ser Arg Cys Arg Met Gly Ser Arg

1 5 10 15

Pro Ser Thr Lys Asn Pro Ala Pro Met Met Leu Thr Ile Arg Val Ala

20 25 30

Leu Val Leu Ser Cys Ile Cys Pro Ala Asn Ser Ile Asp Gly Arg Pro

35 40 45

Leu Ala Ala Ala Gly Ile Val Val Thr Gly Asp Lys Ala Val Asn Ile

50 55 60

Tyr Thr Ser Ser Gln Thr Gly Ser Ile Ile Val Lys Leu Leu Pro Asn

65 70 75 80

Leu Pro Lys Asp Lys Glu Ala Cys Ala Lys Ala Pro Leu Asp Ala Tyr

85 90 95

Asn Arg Thr Leu Thr Thr Leu Leu Thr Pro Leu Gly Asp Ser Ile Arg

100 105 110

Arg Ile Gln Glu Ser Val Thr Thr Ser Gly Gly Gly Arg Gln Gly Arg

115 120 125

Leu Ile Gly Ala Ile Ile Gly Gly Val Ala Leu Gly Val Ala Thr Ala

130 135 140

Ala Gln Ile Thr Ala Ala Ala Ala Leu Ile Gln Ala Lys Gln Asn Ala

145 150 155 160

Ala Asn Ile Leu Arg Leu Lys Glu Ser Ile Ala Ala Thr Asn Glu Ala

165 170 175

Val His Glu Val Thr Asp Gly Leu Ser Gln Leu Ala Val Ala Val Gly

180 185 190

Lys Met Gln Gln Phe Val Asn Asp Gln Phe Asn Lys Thr Ala Gln Glu

195 200 205

Leu Asp Cys Ile Lys Ile Ala Gln Gln Val Gly Val Glu Leu Asn Leu

210 215 220

Tyr Leu Thr Glu Ser Thr Thr Val Phe Gly Pro Gln Ile Thr Ser Pro

225 230 235 240

Ala Leu Asn Lys Leu Thr Ile Gln Ala Leu Tyr Asn Leu Ala Gly Gly

245 250 255

Asn Met Asp Tyr Leu Leu Thr Lys Leu Gly Ile Gly Asn Asn Gln Leu

260 265 270

Ser Ser Leu Ile Gly Ser Gly Leu Ile Thr Gly Asn Pro Ile Leu Tyr

275 280 285

Asp Ser Gln Thr Gln Leu Leu Gly Ile Gln Val Thr Leu Pro Ser Val

290 295 300

Gly Asn Leu Asn Asn Met Arg Ala Thr Tyr Leu Glu Thr Leu Ser Val

305 310 315 320

Ser Thr Thr Arg Gly Phe Ala Ser Ala Leu Val Pro Lys Val Val Thr

325 330 335

Arg Val Gly Ser Val Ile Glu Glu Leu Asp Thr Ser Tyr Cys Ile Glu

340 345 350

Thr Asp Leu Asp Leu Tyr Cys Thr Arg Ile Val Thr Phe Pro Met Ser

355 360 365

Pro Gly Ile Tyr Ser Cys Leu Ser Gly Asn Thr Ser Ala Cys Met Tyr

370 375 380

Ser Lys Thr Glu Gly Ala Leu Thr Thr Pro Tyr Met Thr Ile Lys Gly

385 390 395 400

Ser Val Ile Ala Asn Cys Lys Met Thr Thr Cys Arg Cys Val Asn Pro

405 410 415

Pro Gly Ile Ile Ser Gln Asn Tyr Gly Glu Ala Val Ser Leu Ile Asp

420 425 430

Lys Gln Ser Cys Asn Val Leu Ser Leu Gly Gly Ile Thr Leu Arg Leu

435 440 445

Ser Gly Glu Phe Asp Val Thr Tyr Gln Lys Asn Ile Ser Ile Gln Asp

450 455 460

Ser Gln Val Ile Ile Thr Gly Asn Leu Asp Ile Ser Thr Glu Leu Gly

465 470 475 480

Asn Val Asn Asn Ser Ile Ser Asn Ala Leu Asn Lys Leu Glu Glu Ser

485 490 495

Asn Arg Lys Leu Asp Lys Val Asn Val Lys Leu Thr Ser Thr Ser Ala

500 505 510

Leu Ile Thr Tyr Ile Val Leu Thr Ile Ile Ser Leu Val Phe Gly Ile

515 520 525

Leu Ser Leu Ile Leu Ala Cys Tyr Leu Met Tyr Lys Gln Lys Ala Gln

530 535 540

Gln Lys Thr Leu Leu Trp Leu Gly Asn Asn Thr Leu Asp Gln Met Arg

545 550 555 560

Ala Thr Thr Lys Met

565

<210> 11

<211> 527

<212> DNA

<213> Infectious laryngotracheitis virus

<400> 11

aaacagctgt actacagagt aaccgatgga agaacatcgg tccagctaat gtgcctgtcg 60

tgcacgagcc attctccgga accttactgt cttttcgaca cgtctcttat agcgagggaa 120

aaagatatcg cgccagagtt atactttacc tctgatccgc aaacggcata ctgcacaata 180

actctgccgt ccggcgttgt tccgagattc gaatggagcc ttaataatgt ttcactgccg 240

gaatatttga cggccacgac cgttgtttcg cataccgctg gccaaagtac agtgtggaag 300

agcagcgcga gagcaggcga ggcgtggatt tctggccggg gaggcaatat atacgaatgc 360

accgtcctca tctcagacgg cactcgcgtt actacgcgaa aggagaggtg cttaacaaac 420

acatggattg cggtggaaaa cggtgctgct caggcgcagc tgtattcact cttttctgga 480

cttgtgtcag gattatgcgg gagcatatct gctttgtacg caacgct 527

<210> 12

<211> 264

<212> DNA

<213> Infectious laryngotracheitis virus

<400> 12

tgactattac aatgacatgc ccgccgtgat cccggtggag gagactacta aaagttctaa 60

tgccgtctcc atgcccatat tcgcggcgtt cgtagcctgc gcggtcgcgc tcgtggggct 120

actggtttgg agcatcgtaa aatgcgcgcg tagctaatcg agcctagaat aggtggtttc 180

ttcctacatg ccacgcctca cgctcataat ataaatcaca tggaatagca taccaatgcc 240

tattcattgg gacgttcgaa aagc 264

<210> 13

<211> 1391

<212> DNA

<213> Mouse cytomegalovirus 1

<400> 13

aactccgccc gttttatgac tagaaccaat agtttttaat gccaaatgca ctgaaatccc 60

ctaatttgca aagccaaacg ccccctatgt gagtaatacg gggacttttt acccaatttc 120

ccacgcggaa agccccctaa tacactcata tggcatatga atcagcacgg tcatgcactc 180

taatggcggc ccatagggac tttccacata gggggcgttc accatttccc agcataggggg 240

tggtgactca atggccttta cccaagtaca ttgggtcaat gggaggtaag ccaatgggtt 300

tttcccatta ctggcaagca cactgagtca aatgggactt tccactgggt tttgcccaag 360

tacattgggt caatgggagg tgagccaatg ggaaaaaccc attgctgcca agtacactga 420

ctcaataggg actttccaat gggtttttcc attgttggca agcatataag gtcaatgtgg 480

gtgagtcaat agggactttc cattgtattc tgcccagtac ataaggtcaa tagggggtga 540

atcaacagga aagtcccatt ggagccaagt acactgcgtc aatagggact ttccattggg 600

ttttgcccag tacataaggt caatagggga tgagtcaatg ggaaaaaccc attggagcca 660

agtacactga ctcaataggg actttccatt gggttttgcc cagtacataa ggtcaatagg 720

gggtgagtca acaggaaagt tccattggag ccaagtacat tgagtcaata gggactttcc 780

840

ctggcacgta tactgagtca ttagggactt tccaatgggt tttgcccagt acataaggtc 900

aataggggtg aatcaacagg aaagtcccat tggagccaag tacactgagt caatagggac 960

tttccattgg gttttgccca gtacaaaagg tcaatagggg gtgagtcaat gggtttttcc 1020

cattattggc acgtacataa ggtcaatagg ggtgagtcat tgggtttttc cagccaattt 1080

aattaaaacg ccatgtactt tcccaccatt gacgtcaatg ggctattgaa actaatgcaa 1140

cgtgaccttt aaacggtact ttcccatagc tgattaatgg gaaagtaccg ttctcgagcc 1200

aatacacgtc aatgggaagt gaaagggcag ccaaaacgta acaccgcccc ggttttcccc 1260

tggaaattcc atattggcac gcattctatt ggctgagctg cgttctacgt gggtataaga 1320

ggcgcgacca gcgtcggtac cgtcgcagtc ttcggtctga ccaccgtaga acgcagagct 1380

cctcgctgca g 1391

<210> 14

<211> 301

<212> DNA

<213> Human cytomegalovirus

<400> 14

ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 60

tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 120

tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 180

ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 240

ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata 300

g 301

<210> 15

<211> 360

<212> DNA

<213> Human cytomegalovirus

<400> 15

cgcgccaggt caattccctg gcattatgcc cagtacatga ccttatggga ctttcctact 60

tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 120

atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 180

gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 240

tccgccccat tgacgcaaat gggcggtagc gtgtacggtg ggaggtctat ataagcagag 300

ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata 360

<210> 16

<211> 1191

<212> DNA

<213> Human cytomegalovirus

<400> 16

gtgaataata aaatgtgtgt ttgtccgaaa tacgcgtttg agatttctgt cccgactaaa 60

ttcatgtcgc gcgatagtgg tgtttatcgc cgatagagat ggcgatattg gaaaaatcga 120

tatttgaaaa tatggcatat tgaaaatgtc gccgatgtga gtttctgtgt aactgatatc 180

gccatttttc caaaagttga tttttgggca tacgcgatat ctggcgatac gcttatatcg 240

tttacggggg atggcgatag acgcctttgg tgacttgggc gattctgtgt gtcgcaaata 300

tcgcagtttc gatataggtg acagacgata tgaggctata tcgccgatag aggcgacatc 360

aagctggcac atggccaatg catatcgatc tatacattga atcaatattg gccattagcc 420

atattattca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg 480

tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc gccatgttga 540

cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 600

tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 660

gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 720

ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 780

gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 840

cattatgccc agtacatgac cttatgggac ttttcctactt ggcagtacat ctacgtatta 900

gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 960

tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 1020

caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 1080

ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag 1140

atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg g 1191

<210> 17

<211> 692

<212> DNA

<213> Gallus gallus

<220>

<221> other_features

<222> (141)..(143)

<223> n is a, c, g, or t

<400> 17

cgcgccggat cagatctcca tggtcgaggt gagccccg ttctgcttca ctctccccat 60

120

180

gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc 240

300

360

cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 420

cgggctgtaa ttagcggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc 480

cgccgtcccc ttctccctct cgcctcgg ggctgtccgc ggggggacgg ctgccttcgg 540

gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct 600

gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt 660

gtgctgtctc atcattttgg caaagaattg ca 692

<210> 18

<211> 55

<212> DNA

<213> Feline herpesvirus 1

<400> 18

caataaacat agcatacgtt atgacatggt ctaccgcgtc ttatatgggg acgac 55

<210> 19

<211> 370

<212> DNA

<213> herpes simplex virus

<400> 19

gatccataat tgattgacgg gagatggggg aggctaactg aaacacggaa ggagacaata 60

ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg 120

gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc 180

gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag 240

ttcgggtgaa ggcccaggggc tcgcagccaa cgtcggggcg gcaggccctg ccatagccac 300

360

attattttga 370

<210> 20

<211> 199

<212> DNA

<213> Monkey Virus 40

<400> 20

agcttcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg 60

aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag 120

ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga 180

ggtgtgggag gttttttcg 199

<210> 21

<211> 15252

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 21

ggcgcgccac tggagaacgg catgaccgca aaaggcgttg tagagatcga tcccacgaac 60

tctcaggcga tcgtgtcagt cgccataaac agcgacgatc gtctccagga tctgaacggt 120

tttcttctca acgatcatca gtatatgagg aactgaacct gatatttagc cgagggaaac 180

gcaggttaaa aaccctatca agcgattgcg attttcgcgt atctagtaaa aatagatggg 240

cttcggtact agccttcgcc gccaactctg aatatgccct tcgtggacct catataacat 300

ggcattgttt gttggatgcg gggccggaat taagaagaac attcgaaata cgagcaaaaa 360

tttcggccct ggcatgtgct gcgcgagaat cggtacttcg gggagaaagt tttatcggag 420

ctttgggtag tgcagaggaa actctatctt ggttgaaaat gcatgcgacc ctgcacttga 480

ttctggttaa ccacgatcca atttttaaga cggctggcgc ggtcctagat aacctccgct 540

taaaactagc cccaatattg atgtgcagat ataacacaga aaaacgatca atggaagaca 600

tgctacggcg gtcatctccc gaagacatca ccgattccct aacaatgtgc ctgattatgt 660

tatcgcgcat tcgtcgtacc atgcgcaccg caggaaataa atatagctat atgatagatc 720

caatgaatcg tatgtctaat tacactccag gcgaatgtat gacaggtata ttgcgatata 780

ttgacgaaca tgctagaagg tgtcctgatc acatatgtaa tttgtatatc acatgtacac 840

ttatgccgat gtatgtgcac gggcgatatt tctattgtaa ttcatttttt tgttagtaaa 900

ctaccacagg ctgtccggaa atctaagtta atgaataaag tagatggtta atactcattg 960

cttagaattg gactactttt aattctcttt aatgttcgta ttaaataaaa acatctttaa 1020

1080

tcttcggaga gtgtactcgc cacggtggtt ggaacatcac tatgtctaca cgtcaaattt 1140

aagcacgtca ggtctgtcga ggacaagaaa tggttaacta gtgtttcaat tattcttata 1200

aacgttaagc attgtaagcc ccccggccgt ccgcagcaac aatttactag tatgccgtgg 1260

gctccgggac tatcacggat gtccaattcg cacatgcata taatttttct agggtctctc 1320

1380

aaatgaaggt gctccaacac ggtcttcaaa gcaaccggca taccagcaaa cacagactgc 1440

aactccccgc tgcaatgatt ggttataaac agtaatctgt cttctggaag tatatttcgc 1500

ccgacaatcc acggcgcccc caaagttaaa aaccatccat gtgtatttgc gtcttctctg 1560

ttaaaagaat attgactggc attttcccgt tgaccgccag atatccaaag tacagcacga 1620

tgttgcacgg acgactttgc agtcaccagc cttcctttcc acccccccac caacaaaatg 1680

tttatcgtag gacccatatc cgtaataagg atgggtctgg cagcaacccc ataggcgcct 1740

cggcgtggta gttctcgagg ccttaagctt aaggatcccc caactccgcc cgttttatga 1800

ctagaaccaa tagtttttaa tgccaaatgc actgaaatcc cctaatttgc aaagccaaac 1860

gccccctatg tgagtaatac ggggactttt tacccaattt cccacgcgga aagcccccta 1920

atacactcat atggcatatg aatcagcacg gtcatgcact ctaatggcgg cccataggga 1980

ctttccacat aggggcgtt caccatttcc cagcataggg gtggtgactc aatggccttt 2040

acccaagtac attgggtcaa tgggaggtaa gccaatgggt ttttcccatt actggcaagc 2100

2160

gtgagccaat gggaaaaacc cattgctgcc aagtacactg actcaatagg gactttccaa 2220

tgggtttttc cattgttggc aagcatataa ggtcaatgtg ggtgagtcaa tagggacttt 2280

ccattgtatt ctgcccagta cataaggtca atagggggtg aatcaacagg aaagtcccat 2340

tggagccaag tacactgcgt caatagggac tttccattgg gttttgccca gtacataagg 2400

tcaatagggg atgagtcaat gggaaaaacc cattggagcc aagtacactg actcaatagg 2460

gactttccat tgggttttgc ccagtacata aggtcaatag ggggtgagtc aacaggaaag 2520

ttccattgga gccaagtaca ttgagtcaat agggactttc caatgggttt tgcccagtac 2580

ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt actggcacgt atactgagtc 2640

attagggact ttccaatggg ttttgcccag tacataaggt caataggggt gaatcaacag 2700

gaaagtccca ttggagccaa gtacactgag tcaataggga ctttccattg ggttttgccc 2760

agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc ccattattgg cacgtacata 2820

2880

ttcccaccat tgacgtcaat gggctattga aactaatgca acgtgacctt taaacggtac 2940

tttcccatag ctgattaatg ggaaagtacc gttctcgagc caatacacgt caatgggaag 3000

tgaaagggca gccaaaacgt aacaccgccc cggttttccc ctggaaattc catattggca 3060

3120

ccgtcgcagt cttcggtctg accaccgtag aacgcagagc tcctcgctgc aggcggccgc 3180

tctagaactc gtcgatcgca gcgatgacaa acctgcaaga tcaaacccaa cagattgttc 3240

cgttcatacg gagccttctg atgccaacaa ccggaccggc gtccattccg gacgacaccc 3300

tggagaagca cactctcagg tcagagacct cgacctacaa tttgactgtg ggggacacag 3360

ggtcagggct aattgtcttt ttccctggat tccctggctc aattgtgggt gctcactaca 3420

cactgcagag caatgggaac tacaagttcg atcagatgct cctgactgcc cagaacctac 3480

cggccagcta caactactgc agactagtga gtcggagtct cacagtgagg tcaagcacac 3540

tccctggtgg cgtttatgca ctaaacggca ccataaacgc cgtgaccttc caaggaagcc 3600

tgagtgaact gacagatgtt agctacaatg ggttgatgtc tgcaacagcc aacatcaacg 3660

acaaaattgg gaatgtcctg gtaggggaag gggtcactgt cctcagccta cccacatcat 3720

atgatcttgg gtatgtgagg cttggtgacc ccattcccgc tatagggctt gacccaaaaa 3780

tggtagctac atgcgacagc agtgacaggc ccagagtcta caccataact gcagccgatg 3840

attaccaatt ctcatcacag taccaaccag gtggggtaac aatcacactg ttctcagcca 3900

acattgatgc tatcacaagc ctcagcattg ggggagagct cgtgtttcaa acaagcgtcc 3960

aaggccttgt actgggcgcc accatctacc ttataggctt tgatgggact gcggtaatca 4020

cggagctgt ggccgcagat aatggggctga cggccggcac cgacaatctt atgccattca 4080

atcttgtcat tccaaccaat gagataaccc agccaatcac atccatcaaa ctggagatag 4140

tgacctccaa aagtggtggt caggcagggg atcagatgtc atggtcggca agtgggagcc 4200

tagcagtgac gatccatggt ggcaactatc caggggccct ccgtcccgtc acactagtag 4260

cctacgaaag agtggcaaca ggatccgtcg ttacggtcgc tggggtgagt aacttcgagc 4320

tgattccaaa tcctgaacta gcaaagaacc tggttacaga atacggccga tttgacccag 4380

gagccatgaa ctacacaaaa ttgatactga gtgagaggga ccgtcttggc atcaagaccg 4440

tctggccaac aagggagtac actgattttc gtgagtactt catggaggtg gccgacctca 4500

actctcccct gaagattgca ggagcatttg gcttcaaaga cataatccgg gctataagga 4560

ggtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca 4620

gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat 4680

aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg 4740

ggaggtgtgg gaggtttttt cggatcctct agagtcgacg gcagagtcgc agacgcccct 4800

attggacgtc aaaattgtag aggtgaagtt ttcaaacgat ggcgaagtaa cggcgacttg 4860

cgtttccacc gtcaaatctc cctatagggt agaaactaat tggaaagtag acctcgtaga 4920

tgtaatggat gaaatttctg ggaacagtcc cgccggggtt tttaacagta atgagaaatg 4980

gcagaaacag ctgtactaca gagtaaccga tggaagaaca tcggtccagc taatgtgcct 5040

gtcgtgcacg agccattctc cggaacctta ctgtcttttc gacacgtctc ttatagcgag 5100

ggaaaaagat atcgcgccag agttatactt tacctctgat ccgcaaacgg catactgcac 5160

aataactctg ccgtccggcg ttgttccgag attcgaatgg agccttaata atgtttcact 5220

gccggaatat ttgacggcca cgaccgttgt ttcgcatacc gctggccaaa gtacagtgtg 5280

gaagagcagc gcgagagcag gcgaggcgtg gatttctggc cggggaggca atatatacga 5340

atgcaccgtc ctcatctcag acggcactcg cgttactacg cgaaaggaga ggtgcttaac 5400

aaacacatgg attgcggtgg aaaacggtgc tgctcaggcg cagctgtatt cactcttttc 5460

tggacttgtg tcaggattat gcgggagcat atctgctttg tacgcaacgc tatggaccgc 5520

catttatttt tgaggaatgc ttttttggact atcgtactgc ttttcttcctt cgctagccag 5580

agcaccgccg ccgtcacgta cgactacatt ttaggccgtc gcgcgctcga cgcgctaacc 5640

ataccggcgg ttggcccgta taacagatac ctcactaggg tatcaagagg ctgcgacgtt 5700

gtcgagctca acccgatttc taacgtggac gacatgatat cggcggccaa agaaaaagag 5760

aaggggggcc ctttcgaggc ctccgtcgtc tggttctacg tgattaaggg cgacgacggc 5820

gaggacaagt actgtccaat ctatagaaaa gagtacaggg aatgtggcga cgtacaactg 5880

ctatctgaat gcgccgttca atctgcacag atgtgggcag tggactatgt tcctagcacc 5940

cttgtatcgc gaaatggcgc gggactgact atattctccc ccactgctgc gctctctggc 6000

6060

gttaacgatc gctgtttaaa gatcgggtcg cagcttaact ttttaccgtc gaaatgctgg 6120

acaacagaac agtatcagac tggatttcaa ggcgaacacc tttatccgat cgcagacacc 6180

aatacacgac acgcggacga cgtatatcgg ggatacgaag atattctgca gcgctggaat 6240

aatttgctga ggaaaaagaa tcctagcgcg ccagaccctc gtccagatag cgtcccgcaa 6300

6360

aagaaggccc ctccagaaga ctcggaggac gacatgcagg cagaggcttc tggagaaaat 6420

6480

tcggatcctg actattacaa tgacatgccc gccgtgatcc cggtggagga gactactaaa 6540

agttctaatg ccgtctccat gcccatattc gcggcgttcg tagcctgcgc ggtcgcgctc 6600

gtggggctac tggtttggag catcgtaaaa tgcgcgcgta gctaatcgag cctagaatag 6660

gtggtttctt cctacatgcc acgcctcacg ctcataatat aaatcacatg gaatagcata 6720

ccaatgccta ttcattggga cgttcgaaaa gcatggcatc gctacttgga actctggctc 6780

tccttgccgc gacgctcgca cccttcggcg cgatgggaat cgtgatcact ggaaatcacg 6840

tctccgccag gattgacgac gatcacatcg tgatcgtcgc gcctcgcccc gaagctacaa 6900

ttcaactgca gctatttttc atgcctggcc agagacccca caaaccctac tcaggaaccg 6960

7020

gctttgaaaa ttgcactcat cgatcgtctt ctgtttttgt cggctgtaaa gtgaccgagt 7080

acacgttctc cgcctcgaac agactaaccg gacctccaca cccgtttaag ctcactatac 7140

gaaatcctcg tccgaacgac agcgggatgt tctacgtaat tgttcggcta gacgacacca 7200

aagaacccat tgacgtcttc gcgatccaac tatcggtgta tcaattcgcg aacaccgccg 7260

cgactcgcgg actctattcc aaggcttcgt gtcgcacctt cggattacct accgtccaac 7320

ttgaggccta tctcaggacc gaggaaagtt ggcgcaactg gcaagcgtac gttgccacgg 7380

aggccacgac gaccagcgcc gaggcgacaa ccccgacgcc cgtcactgca accagcgcct 7440

ccgaacttga agcggaacac tttacctttc cctggctaga aaatggcgtg gatcattacg 7500

aaccgacacc cgcaaacgaa aattcaaacg ttactgtccg tctcgggaca atgagcccta 7560

cgctaattgg ggtaaccgtg gctgccgtcg tgagcgcaac gatcggcctc gtcattgtaa 7620

tttccatcgt caccagaaac atgtgcaccc cgcaccgaaa attagacacg gtctcgcaag 7680

acgacgaaga acgttcccaa actagaaggg aatcgcgaaa atttggaccc atggttgcgt 7740

gcgaaataaa caagggggct gaccaggata gtgaacttgt ggaactggtt gcgattgtta 7800

acccgtctgc gctaagctcg cccgactcaa taaaaatgtg attaagtctg aatgtggctc 7860

tccaatcatt tcgattctct aatctcccaa tcctctcaaa aggggcagta tcggacacgg 7920

actgggaggg gcgtacacga tagttatg gtacagcaga ggcctctgaa cacttaggag 7980

gagaattcag ccggggagag cccctgttga gtaggcttgg gagcatattg caggatgaac 8040

atgttagtga tagttctcgc ctcttgtctt gcgcgcctaa cttttgcgac gcgacacgtc 8100

ctctttttgg aaggcactca ggctgtcctc ggggaagatg atcccagaaa cgttccggaa 8160

gggactgtaa tcaaatggac aaaagtcctg cggaacgcgt gcaagatgaa ggcggccgat 8220

gtctgctctt cgcctaacta ttgctttcat gatttaattt acgacgggagg aaagaaagac 8280

tgcccgcccg cgggacccct gtctgcaaac ctggtaattt tactaaagcg cggcgaaagc 8340

ttcccggtt aattaaggcc ctcgaggata catccaaaga ggttgagtat tctctctaca 8400

cttcttgtta aatggaaagt gcatttgctt gttcttacaa tcggcccgag tctcgttcac 8460

agcgcctcgt tcacacttaa accacaaata gtctacaggc tatatgggag ccagactgaa 8520

actcacatat gactaatatt cgggggtgtt agtcacgtgt agcccattgt gtgcatataa 8580

cgatgttgga cgcgtcctta ttcgcggtgt acttgatact atggcagcga gcatgggata 8640

ttcatcctcg tcatcgttaa catctctacg ggttcagaat gtttggcatg tcgtcgatcc 8700

tttgcccatc gttgcaaatt acaagtccga tcgccatgac cgcgataagc ctgtaccatg 8760

tggcattagg gtgacatctc gatcatacat tataagacca acgtgcgagt cttccaaaga 8820

cctgcacgcc ttcttcttcg gattgtcaac gggttcttca gaatctatgc ccatatctgg 8880

8940

cagatctcca ttttctcacg ccactatcct ggacgctgta gacgataatt ataccatgaa 9000

tatagagggg gtatgtttcc actgccactg tgatgataag ttttctccag attgttggat 9060

atctgcattt tctgctgccg aacaaacttc atcgctatgc aaagagatgc gtgtgtacac 9120

gcgccggtgg agtatacggg aaactaaatg ttcatagagg tctttgggct atatgttatt 9180

aaataaaata attgaccagt gaacaatttg tttaatgtta gtttattcaa tgcattggtt 9240

gcaaatattc attacttctc caatcccagg tcattcttta gcgagatgat gttatgacat 9300

tgctgtgaaa attactacag gatatatttt taagatgcag gagtaacaat gtgcatagta 9360

ggcgtagtta tcgcagacgt gcaacgcttc gcatttgagt taccgaagtg cccaacagtg 9420

ctgcggttat ggtttatgcg cacagaatcc atgcatgtcc taattgaacc atccgatttt 9480

tcttttaatc gcgatcgatg tttgggcaac tgcgttattt cagatctaaa aaatttaccc 9540

tttatgacca tcacatctct ctggctcata ccccgcttgg ataagatatc atgtagattc 9600

cgccctaaga aatgcaaact aacattattg tcggttccat atacacttcc atcttgtcct 9660

tcgaaaataa caaactcgcg caatagaccg tccgtacatg catggccgat gtgtgtcaac 9720

atcattggtc tgctagatcc cgatgggacg aatcgtacag tcgtcgctcc agcattggca 9780

aaaatcccca gataccctcc atgcggcaaa tctaaattgc gaccccgaag agactgcacc 9840

aaagtcttat cgacgcacgc tgattttttt gaacagcggg agcccattat cttcagtgga 9900

gcgtagacgg gcgaggctaa ttatgtgaca tagcaacact gcatgtatgt ttttataaat 9960

10020

10080

ctatattgtc cgatatttac acataacatt tcatcgacat gattaaatac ctaagtactg 10140

cacacagatg tttaatgtat atcgtcatat aaattatatc gctaggacag acccaaacga 10200

cctttatccc aaacagtcag atcctcttct caagtgtcga tttctgttat ggaatatgca 10260

taccctggcc cagaaattgc acgcacgagc gtagtgaatg cgtcattggt tttacattta 10320

aaggctaaat gcacaaattc tttagacgac agcacatcgt taaatagcat ctctagcgtt 10380

cttatgaatg ctaagcattg gagtcctcct ggtcggccac aataacagct gagtatcata 10440

ccctgagctc cggggttgtc gcacatagcg gattcgtata aacataggat tttccgcgaa 10500

tccatcagtt gcaaaaatct gttaggctcc atcaacaacg ctggatttac ttcagatcca 10560

cgcgtaaagt aatggtgctc gaataccgtt tttagagttg tcggcatttc aaggaacaaa 10620

gaattcattt cttcattgca acgacgcgcc agaaatccca agacctcttt gggtagtatg 10680

ttcttgccta taaaacacgg cgttccaagt gccaggaacc acgcatgtgt tactgttggg 10740

gcgtattcag aaataaagcg gggtttatgc ggcttttgaa gctcggatat ccaaagtatc 10800

gcttgctgat gaacgagcga tgtagctgtt acaaaacctc ctttccatcc tccagtcaac 10860

ataatttta tcggcctacc tatgtccgta ataagtattg gtcgggcaat tattccgtat 10920

gaggtcttgc aggaataagc tcttagggac agccagcttg gatatggtgc gaaacagacc 10980

ttctcggctt cagaatgtcg ctccgcagtc tcttcgtgtc ggtgcatctt agatccacca 11040

tcaatgtgtg cagcattgac tcccgcccgt cgaatattcc ttttgttacg atgcagtaat 11100

gagcacgatc atgggcgggg cgatgacgtt ctatttgcat gtctgcgaac aatttgcgtc 11160

agtcatacag ctatggagtg ggccatttct ggccgtcaac ttaaaaacgc gaaccgcaga 11220

catatgtatt tgcatgcaaa gacgtatctt cgtatttctg ggcatcttca aatgctctgg 11280

ccaatatggc aatgaatttg gattcgtttg acgccgatgg tatgcagtgc aaatgtgcca 11340

atagcccaca tccgaaaaag ttatttgtca tacaagcagg tgttaagtag caatcacata 11400

aaggcaccag acgcctcatg gcatcataat gaatagctcc ttctccccac tggaaccact 11460

gacaaaatct gcgagtatat tccgcaaacc acattttatt tctcatagaa actaccctaa 11520

atccttttaa cgggaagaag aatcctagat agtgcttgaa gtcatgactg ttactgctgc 11580

11640

atccctttac tattgcgtct tcacgaccaa atgggaatgc gccaaaatcc ccacacctca 11700

tcaccctgga ggcagattgt gtattattaa tatccgccga ttgaagcaca aaacggtacg 11760

gtactgttcc taattctggt atagattcta tggtcaaaag tctgcatatc cccgacattg 11820

11880

11940

cgctagcagc aatcgcgacg caaaacataa aggccatgtt gggatttgaa ctctctgggg 12000

ggcttgttat cttctgcacc gtcgcagtcg cagttttccg aaatttatgt ctaatatatt 12060

ttccggccgt gctccaatcg gccgaaaaga atctgcgtat taccagactc attgacggggc 12120

cgataaagac cataaaacaa aattcctgtg cactccctcc tccagttttg ccatcgtcca 12180

12240

ttttccaccg ttttcttatt ataagcggtt gtatcgccaa cgcgtcaccg caggttgtca 12300

catacagtga tggcatactt gaacgtgcaa caacgcgctc gctttgcaaa tctaagtcat 12360

tgaccatcaa atcgcgttga gaggatagcc aggcatcttt tttcctagta tggtgacggt 12420

gcagccaccc caactcagtt cttgtaaaaa aagctattgg cgggaattta tgttctgagg 12480

12540

acccggcatc actatggata caatacctttt ctatggccca tttcagctct cgaaccaacc 12600

12660

atttatggca tccgagcgct cttactgtac ggtcggatac acccatggtt tttcctttat 12720

atagtcgggt tatagtctgt cgggtttggc ggtagcacgg agtagtttga tttttaagaa 12780

tcgaaaaccg gcttggagag accactgtcg aatatttgtc cgtatactct acacgtgagt 12840

12900

ccttaaagca tatgttatgt tgtacatcaa aacttggtga gttatgttcg attgccgcgc 12960

ataaagaatc gtacatgagc gtttctgcta acatactatc tatattctca cacgcccctg 13020

catatactgt tcctattcca aattcacgtt ttgccccatc ggctatctgc tcccaaaaag 13080

ttgtaatata ggtgccgctg ggtgcgaaat tttcatcagt tgtattcctg ataaactgaa 13140

13200

13260

agttaggtcc gtacctagac cttaatgttt ccaattcttc tgaatcactg cactctcgta 13320

ggggagtaac ggtaataatt tcgtctctga gccccgtttt gcgttgaaaa ctaatcacat 13380

tagataatgt gcaatcggtt tcttttatcc ggatacatct aagtattatg acatcggtgg 13440

tcattgtttc catcaacgac catcttttac gatcgcccat actactcatg gacgttgtcg 13500

gtgttgaaaa atcaccagaa ttgcaacgga tctctgggta ccatgctgct gatggaattg 13560

gcggttttaa ttgttgtttc agtctattat tgctatcttt ggcggggttg aataatgtgg 13620

ggggagagtg attgcaggaa tccgaatggg tcaataaaac gaccgtgctc cgttctgccg 13680

gcgccgatcc gattgaagct atatacttcg cttctctccc cacttttcca atttgatccg 13740

gaaataaaac ggccccggac aacagtatcg tacgatccgg atccggatcc tgcttgccta 13800

cagaagaatc aacatctcgc cccaatattc tggtcaaaac tggctcgctc atggcaacgc 13860

ggacgtttcc cccggtggcc agtcttaatg gttaatgttc ttttcggcaa tcttatacat 13920

cagcgggttg cgtgaatact ggtcacagtt cagtcattta ctacacacca gcaatacgac 13980

14040

ggcgatgtcg tacgggcggt gcggggttgg atcctcggca aagagatcct cgtaattcgg 14100

cggtgggagc ggagggtaaa gacgcgggtg gggatctccc tccggaccgc gcgccgggcg 14160

cggttcgaaa atgctttccg cctcgctcag tgtcaacgcc aagtattcgg gcgggctggg 14220

ggccggaata tctcccgcga cttcttctat cggcgcggaa ttggagtcgc ggtcgtggcg 14280

cgcttctagc gtcgtcaacg gaagtccatt ttcggggtct cccggtgggc gttcagcgtc 14340

catcgtcgta tatgctctaa cacacgtctc gctatattaa aaaaaagaag agtatcggtc 14400

agtgtcgagt gtcgccgaca atgtcgcgag ttctcggcga tttaattttt ggaactgctc 14460

cctatgaatc ccgtaactgt agcgcccgcg cagaaagccg ccatcagacc aactacgtgt 14520

ctgttcgatg tttgcccgcc gatcgcttta ccgattaagg ttccggcgag aaatgacatg 14580

14640

gaaaccacat ctcccgaatt agtagaggaa agcccgcgct gtcggtttgg ggacatatcg 14700

atcttttttg tgtttttcct aggacccttt tgccagatcg tacaaagtcg cgtcttatga 14760

14820

ggcccccgta tgcgccgcgc caccctcttc gccgagctct ttatgcgcgg tgggggtgag 14880

cgcttccgga gttgcgatct ccgatctcga gccgcagccc ggcggtgtct ctttcagtgg 14940

agcgttagcg ccatcatgtg gttcgtggcg gtggaaaggc tattatgtgt taggggagag 15000

accacgtgat cggcatgcaa atgagcaagg cgaacgcgtc agcgttcgca ctgcgaacca 15060

ataatatata tattatacta ttggctttag gtgcgaacgt ccggctagtc caatagcggg 15120

gtcgcgtttc gtaccacgtg ttatagaccg ccctaaactc gcactcgggg gtccggccgc 15180

gcccagacag ggcggagacg tgccacaggg gctttaaaac accgcttcgg gcaccgttca 15240

tctcggcgcg cc 15252

<210> 22

<211> 12692

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 22

ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 60

acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 120

gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 180

caccatatcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt 240

tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca 300

gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga 360

agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac 420

ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg ctagcgatga 480

ccctgctgat tggttcgctg accatttccg gggtgcggaa cggcgttacc agaaactcag 540

aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga tagggtctgc 600

ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga acgcggctac 660

aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaatacaa 720

gctagcttgg gctgcaggtc gactctagag gatcgttaat taacgatccc cgggcgagct 780

cgaattccag actaaatgcc ccggcccaat ttgtcaagtg tgcagtcacg gaggcgtcga 840

ccgtgtcccc ggcattaaac aggaaagcgt taaagttttt gaatgttagg tcacaggtac 900

aaacataaat gtttgtacaa acaggtaaca ggtacaaaca taaatgcccc ggcataaatg 960

tcccttacgg cggatcgaaa cgacattagg catactcggg taccattttg cattccgatc 1020

agcacggatg aaattaggca ggaatgcggt ttatattatg cggcattgga caaacgatat 1080

ggcattgatt ggcagtttat gaatgtcttc atgttgggcg taaacggatt cctattggtt 1140

cagaagacaa cgacgatata tttagagaga aaaagctacc cagcatagga taaacacaca 1200

ttgagcattg agagacatag gtatcggtat ggatgggaaa actacacacg tgaacaccaa 1260

acgacttata tactcgagcg gtgatactac tgagcaagaa tgcactgcat ctgagccact 1320

1380

cttcatgtcc ccgcaatggg tcccaaatct acatcgcttg agcgaggata ccaaaaaggt 1440

ataccgatgt atggtttcca acagactcaa ttatttttccc tattatgagg cgttcaggcg 1500

gtctttgttt gatatgtata tgctaggtcg gttggggcgt cgacttaagc gatctgactg 1560

ggagactatt atgcatctgt caccaacgca aagtcggcgt ctacatagaa ctttaagatt 1620

tgtggagcgt agaattatcc catctaacag ttatatacgc acatcgggcc acgttccgcc 1680

ttcgagggca cttccgacag atacgaattt aaagatggat gaataattaa attggaaaga 1740

gtaactacat taatcgagcg tcatgacggc gtcccgtgaa aatgggaatt ttctactcga 1800

aacaccgtga catttgacag acctggaatt gttattctga tatatagtgg gtgtgtctgg 1860

ccggcaacat acataatgtg catgcgaaac cactttttca gtgtacgctg acattgtgca 1920

acacggaggg gtagcatcta catacaatat atgttgatta atgattggag aaaaaactat 1980

gcagctcgcc gatcatatgg ctaactcgcc ttcgtctata tggcggaccc cgcgggaaaa 2040

atcgacgtac catctgattt acaacaccag taatgaacat gtcgcatccc tgcccagatc 2100

tgtgcgccca ttggcgcgga tcgttgtgaa tgccgccgaa acacttcagg tcggtatgag 2160

2220

cttccgtgac cacgagccta ctgcgacatt taatgctgca aatcccatta gaaaaatggt 2280

cgagacagtt ctacagaata atgaagagcc cccgcggacg catgctgaaa tgggtaatcg 2340

ccttatgaac attatgtact ggtgttgctt gggacacgca ggacaatgct cgatatggca 2400

gttgtacgag acgaatcagg ccattttaag tttattagat gaagtggtta tcggcacaac 2460

aaatcccttt tgcaccctcg agcaatactg gaagccatta tgcaccgcaa tcgccaacaa 2520

ggggacctca tcgcttgttg aggatgccaa agtggccgag tacctggtta gcatgcgcaa 2580

2640

aaggttgtta ataaaggttt attctatgta agactacaat actttcgaca ttgcttgtat 2700

acatattaaa tactttctca agttcctatt acataaaatg ggatctatca ttacattcgt 2760

2820

gccttacttg gaatcgtgaa aatttgaaac gtccattatt tggatatctt ccggttgtcc 2880

catatcccgc cctggtaccg ctcggatacc ttgcccgtat ggattcgtat tgacagtcgc 2940

gcaatcgggg accaacaacg cgtgggtcca cactcattcg gaaattttcc gatgattctg 3000

aatatttatt gccgctcgtt acgagtcgtt ggacatatct gtaatacatt tcttcttctg 3060

aaggatcgct gcacatttga tctatacatt ggccaggatg ttcaagtctc agatgttgca 3120

ttctggcaca gcacaacttt atggcatttc cgatgtaatc gtccggcagc cctgggggag 3180

ttctatattc gcatattgggg atggtaagga caatagcaga tctcgcaacc tccagggagg 3240

ctataataac gtttttaaag gatggatttc tcataaaaat ctgtcgcaaa ttacactgag 3300

aatatccttt actagcgccg attgagagca tcgtcgtcca attttctaaa tggaaagaaa 3360

acaaggcggg caagagtgtt ccaaacattt tcattttcgg cgaatctctc aaatcccatg 3420

gcgtgcaatt gattgcaaaa ttggcacttc cgttcacgtt tgtatctcca aactctaaga 3480

3540

ctatttgaca ccacatatct ttttgtatgt caaactgacc atgatcgtat gttgctgaat 3600

gcactagggc aattcgctcg cgcgactcca tacattgaat aattccacac gtcagctcat 3660

cggttagcaa ggtccagtag ttgaagtcat ttatttttcc ccgcggctgg ccaaatctac 3720

ctctgggaat atccaagttg tcgaatatga tcgcaccggc tctggtcatg gtgaaggaac 3780

tgtagcataa agacgcaggt atcatagggg taatattttt ttattcactc acatactaaa 3840

agtaacgcat attagcacca tgtatgggct atcaattgac atttgcgtag cactacatca 3900

gattatgta caacataatg ggacaacata tggcaagtag atgcaatttc ctcacactag 3960

ttgggtttat ctactattga attttcccct atctgtgata cacttgggag cctctacaag 4020

catattgcca tcatgtacgt ttttatctac tgtcttaacg cccatgggaa cggaggcgtc 4080

gtcgtcatgt attggacggc aacataggca gcaacacaaa ttgcgtttag gtggggtgca 4140

tgtggactcg ataccaagcc cctgcagctg gggaacgtct ggtggagagc cgataatttg 4200

atatacgcac gccatattac tgtcgttgaa gtacgcctta tcttctatgt tttcaaattt 4260

aggttcccaa gtggacgtga gaagtgtttg tatctcacat ggaatggccc aaggcattcc 4320

agcccaggtg cctggtactt taatggcaaa caaacgtttt ggtagaggta ttgattctat 4380

tgcagttctg cagatatctg cagccccgag tatccacagg ctatacgata cgttatcgga 4440

ggcaagcttc gcgccaggtc aattccctgg cattatgccc agtacatgac cttatgggac 4500

tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 4560

tggcagtaca tcaatggggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 4620

4680

cgtaacaact ccgccccatt gacgcaaatg ggcggtagcg tgtacggtgg gaggtctata 4740

taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg 4800

acctccatag aagacaccgg ttgcgccgcc accatgggcc ccagaccttc taccaagaac 4860

ccagtaccta tgatgctgac tgtccgagtc gcgctggtac tgagttgcat ctgtccggca 4920

aactccattg atggcaggcc tcttgcggct gcaggaattg tggttacagg agacaaagcc 4980

gtcaacatat acacctcatc ccagacagga tcaatcatag ttaagctcct cccgaatctg 5040

cccaaggata aggaggcatg tgcgaaagcc cccttggatg catacaacag gacattgacc 5100

actttgctca ccccccttgg tgactctatc cgtaggatac aagagtctgt gactacatct 5160

ggagggggga gacaggggcg ccttataggc gccattattg gcggtgtggc tcttggggtt 5220

gcaactgccg cacaaataac agcggccgca gctctgatac aagccaaaca aaatgctgcc 5280

aacatcctcc gacttaaaga gagcattgcc gcaaccaatg aggctgtgca tgaggtcact 5340

gacggattat cgcaactagc agtggcagtt gggaagatgc agcagtttgt taatgaccaa 5400

tttaataaaa cagctcagga attagactgc atcaaaattg cacagcaagt tggtgtagag 5460

ctcaacctgt acctaaccga attgactaca gtattcggac cacaaatcac ttcacctgct 5520

ttaaacaagc tgactattca ggcactttac aatctagctg gtggaaatat ggattactta 5580

ttgactaagt taggtgtagg gaacaatcaa ctcagctcat taatcggtag cggcttaatc 5640

accggtaacc ctattctata cgactcacag actcaactct tgggtataca ggtaactcta 5700

ccttcagtcg ggaagctaaa taatatgcgt gccacctact tggaaacctt atccgtaagc 5760

acaaccaggg gatttgcctc ggcacttgtc ccaaaagtgg tgacacaggt cggttctgtg 5820

atagaagaac ttgacacctc atactgtata gaaactgact tacatttata ttgtacaaga 5880

atagtaacgt tccctatgtc ccctggtatt tattcctgct tgagcggcaa tacgtcggcc 5940

tgtatgtact caaagaccga aggcgcactt actacaccat acatgactat caaaggttca 6000

gtcatcgcca actgcaagat gacaacatgt agatgtgtaa accccccgggg tatcatatcg 6060

6120

ggcgggataa ctttaaggct cagtggggaa ttcgatgtaa cttatcagaa gaatatctca 6180

atacaagatt ctcaagtaat aataacaggc aatcttgata tctcaactga gcttgggaat 6240

gtcaacaact cgatcagtaa tgctttgaat aagttagagg aaagcaacag aaaactagac 6300

aaagtcaatg tcaaactgac tagcacatct gctctcatta cctatatcgt gttgactatc 6360

atatctcttg tttttggtat acttagcctg attctagcat gctacctaat gtacaagcaa 6420

aaggcgcaac aaaagacctt attatggctt gggaataata ctctagatca gatgagagcc 6480

actacaaaaa tgtgaggatc tctcgaggaa ttctagatcc cacgtcacta ttgtatactc 6540

tatattatac tctatgttat actctgtaat cctactcaat aaacgtgtca cgcctgtgaa 6600

accgtactaa gtctcccgtg tcttcttatc accatcaggt gacatcctcg cccaggctgt 6660

caatcatgcc ggtatcgatt ccagtagcac cggccccacg ctgacaaccc actcttgcag 6720

cgttagcagc gcccctctta acaagccgac ccccaccagc gtcgcggtta ctaacactcc 6780

tctccccgac ctgcaactag taagcttgcc tccgattcta gcattacata gccggtcagt 6840

agatcctgcc attcggtagc gcaaccggct acatcttcaa acagtctcac aataaatgca 6900

tctctcgttc ctgccaatcc ggaaccgggc ataccactcc cgcctgccga tttaattctc 6960

acaattgggc gatgccggcg gggcaaaacg aatgtggatt tggcaaaccg acacaggtct 7020

gctgtacgga ctaatatgggg cacacccaca tcattcttca gatgctccat gcattgttct 7080

atgagaaaga tccatagggt ggaggcagcg tcacgagatc gcccaggcaa tcgatcgcat 7140

tcgtctagta aagtgacgag agttatcatg cacacaccca tgccccgcc ttccgaataa 7200

ctggagctgt ggaagatcgg aaacgtcttt ttgactgccg gtctcgtact actttcgcac 7260

aggtgtatac ccggacgcgt actatatatt ttatatcatc caacgtccga aattacatac 7320

7380 gtggcggcga tggaagtaga tgttgagtct tcgaaagtaa

gtctgtgaaa atatcgaaag cggtacgacg gttgcagaac cgtcgatgtc gccagatact 7440

agtaacaata gcttcgataa cgaagacttc cgtgggcctg aatacgatgt ggagataaat 7500

accagaaaat ctgctaatct tgatcgtatg gaatcttcgt gccgtgaaca acgagcggcg 7560

tgcgaacttc gaaagtgttc gtgtcctacg tctgccgtgc gcatgcaata cagtattctt 7620

tcatctctcg ctccgggttc agagggtcat gtatatatat gtactagata cggggacgcg 7680

gaccaaaaaa aatgcatagt gaaggcagtc gttggaggaa agaatcccgg gagggaagtg 7740

gatatttttaa aaaccatctc acataaatca attataaaat taatccatgc ctataaatgg 7800

aaaaatgttg tgtgtatggc aatgcgtgta tatcgttatg atcttttcac atatattgac 7860

ggagtcggcc ctatgcccct tcaacagatg atctatattc aacgtggact actagaggcg 7920

ctagcataca tacatgaaag gggcatcatt caccgagacg taaagacggga gaatatattc 7980

ttggataatc acgaaaatgc agttttggggt gacttcggtg ctgcatgcca actaggagat 8040

tgtatagata cgccccaatg ttacggttgg agcggaactg tggaaacaaa ttcgccggaa 8100

ttatctgcac ttgatccgta ttgcacaaaa acagatattt ggagtgccgg attggttcta 8160

tatgagatgg caattaaaaa tgtaccattg tttagtaagc aggtgaaaag ttcgggatct 8220

cagctgagat ccataatacg gtgcatgcaa gtgcatgaac tggagtttcc ccgcaacgat 8280

tctaccaacc tctgtaaaca tttcaaacaa tatgcggttc gtgtacgacc gccttatacc 8340

attcctcgag ttataagaaa tggggggatg ccaatggatg ttgaatatgt catttctaaa 8400

atgcttacgt ttgaccagga gttcagacct tctgctaagg aaatattgaa tatgccccta 8460

tttactaagg cgccgattaa cctgcttaat atcacaccct ctgacagtgt ctaacggtat 8520

acaggcggga gcgggtcgtg gcgtcatcat caccacttga gaatttatat tttgaattgt 8580

tgattgataa attaacctga ttcattgaga actgaaacgc catattggtt tcttggatat 8640

gtctacaaca attagttaaa ttgctatgtt ctactgcgag taacatttga taagttgtaa 8700

gagacgggcg actcatgtcg aagttgacga atataaagta cataacgtgt ttagaatacc 8760

cagaatccga atagtccgcg ggggcgtctt ctcgcgtgag taccaaatac tgagttgaac 8820

ttgaaaatgc taaatctgtg acactctttg tgtgatgatt attgtcacca cttcgaagat 8880

ggcttcgaca ttcatgatgt tctggtgttt gtttggaatc gtaatagcgc ttgtttcgtc 8940

caagtctgac aacaaagaaa atctgaagaa ttatatcacg gataagtcaa ccaatattag 9000

aatacccacg ccattatttg tatcaacgga aaactcttat cccacaaaac atgtaatcta 9060

cgatgaaaac tgtggcttcg ctgtactcaa tcctataagt gaccccaaat atgtcctttt 9120

gagccagctt ctaatgggaa ggcgcaaata tgatgcgacg gtcgcgtggt ttgttctcgg 9180

taaaatgtgt gccagattaa tatatttgcg cgaattttat aactgctcga caaatgagcc 9240

ttttggcaca tgttctatga gctctcctgg atggtgggac aggcgctacg tctcaaccag 9300

tttcatttct cgcgacgaat tacagctggt ttttgcagcg ccgtcccgag aattagatgg 9360

tttatatacg cgcgtagtag ttgtcaacgg ggactttact acggccgata taatgtttaa 9420

tgttaaagtg gcatgtgcct tttcaaagac tggaatagaa gatgatacat tatgcaaacc 9480

ctttcatttc tttgccaatg caacattgca caatttaacc atgattagat cggtaactct 9540

tcgagcgcac gaaagccatt taaaggaatg ggtggcacgg agaggtggta acgtccctgc 9600

agtgctactt gagtctacca tgtatcatgc atccaatctg cctagaaatt tcagggattt 9660

ctacataaag tctccagatg attataagta taatcaccta gatgggccat ctgtaatgct 9720

catcactgac agacctagtg aagatttgga tgggaggctc gttcaccaaa gtgacatttt 9780

tactactaca agtcctataa aacaggtccg gtatgaagag catcagtcac atacaaagca 9840

gtatcctgta aacaaaatac aagctataat ttttttgata gggttaggct cgttcattgg 9900

aagcatattc gtagttttgg tagtatggat tatacgcaga tattgcaatg gagcgcggag 9960

tgggggaacg ccccccagtc ctcgccggta tgtgtatacc aggctatgat cacgtgtgaa 10020

acttgggcgg acctgtatca tatgtacacc gtccctattc gtttatagcc agtacgtgtt 10080

atctgcacat agaggaacat gtgtcatact gggatcgcat gcatggtatg tgtgactcta 10140

atattattct gtatcataat aaaaacacag tgcatggtat atagaggatc gctggtaagc 10200

actacggtag accaatcggc tcagattgca ttctttggca tcgataccgt tgttaattta 10260

tatggcaaag tcttgttcat gggagatcag tatttggagg aaatatactc tggaacgatg 10320

gaaatactca aatggaatca agctaaccgc tgctattcta ttgcgcatgc aacatattac 10380

gccgactgtc ctataatcag ttctacggta ttcagaggat gccgggacgc cgttgtttat 10440

actaggcccc acagcagaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 10500

atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 10560

cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 10620

gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 10680

10740

ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 10800

acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 10860

cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 10920

caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 10980

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 11040

tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 11100

aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 11160

ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 11220

cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 11280

11340

tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 11400

ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 11460

aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 11520

aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 11580

11640

gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 11700

gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 11760

11820

11880

attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 11940

ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 12000

gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 12060

ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 12120

tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 12180

gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 12240

cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 12300

gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 12360

tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 12420

ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 12480

gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 12540

tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 12600

catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 12660

ataaaaatag gcgtatcacg aggccctttc gt 12692

<210> 23

<211> 14113

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 23

gaattccaga ctaaatgccc cggcccaatt tgtcaagtgt gcagtcacgg aggcgtcgac 60

cgtgtccccg gcattaaaca ggaaagcgtt aaagtttttg aatgttaggt cacaggtaca 120

aacataaatg tttgtacaaa caggtaacag gtacaaacat aaatgccccg gcataaatgt 180

cccttacggc ggatcgaaac gacattaggc atactcgggt accattttgc attccgatca 240

gcacggatga aattaggcag gaatgcggtt tatattatgc ggcattggac aaacgatatg 300

gcattgattg gcagtttatg aatgtcttca tgttgggcgt aaacggattc ctattggttc 360

agaagacaac gacgatatat ttagagagaa aaagctaccc agcataggat aaacacacat 420

tgagcattga gagacatagg tatcggtatg gatgggaaaa ctacacacgt gaacaccaaa 480

cgacttatat actcgagcgg tgatactact gagcaagaat gcactgcatc tgagccactg 540

aatgaagact gtgatgaaaa tgtgaccatc gatggaattg gagaagaata tgcgcagttc 600

ttcatgtccc cgcaatgggt cccaaatcta catcgcttga gcgaggatac caaaaaggta 660

taccgatgta tggtttccaa cagactcaat tattttccct attatgaggc gttcaggcgg 720

tctttgtttg atatgtatat gctaggtcgg ttggggcgtc gacttaagcg atctgactgg 780

gagactatta tgcatctgtc accaacgcaa agtcggcgtc tacatagaac tttaagattt 840

gtggagcgta gaattatccc atctaacagt tatatacgca catcgggcca cgttccgcct 900

tcgagggcac ttccgacaga tacgaattta aagatggatg aataattaaa ttggaaagag 960

taactacatt aatcgagcgt catgacggcg tcccgtgaaa atgggaattt tctactcgaa 1020

1080

cggcaacata cataatgtgc atgcgaaacc actttttcag tgtacgctga cattgtgcaa 1140

cacggagggg tagcatctac atacaatata tgttgattaa tgattggaga aaaaactatg 1200

cagctcgccg atcatatggc taactcgcct tcgtctatat ggcggacccc gcgggaaaaa 1260

tcgacgtacc atctgattta caacaccagt aatgaacatg tcgcatccct gccagatct 1320

gtgcgcccat tggcgcggat cgttgtgaat gccgccgaaa cacttcaggt cggtatgaga 1380

gccgggaggc cgccatcagc aggagtttgg cgagaggtgt ttgatagaat gatgacagcc 1440

ttccgtgacc acgagcctac tgcgacattt aatgctgcaa atcccattag aaaaatggtc 1500

gagacagttc tacagaataa tgaagagccc ccgcggacgc atgctgaaat gggtaatcgc 1560

cttatgaaca ttatgtactg gtgttgcttg ggacacgcag gacaatgctc gatatggcag 1620

ttgtacgaga cgaatcaggc cattttaagt ttattagatg aagtggttat cggcacaaca 1680

aatccctttt gcaccctcga gcaatactgg aagccattat gcaccgcaat cgccaacaag 1740

gggacctcat cgcttgttga ggatgccaaa gtggccgagt acctggttag catgcgcaaa 1800

ttgatataac ataggcacgc tctgatgtta cagaccacaa taccgcatac atttattgta 1860

aggttgttaa taaaggttta ttctatgtaa gactacaata ctttcgacat tgcttgtata 1920

catattaaat actttctcaa gttcctatta cataaaatgg gatctatcat tacattcgtt 1980

aagagtctgg ataattttac tgtttgccag cttcgatctt ggaacgtact gtggatagtg 2040

ccttacttgg aatcgtgaaa atttgaaacg tccattattt ggatatcttc cggttgtccc 2100

2160

caatcgggga ccaacaacgc gtgggtccac actcattcgg aaattttccg atgattctga 2220

atatttattg ccgctcgtta cgagtcgttg gacatatctg taatacattt cttcttctga 2280

aggatcgctg cacatttgat ctatacattg gccaggatgt tcaagtctca gatgttgcat 2340

tctggcacag cacaacttta tggcatttcc gatgtaatcg tccggcagcc ctgggggagt 2400

tctatattcg catattggga tggtaaggac aatagcagat ctcgcaacct ccagggaggc 2460

tataataacg tttttaaagg atggatttct cataaaaatc tgtcgcaaat tacactgaga 2520

atatccttta ctagcgccga ttgagagcat cgtcgtccaa ttttctaaat ggaaagaaaa 2580

caaggcgggc aagagtgttc caaacatttt cattttcggc gaatctctca aatcccatgg 2640

cgtgcaattg attgcaaaat tggcacttcc gttcacgttt gtatctccaa actctaagac 2700

acttttaatt gaaaaactac gttctagtgt ggaaagaaac ctataggcag accatagaac 2760

tattgacac cacatatctt tttgtatgtc aaactgacca tgatcgtatg ttgctgaatg 2820

cactagggca attcgctcgc gcgactccat acattgaata attccacacg tcagctcatc 2880

ggttagcaag gtccagtagt tgaagtcatt tatttttccc cgcggctggc caaatctacc 2940

tctgggaata tccaagttgt cgaatatgat cgcaccggct ctggtcatgg tgaaggaact 3000

3060

gtaacgcata ttagcaccat gtatgggcta tcaattgaca tttgcgtagc actacatcac 3120

gattatgtac aacataatgg gacaacatat ggcaagtaga tgcaatttcc tcacactagt 3180

tgggtttatc tactattgaa ttttccccta tctgtgatac acttgggagc ctctacaagc 3240

atattgccat catgtacgtt tttatctact gtcttaacgc ccatgggaac ggaggcgtcg 3300

3360

gtggactcga taccaagccc ctgcagctgg ggaacgtctg gtggagagcc gataatttga 3420

tatacgcacg ccatattact gtcgttgaag tacgccttat cttctatgtt ttcaaattta 3480

ggttcccaag tggacgtgag aagtgtttgt atctcacatg gaatggccca aggcattcca 3540

gcccaggtgc ctggtacttt aatggcaaac aaacgttttg gtagaggtat tgattctatt 3600

3660

gcaagctgcg gccgctctag aactagtgga tcccccgggc tgcagcccaa tgtggaattc 3720

gcccttgcac attgttactc ctgcatctta aaaatatatc ctgtagtaat tttcacagca 3780

atgtcataac atcatctcgc taaagaatga cctgggattg gagaagtaat gaatatttgc 3840

aaccaatgca ttgaataaac taacattaaa cgaattcact agtggatccc ccaactccgc 3900

ccgttttatg actagaacca atagttttta atgccaaatg cactgaaatc ccctaatttg 3960

caaagccaaa cgccccctat gtgagtaata cggggacttt ttacccaatt tcccacgcgg 4020

aaagccccct aatacactca tatggcatat gaatcagcac ggtcatgcac tctaatggcg 4080

gcccataggg actttccaca tagggggcgt tcaccatttc ccagcatagg ggtggtgact 4140

4200

tactggcaag cacactgagt caaatgggac tttccactgg gttttgccca agtacattgg 4260

gtcaatggga ggtgagccaa tgggaaaaac ccattgctgc caagtacact gactcaatag 4320

ggactttcca atgggttttt ccattgttgg caagcatata aggtcaatgt gggtgagtca 4380

atagggactt tccattgtat tctgcccagt acataaggtc aatagggggt gaatcaacag 4440

gaaagtccca ttggagccaa gtacactgcg tcaataggga ctttccattg ggttttgccc 4500

agtacataag gtcaataggg gatgagtcaa tgggaaaaac ccattggagc caagtacact 4560

gactcaatag ggactttcca ttgggttttg cccagtacat aaggtcaata gggggtgagt 4620

caacaggaaa gttccattgg agccaagtac attgagtcaa tagggacttt ccaatgggtt 4680

ttgcccagta cataaggtca atgggaggta agccaatggg tttttcccat tactggcacg 4740

tatactgagt cattagggac tttccaatgg gttttgccca gtacataagg tcaatagggg 4800

tgaatcaaca ggaaagtccc attggagcca agtacactga gtcaataggg actttccatt 4860

gggttttgcc cagtacaaaa ggtcaatagg gggtgagtca atgggttttt cccattattg 4920

gcacgtacat aaggtcaata ggggtgagtc attgggtttt tccagccaat ttaattaaaa 4980

cgccatgtac tttcccacca ttgacgtcaa tgggctattg aaactaatgc aacgtgacct 5040

ttaaacggta ctttcccata gctgattaat gggaaagtac cgttctcgag ccaatacacg 5100

tcaatgggaa gtgaaagggc agccaaaacg taacaccgcc ccggttttcc cctggaaatt 5160

ccatattggc acgcattcta ttggctgagc tgcgttctac gtgggtataa gaggcgcgac 5220

cagcgtcggt accgtcgcag tcttcggtct gaccaccgta gaacgcagag ctcctcgctg 5280

caggcggccg ctctagaact cgtcgatcgc agcgatgaca aacctgcaag atcaaaccca 5340

acagattgtt ccgttcatac ggagccttct gatgccaaca accggaccgg cgtccattcc 5400

ggacgacacc ctggagaagc acactctcag gtcagagacc tcgacctaca atttgactgt 5460

gggggacaca gggtcagggc taattgtctt ttttccctggga ttccctggct caattgtggg 5520

tgctcactac acactgcaga gcaatgggaa ctacaagttc gatcagatgc tcctgactgc 5580

ccagaaccta ccggccagct acaactactg cagactagtg agtcggagtc tcacagtgag 5640

gtcaagcaca ctccctggtg gcgtttatgc actaaacggc accataaacg ccgtgacctt 5700

ccaaggaagc ctgagtgaac tgacagatgt tagctacaat gggttgatgt ctgcaacagc 5760

caacatcaac gacaaaattg ggaatgtcct ggtaggggaa ggggtcactg tcctcagcct 5820

acccacatca tatgatcttg ggtatgtgag gcttggtgac cccattcccg ctatagggct 5880

tgacccaaaa atggtagcta catgcgacag cagtgacagg cccagagtct acaccataac 5940

tgcagccgat gattaccaat tctcatcaca gtaccaacca ggtggggtaa caatcacact 6000

gttctcagcc aacattgatg ctatcacaag cctcagcatt gggggagagc tcgtgtttca 6060

6120

tgcggtaatc accagagctg tggccgcaga taatgggctg acggccggca ccgacaatct 6180

tatgccattc aatcttgtca ttccaaccaa tgagataacc cagccaatca catccatcaa 6240

actggagata gtgacctcca aaagtggtgg tcaggcaggg gatcagatgt catggtcggc 6300

aagtgggagc ctagcagtga cgatccatgg tggcaactat ccaggggccc tccgtcccgt 6360

cacactagta gcctacgaaa gagtggcaac aggatccgtc gttacggtcg ctggggtgag 6420

taacttcgag ctgattccaa atcctgaact agcaaagaac ctggttacag aatacggccg 6480

atttgaccca ggagccatga actacacaaa attgatactg agtgagaggg accgtcttgg 6540

catcaagacc gtctggccaa caagggagta cactgatttt cgtgagtact tcatggaggt 6600

ggccgacctc aactctcccc tgaagattgc aggagcattt ggcttcaaag acataatccg 6660

ggctataagg aggtaagctt cagacatgat aagatacatt gatgagtttg gacaaaccac 6720

aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgcttttatt 6780

6840

tcaggttcag ggggaggtgt gggaggtttt ttcggatcct ctagagtcga cggcagagtc 6900

6960 gcagacgccc ctattggacg tcaaaattgt

aacggcgact tgcgtttcca ccgtcaaatc tccctatagg gtagaaacta attggaaagt 7020

agacctcgta gatgtaatgg atgaaatttc tgggaacagt cccgccgggg tttttaacag 7080

taatgagaaa tggcagaaac agctgtacta cagagtaacc gatggaagaa catcggtcca 7140

gctaatgtgc ctgtcgtgca cgagccattc tcgggaacct tactgtcttt tcgacacgtc 7200

tcttatagcg agggaaaaag atatcgcgcc agagttatac tttacctctg atccgcaaac 7260

ggcatactgc acaataactc tgccgtccgg cgttgttccg agattcgaat ggagccttaa 7320

taatgtttca ctgccggaat atttgacggc cacgaccgtt gtttcgcata ccgctggcca 7380

aagtacagtg tggaagagca gcgcgagagc aggcgaggcg tggatttctg gccgggggagg 7440

caatatatac gaatgcaccg tcctcatctc agacggcact cgcgttacta cgcgaaagga 7500

gaggtgctta acaaacacat ggattgcggt ggaaaacggt gctgctcagg cgcagctgta 7560

ttcactcttt tctggacttg tgtcaggatt atgcgggagc atatctgctt tgtacgcaac 7620

gctatggacc gccatttatt tttgaggaat gctttttgga ctatcgtact gctttcttcc 7680

7740

gacgcgctaa ccataccggc ggttggcccg tataacagat acctcactag ggtatcaaga 7800

ggctgcgacg ttgtcgagct caacccgatt tctaacgtgg acgacatgat atcggcggcc 7860

aaagaaaaag agaagggggg ccctttcgag gcctccgtcg tctggttcta cgtgattaag 7920

ggcgacgacg gcgaggacaa gtactgtcca atctatagaa aagagtacag ggaatgtggc 7980

gacgtacaac tgctatctga atgcgccgtt caatctgcac agatgtgggc agtggactat 8040

gttcctagca cccttgtatc gcgaaatggc gcgggactga ctatattctc ccccactgct 8100

gcgctctctg gccaatactt gctgaccctg aaaatcggga gatttgcgca aacagctctc 8160

gtaactctag aagttaacga tcgctgttta aagatcgggt cgcagcttaa ctttttaccg 8220

tcgaaatgct ggacaacaga acagtatcag actggatttc aaggcgaaca cctttatccg 8280

atcgcagaca ccaatacacg acacgcggac gacgtatatc ggggatacga agatattctg 8340

cagcgctgga ataatttgct gaggaaaaag aatcctagcg cgccagaccc tcgtccagat 8400

agcgtcccgc aagaaattcc cgctgtaacc aagaaagcgg aagggcgcac cccggacgca 8460

gaaagcagcg aaaagaaggc ccctccagaa gactcggagg acgacatgca ggcagaggct 8520

tctggagaaa atcctgccgc cctccccgaa gacgacgaag tccccgagga caccgagcac 8580

gatgatccaa actcggatcc tgactattac aatgacatgc ccgccgtgat cccggtggag 8640

gagactacta aaagttctaa tgccgtctcc atgcccatat tcgcggcgtt cgtagcctgc 8700

gcggtcgcgc tcgtggggct actggtttgg agcatcgtaa aatgcgcgcg tagctaatcg 8760

agcctagaat aggtggtttc ttcctacatg ccacgcctca cgctcataat ataaatcaca 8820

tggaatagca taccaatgcc tattcattgg gacgttcgaa aagcatggca tcgctacttg 8880

gaactctggc tctccttgcc gcgacgctcg cacccttcgg cgcgatggga atcgtgatca 8940

ctggaaatca cgtctccgcc aggattgacg acgatcacat cgtgatcgtc gcgcctcgcc 9000

ccgaagctac aattcaactg cagctatttt tcatgcctgg ccagagaccc cacaaaccct 9060

actcaggaac cgtccgcgtc gcgtttcggt ctgatataac aaaccagtgc taccaggaac 9120

ttagcgagga gcgctttgaa aattgcactc atcgatcgtc ttctgtttt gtcggctgta 9180

aagtgaccga gtacacgttc tccgcctcga acagactaac cggacctcca cacccgttta 9240

agctcactat acgaaatcct cgtccgaacg acagcgggat gttctacgta attgttcggc 9300

tagacgacac caaagaaccc attgacgtct tcgcgatcca actatcggtg tatcaattcg 9360

cgaacaccgc cgcgactcgc ggactctatt ccaaggcttc gtgtcgcacc ttcggattac 9420

ctaccgtcca acttgaggcc tatctcagga ccgaggaaag ttggcgcaac tggcaagcgt 9480

acgttgccac ggaggccacg acgaccagcg ccgaggcgac aaccccgacg cccgtcactg 9540

caaccagcgc ctccgaactt gaagcggaac actttacctt tccctggcta gaaaatggcg 9600

tggatcatta cgaaccgaca cccgcaaacg aaaattcaaa cgttactgtc cgtctcggga 9660

caatgagccc tacgctaatt ggggtaaccg tggctgccgt cgtgagcgca acgatcggcc 9720

tcgtcattgt aatttccatc gtcaccagaa acatgtgcac cccgcaccga aaattagaca 9780

cggtctcgca agacgacgaa gaacgttccc aaactagaag ggaatcgcga aaatttggac 9840

ccatggttgc gtgcgaaata aacaaggggg ctgacgga tagtgaactt gtggaactgg 9900

ttgcgattgt taacccgtct gcgctaagct cgcccgactc aataaaaatg tgattaagtc 9960

tgaatgtggc tctccaatca tttcgattct ctaatctccc aatcctctca aaaggggcag 10020

10080

aacacttagg aggagaattc agccggggag agcccctgtt gagtaggctt gggagcatat 10140

tgcaggatga acatgttagt gatagttctc gcctcttgtc ttgcgcgcct aacttttgcg 10200

10260

aacgttccgg aagggactgt aatcaaatgg acaaaagtcc tgcggaacgc gtgcaagatg 10320

aaggcggccg atgtctgctc ttcgcctaac tattgctttc atgatttaat ttacgacgga 10380

ggaaagaaag actgcccgcc cgcgggaccc ctgtctgcaa acctggtaat tttactaaag 10440

cgcggcgaag cttagcttgc ctccgattct agcattacat agccggtcag tagatcctgc 10500

cattcggtag cgcaaccggc tacatcttca aacagtctca cgataaatgc atctctcgtt 10560

cctgccaatc cggaaccggg cataccactc ccgcctgccg atttaattct cacaattggg 10620

cgatgccggc ggggcaaaac gaatgtggat ttggcaaacc gacacaggtc tgctgtacgg 10680

actaatatgg gcacacccac atcattcttc agatgctcca tgcattgttc tatgagaaag 10740

atccataggg tggaggcagc gtcacgagat cgcccaggca atcgatcgca ttcgtctagt 10800

aaagtgacga gagttatcat gcacacaccc atgccccgc cttccgaata actggagctg 10860

tggaagatcg gaaacgtctt tttgactgcc ggtctcgtac tactttcgca caggtgtata 10920

cccggacgcg tactatatat tttatatcat ccaacgtccg aaattacata cgtggcggcg 10980

11040

aatatcgaaa gcggtacgac ggttgcagaa ccgtcgatgt cgccagatac tagtaacaat 11100

agcttcgata acgaagactt ccgtgggcct gaatacgatg tggagataaa taccagaaaa 11160

tctgctaatc ttgatcgtat ggaatcttcg tgccgtgaac aacgagcggc gtgcgaactt 11220

cgaaagtgtt cgtgtcctac gtctgccgtg cgcatgcaat acagtattct ttcatctctc 11280

gctccggggtt cagagggtca tgtatatata tgtactagat acggggacgc ggaccaaaaa 11340

aaatgcatag tgaaggcagt cgttggagga aagaatcccg ggagggaagt ggatatttta 11400

11460

gtgtgtatgg caatgcgtgt atatcgttat gatcttttca catatattga cggagtcggc 11520

cctatgcccc ttcaacagat gatctatatt caacgtggac tactagaggc gctagcatac 11580

atacatgaaa ggggcatcat tcaccgagac gtaaagacgg agaatatatt cttggataat 11640

11700

acgccccaat gttacggttg gagcggaact gtggaaacaa attcgccgga attatctgca 11760

attgcacaaa aacagatatt tggagtgccg gattggttct atatgagatg 11820

gcaattaaaa atgtaccatt gtttagtaag caggtgaaaa gttcgggatc tcagctgaga 11880

tccataatac ggtgcatgca agtgcatgaa ctggagtttc cccgcaacga ttctaccaac 11940

ctctgtaaac atttcaaaca atatgcggtt cgtgtacgac cgccttatac cattcctcga 12000

gtttataagaa atggggggat gccaatggat gttgaatatg tcatttctaa aatgcttacg 12060

12120

12180

agcgggtcgt ggcgtcatca tcaccacttg agaatttata ttttgaattg ttgattgata 12240

aattaacctg attcattgag aactgaaacg ccatattggt ttcttggata tgtctacaac 12300

12360

12420

aatagtccgc gggggcgtct tctcgcgtga gtaccaaata ctgagttgaa cttgaaaatg 12480

ctaaatctgt gacactcttt gtgtgatgat tattgtcacc acttcgaaga tggcttcgac 12540

12600

12660

gccattattt gtatcaacgg aaaactctta tcccacaaaa catgtaatct acgatgaaaa 12720

12780

tctaatggga aggcgcaaat atgatgcgac ggtcgcgtgg tttgttctcg gtaaaatgtg 12840

tgccagatta atatatttgc gcgaatttta taactgctcg acaaatgagc cttttggcac 12900

atgttctatg agctctcctg gatggtggga caggcgctac gtctcaacca gtttcatttc 12960

13020

gcgcgtagta gttgtcaacg gggactttac tacggccgat ataatgttta atgttaaagt 13080

ggcatgtgcc ttttcaaaga ctggaataga agatgataca ttatgcaaac cctttcattt 13140

ctttgccaat gcaacattgc acaatttaac catgattaga tcggtaactc ttcgagcgca 13200

cgaaagccat ttaaaggaat gggtggcacg gagaggtggt aacgtccctg cagtgctact 13260

tgagtctacc atgtatcatg catccaatct gcctagaaat ttcagggatt tctacataaa 13320

gtctccagat gattataagt ataatcacct agatgggcca tctgtaatgc tcatcactga 13380

cagacctagt gaagatttgg atgggaggct cgttcaccaa agtgacattt ttactactac 13440

13500

13560

13620

gccccccagt cctcgccggt atgtgtatac caggctatga tcacgtgtga aacttgggcg 13680

13740

tagaggaaca tgtgtcatac tgggatcgca tgcatggtat gtgtgactct aatattattc 13800

13860

gaccaatcgg ctcagattgc attctttggc atcgataccg ttgttaattt atatggcaaa 13920

13980

aaatggaatc aagctaaccg ctgctattct attgcgcatg caacatatta cgccgactgt 14040

cctataatca gttctacggt attcagagga tgccgggacg ccgttgttta tactaggccc 14100

cacagcagaa ttc 14113

<210> 24

<211> 13064

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<220>

<221> other_features

<222> (7401)..(7403)

<223> n is a, c, g, or t

<400> 24

gaattccaga ctaaatgccc cggcccaatt tgtcaagtgt gcagtcacgg aggcgtcgac 60

cgtgtccccg gcattaaaca ggaaagcgtt aaagtttttg aatgttaggt cacaggtaca 120

aacataaatg tttgtacaaa caggtaacag gtacaaacat aaatgccccg gcataaatgt 180

cccttacggc ggatcgaaac gacattaggc atactcgggt accattttgc attccgatca 240

gcacggatga aattaggcag gaatgcggtt tatattatgc ggcattggac aaacgatatg 300

gcattgattg gcagtttatg aatgtcttca tgttgggcgt aaacggattc ctattggttc 360

agaagacaac gacgatatat ttagagagaa aaagctaccc agcataggat aaacacacat 420

tgagcattga gagacatagg tatcggtatg gatgggaaaa ctacacacgt gaacaccaaa 480

cgacttatat actcgagcgg tgatactact gagcaagaat gcactgcatc tgagccactg 540

aatgaagact gtgatgaaaa tgtgaccatc gatggaattg gagaagaata tgcgcagttc 600

ttcatgtccc cgcaatgggt cccaaatcta catcgcttga gcgaggatac caaaaaggta 660

taccgatgta tggtttccaa cagactcaat tattttccct attatgaggc gttcaggcgg 720

tctttgtttg atatgtatat gctaggtcgg ttggggcgtc gacttaagcg atctgactgg 780

gagactatta tgcatctgtc accaacgcaa agtcggcgtc tacatagaac tttaagattt 840

gtggagcgta gaattatccc atctaacagt tatatacgca catcgggcca cgttccgcct 900

tcgagggcac ttccgacaga tacgaattta aagatggatg aataattaaa ttggaaagag 960

taactacatt aatcgagcgt catgacggcg tcccgtgaaa atgggaattt tctactcgaa 1020

1080

cggcaacata cataatgtgc atgcgaaacc actttttcag tgtacgctga cattgtgcaa 1140

cacggagggg tagcatctac atacaatata tgttgattaa tgattggaga aaaaactatg 1200

cagctcgccg atcatatggc taactcgcct tcgtctatat ggcggacccc gcgggaaaaa 1260

tcgacgtacc atctgattta caacaccagt aatgaacatg tcgcatccct gccagatct 1320

gtgcgcccat tggcgcggat cgttgtgaat gccgccgaaa cacttcaggt cggtatgaga 1380

gccgggaggc cgccatcagc aggagtttgg cgagaggtgt ttgatagaat gatgacagcc 1440

ttccgtgacc acgagcctac tgcgacattt aatgctgcaa atcccattag aaaaatggtc 1500

gagacagttc tacagaataa tgaagagccc ccgcggacgc atgctgaaat gggtaatcgc 1560

cttatgaaca ttatgtactg gtgttgcttg ggacacgcag gacaatgctc gatatggcag 1620

ttgtacgaga cgaatcaggc cattttaagt ttattagatg aagtggttat cggcacaaca 1680

aatccctttt gcaccctcga gcaatactgg aagccattat gcaccgcaat cgccaacaag 1740

gggacctcat cgcttgttga ggatgccaaa gtggccgagt acctggttag catgcgcaaa 1800

ttgatataac ataggcacgc tctgatgtta cagaccacaa taccgcatac atttattgta 1860

aggttgttaa taaaggttta ttctatgtaa gactacaata ctttcgacat tgcttgtata 1920

catattaaat actttctcaa gttcctatta cataaaatgg gatctatcat tacattcgtt 1980

aagagtctgg ataattttac tgtttgccag cttcgatctt ggaacgtact gtggatagtg 2040

ccttacttgg aatcgtgaaa atttgaaacg tccattattt ggatatcttc cggttgtccc 2100

2160

caatcgggga ccaacaacgc gtgggtccac actcattcgg aaattttccg atgattctga 2220

atatttattg ccgctcgtta cgagtcgttg gacatatctg taatacattt cttcttctga 2280

aggatcgctg cacatttgat ctatacattg gccaggatgt tcaagtctca gatgttgcat 2340

tctggcacag cacaacttta tggcatttcc gatgtaatcg tccggcagcc ctgggggagt 2400

tctatattcg catattggga tggtaaggac aatagcagat ctcgcaacct ccagggaggc 2460

tataataacg tttttaaagg atggatttct cataaaaatc tgtcgcaaat tacactgaga 2520

atatccttta ctagcgccga ttgagagcat cgtcgtccaa ttttctaaat ggaaagaaaa 2580

caaggcgggc aagagtgttc caaacatttt cattttcggc gaatctctca aatcccatgg 2640

cgtgcaattg attgcaaaat tggcacttcc gttcacgttt gtatctccaa actctaagac 2700

acttttaatt gaaaaactac gttctagtgt ggaaagaaac ctataggcag accatagaac 2760

tattgacac cacatatctt tttgtatgtc aaactgacca tgatcgtatg ttgctgaatg 2820

cactagggca attcgctcgc gcgactccat acattgaata attccacacg tcagctcatc 2880

ggttagcaag gtccagtagt tgaagtcatt tatttttccc cgcggctggc caaatctacc 2940

tctgggaata tccaagttgt cgaatatgat cgcaccggct ctggtcatgg tgaaggaact 3000

3060

gtaacgcata ttagcaccat gtatgggcta tcaattgaca tttgcgtagc actacatcac 3120

gattatgtac aacataatgg gacaacatat ggcaagtaga tgcaatttcc tcacactagt 3180

tgggtttatc tactattgaa ttttccccta tctgtgatac acttgggagc ctctacaagc 3240

atattgccat catgtacgtt tttatctact gtcttaacgc ccatgggaac ggaggcgtcg 3300

3360

gtggactcga taccaagccc ctgcagctgg ggaacgtctg gtggagagcc gataatttga 3420

tatacgcacg ccatattact gtcgttgaag tacgccttat cttctatgtt ttcaaattta 3480

ggttcccaag tggacgtgag aagtgtttgt atctcacatg gaatggccca aggcattcca 3540

gcccaggtgc ctggtacttt aatggcaaac aaacgttttg gtagaggtat tgattctatt 3600

3660

gcaagcttaa ttaagtaccg agctcgaatt ggcgcgcccg acggcagagt cgcagacgcc 3720

cctattggac gtcaaaattg tagaggtgaa gttttcaaac gatggcgaag taacggcgac 3780

ttgcgtttcc accgtcaaat ctccctatag ggtagaaact aattggaaag tagacctcgt 3840

agatgtaatg gatgaaattt ctgggaacag tcccgccggg gtttttaaca gtaatgagaa 3900

atggcagaaa cagctgtact acagagtaac cgatggaaga acatcggtcc agctaatgtg 3960

cctgtcgtgc acgagccatt ctcgggaacc ttactgtctt ttcgacacgt ctcttatagc 4020

gagggaaaaa gatatcgcgc cagagttata ctttacctct gatccgcaaa cggcatactg 4080

cacaataact ctgccgtccg gcgttgttcc gagattcgaa tggagcctta ataatgtttc 4140

actgccggaa tatttgacgg ccacgaccgt tgtttcgcat accgctggcc aaagtacagt 4200

gtggaagagc agcgcgagag caggcgaggc gtggatttct ggccggggag gcaatatata 4260

cgaatgcacc gtcctcatct cagacggcac tcgcgttact acgcgaaagg agaggtgctt 4320

aacaaacaca tggattgcgg tggaaaacgg tgctgctcag gcgcagctgt attcactctt 4380

ttctggactt gtgtcaggat tatgcgggag catatctgct ttgtacgcaa cgctatggac 4440

cgccatttat ttttgaggaa tgctttttgg actatcgtac tgctttcttc cttcgctagc 4500

cagagcaccg ccgccgtcac gtacgactac attttaggcc gtcgcgcgct cgacgcgcta 4560

accataccgg cggttggccc gtataacaga tacctcacta gggtatcaag aggctgcgac 4620

gttgtcgagc tcaacccgat ttctaacgtg gacgacatga tatcggcggc caaagaaaaa 4680

gagaaggggg gccctttcga ggcctccgtc gtctggttct acgtgattaa gggcgacgac 4740

ggcgaggaca agtactgtcc aatctataga aaagagtaca gggaatgtgg cgacgtacaa 4800

ctgctatctg aatgcgccgt tcaatctgca cagatgtggg cagtggacta tgttcctagc 4860

acccttgtat cgcgaaatgg cgcgggactg actatattct cccccactgc tgcgctctct 4920

ggccaatact tgctgaccct gaaaatcggg agatttgcgc aaacagctct cgtaactcta 4980

gaagttaacg atcgctgttt aaagatcggg tcgcagctta actttttacc gtcgaaatgc 5040

tggacaacag aacagtatca gactggattt caaggcgaac acctttatcc gatcgcagac 5100

accaatacac gacacgcgga cgacgtatat cggggatacg aagatattct gcagcgctgg 5160

aataatttgc tgaggaaaaa gaatcctagc gcgccagacc ctcgtccaga tagcgtcccg 5220

caagaaattc ccgctgtaac caagaaagcg gaagggcgca ccccggacgc agaaagcagc 5280

gaaaagaagg cccctccaga agactcggag gacgacatgc aggcagaggc ttctggagaa 5340

aatcctgccg ccctccccga agacgacgaa gtccccgagg acaccgagca cgatgatcca 5400

aactcggatc ctgactatta caatgacatg cccgccgtga tcccggtgga ggagactact 5460

aaaagttcta atgccgtctc catgcccata ttcgcggcgt tcgtagcctg cgcggtcgcg 5520

ctcgtggggc tactggtttg gagcatcgta aaatgcgcgc gtagctaatc gagcctagaa 5580

taggtggttt cttcctacat gccacgcctc acgctcataa tataaatcac atggaatagc 5640

ataccaatgc ctattcattg ggacgttcga aaagcatggc atcgctactt ggaactctgg 5700

ctctccttgc cgcgacgctc gcacccttcg gcgcgatggg aatcgtgatc actggaaatc 5760

acgtctccgc caggattgac gacgatcaca tcgtgatcgt cgcgcctcgc cccgaagcta 5820

caattcaact gcagctattt ttcatgcctg gccagagacc ccacaaaccc tactcaggaa 5880

cgtccgcgt cgcgtttcgg tctgatataa caaaccagtg ctaccaggaa cttagcgagg 5940

agcgctttga aaattgcact catcgatcgt cttctgtttt tgtcggctgt aaagtgaccg 6000

agtacacgtt ctccgcctcg aacagactaa ccggacctcc acacccgttt aagctcacta 6060

tacgaaatcc tcgtccgaac gacagcggga tgttctacgt aattgttcgg ctagacgaca 6120

ccaaagaacc cattgacgtc ttcgcgatcc aactatcggt gtatcaattc gcgaacaccg 6180

ccgcgactcg cggactctat tccaaggctt cgtgtcgcac cttcggatta cctaccgtcc 6240

aacttgaggc ctatctcagg accgaggaaa gttggcgcaa ctggcaagcg tacgttgcca 6300

6360

cctccgaact tgaagcggaa cactttacct ttccctggct agaaaatggc gtggatcatt 6420

acgaaccgac acccgcaaac gaaaattcaa acgttactgt ccgtctcggg acaatgagcc 6480

ctacgctaat tggggtaacc gtggctgccg tcgtgagcgc aacgatcggc ctcgtcattg 6540

taatttccat cgtcaccaga aacatgtgca ccccgcaccg aaaattagac acggtctcgc 6600

aagacgacga agaacgttcc caaactagaa gggaatcgcg aaaatttgga cccatggttg 6660

cgtgcgaaat aaacaagggg gctgaccagg atagtgaact tgtggaactg gttgcgattg 6720

ttaacccgtc tgcgctaagc tcgcccgact caataaaaat gtgattaagt ctgaatgtgg 6780

ctctccaatc atttcgattc tctaatctcc caatcctctc aaaaggggca gtatcggaca 6840

cggactggga ggggcgtaca cgatagttat atggtacagc agaggcctct gaacacttag 6900

6960

aacatgttag tgatagttct cgcctcttgt cttgcgcgcc taacttttgc gacgcgacac 7020

gtcctctttt tggaaggcac tcaggctgtc ctcggggaag atgatcccag aaacgttccg 7080

gaagggactg taatcaaatg gacaaaagtc ctgcggaacg cgtgcaagat gaaggcggcc 7140

gatgtctgct cttcgcctaa ctattgcttt catgatttaa tttacgacgg aggaaagaaa 7200

7260

cgcgccggat cagatctcca tggtcgaggt gagccccg ttctgcttca ctctccccat 7320

7380

gatggggggcg gggggggggg nnncgcgcgc caggcggggc ggggcggggc gaggggcggg 7440

gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc 7500

cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg 7560

7620

cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 7680

cgggctgtaa ttagcggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc 7740

cgccgtcccc ttctccctct cgcctcgg ggctgtccgc ggggggacgg ctgccttcgg 7800

gggggacgggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct 7860

gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt 7920

gtgctgtctc atcattttgg caaagaattg cagatctgga tctatgacaa acctgcaaga 7980

tcaaacccaa cagattgttc cgttcatacg gagccttctg atgccaacaa ccggaccggc 8040

gtccattccg gacgacaccc tggagaagca cactctcagg tcagagacct cgacctacaa 8100

tttgactgtg ggggacacag ggtcaggggct aattgtcttt ttccctggat tccctggctc 8160

aattgtgggt gctcactaca cactgcagag caatgggaac tacaagttcg atcagatgct 8220

cctgactgcc cagaacctac cggccagcta caactactgc agactagtga gtcggagtct 8280

cacagtgagg tcaagcacac tccctggtgg cgtttatgca ctaaacggca ccataaacgc 8340

cgtgaccttc caaggaagcc tgagtgaact gacagatgtt agctacaatg ggttgatgtc 8400

tgcaacagcc aacatcaacg acaaagttgg gaatgtcctg gtaggggaag gggtcactgt 8460

cctcagccta cccacatcat atgatcttgg gtatgtgagg cttggtgacc ccattcccgc 8520

tatagggctt gacccaaaaa tggtagctac atgcgacagc agtgacaggc ccagagtcta 8580

caccataact gcagccgatg attaccaatt ctcatcacag taccaaccag gtggggtaac 8640

aatcacactg ttctcagcca acattgatgc tatcacaagc ctcagcattg ggggagagct 8700

cgtgtttcaa acaagcgtcc aaggccttgt actgggcgcc accatctacc ttataggctt 8760

tgatgggact gcggtaatca ccagagctgt ggccgcagat aatggggctga cggccggcac 8820

cgacaatctt atgccattca atcttgtcat tccaaccaat gagataaccc agccgatcac 8880

atccatcaaa ctggagatag tgacctccaa aagtggtggt caggcagggg atcagatgtc 8940

atggtcggca agtgggagcc tagcagtgac gatccatggt ggcaactatc caggggccct 9000

ccgtcccgtc acactagtag cctacgaaag agtggcaaca ggatccgtcg ttacggtcgc 9060

tggggtgagt aacttcgagc tgatcccaaa tcctgaacta gcaaagaacc tggttacaga 9120

atacggccga tttgacccag gagccatgaa ctacacaaaa ttgatactga gtgagaggga 9180

ccgtcttggc atcaagaccg tctggccaac aagggagtac actgattttc gtgagtactt 9240

catggaggtg gccgacctca actctcccct gaagattgca ggagcatttg gcttcaaaga 9300

cataatccgg gctataagga ggtaagatcc gatctctcga ttaattaaca ataaacatag 9360

catacgttat gacatggtct accgcgtctt atatggggac gacaagcttg cctccgattc 9420

tagcattaca tagccggtca gtagatcctg ccattcggta gcgcaaccgg ctacatcttc 9480

aaacagtctc acgataaatg catctctcgt tcctgccaat cgggaaccgg gcataccact 9540

cccgcctgcc gatttaattc tcacaattgg gcgatgccgg cggggcaaaa cgaatgtgga 9600

tttggcaaac cgacacaggt ctgctgtacg gactaatatg ggcacaccca catcattctt 9660

cagatgctcc atgcattgtt ctatgagaaa gatccatagg gtggaggcag cgtcacgaga 9720

tcgcccaggc aatcgatcgc attcgtctag taaagtgacg agagttatca tgcacacacc 9780

catgcccacg ccttccgaat aactggagct gtggaagatc ggaaacgtct ttttgactgc 9840

cggtctcgta ctactttcgc acaggtgtat acccggacgc gtactatata ttttatatca 9900

tccaacgtcc gaaattacat acgtggcggc gatggaagta gatgttgagt cttcgaaagt 9960

aagtgcctcg aatatgggta ttgtctgtga aaatatcgaa agcggtacga cggttgcaga 10020

accgtcgatg tcgccagata ctagtaacaa tagcttcgat aacgaagact tccgtgggcc 10080

10140

gtgccgtgaa caacgagcgg cgtgcgaact tcgaaagtgt tcgtgtccta cgtctgccgt 10200

gcgcatgcaa tacagtattc tttcatctct cgctccgggt tcagagggtc atgtatatat 10260

atgtactaga tacggggacg cggaccaaaa aaaatgcata gtgaaggcag tcgttggagg 10320

aaagaatccc gggagggaag tggatatttt aaaaaccatc tcacataaat caattataaa 10380

attaatccat gcctataaat ggaaaaatgt tgtgtgtatg gcaatgcgtg tatatcgtta 10440

tgatcttttc acatatattg acggagtcgg ccctatgccc cttcaacaga tgatctatat 10500

tcaacgtgga ctactagagg cgctagcata catacatgaa aggggcatca ttcaccgaga 10560

cgtaaagacg gagaatat tcttggataa tcacgaaaat gcagttttgg gtgacttcgg 10620

10680

tgtggaaaca aattcgccgg aattatctgc acttgatccg tattgcacaa aaacagatat 10740

ttggagtgcc ggattggttc tatatgagat ggcaattaaa aatgtaccat tgtttagtaa 10800

gcaggtgaaa agttcgggat ctcagctgag atccataata cggtgcatgc aagtgcatga 10860

actggagttt ccccgcaacg attctaccaa cctctgtaaa catttcaaac aatatgcggt 10920

tcgtgtacga ccgccttata ccattcctcg agttataaga aatgggggga tgccaatgga 10980

tgttgaatat gtcatttcta aaatgcttac gtttgaccag gagttcagac cttctgctaa 11040

ggaaatattg aatatgcccc tatttactaa ggcgccgatt aacctgctta atatcacacc 11100

11160

gagaatttat attttgaatt gttgattgat aaattaacct gattcattga gaactgaaac 11220

gccatattgg tttcttggat atgtctacaa caattagtta aattgctatg ttctactgcg 11280

11340

tacataacgt gtttagaata cccagaatcc gaatagtccg cggggggcgtc ttctcgcgtg 11400

agtaccaaat actgagttga acttgaaaat gctaaatctg tgacactctt tgtgtgatga 11460

ttattgtcac cacttcgaag atggcttcga cattcatgat gttctggtgt ttgtttggaa 11520

tcgtaatagc gcttgtttcg tccaagtctg acaacaaaga aaatctgaag aattatatca 11580

11640

atcccacaaa acatgtaatc tacgatgaaa actgtggctt cgctgtactc aatcctataa 11700

gtgaccccaa atatgtcctt ttgagccagc ttctaatggg aaggcgcaaa tatgatgcga 11760

11820

ataactgctc gacaaatgag ccttttggca catgttctat gagctctcct ggatggtggg 11880

acaggcgcta cgtctcaacc agtttcattt ctcgcgacga attacagctg gtttttgcag 11940

12000

ctacggccga tataatgttt aatgttaaag tggcatgtgc cttttcaaag actggaatag 12060

aagatgatac attatgcaaa ccctttcatt tctttgccaa tgcaacattg cacaatttaa 12120

ccatgattag atcggtaact cttcgagcgc acgaaagcca tttaaaggaa tgggtggcac 12180

ggagaggtgg taacgtccct gcagtgctac ttgagtctac catgtatcat gcatccaatc 12240

tgcctagaaa tttcagggat ttctacataa agtctccaga tgattataag tataatcacc 12300

tagatgggcc atctgtaatg ctcatcactg acagacctag tgaagatttg gatgggaggc 12360

tcgttcacca aagtgacatt tttactacta caagtcctat aaaacaggtc cggtatgaag 12420

12480

tagggttagg ctcgttcatt ggaagcatat tcgtagtttt ggtagtatgg attatacgca 12540

gatattgcaa tggagcgcgg agtgggggaa cgccccccag tcctcgccgg tatgtgtata 12600

ccaggctatg atcacgtgtg aaacttgggc ggacctgtat catatgtaca ccgtccctat 12660

tcgtttatag ccagtacgtg ttatctgcac atagaggaac atgtgtcata ctgggatcgc 12720

atgcatggta tgtgtgactc taatattatt ctgtatcata ataaaaacac agtgcatggt 12780

atatagagga tcgctggtaa gcactacggt agaccaatcg gctcagattg cattctttgg 12840

12900

ggaaatatac tctggaacga tggaaatact caaatggaat caagctaacc gctgctattc 12960

tattgcgcat gcaacatatt acgccgactg tcctataatc agttctacgg tattcagagg 13020

atgccgggac gccgttgttt atactaggcc ccacagcaga attc 13064

<210> 25

<211> 13017

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 25

gaattccaga ctaaatgccc cggcccaatt tgtcaagtgt gcagtcacgg aggcgtcgac 60

cgtgtccccg gcattaaaca ggaaagcgtt aaagtttttg aatgttaggt cacaggtaca 120

aacataaatg tttgtacaaa caggtaacag gtacaaacat aaatgccccg gcataaatgt 180

cccttacggc ggatcgaaac gacattaggc atactcgggt accattttgc attccgatca 240

gcacggatga aattaggcag gaatgcggtt tatattatgc ggcattggac aaacgatatg 300

gcattgattg gcagtttatg aatgtcttca tgttgggcgt aaacggattc ctattggttc 360

agaagacaac gacgatatat ttagagagaa aaagctaccc agcataggat aaacacacat 420

tgagcattga gagacatagg tatcggtatg gatgggaaaa ctacacacgt gaacaccaaa 480

cgacttatat actcgagcgg tgatactact gagcaagaat gcactgcatc tgagccactg 540

aatgaagact gtgatgaaaa tgtgaccatc gatggaattg gagaagaata tgcgcagttc 600

ttcatgtccc cgcaatgggt cccaaatcta catcgcttga gcgaggatac caaaaaggta 660

taccgatgta tggtttccaa cagactcaat tattttccct attatgaggc gttcaggcgg 720

tctttgtttg atatgtatat gctaggtcgg ttggggcgtc gacttaagcg atctgactgg 780

gagactatta tgcatctgtc accaacgcaa agtcggcgtc tacatagaac tttaagattt 840

gtggagcgta gaattatccc atctaacagt tatatacgca catcgggcca cgttccgcct 900

tcgagggcac ttccgacaga tacgaattta aagatggatg aataattaaa ttggaaagag 960

taactacatt aatcgagcgt catgacggcg tcccgtgaaa atgggaattt tctactcgaa 1020

1080

cggcaacata cataatgtgc atgcgaaacc actttttcag tgtacgctga cattgtgcaa 1140

cacggagggg tagcatctac atacaatata tgttgattaa tgattggaga aaaaactatg 1200

cagctcgccg atcatatggc taactcgcct tcgtctatat ggcggacccc gcgggaaaaa 1260

tcgacgtacc atctgattta caacaccagt aatgaacatg tcgcatccct gccagatct 1320

gtgcgcccat tggcgcggat cgttgtgaat gccgccgaaa cacttcaggt cggtatgaga 1380

gccgggaggc cgccatcagc aggagtttgg cgagaggtgt ttgatagaat gatgacagcc 1440

ttccgtgacc acgagcctac tgcgacattt aatgctgcaa atcccattag aaaaatggtc 1500

gagacagttc tacagaataa tgaagagccc ccgcggacgc atgctgaaat gggtaatcgc 1560

cttatgaaca ttatgtactg gtgttgcttg ggacacgcag gacaatgctc gatatggcag 1620

ttgtacgaga cgaatcaggc cattttaagt ttattagatg aagtggttat cggcacaaca 1680

aatccctttt gcaccctcga gcaatactgg aagccattat gcaccgcaat cgccaacaag 1740

gggacctcat cgcttgttga ggatgccaaa gtggccgagt acctggttag catgcgcaaa 1800

ttgatataac ataggcacgc tctgatgtta cagaccacaa taccgcatac atttattgta 1860

aggttgttaa taaaggttta ttctatgtaa gactacaata ctttcgacat tgcttgtata 1920

catattaaat actttctcaa gttcctatta cataaaatgg gatctatcat tacattcgtt 1980

aagagtctgg ataattttac tgtttgccag cttcgatctt ggaacgtact gtggatagtg 2040

ccttacttgg aatcgtgaaa atttgaaacg tccattattt ggatatcttc cggttgtccc 2100

2160

caatcgggga ccaacaacgc gtgggtccac actcattcgg aaattttccg atgattctga 2220

atatttattg ccgctcgtta cgagtcgttg gacatatctg taatacattt cttcttctga 2280

aggatcgctg cacatttgat ctatacattg gccaggatgt tcaagtctca gatgttgcat 2340

tctggcacag cacaacttta tggcatttcc gatgtaatcg tccggcagcc ctgggggagt 2400

tctatattcg catattggga tggtaaggac aatagcagat ctcgcaacct ccagggaggc 2460

tataataacg tttttaaagg atggatttct cataaaaatc tgtcgcaaat tacactgaga 2520

atatccttta ctagcgccga ttgagagcat cgtcgtccaa ttttctaaat ggaaagaaaa 2580

caaggcgggc aagagtgttc caaacatttt cattttcggc gaatctctca aatcccatgg 2640

cgtgcaattg attgcaaaat tggcacttcc gttcacgttt gtatctccaa actctaagac 2700

acttttaatt gaaaaactac gttctagtgt ggaaagaaac ctataggcag accatagaac 2760

tattgacac cacatatctt tttgtatgtc aaactgacca tgatcgtatg ttgctgaatg 2820

cactagggca attcgctcgc gcgactccat acattgaata attccacacg tcagctcatc 2880

ggttagcaag gtccagtagt tgaagtcatt tatttttccc cgcggctggc caaatctacc 2940

tctgggaata tccaagttgt cgaatatgat cgcaccggct ctggtcatgg tgaaggaact 3000

3060

gtaacgcata ttagcaccat gtatgggcta tcaattgaca tttgcgtagc actacatcac 3120

gattatgtac aacataatgg gacaacatat ggcaagtaga tgcaatttcc tcacactagt 3180

tgggtttatc tactattgaa ttttccccta tctgtgatac acttgggagc ctctacaagc 3240

atattgccat catgtacgtt tttatctact gtcttaacgc ccatgggaac ggaggcgtcg 3300

3360

gtggactcga taccaagccc ctgcagctgg ggaacgtctg gtggagagcc gataatttga 3420

tatacgcacg ccatattact gtcgttgaag tacgccttat cttctatgtt ttcaaattta 3480

ggttcccaag tggacgtgag aagtgtttgt atctcacatg gaatggccca aggcattcca 3540

gcccaggtgc ctggtacttt aatggcaaac aaacgttttg gtagaggtat tgattctatt 3600

3660

3720

gtagaggtga agttttcaaa cgatggcgaa gtaacggcga cttgcgtttc caccgtcaaa 3780

tctccctata gggtagaaac taattggaaa gtagacctcg tagatgtaat ggatgaaatt 3840

tctgggaaca gtcccgccgg ggtttttaac agtaatgaga aatggcagaa acagctgtac 3900

tacagagtaa ccgatggaag aacatcggtc cagctaatgt gcctgtcgtg cacgagccat 3960

tctccggaac cttactgtct tttcgacacg tctcttatag cgagggaaaa agatatcgcg 4020

ccagagttat actttacctc tgatccgcaa acggcatact gcacaataac tctgccgtcc 4080

ggcgttgttc cgagattcga atggagcctt aataatgttt cactgccgga atatttgacg 4140

gccacgaccg ttgtttcgca taccgctggc caaagtacag tgtggaagag cagcgcgaga 4200

gcaggcgagg cgtggatttc tggccggggga ggcaatatat acgaatgcac cgtcctcatc 4260

tcagacggca ctcgcgttac tacgcgaaag gagaggtgct taacaaacac atggattgcg 4320

gtggaaaacg gtgctgctca ggcgcagctg tattcactct tttctggact tgtgtcagga 4380

ttatgcggga gcatatctgc tttgtacgca acgctatgga ccgccattta tttttgagga 4440

atgctttttg gactatcgta ctgctttctt ccttcgctag ccagagcacc gccgccgtca 4500

cgtacgacta cattttaggc cgtcgcgcgc tcgacgcgct aaccataccg gcggttggcc 4560

cgtataacag atacctcact agggtatcaa gaggctgcga cgttgtcgag ctcaacccga 4620

tttctaacgt ggacgacatg atatcggcgg ccaaagaaaa agagaagggg ggccctttcg 4680

aggcctccgt cgtctggttc tacgtgatta agggcgacga cggcgaggac aagtactgtc 4740

caatctatag aaaagagtac agggaatgtg gcgacgtaca actgctatct gaatgcgccg 4800

ttcaatctgc acagatgtgg gcagtggact atgttcctag cacccttgta tcgcgaaatg 4860

gcgcgggact gactatattc tcccccactg ctgcgctctc tggccaatac ttgctgaccc 4920

tgaaaatcgg gagatttgcg caaacagctc tcgtaactct agaagttaac gatcgctgtt 4980

taaagatcgg gtcgcagctt aactttttac cgtcgaaatg ctggacaaca gaacagtatc 5040

agactggatt tcaaggcgaa cacctttatc cgatcgcaga caccaataca cgacacgcgg 5100

5160

agaatcctag cgcgccagac cctcgtccag atagcgtccc gcaagaaatt cccgctgtaa 5220

ccaagaaagc ggaagggcgc accccggacg cagaaagcag cgaaaagaag gcccctccag 5280

aagactcgga ggacgacatg caggcagagg cttctggaga aaatcctgcc gccctccccg 5340

aagacgacga agtccccgag gacaccgagc acgatgatcc aaactcggat cctgactatt 5400

acaatgacat gcccgccgtg atcccggtgg aggagactac taaaagttct aatgccgtct 5460

ccatgcccat attcgcggcg ttcgtagcct gcgcggtcgc gctcgtgggg ctactggttt 5520

ggagcatcgt aaaatgcgcg cgtagctaat cgagcctaga ataggtggtt tcttcctaca 5580

tgccacgcct cacgctcata atataaatca catggaatag cataccaatg cctattcatt 5640

gggacgttcg aaaagcatgg catcgctact tggaactctg gctctccttg ccgcgacgct 5700

cgcacccttc ggcgcgatgg gaatcgtgat cactggaaat cacgtctccg ccaggattga 5760

cgacgatcac atcgtgatcg tcgcgcctcg ccccgaagct acaattcaac tgcagctatt 5820

tttcatgcct ggccagagac cccacaaacc ctactcagga accgtccgcg tcgcgtttcg 5880

gtctgatata acaaaccagt gctacgga acttagcgag gagcgctttg aaaattgcac 5940

tcatcgatcg tcttctgttt ttgtcggctg taaagtgacc gagtacacgt tctccgcctc 6000

gaacagacta accggacctc cacacccgtt taagctcact atacgaaatc ctcgtccgaa 6060

cgacagcggg atgttctacg taattgttcg gctagacgac accaaagaac ccattgacgt 6120

6180

ttccaaggct tcgtgtcgca ccttcggatt acctaccgtc caacttgagg cctatctcag 6240

gaccgaggaa agttggcgca actggcaagc gtacgttgcc acggaggcca cgacgaccag 6300

6360

acactttacc tttccctggc tagaaaatgg cgtggatcat tacgaaccga cacccgcaaa 6420

6480 cgaaaattca aacgttactg tccgtctcgg gacaatgagc

cgtggctgcc gtcgtgagcg caacgatcgg cctcgtcatt gtaatttcca tcgtcaccag 6540

aaacatgtgc accccgcacc gaaaattaga cacggtctcg caagacgacg aagaacgttc 6600

ccaaactaga agggaatcgc gaaaatttgg acccatggtt gcgtgcgaaa taaacaaggg 6660

ggctgaccag gatagtgaac ttgtggaact ggttgcgatt gttaacccgt ctgcgctaag 6720

ctcgccgac tcaataaaaa tgtgattaag tctgaatgtg gctctccaat catttcgatt 6780

ctctaatctc ccaatcctct caaaaggggc agtatcggac acggactggg aggggcgtac 6840

acgatagtta tatggtacag cagaggcctc tgaacactta ggaggagaat tcagccgggg 6900

agagcccctg ttgagtaggc ttgggagcat attgcaggat gaacatgtta gtgatagttc 6960

7020

ctcaggctgt cctcggggaa gatgatccca gaaacgttcc ggaagggact gtaatcaaat 7080

ggacaaaagt cctgcggaac gcgtgcaaga tgaaggcggc cgatgtctgc tcttcgccta 7140

actattgctt tcatgattta atttacgacg gaggaaagaa agactgcccg cccgcgggac 7200

ccctgtctgc aaacctggta attttactaa agcgcggcga aagcttaggt caattccctg 7260

gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 7320

agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 7380

gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 7440

gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 7500

gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca 7560

gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggcgcgccg 7620

gatctatgac aaacctgcaa gatcaaaccc aacagattgt tccgttcata cggagccttc 7680

tgatgccaac aaccggaccg gcgtccattc cggacgacac cctggagaag cacactctca 7740

ggtcagagac ctcgacctac aatttgactg tgggggacac agggtcaggg ctaattgtct 7800

ttttccctgg attccctggc tcaattgtgg gtgctcacta cacactgcag agcaatggga 7860

actacaagtt cgatcagatg ctcctgactg cccagaacct accggccagc tacaactact 7920

gcagactagt gagtcggagt ctcacagtga ggtcaagcac actccctggt ggcgtttatg 7980

8040 cactaaacgg caccataaac gccgtgacct tccaaggaag

ttagctacaa tgggttgatg tctgcaacag ccaacatcaa cgacaaagtt gggaatgtcc 8100

tggtagggga aggggtcact gtcctcagcc tacccacatc atatgatctt gggtatgtga 8160

ggcttggtga ccccattccc gctatagggc ttgacccaaa aatggtagct acatgcgaca 8220

gcagtgacag gccgagtc tacaccataa ctgcagccga tgattaccaa ttctcatcac 8280

agtaccaacc aggtggggta acaatcacac tgttctcagc caacattgat gctatcacaa 8340

gcctcagcat tgggggagag ctcgtgtttc aaacaagcgt ccaaggcctt gtactgggcg 8400

ccaccatcta ccttataggc tttgatggga ctgcggtaat caccagagct gtggccgcag 8460

ataatgggct gacggccggc accgacaatc ttatgccatt caatcttgtc attccaacca 8520

8580

gtcaggcagg ggatcagatg tcatggtcgg caagtgggag cctagcagtg acgatccatg 8640

gtggcaacta tccaggggcc ctccgtcccg tcacactagt agcctacgaa agagtggcaa 8700

caggatccgt cgttacggtc gctggggtga gtaacttcga gctgatccca aatcctgaac 8760

tagcaaagaa cctggttaca gaatacggcc gatttgaccc aggagccatg aactacacaa 8820

aattgatact gagtgagagg gaccgtcttg gcatcaagac cgtctggcca acaagggagt 8880

acactgattt tcgtgagtac ttcatggagg tggccgacct caactctccc ctgaagattg 8940

caggagcatt tggcttcaaa gacataatcc gggctataag gaggtaagat ccataattga 9000

ttgacgggag atgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 9060

gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttggggtc gtttgttcat 9120

aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 9180

ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 9240

cggggctcg cagccaacgt cggggcggca ggccctgcca tagccactgg ccccgtgggt 9300

tagggacggg gtcccccatg gggaatggtt tatggttcgt gggggtttatt attttgaagc 9360

ttgcctccga ttctagcatt acatagccgg tcagtagatc ctgccattcg gtagcgcaac 9420

cggctacatc ttcaaacagt ctcacaataa atgcatctct cgttcctgcc aatccggaac 9480

cgggcatacc actcccgcct gccgatttaa ttctcacaat tgggcgatgc cggcggggca 9540

aaacgaatgt ggatttggca aaccgacaca ggtctgctgt acggactaat atgggcacac 9600

ccacatcatt cttcagatgc tccatgcatt gttctatgag aaagatccat agggtggagg 9660

cagcgtcacg agatcgccca ggcaatcgat cgcattcgtc tagtaaagtg acgagagtta 9720

tcatgcacac acccatgccc acgccttccg aataactgga gctgtggaag atcggaaacg 9780

tctttttgac tgccggtctc gtactacttt cgcacaggtg tatacccggga cgcgtactat 9840

atattttata tcatccaacg tccgaaatta catacgtggc ggcgatggaa gtagatgttg 9900

agtcttcgaa agtaagtgcc tcgaatatgg gtattgtctg tgaaaatatc gaaagcggta 9960

cgacggttgc agaaccgtcg atgtcgccag atactagtaa caatagcttc gataacgaag 10020

acttccgtgg gcctgaatac gatgtggaga taaataccag aaaatctgct aatcttgatc 10080

gtatggaatc ttcgtgccgt gaacaacgag cggcgtgcga acttcgaaag tgttcgtgtc 10140

ctacgtctgc cgtgcgcatg caatacagta ttctttcatc tctcgctccg ggttcagagg 10200

gtcatgtata tatatgtact agatacgggg acgcggacca aaaaaaatgc atagtgaagg 10260

cagtcgttgg aggaaagaat cccgggaggg aagtggatat tttaaaaacc atctcacata 10320

aatcaattat aaaattaatc catgcctata aatggaaaaa tgttgtgtgt atggcaatgc 10380

gtgtatatcg ttatgatctt ttcacatata ttgacggagt cggccctatg ccccttcaac 10440

agatgatcta tattcaacgt ggactactag aggcgctagc atacatacat gaaaggggca 10500

10560

tgggtgactt cggtgctgca tgccaactag gagattgtat agatacgccc caatgttacg 10620

10680

caaaaacaga tatttggagt gccggattgg ttctatatga gatggcaatt aaaaatgtac 10740

cattgtttag taagcaggtg aaaagttcgg gatctcagct gagatccata atacggtgca 10800

tgcaagtgca tgaactggag tttccccgca acgattctac caacctctgt aaacatttca 10860

aacaatatgc ggttcgtgta cgaccgcctt ataccattcc tcgagttata agaaatgggg 10920

ggatgccaat ggatgttgaa tatgtcattt ctaaaatgct tacgtttgac caggagttca 10980

gaccttctgc taaggaaata ttgaatatgc ccctatttac taaggcgccg attaacctgc 11040

ttaatatcac accctctgac agtgtctaac ggtatacagg cgggagcggg tcgtggcgtc 11100

11160

11220

atgttctact gcgagtaaca tttgataagt tgtaagagac gggcgactca tgtcgaagtt 11280

gacgaatata aagtacataa cgtgtttaga atacccagaa tccgaatagt ccgcgggggc 11340

gtcttctcgc gtgagtacca aatactgagt tgaacttgaa aatgctaaat ctgtgacact 11400

ctttgtgtga tgattattgt caccacttcg aagatggctt cgacattcat gatgttctgg 11460

11520

11580

acggaaaact cttatcccac 11640

ctcaatccta taagtgaccc caaatatgtc cttttgagcc agcttctaat gggaaggcgc 11700

11760

ttgcgcgaat tttataactg ctcgacaaat gagccttttg gcacatgttc tatgagctct 11820

cctggatggt gggacaggcg ctacgtctca accagtttca tttctcgcga cgaattacag 11880

ctggtttttg cagcgccgtc ccgagaatta gatggtttat atacgcgcgt agtagttgtc 11940

aacggggact ttactacggc cgatataatg tttaatgtta aagtggcatg tgccttttca 12000

12060

ttgcacaatt taaccatgat tagatcggta actcttcgag cgcacgaaag ccatttaaag 12120

12180

catgcatcca atctgcctag aaatttcagg gatttctaca taaagtctcc agatgattat 12240

aagtataatc acctagatgg gccatctgta atgctcatca ctgacagacc tagtgaagat 12300

ttggatggga ggctcgttca ccaaagtgac atttttacta ctacaagtcc tataaaacag 12360

gtccggtatg aagagcatca gtcacataca aagcagtatc ctgtaaacaa aatacaagct 12420

ataatttttt tgatagggtt aggctcgttc attggaagca tattcgtagt tttggtagta 12480

tggattatac gcagatattg caatggagcg cggagtgggg gaacgccccc cagtcctcgc 12540

cggtatgtgt ataccaggct atgatcacgt gtgaaacttg ggcggacctg tatcatatgt 12600

acaccgtccc tattcgttta tagccagtac gtgttatctg cacatagagg aacatgtgtc 12660

atactgggat cgcatgcatg gtatgtgtga ctctaatatt attctgtatc ataataaaaa 12720

cacagtgcat ggtatataga ggatcgctgg taagcactac ggtagaccaa tcggctcaga 12780

ttgcattctt tggcatcgat accgttgtta atttatatgg caaagtcttg ttcatggggag 12840

atcagtattt ggaggaaata tactctggaa cgatggaaat actcaaatgg aatcaagcta 12900

accgctgcta ttctattgcg catgcaacat attacgccga ctgtcctata atcagttcta 12960

cggtattcag aggatgccgg gacgccgttg tttatactag gccccacagc agaattc 13017

<210> 26

<211> 11017

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 26

ggcgcgccac tggagaacgg catgaccgca aaaggcgttg tagagatcga tcccacgaac 60

tctcaggcga tcgtgtcagt cgccataaac agcgacgatc gtctccagga tctgaacggt 120

tttcttctca acgatcatca gtatatgagg aactgaacct gatatttagc cgagggaaac 180

gcaggttaaa aaccctatca agcgattgcg attttcgcgt atctagtaaa aatagatggg 240

cttcggtact agccttcgcc gccaactctg aatatgccct tcgtggacct catataacat 300

ggcattgttt gttggatgcg gggccggaat taagaagaac attcgaaata cgagcaaaaa 360

tttcggccct ggcatgtgct gcgcgagaat cggtacttcg gggagaaagt tttatcggag 420

ctttgggtag tgcagaggaa actctatctt ggttgaaaat gcatgcgacc ctgcacttga 480

ttctggttaa ccacgatcca atttttaaga cggctggcgc ggtcctagat aacctccgct 540

taaaactagc cccaatattg atgtgcagat ataacacaga aaaacgatca atggaagaca 600

tgctacggcg gtcatctccc gaagacatca ccgattccct aacaatgtgc ctgattatgt 660

tatcgcgcat tcgtcgtacc atgcgcaccg caggaaataa atatagctat atgatagatc 720

caatgaatcg tatgtctaat tacactccag gcgaatgtat gacaggtata ttgcgatata 780

ttgacgaaca tgctagaagg tgtcctgatc acatatgtaa tttgtatatc acatgtacac 840

ttatgccgat gtatgtgcac gggcgatatt tctattgtaa ttcatttttt tgttagtaaa 900

ctaccacagg ctgtccggaa atctaagtta atgaataaag tagatggtta atactcattg 960

cttagaattg gactactttt aattctcttt aatgttcgta ttaaataaaa acatctttaa 1020

1080

tcttcggaga gtgtactcgc cacggtggtt ggaacatcac tatgtctaca cgtcaaattt 1140

aagcacgtca ggtctgtcga ggacaagaaa tggttaacta gtgtttcaat tattcttata 1200

aacgttaagc attgtaagcc ccccggccgt ccgcagcaac aatttactag tatgccgtgg 1260

gctccgggac tatcacggat gtccaattcg cacatgcata taatttttct agggtctctc 1320

1380

aaatgaaggt gctccaacac ggtcttcaaa gcaaccggca taccagcaaa cacagactgc 1440

aactccccgc tgcaatgatt ggttataaac agtaatctgt cttctggaag tatatttcgc 1500

ccgacaatcc acggcgcccc caaagttaaa aaccatccat gtgtatttgc gtcttctctg 1560

ttaaaagaat attgactggc attttcccgt tgaccgccag atatccaaag tacagcacga 1620

tgttgcacgg acgactttgc agtcaccagc cttcctttcc acccccccac caacaaaatg 1680

tttatcgtag gacccatatc cgtaataagg atgggtctgg cagcaacccc ataggcgcct 1740

cggcgtggta gttctcgagg ccttaattaa gtaccgagct cgaattggcg cgccaggtca 1800

attccctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 1860

tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 1920

ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 1980

ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 2040

acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 2100

aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 2160

gcgcgccgga tccatgggcc ccagaccttc taccaagaac ccagtaccta tgatgctgac 2220

tgtccgagtc gcgctggtac tgagttgcat ctgtccggca aactccattg atggcaggcc 2280

tcttgcggct gcaggaattg tggttacagg agacaaagcc gtcaacatat acacctcatc 2340

ccagacagga tcaatcatag ttaagctcct cccgaatctg cccaaggata aggaggcatg 2400

tgcgaaagcc cccttggatg catacaacag gacattgacc actttgctca ccccccttgg 2460

tgactctatc cgtaggatac aagagtctgt gactacatct ggagggggga gacaggggcg 2520

ccttataggc gccattattg gcggtgtggc tcttggggtt gcaactgccg cacaaataac 2580

agcggccgca gctctgatac aagccaaaca aaatgctgcc aacatcctcc gacttaaaga 2640

gagcattgcc gcaaccaatg aggctgtgca tgaggtcact gacggattat cgcaactagc 2700

agtggcagtt gggaagatgc agcagtttgt taatgaccaa tttaataaaa cagctcagga 2760

attagactgc atcaaaattg cacagcaagt tggtgtagag ctcaacctgt acctaaccga 2820

attgactaca gtattcggac cacaaatcac ttcacctgct ttaaacaagc tgactattca 2880

ggcactttac aatctagctg gtggaaatat ggattactta ttgactaagt taggtgtagg 2940

gaacaatcaa ctcagctcat taatcggtag cggcttaatc accggtaacc ctattctata 3000

3060

taatatgcgt gccacctact tggaaacctt atccgtaagc acaaccaggg gatttgcctc 3120

ggcacttgtc ccaaaagtgg tgacacaggt cggttctgtg atagaagaac ttgacacctc 3180

atactgtata gaaactgact tagatttata ttgtacaaga atagtaacgt tccctatgtc 3240

ccctggtatt tattcctgct tgagcggcaa tacgtcggcc tgtatgtact caaagaccga 3300

aggcgcactt actacaccat acatgactat caaaggttca gtcatcgcca actgcaagat 3360

gacaacatgt agatgtgtaa accccccggg tatcatatcg caaaactatg gagaagccgt 3420

3480

3540

aataacaggc aatcttgata tctcaactga gcttgggaat gtcaacaact cgatcagtaa 3600

3660

3720

acttagcccg attctagcat gctacctaat gtacaagcaa aaggcgcaac aaaagacctt 3780

attatggctt gggaataata ctctagatca gatgagagcc actacaaaaa tgtgaggatc 3840

tctcgaggaa ttctagatcc cacgtcacta ttgtatactc tatattatac tctatgttat 3900

actctgtaat cctactcaat aaacgtgtca cgcctgtgaa accgtactaa gtctcccgtg 3960

tcttcttatc accatcaggt gacatcctcg cccaggctgt caatcatgcc ggtatcgatt 4020

ccagtagcac cggccccg ctgacaaccc actcttgcag cgttagcagc gcccctctta 4080

acaagccgac ccccaccagc gtcgcggtta ctaacactcc tctcccctcg aggatacatc 4140

caaagaggtt gagtattctc tctacacttc ttgttaaatg gaaagtgcat ttgcttgttc 4200

ttacaatcgg cccgagtctc gttcacagcg cctcgttcac acttaaacca caaatagtct 4260

acaggctata tgggagccag actgaaactc acatatgact aatattcggg ggtgttagtc 4320

acgtgtagcc cattgtgtgc atataacgat gttggacgcg tcctttattcg cggtgtactt 4380

gatactatgg cagcgagcat gggatattca tcctcgtcat cgttaacatc tctacgggtt 4440

cagaatgttt ggcatgtcgt cgatcctttg cccatcgttg caaattacaa gtccgatcgc 4500

catgaccgcg ataagcctgt accatgtggc attagggtga catctcgatc atacattata 4560

agaccaacgt gcgagtcttc caaagacctg cacgccttct tcttcggatt gtcaacgggt 4620

tcttcagaat ctatgcccat atctggcgtt gagaccattg tgcgtttaat gaacaataaa 4680

gcggcatgcc atggaaagga gggctgcaga tctccatttt ctcacgccac tatcctggac 4740

gctgtagacg ataattatac catgaatata gagggggtat gtttccactg ccactgtgat 4800

gataagtttt ctccagattg ttggatatct gcattttctg ctgccgaaca aacttcatcg 4860

ctatgcaaag agatgcgtgt gtacacgcgc cgttgagtat acgggaaact aaatgttcat 4920

agaggtcttt gggctatatg ttattaaata aaataattga ccagtgaaca atttgtttaa 4980

tgttagttta ttcaatgcat tggttgcaaa tattcattac ttctccaatc ccaggtcatt 5040

ctttagcgag atgatgttat gacattgctg tgaaaattac tacaggatat atttttaaga 5100

tgcaggagta acaatgtgca tagtaggcgt agttatcgca gacgtgcaac gcttcgcatt 5160

tgagttaccg aagtgcccaa cagtgctgcg gttatggttt atgcgcacag aatccatgca 5220

tgtcctaatt gaaccatccg atttttcttt taatcgcgat cgttgtttgg gcaactgcgt 5280

tatttcagat ctaaaaaatt taccctttat gaccatcaca tctctctggc tcatacccg 5340

cttggataag atatcatgta gattccgccc taagaaatgc aaactaacat tattgtcggt 5400

tccatataca cttccatctt gtccttcgaa aataacaaac tcgcgcaata gaccgtccgt 5460

acatgcatgg ccgatgtgtg tcaacatcat tggtctgcta gatcccgatg ggacgaatcg 5520

tacagtcgtc gctccagcat tggcaaaaat ccccagatac cctccatgcg gcaaatctaa 5580

attgcgaccc cgaagagact gcaccaaagt cttatcgacg cacgctgatt tttttgaaca 5640

gcgggagccc attatcttca gtggagcgta gacgggcgag gctaattatg tgacatagca 5700

acactgcatg tatgttttta taaatcaata agagtacata atttattacg tatcatttcc 5760

gtttgtaata tactgtatac atcatccaca ctattagtca gcactagcgc gcgggcgcac 5820

gttacaatag cagcgtgccc gttatctata ttgtccgata tttacacata acatttcatc 5880

gacatgatta aatacctaag tactgcacac agatgtttaa tgtatatcgt catataaatt 5940

atatcgctag gacagaccca aacgaccttt atcccaaaca gtcagatcct cttctcaagt 6000

gtcgatttct gttatggaat atgcataccc tggcccagaa attgcacgca cgagcgtagt 6060

gaatgcgtca ttggttttac atttaaaggc taaatgcaca aattctttag acgacagcac 6120

atcgttaaat agcatctcta gcgttcttat gaatgctaag cattggagtc ctcctggtcg 6180

gccacaataa cagctgagta tcataccctg agctccgggg ttgtcgcaca tagcggattc 6240

gtataaacat aggattttcc gcgaatccat cagttgcaaa aatctgttag gctccatcaa 6300

6360

agttgtcggc atttcaagga acaaagaatt catttcttca ttgcaacgac gcgccagaaa 6420

tcccaagacc tctttgggta gtatgttctt gcctataaaa cacggcgttc caagtgccag 6480

gaaccacgca tgtgttactg ttggggcgta ttcagaaata aagcggggtt tatgcggctt 6540

ttgaagctcg gatatccaaa gtatcgcttg ctgatgaacg agcgatgtag ctgttacaaa 6600

acctcctttc catcctccag tcaacataat atttatcggc ctacctatgt ccgtaataag 6660

tattggtcgg gcaattattc cgtatgaggt cttgcaggaa taagctctta gggacagcca 6720

gcttggatat ggtgcgaaac agaccttctc ggcttcagaa tgtcgctccg cagtctcttc 6780

gtgtcggtgc atcttagatc caccatcaat gtgtgcagca ttgactcccg cccgtcgaat 6840

attccttttg ttacgatgca gtaatgagca cgatcatggg cggggcgatg acgttctatt 6900

tgcatgtctg cgaacaattt gcgtcagtca tacagctatg gagtgggcca tttctggccg 6960

tcaacttaaa aacgcgaacc gcagacatat gtatttgcat gcaaagacgt atcttcgtat 7020

ttctgggcat cttcaaatgc tctggccaat atggcaatga atttggattc gtttgacgcc 7080

gatggtatgc agtgcaaatg tgccaatagc ccacatccga aaaagtttatt tgtcatacaa 7140

gcaggtgtta agtagcaatc acataaaggc accagacgcc tcatggcatc ataatgaata 7200

gctccttctc cccactggaa ccactgacaa aatctgcgag tatattccgc aaaccacatt 7260

ttatttctca tagaaactac cctaaatcct tttaacggga agaagaatcc tagatagtgc 7320

ttgaagtcat gactgttact gctgcaataa cactgtatat tatttataaa ttccgtttgt 7380

ctaggtatct gatgtaggca ttccgatccc tttactattg cgtcttcacg accaaatggg 7440

aatgcgccaa aatccccaca cctcatcacc ctggaggcag attgtgtatt attaatatcc 7500

gccgattgaa gcacaaaacg gtacggtact gttcctaatt ctggtataga ttctatggtc 7560

aaaagtctgc atatccccga cattgccatg agatcacaca gtccaagtag catgtttatt 7620

gagtcactca gactgtcaac gtccctcgcc gcaccaccaa tcgaaaataa agtatctacg 7680

caagttatag ctccgcattt tctatcgcta gcagcaatcg cgacgcaaaa cataaaggcc 7740

atgttgggat ttgaactctc tggggggctt gttatcttct gcaccgtcgc agtcgcagtt 7800

ttccgaaatt tatgtctaat atattttccg gccgtgctcc aatcggccga aaagaatctg 7860

cgtattacca gactcattga cgggccgata aagaccataa aacaaaattc ctgtgcactc 7920

cctcctccag ttttgccatc gtccaagtcc cgtaactttt tttgcgtttc gaggagcaag 7980

cgttcgttat ccctacccac acttgttttc caccgttttc ttattataag cggttgtatc 8040

gccaacgcgt caccgcaggt tgtcacatac agtgatggca tacttgaacg tgcaacaacg 8100

cgctcgcttt gcaaatctaa gtcattgacc atcaaatcgc gttgagagga tagccaggca 8160

tcttttttcc tagtatggtg acggtgcagc caccccaact cagttcttgt aaaaaaagct 8220

attggcggga atttatgttc tgaggtgcat tctatattta tgagtccatc aaatgccatt 8280

aaccagattc gtattttttc gctcgacccg gcatcactat ggatacaata cctttctatg 8340

gcccatttca gctctcgaac caaccacacg gacaattgac taacataagt atgatcttta 8400

tcacagtcgc acccatctga gttatattta tggcatccga gcgctcttac tgtacggtcg 8460

gatacaccca tggtttttcc tttatatagt cgggttatag tctgtcgggt ttggcggtag 8520

cacggagtag tttgattttt aagaatcgaa aaccggcttg gagagaccac tgtcgaatat 8580

ttgtccgtat actctacacg tgagtgttgt ccattcctag gtatattcat ctgttcggat 8640

accttcaatt gctgttcagg cataacctta aagcatatgt tatgttgtac atcaaaactt 8700

ggtgagttat gttcgattgc cgcgcataaa gaatcgtaca tgagcgtttc tgctaacata 8760

ctatctatat tctcacacgc ccctgcatat actgttccta ttccaaattc acgttttgcc 8820

ccatcggcta tctgctccca aaaagttgta atataggtgc cgctgggtgc gaaattttca 8880

tcagttgtat tcctgataaa ctgaatcact ttacataatt tttgccacat atctgcgtgc 8940

agccatagta tcgaacccgt gggctcggag acgacagtgc gtacaatggg tattttacct 9000

ttccccaaca aaataatggt atacaagtta ggtccgtacc tagaccttaa tgtttccaat 9060

tcttctgaat cactgcactc tcgtagggga gtaacggtaa taatttcgtc tctgagcccc 9120

gttttgcgtt gaaaactaat cacattagat aatgtgcaat cggtttcttt tatccggata 9180

catctaagta ttatgacatc ggtggtcatt gtttccatca acgaccatct tttacgatcg 9240

cccatactac tcatggacgt tgtcggtgtt gaaaaatcac cagaattgca acggatctct 9300

gggtaccatg ctgctgatgg aattggcggt tttaattgtt gtttcagtct attattgcta 9360

tctttggcgg ggttgaataa tgtggggggga gagtgattgc aggaatccga atgggtcaat 9420

aaaacgaccg tgctccgttc tgccggcgcc gatccgattg aagctatata cttcgcttct 9480

ctccccactt ttccaatttg atccggaaat aaaacggccc cggacaacag tatcgtacga 9540

tccggatccg gatcctgctt gcctacagaa gaatcaacat ctcgccccaa tattctggtc 9600

aaaactggct cgctcatggc aacgcggacg tttcccccgg tggccagtct taatggttaa 9660

tgttcttttc ggcaatctta tacatcagcg ggttgcgtga atactggtca cagttcagtc 9720

atttactaca caccagcaat acgacgacgg acagtaccgt cccgacgaac gcgacgccca 9780

aaattgctat cgcgaccgcg tccgaggcga tgtcgtacgg gcggtgcggg gttggatcct 9840

cggcaaagag atcctcgtaa ttcggcggtg ggagcggagg gtaaagacgc gggtggggat 9900

ctccctccgg accgcgcgcc gggcgcggtt cgaaaatgct ttccgcctcg ctcagtgtca 9960

acgccaagta ttcgggcggg ctgggggccg gaatatctcc cgcgacttct tctatcggcg 10020

cggaattgga gtcgcggtcg tggcgcgctt ctagcgtcgt caacggaagt ccattttcgg 10080

ggtctcccgg tgggcgttca gcgtccatcg tcgtatatgc tctaacacac gtctcgctat 10140

attaaaaaaa agaagagtat cggtcagtgt cgagtgtcgc cgacaatgtc gcgagttctc 10200

ggcgatttaa ttttttggaac tgctccctat gaatcccgta actgtagcgc ccgcgcagaa 10260

agccgccatc agaccaacta cgtgtctgtt cgatgtttgc ccgccgatcg ctttaccgat 10320

taaggttccg gcgagaaatg acatgctcga tccaagaaca aagtttttcg cggtaaacaa 10380

caacatagtt accgtgcgag atggagaaac cacatctccc gaattagtag aggaaagccc 10440

gcgctgtcgg tttggggaca tatcgatctt ttttgtgttt ttcctaggac ccttttgcca 10500

gatcgtacaa agtcgcgtct tatgagcgga cgttcttact gcagctcggt aggagtgggg 10560

cagggttaga tttcgtcggc gtttcggccc ccgtatgcgc cgcgccaccc tcttcgccga 10620

gctctttatg cgcggtgggg gtgagcgctt ccggagttgc gatctccgat ctcgagccgc 10680

agcccggcgg tgtctctttc agtggagcgt tagcgccatc atgtggttcg tggcggtgga 10740

aaggctatta tgtgttaggg gagagaccac gtgatcggca tgcaaatgag caaggcgaac 10800

gcgtcagcgt tcgcactgcg aaccaataat atatatatta tactattggc tttaggtgcg 10860

aacgtccggc tagtccaata gcggggtcgc gtttcgtacc acgtgttata gaccgcccta 10920

aactcgcact cgggggtccg gccgcgccca gacaggggcgg agacgtgcca caggggcttt 10980

aaaacaccgc ttcgggcacc gttcatctcg gcgcgcc 11017

<210> 27

<211> 11665

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 27

ggcgcgccac tggagaacgg catgaccgca aaaggcgttg tagagatcga tcccacgaac 60

tctcaggcga tcgtgtcagt cgccataaac agcgacgatc gtctccagga tctgaacggt 120

tttcttctca acgatcatca gtatatgagg aactgaacct gatatttagc cgagggaaac 180

gcaggttaaa aaccctatca agcgattgcg attttcgcgt atctagtaaa aatagatggg 240

cttcggtact agccttcgcc gccaactctg aatatgccct tcgtggacct catataacat 300

ggcattgttt gttggatgcg gggccggaat taagaagaac attcgaaata cgagcaaaaa 360

tttcggccct ggcatgtgct gcgcgagaat cggtacttcg gggagaaagt tttatcggag 420

ctttgggtag tgcagaggaa actctatctt ggttgaaaat gcatgcgacc ctgcacttga 480

ttctggttaa ccacgatcca atttttaaga cggctggcgc ggtcctagat aacctccgct 540

taaaactagc cccaatattg atgtgcagat ataacacaga aaaacgatca atggaagaca 600

tgctacggcg gtcatctccc gaagacatca ccgattccct aacaatgtgc ctgattatgt 660

tatcgcgcat tcgtcgtacc atgcgcaccg caggaaataa atatagctat atgatagatc 720

caatgaatcg tatgtctaat tacactccag gcgaatgtat gacaggtata ttgcgatata 780

ttgacgaaca tgctagaagg tgtcctgatc acatatgtaa tttgtatatc acatgtacac 840

ttatgccgat gtatgtgcac gggcgatatt tctattgtaa ttcatttttt tgttagtaaa 900

ctaccacagg ctgtccggaa atctaagtta atgaataaag tagatggtta atactcattg 960

cttagaattg gactactttt aattctcttt aatgttcgta ttaaataaaa acatctttaa 1020

1080

tcttcggaga gtgtactcgc cacggtggtt ggaacatcac tatgtctaca cgtcaaattt 1140

aagcacgtca ggtctgtcga ggacaagaaa tggttaacta gtgtttcaat tattcttata 1200

aacgttaagc attgtaagcc ccccggccgt ccgcagcaac aatttactag tatgccgtgg 1260

gctccgggac tatcacggat gtccaattcg cacatgcata taatttttct agggtctctc 1320

1380

aaatgaaggt gctccaacac ggtcttcaaa gcaaccggca taccagcaaa cacagactgc 1440

aactccccgc tgcaatgatt ggttataaac agtaatctgt cttctggaag tatatttcgc 1500

ccgacaatcc acggcgcccc caaagttaaa aaccatccat gtgtatttgc gtcttctctg 1560

ttaaaagaat attgactggc attttcccgt tgaccgccag atatccaaag tacagcacga 1620

tgttgcacgg acgactttgc agtcaccagc cttcctttcc acccccccac caacaaaatg 1680

tttatcgtag gacccatatc cgtaataagg atgggtctgg cagcaacccc ataggcgcct 1740

cggcgtggta gttctcgagg ccttaagctt aaggatcccc caactccgcc cgttttatga 1800

ctagaaccaa tagtttttaa tgccaaatgc actgaaatcc cctaatttgc aaagccaaac 1860

gccccctatg tgagtaatac ggggactttt tacccaattt cccacgcgga aagcccccta 1920

atacactcat atggcatatg aatcagcacg gtcatgcact ctaatggcgg cccataggga 1980

ctttccacat aggggcgtt caccatttcc cagcataggg gtggtgactc aatggccttt 2040

acccaagtac attgggtcaa tgggaggtaa gccaatgggt ttttcccatt actggcaagc 2100

2160

gtgagccaat gggaaaaacc cattgctgcc aagtacactg actcaatagg gactttccaa 2220

tgggtttttc cattgttggc aagcatataa ggtcaatgtg ggtgagtcaa tagggacttt 2280

ccattgtatt ctgcccagta cataaggtca atagggggtg aatcaacagg aaagtcccat 2340

tggagccaag tacactgcgt caatagggac tttccattgg gttttgccca gtacataagg 2400

tcaatagggg atgagtcaat gggaaaaacc cattggagcc aagtacactg actcaatagg 2460

gactttccat tgggttttgc ccagtacata aggtcaatag ggggtgagtc aacaggaaag 2520

ttccattgga gccaagtaca ttgagtcaat agggactttc caatgggttt tgcccagtac 2580

ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt actggcacgt atactgagtc 2640

attagggact ttccaatggg ttttgcccag tacataaggt caataggggt gaatcaacag 2700

gaaagtccca ttggagccaa gtacactgag tcaataggga ctttccattg ggttttgccc 2760

agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc ccattattgg cacgtacata 2820

2880

ttcccaccat tgacgtcaat gggctattga aactaatgca acgtgacctt taaacggtac 2940

tttcccatag ctgattaatg ggaaagtacc gttctcgagc caatacacgt caatgggaag 3000

tgaaagggca gccaaaacgt aacaccgccc cggttttccc ctggaaattc catattggca 3060

3120

ccgtcgcagt cttcggtctg accaccgtag aacgcagagc tcctcgctgc aggcggccgc 3180

tctagaactc gtcgatcgca gcgatgacaa acctgcaaga tcaaacccaa cagattgttc 3240

cgttcatacg gagccttctg atgccaacaa ccggaccggc gtccattccg gacgacaccc 3300

tggagaagca cactctcagg tcagagacct cgacctacaa tttgactgtg ggggacacag 3360

ggtcagggct aattgtcttt ttccctggat tccctggctc aattgtgggt gctcactaca 3420

cactgcagag caatgggaac tacaagttcg atcagatgct cctgactgcc cagaacctac 3480

cggccagcta caactactgc agactagtga gtcggagtct cacagtgagg tcaagcacac 3540

tccctggtgg cgtttatgca ctaaacggca ccataaacgc cgtgaccttc caaggaagcc 3600

tgagtgaact gacagatgtt agctacaatg ggttgatgtc tgcaacagcc aacatcaacg 3660

acaaaattgg gaatgtcctg gtaggggaag gggtcactgt cctcagccta cccacatcat 3720

atgatcttgg gtatgtgagg cttggtgacc ccattcccgc tatagggctt gacccaaaaa 3780

tggtagctac atgcgacagc agtgacaggc ccagagtcta caccataact gcagccgatg 3840

attaccaatt ctcatcacag taccaaccag gtggggtaac aatcacactg ttctcagcca 3900

acattgatgc tatcacaagc ctcagcattg ggggagagct cgtgtttcaa acaagcgtcc 3960

aaggccttgt actgggcgcc accatctacc ttataggctt tgatgggact gcggtaatca 4020

cggagctgt ggccgcagat aatggggctga cggccggcac cgacaatctt atgccattca 4080

atcttgtcat tccaaccaat gagataaccc agccaatcac atccatcaaa ctggagatag 4140

tgacctccaa aagtggtggt caggcagggg atcagatgtc atggtcggca agtgggagcc 4200

tagcagtgac gatccatggt ggcaactatc caggggccct ccgtcccgtc acactagtag 4260

cctacgaaag agtggcaaca ggatccgtcg ttacggtcgc tggggtgagt aacttcgagc 4320

tgattccaaa tcctgaacta gcaaagaacc tggttacaga atacggccga tttgacccag 4380

gagccatgaa ctacacaaaa ttgatactga gtgagaggga ccgtcttggc atcaagaccg 4440

tctggccaac aagggagtac actgattttc gtgagtactt catggaggtg gccgacctca 4500

actctcccct gaagattgca ggagcatttg gcttcaaaga cataatccgg gctataagga 4560

ggtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca 4620

gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat 4680

aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg 4740

ggaggtgtgg gaggtttttt cggatcctct agagtcgagg atacatccaa agaggttgag 4800

tattctctct acacttcttg ttaaatggaa agtgcatttg cttgttctta caatcggccc 4860

gagtctcgtt cacagcgcct cgttcacact taaaccacaa atagtctaca ggctatatgg 4920

gagccagact gaaactcaca tatgactaat attcgggggt gttagtcacg tgtagcccat 4980

tgtgtgcata taacgatgtt ggacgcgtcc ttattcgcgg tgtacttgat actatggcag 5040

cgagcatggg atattcatcc tcgtcatcgt taacatctct acgggttcag aatgtttggc 5100

atgtcgtcga tcctttgccc atcgttgcaa attacaagtc cgatcgccat gaccgcgata 5160

agcctgtacc atgtggcatt agggtgacat ctcgatcata cattataaga ccaacgtgcg 5220

agtcttccaa agacctgcac gccttcttct tcggattgtc aacgggttct tcagaatcta 5280

tgcccatatc tggcgttgag accattgtgc gtttaatgaa caataaagcg gcatgccatg 5340

gaaaggaggg ctgcagatct ccattttctc acgccactat cctggacgct gtagacgata 5400

attataccat gaatatagag ggggtatgtt tccactgcca ctgtgatgat aagttttctc 5460

cagattgttg gatatctgca ttttctgctg ccgaacaaac ttcatcgcta tgcaaagaga 5520

tgcgtgtgta cacgcgccgg tggagtatac gggaaactaa atgttcatag aggtctttgg 5580

gctatatgtt attaaataaa ataattgacc agtgaacaat ttgtttaatg ttagtttatt 5640

caatgcattg gttgcaaata ttcattactt ctccaatccc aggtcattct ttagcgagat 5700

gatgttatga cattgctgtg aaaattacta caggatatat ttttaagatg caggagtaac 5760

aatgtgcata gtaggcgtag ttatcgcaga cgtgcaacgc ttcgcatttg agttaccgaa 5820

gtgcccaaca gtgctgcggt tatggtttat gcgcacagaa tccatgcatg tcctaattga 5880

accatccgat ttttctttta atcgcgatcg atgtttgggc aactgcgtta tttcagatct 5940

aaaaaattta ccctttatga ccatcacatc tctctggctc ataccccgct tggataagat 6000

atcatgtaga ttccgcccta agaaatgcaa actaacatta ttgtcggttc catatacact 6060

tccatcttgt ccttcgaaaa taacaaactc gcgcaataga ccgtccgtac atgcatggcc 6120

gatgtgtgtc aacatcattg gtctgctaga tcccgatggg acgaatcgta cagtcgtcgc 6180

tccagcattg gcaaaaatcc ccagataccc tccatgcggc aaatctaaat tgcgaccccg 6240

aagagactgc accaaagtct tatcgacgca cgctgatttt tttgaacagc gggagcccat 6300

tatcttcagt ggagcgtaga cgggcgaggc taattatgtg acatagcaac actgcatgta 6360

tgtttttata aatcaataag agtacataat ttattacgta tcatttccgt ttgtaatata 6420

ctgtatacat catccacact attagtcagc actagcgcgc gggcgcacgt tacaatagca 6480

gcgtgcccgt tatctatatt gtccgatatt tacacataac atttcatcga catgattaaa 6540

6600

cagacccaaa cgacctttat cccaaacagt cagatcctct tctcaagtgt cgatttctgt 6660

tatggaatat gcataccctg gcccagaaat tgcacgcacg agcgtagtga atgcgtcatt 6720

ggttttacat ttaaaggcta aatgcacaaa ttctttagac gacagcacat cgttaaaatag 6780

catctctagc gttcttatga atgctaagca ttggagtcct cctggtcggc cacaataaca 6840

gctgagtatc ataccctgag ctccggggtt gtcgcacata gcggattcgt ataaacatag 6900

gattttccgc gaatccatca gttgcaaaaa tctgttaggc tccatcaaca acgctggatt 6960

tacttcagat ccacgcgtaa agtaatggtg ctcgaatacc gtttttagag ttgtcggcat 7020

ttcaaggaac aaagaattca ttttcttcatt gcaacgacgc gccagaaatc ccaagacctc 7080

tttggggtagt atgttcttgc ctataaaaca cggcgttcca agtgccagga accacgcatg 7140

tgttactgtt ggggcgtatt cagaaataaa gcggggttta tgcggctttt gaagctcgga 7200

tatccaaagt atcgcttgct gatgaacgag cgatgtagct gttacaaaac ctcctttcca 7260

tcctccagtc aacataatat ttatcggcct acctatgtcc gtaataagta ttggtcgggc 7320

aattattccg tatgaggtct tgcaggaata agctcttagg gacagccagc ttggatatgg 7380

tgcgaaacag accttctcgg cttcagaatg tcgctccgca gtctcttcgt gtcggtgcat 7440

cttagatcca ccatcaatgt gtgcagcatt gactcccgcc cgtcgaatat tccttttgtt 7500

acgatgcagt aatgagcacg atcatgggcg gggcgatgac gttctatttg catgtctgcg 7560

aacaatttgc gtcagtcata cagctatgga gtgggccatt tctggccgtc aacttaaaaa 7620

cgcgaaccgc agacatatgt atttgcatgc aaagacgtat cttcgtattt ctgggcatct 7680

tcaaatgctc tggccaatat ggcaatgaat ttggattcgt ttgacgccga tggtatgcag 7740

tgcaaatgtg ccaatagccc acatccgaaa aagttatttg tcatacaagc aggtgttaag 7800

tagcaatcac ataaaggcac cagacgcctc atggcatcat aatgaatagc tccttctccc 7860

cactggaacc actgacaaaa tctgcgagta tattccgcaa accacatttt atttctcata 7920

gaaactaccc taaatccttt taacgggaag aagaatccta gatagtgctt gaagtcatga 7980

ctgttactgc tgcaataaca ctgtatatta tttataaatt ccgtttgtct aggtatctga 8040

tgtaggcatt ccgatccctt tactattgcg tcttcacgac caaatgggaa tgcgccaaaa 8100

tccccacacc tcatcaccct ggaggcagat tgtgtattat taatatccgc cgattgaagc 8160

acaaaacggt acggtactgt tcctaattct ggtatagatt ctatggtcaa aagtctgcat 8220

atccccgaca ttgccatgag atcacacagt ccaagtagca tgtttattga gtcactcaga 8280

ctgtcaacgt ccctcgccgc accaccaatc gaaaataaag tatctacgca agttatagct 8340

ccgcattttc tatcgctagc agcaatcgcg acgcaaaaca taaaggccat gttgggattt 8400

gaactctctg gggggcttgt tatcttctgc accgtcgcag tcgcagtttt ccgaaattta 8460

tgtctaatat attttccggc cgtgctccaa tcggccgaaa agaatctgcg tattaccaga 8520

ctcattgacg ggccgataaa gaccataaaa caaaattcct gtgcactccc tcctccagtt 8580

ttgccatcgt ccaagtcccg taactttttt tgcgtttcga ggagcaagcg ttcgttatcc 8640

ctacccacac ttgttttcca ccgttttctt attataagcg gttgtatcgc caacgcgtca 8700

ccgcaggttg tcacatacag tgatggcata cttgaacgtg caacaacgcg ctcgctttgc 8760

aaatctaagt cattgaccat caaatcgcgt tgagaggata gccaggcatc ttttttccta 8820

gtatggtgac ggtgcagcca ccccaactca gttcttgtaa aaaaagctat tggcgggaat 8880

ttatgttctg aggtgcattc tatatttatg agtccatcaa atgccattaa ccagattcgt 8940

attttttcgc tcgacccggc atcactatgg atacaatacc tttctatggc ccatttcagc 9000

tctcgaacca accacacgga caattgacta acataagtat gatctttatc acagtcgcac 9060

9120

gtttttcctt tatatagtcg ggttatagtc tgtcgggttt ggcggtagca cggagtagtt 9180

tgatttttaa gaatcgaaaa ccggcttgga gagaccactg tcgaatattt gtccgtatac 9240

tctacacgtg agtgttgtcc attcctaggt atattcatct gttcggatac cttcaattgc 9300

tgttcaggca taaccttaaa gcatatgtta tgttgtacat caaaacttgg tgagttatgt 9360

tcgattgccg cgcataaaga atcgtacatg agcgtttctg ctaacatact atctatattc 9420

tcacacgccc ctgcatatac tgttcctatt ccaaattcac gttttgcccc atcggctatc 9480

tgctcccaaa aagttgtaat ataggtgccg ctgggtgcga aattttcatc agttgtattc 9540

ctgataaact gaatcacttt acataatttt tgccacatat ctgcgtgcag ccatagtatc 9600

gaacccgtgg gctcggagac gacagtgcgt acaatgggta ttttaccttt ccccaacaaa 9660

ataatggtat acaagttagg tccgtaccta gaccttaatg tttccaattc ttctgaatca 9720

ctgcactctc gtaggggagt aacggtaata atttcgtctc tgagccccgt tttgcgttga 9780

aaactaatca cattagataa tgtgcaatcg gtttctttta tccggataca tctaagtatt 9840

atgacatcgg tggtcattgt ttccatcaac gaccatcttt tacgatcgcc catactactc 9900

atggacgttg tcggtgttga aaaatcacca gaattgcaac ggatctctgg gtaccatgct 9960

10020

ttgaataatg tggggggaga gtgattgcag gaatccgaat gggtcaataa aacgaccgtg 10080

ctccgttctg ccggcgccga tccgattgaa gctatatact tcgcttctct ccccactttt 10140

ccaatttgat ccggaaataa aacggccccg gacaacagta tcgtacgatc cggatccgga 10200

tcctgcttgc ctacagaaga atcaacatct cgccccaata ttctggtcaa aactggctcg 10260

ctcatggcaa cgcggacgtt tcccccggtg gccagtctta atggttaatg ttcttttcgg 10320

caatcttata catcagcggg ttgcgtgaat actggtcaca gttcagtcat ttactacaca 10380

ccagcaatac gacgacggac agtaccgtcc cgacgaacgc gacgcccaaa attgctatcg 10440

cgaccgcgtc cgaggcgatg tcgtacggggc ggtgcggggt tggatcctcg gcaaagagat 10500

cctcgtaatt cggcggtggg agcggagggt aaagacgcgg gtggggatct ccctccggac 10560

cgcgcgccgg gcgcggttcg aaaatgcttt ccgcctcgct cagtgtcaac gccaagtatt 10620

cgggcgggct gggggccgga atatctcccg cgacttcttc tatcggcgcg gaattggagt 10680

10740 cgcggtcgtg gcgcgcttct agcgtcgtca acggaagtcc

ggcgttcagc gtccatcgtc gtatatgctc taacacacgt ctcgctatat taaaaaaaag 10800

aagagtatcg gtcagtgtcg agtgtcgccg acaatgtcgc gagttctcgg cgatttaatt 10860

tttggaactg ctccctatga atcccgtaac tgtagcgccc gcgcagaaag ccgccatcag 10920

accaactacg tgtctgttcg atgtttgccc gccgatcgct ttaccgatta aggttccggc 10980

gagaaatgac atgctcgatc caagaacaaa gtttttcgcg gtaaacaaca acatagttac 11040

cgtgcgagat ggagaaacca catctcccga attagtagag gaaagcccgc gctgtcggtt 11100

11160

tcgcgtctta tgagcggacg ttcttactgc agctcggtag gagtggggca gggttagatt 11220

tcgtcggcgt ttcggcccc gtatgcgccg cgccaccctc ttcgccgagc tctttatgcg 11280

cggtgggggt gagcgcttcc ggagttgcga tctccgatct cgagccgcag cccggcggtg 11340

tctctttcag tggagcgtta gcgccatcat gtggttcgtg gcggtggaaa ggctattatg 11400

tgttagggga gagaccacgt gatcggcatg caaatgagca aggcgaacgc gtcagcgttc 11460

gcactgcgaa ccaataatat atatattata ctattggctt taggtgcgaa cgtccggcta 11520

gtccaatagc ggggtcgcgt ttcgtaccac gtgttataga ccgccctaaa ctcgcactcg 11580

ggggtccggc cgcgcccaga cagggcggag acgtgccaca ggggctttaa aacaccgctt 11640

cgggcaccgt tcatctcggc gcgcc 11665

<210> 28

<211> 13253

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 28

aattccagac taaatgcccc ggcccaattt gtcaagtgtg cagtcacgga ggcgtcgacc 60

gtgtccccgg cattaaacag gaaagcgtta aagtttttga atgttaggtc acaggtacaa 120

acataaatgt ttgtacaaac aggtaacagg tacaaacata aatgccccgg cataaatgtc 180

ccttacggcg gatcgaaacg acattaggca tactcgggta ccattttgca ttccgatcag 240

cacggatgaa attaggcagg aatgcggttt atattatgcg gcattggaca aacgatatgg 300

cattgattgg cagtttatga atgtcttcat gtggggcgta aacggattcc tattggttca 360

gaagacaacg acgatatatt tagagagaaa aagctaccca gcataggata aacacacatt 420

gagcattgag agacataggt atcggtatgg atgggaaaac tacacacgtg aacaccaaac 480

gacttatata ctcgagcggt gatactactg agcaagaatg cactgcatct gagccactga 540

atgaagactg tgatgaaaat gtgaccatcg atggaattgg agaagaatat gcgcagttct 600

tcatgtcccc gcaatgggtc ccaaatctac atcgcttgag cgaggatacc aaaaaggtat 660

accgatgtat ggtttccaac agactcaatt attttcccta ttatgaggcg ttcaggcggt 720

ctttgtttga tatgtatatg ctaggtcggt tggggcgtcg acttaagcga tctgactgggg 780

agactattat gcatctgtca ccaacgcaaa gtcggcgtct acatagaact ttaagatttg 840

tggagcgtag aattatccca tctaacagtt atatacgcac atcgggccac gttccgcctt 900

cgagggcact tccgacagat acgaatttaa agatggatga ataattaaat tggaaagagt 960

aactacatta atcgagcgtc atgacggcgt cccgtgaaaa tgggaatttt ctactcgaaa 1020

1080

ggcaacatac ataatgtgca tgcgaaacca ctttttcagt gtacgctgac attgtgcaac 1140

acggaggggt agcatctaca tacaatatat gttgattaat gattggagaa aaaactatgc 1200

agctcgccga tcatatggct aactcgcctt cgtctatatg gcggaccccg cgggaaaaat 1260

cgacgtacca tctgatttac aacaccagta atgaacatgt cgcatccctg cccagatctg 1320

tgcgcccatt ggcgcggatc gttgtgaatg ccgccgaaac acttcaggtc ggtatgagag 1380

ccgggaggcc gccatcagca ggagtttggc gagaggtgtt tgatagaatg atgacagcct 1440

tccgtgacca cgagcctact gcgacattta atgctgcaaa tcccattaga aaaatggtcg 1500

agacagttct acagaataat gaagagcccc cgcggacgca tgctgaaatg ggtaatcgcc 1560

ttatgaacat tatgtactgg tgttgcttgg gacacgcagg acaatgctcg atatggcagt 1620

tgtacgagac gaatcaggcc attttaagtt tattagatga agtggttatc ggcacaacaa 1680

atcccttttg caccctcgag caatactgga agccattatg caccgcaatc gccaacaagg 1740

ggacctcatc gcttgttgag gatgccaaag tggccgagta cctggttagc atgcgcaaat 1800

tgatataaca taggcacgct ctgatgttac agaccacaat accgcataca tttattgtaa 1860

ggttgttaat aaaggtttat tctatgtaag actacaatac tttcgacatt gcttgtatac 1920

atattaaata ctttctcaag ttcctattac ataaaatggg atctatcatt acattcgtta 1980

agagtctgga taattttact gtttgccagc ttcgatcttg gaacgtactg tggatagtgc 2040

cttacttgga atcgtgaaaa tttgaaacgt ccattatttg gatatcttcc ggttgtccca 2100

2160

aatcggggac caacaacgcg tgggtccaca ctcattcgga aattttccga tgattctgaa 2220

tatttattgc cgctcgttac gagtcgttgg acatatctgt aatacatttc ttcttctgaa 2280

ggatcgctgc acatttgatc tatacattgg ccaggatgtt caagtctcag atgttgcatt 2340

ctggcacagc acaactttat ggcatttccg atgtaatcgt ccggcagccc tgggggagtt 2400

ctatattcgc atattgggat ggtaaggaca atagcagatc tcgcaacctc cagggaggct 2460

2520

tatcctttac tagcgccgat tgagagcatc gtcgtccaat tttctaaatg gaaagaaaac 2580

aaggcgggca agagtgttcc aaacattttc attttcggcg aatctctcaa atcccatggc 2640

gtgcaattga ttgcaaaatt ggcacttccg ttcacgtttg tatctccaaa ctctaagaca 2700

cttttaattg aaaaactacg ttctagtgtg gaaagaaacc tataggcaga ccatagaact 2760

atttgacacc acatatcttt ttgtatgtca aactgaccat gatcgtatgt tgctgaatgc 2820

actagggcaa ttcgctcgcg cgactccata cattgaataa ttccacacgt cagctcatcg 2880

gttagcaagg tccagtagtt gaagtcattt atttttcccc gcggctggcc aaatctacct 2940

ctgggaatat ccaagttgtc gaatatgatc gcaccggctc tggtcatggt gaaggaactg 3000

tagcataaag acgcaggtat cataggggta atattttttt attcactcac atactaaaag 3060

taacgcatat tagcaccatg tatgggctat caattgacat ttgcgtagca ctacatcacg 3120

3180

gggtttatct actattgaat ttttcccctat ctgtgataca cttgggagcc tctacaagca 3240

tattgccatc atgtacgttt ttatctactg tcttaacgcc catgggaacg gaggcgtcgt 3300

3360

tggactcgat accaagcccc tgcagctggg gaacgtctgg tggagagccg ataatttgat 3420

atacgcacgc catattactg tcgttgaagt acgccttatc ttctatgttt tcaaatttag 3480

gttcccaagt ggacgtgaga agtgtttgta tctcacatgg aatggcccaa ggcattccag 3540

cccaggtgcc tggtacttta atggcaaaca aacgttttgg tagaggtatt gattctattg 3600

3660

caagcttgtt aattaagtcg acggcagagt cgcagacgcc cctattggac gtcaaaattg 3720

tagaggtgaa gttttcaaac gatggcgaag taacggcgac ttgcgtttcc accgtcaaat 3780

ctccctatag ggtagaaact aattggaaag tagacctcgt agatgtaatg gatgaaattt 3840

ctgggaacag tcccgccggg gtttttaaca gtaatgagaa atggcagaaa cagctgtact 3900

acagagtaac cgatggaaga acatcggtcc agctaatgtg cctgtcgtgc acgagccatt 3960

ctcgggaacc ttactgtctt ttcgacacgt ctcttatagc gagggaaaaa gatatcgcgc 4020

cagagttata ctttacctct gatccgcaaa cggcatactg cacaataact ctgccgtccg 4080

gcgttgttcc gagattcgaa tggagcctta ataatgtttc actgccggaa tatttgacgg 4140

ccacgaccgt tgtttcgcat accgctggcc aaagtacagt gtggaagagc agcgcgagag 4200

caggcgaggc gtggatttct ggccggggag gcaatatata cgaatgcacc gtcctcatct 4260

cagacggcac tcgcgttact acgcgaaagg agaggtgctt aacaaacaca tggattgcgg 4320

tggaaaacgg tgctgctcag gcgcagctgt attcactctt ttctggactt gtgtcaggat 4380

tatgcgggag catatctgct ttgtacgcaa cgctatggac cgccatttat ttttgaggaa 4440

tgctttttgg actatcgtac tgctttcttc cttcgctagc cagagcaccg ccgccgtcac 4500

gtacgactac attttaggcc gtcgcgcgct cgacgcgcta accataccgg cggttggccc 4560

gtataacaga tacctcacta gggtatcaag aggctgcgac gttgtcgagc tcaacccgat 4620

ttctaacgtg gacgacatga tatcggcggc caaagaaaaa gagaaggggg gccctttcga 4680

ggcctccgtc gtctggttct acgtgattaa gggcgacgac ggcgaggaca agtactgtcc 4740

aatctataga aaagagtaca gggaatgtgg cgacgtacaa ctgctatctg aatgcgccgt 4800

tcaatctgca cagatgtggg cagtggacta tgttcctagc acccttgtat cgcgaaatgg 4860

cgcgggactg actatattct cccccactgc tgcgctctct ggccaatact tgctgaccct 4920

gaaaatcggg agatttgcgc aaacagctct cgtaactcta gaagttaacg atcgctgttt 4980

aaagatcggg tcgcagctta actttttacc gtcgaaatgc tggacaacag aacagtatca 5040

gactggattt caaggcgaac acctttatcc gatcgcagac accaatacac gacacgcgga 5100

cgacgtatat cggggatacg aagatattct gcagcgctgg aataatttgc tgaggaaaaa 5160

gaatcctagc gcgccagacc ctcgtccaga tagcgtcccg caagaaattc ccgctgtaac 5220

caagaaagcg gaagggcgca ccccggacgc agaaagcagc gaaaagaagg cccctccaga 5280

agactcggag gacgacatgc aggcagaggc ttctggagaa aatcctgccg ccctccccga 5340

agacgacgaa gtccccgagg acaccgagca cgatgatcca aactcggatc ctgactatta 5400

caatgacatg cccgccgtga tcccggtgga ggagactact aaaagttcta atgccgtctc 5460

catgcccata ttcgcggcgt tcgtagcctg cgcggtcgcg ctcgtggggc tactggtttg 5520

gagcatcgta aaatgcgcgc gtagctaatc gagcctagaa taggtggttt cttcctacat 5580

gccacgcctc acgctcataa tataaatcac atggaatagc ataccaatgc ctattcattg 5640

ggacgttcga aaagcatggc atcgctactt ggaactctgg ctctccttgc cgcgacgctc 5700

gcacccttcg gcgcgatggg aatcgtgatc actggaaatc acgtctccgc caggattgac 5760

gacgatcaca tcgtgatcgt cgcgcctcgc cccgaagcta caattcaact gcagctattt 5820

ttcatgcctg gccagagacc ccacaaaccc tactcaggaa ccgtccgcgt cgcgtttcgg 5880

tctgatataa caaaccagtg ctaccaggaa cttagcgagg agcgctttga aaattgcact 5940

catcgatcgt cttctgtttt tgtcggctgt aaagtgaccg agtacacgtt ctccgcctcg 6000

6060 aacagactaa ccggacctcc acacccgttt

gacagcggga tgttctacgt aattgttcgg ctagacgaca ccaaagaacc cattgacgtc 6120

ttcgcgatcc aactatcggt gtatcaattc gcgaacaccg ccgcgactcg cggactctat 6180

tccaaggctt cgtgtcgcac cttcggatta cctaccgtcc aacttgaggc ctatctcagg 6240

accgaggaaa gttggcgcaa ctggcaagcg tacgttgcca cggaggccac gacgaccagc 6300

6360

cactttacct ttccctggct agaaaatggc gtggatcatt acgaaccgac acccgcaaac 6420

gaaaattcaa acgttactgt ccgtctcggg acaatgagcc ctacgctaat tggggtaacc 6480

gtggctgccg tcgtgagcgc aacgatcggc ctcgtcattg taatttccat cgtcaccaga 6540

aacatgtgca ccccgcaccg aaaattagac acggtctcgc aagacgacga agaacgttcc 6600

6660

gctgaccagg atagtgaact tgtggaactg gttgcgattg ttaacccgtc tgcgctaagc 6720

tcgcccgact caataaaaat gtgattaagt ctgaatgtgg ctctccaatc atttcgattc 6780

tctaatctcc caatcctctc aaaaggggca gtatcggaca cggactggga ggggcgtaca 6840

cgatagttat atggtacagc agaggcctct gaacacttag gaggagaatt cagccgggga 6900

gagcccctgt tgagtaggct tgggagcata ttgcaggatg aacatgttag tgatagttct 6960

cgcctcttgt cttgcgcgcc taacttttgc gacgcgacac gtcctctttt tggaaggcac 7020

tcaggctgtc ctcggggaag atgatcccag aaacgttccg gaagggactg taatcaaatg 7080

gacaaaagtc ctgcggaacg cgtgcaagat gaaggcggcc gatgtctgct cttcgcctaa 7140

ctattgcttt catgatttaa tttacgacgg aggaaagaaa gactgcccgc ccgcgggacc 7200

cctgtctgca aacctggtaa ttttactaaa gcgcggcgaa agcttcgcgc caggtcaatt 7260

ccctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 7320

gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 7380

tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 7440

ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 7500

caaatgggcg gtagcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc 7560

gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccggttgc 7620

gccgccacca tgggccccag accttctacc aagaacccag tacctatgat gctgactgtc 7680

cgagtcgcgc tggtactgag ttgcatctgt ccggcaaact ccattgatgg caggcctctt 7740

gcggctgcag gaattgtggt tacaggagac aaagccgtca acatatacac ctcatcccag 7800

acaggatcaa tcatagttaa gctcctcccg aatctgccca aggataagga ggcatgtgcg 7860

aaagccccct tggatgcata caacaggaca ttgaccactt tgctcacccc ccttggtgac 7920

tctatccgta ggatacaaga gtctgtgact acatctggag gggggagaca ggggcgcctt 7980

ataggcgcca ttattggcgg tgtggctctt ggggttgcaa ctgccgcaca aataacagcg 8040

gccgcagctc tgatacaagc caaacaaaat gctgccaaca tcctccgact taaagagagc 8100

attgccgcaa ccaatgaggc tgtgcatgag gtcactgacg gattatcgca actagcagtg 8160

gcagttggga agatgcagca gtttgttaat gaccaattta ataaaacagc tcaggaatta 8220

gactgcatca aaattgcaca gcaagttggt gtagagctca acctgtacct aaccgaattg 8280

actacagtat tcggaccaca aatcacttca cctgctttaa acaagctgac tattcaggca 8340

ctttacaatc tagctggtgg aaatatggat tacttattga ctaagttagg tgtagggaac 8400

aatcaactca gctcattaat cggtagcggc ttaatcaccg gtaaccctat tctatacgac 8460

tcacagactc aactcttggg tatacaggta actctacctt cagtcgggaa gctaaataat 8520

atgcgtgcca cctacttgga aaccttatcc gtaagcacaa ccaggggatt tgcctcggca 8580

cttgtcccaa aagtggtgac acaggtcggt tctgtgatag aagaacttga cacctcatac 8640

tgtatagaaa ctgacttaca tttatattgt acaagaatag taacgttccc tatgtcccct 8700

ggtatttatt cctgcttgag cggcaatacg tcggcctgta tgtactcaaa gaccgaaggc 8760

gcacttacta caccatacat gactatcaaa ggttcagtca tcgccaactg caagatgaca 8820

acatgtagat gtgtaaaccc cccgggtatc atatcgcaaa actatggaga agccgtgtct 8880

ctaatagata aacaatcatg caatgtttta tccttaggcg ggataacttt aaggctcagt 8940

ggggaattcg atgtaactta tcagaagaat atctcaatac aagattctca agtaataata 9000

acaggcaatc ttgatatctc aactgagctt gggaatgtca acaactcgat cagtaatgct 9060

ttgaataagt tagaggaaag caacagaaaa ctagacaaag tcaatgtcaa actgactagc 9120

acatctgctc tcattaccta tatcgtgttg actatcatat ctcttgtttt tggtatactt 9180

agcctgattc tagcatgcta cctaatgtac aagcaaaagg cgcaacaaaa gaccttatta 9240

tggcttggga ataatactct agatcagatg agagccacta caaaaatgtg aggatctctc 9300

gaggaattct agatcccacg tcactattgt atactctata ttatactcta tgttatactc 9360

tgtaatccta ctcaataaac gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtctt 9420

cttatcacca tcaggtgaca tcctcgccca ggctgtcaat catgccggta tcgattccag 9480

tagcaccggc cccacgctga caacccactc ttgcagcgtt agcagcgccc ctcttaacaa 9540

gccgaccccc accagcgtcg cggttactaa cactcctctc cccgacctgc aactagtaag 9600

cttgcctccg attctagcat tacatagccg gtcagtagat cctgccattc ggtagcgcaa 9660

ccggctacat cttcaaacag tctcacaata aatgcatctc tcgttcctgc caatccggaa 9720

ccgggcatac cactcccgcc tgccgattta attctcacaa ttgggcgatg ccggcggggc 9780

aaaacgaatg tggatttggc aaaccgacac aggtctgctg tacggactaa tatgggcaca 9840

cccacatcat tcttcagatg ctccatgcat tgttctatga gaaagatcca tagggtggag 9900

gcagcgtcac gagatcgccc aggcaatcga tcgcattcgt ctagtaaagt gacgagagtt 9960

atcatgcaca cacccatgcc cacgccttcc gaataactgg agctgtggaa gatcggaaac 10020

gtctttttga ctgccggtct cgtactactt tcgcacaggt gtatacccgg acgcgtacta 10080

tatattttat atcatccaac gtccgaaatt acatacgtgg cggcgatgga agtagatgtt 10140

gagtcttcga aagtaagtgc ctcgaatatg ggtattgtct gtgaaaatat cgaaagcggt 10200

acgacggttg cagaaccgtc gatgtcgcca gatactagta acaatagctt cgataacgaa 10260

10320

10380 cgtatggaat cttcgtgccg tgaacaacga gcggcgtgcg

cctacgtctg ccgtgcgcat gcaatacagt attctttcat ctctcgctcc gggttcagag 10440

ggtcatgtat atatatgtac tagatacggg gacgcggacc aaaaaaaatg catagtgaag 10500

gcagtcgttg gaggaaagaa tcccgggagg gaagtggata ttttaaaaac catctcacat 10560

aaatcaatta taaaattaat ccatgcctat aaatggaaaa atgttgtgtg tatggcaatg 10620

cgtgtatatc gttatgatct tttcacatat attgacggag tcggccctat gccccttcaa 10680

cagatgatct atattcaacg tggactacta gaggcgctag catacataca tgaaaaggggc 10740

atcattcacc gagacgtaaa gacggagaat atattcttgg ataatcacga aaatgcagtt 10800

ttgggtgact tcggtgctgc atgccaacta ggagattgta tagatacgcc ccaatgttac 10860

ggttggagcg gaactgtgga aacaaattcg ccggaattat ctgcacttga tccgtattgc 10920

10980

ccattgttta gtaagcaggt gaaaagttcg ggatctcagc tgagatccat aatacggtgc 11040

atgcaagtgc atgaactgga gtttccccgc aacgattcta ccaacctctg taaacatttc 11100

aaacaatatg cggttcgtgt acgaccgcct tataccattc ctcgagttat aagaaatggg 11160

gggatgccaa tggatgttga atatgtcatt tctaaaatgc ttacgtttga ccaggagttc 11220

agaccttctg ctaaggaaat attgaatatg cccctattta ctaaggcgcc gattaacctg 11280

cttaatatca caccctctga cagtgtctaa cggtatacag gcgggagcgg gtcgtggcgt 11340

catcatcacc acttgagaat ttatattttg aattgttgat tgataaatta acctgattca 11400

ttgagaactg aaacgccata ttggtttctt ggatatgtct acaacaatta gttaaattgc 11460

11520

11580

aaatactgag ttgaacttga aaatgctaaa tctgtgacac 11640

tctttgtgtg atgattattg tcaccacttc gaagatggct tcgacattca tgatgttctg 11700

11760

gaagaattat atcacggata agtcaaccaa tattagaata cccacgccat tatttgtatc 11820

11880

actcaatcct ataagtgacc ccaaatatgt ccttttgagc cagcttctaa tgggaaggcg 11940

caaatatgat gcgacggtcg cgtggtttgt tctcggtaaa atgtgtgcca gattaatata 12000

tttgcgcgaa ttttataact gctcgacaaa tgagcctttt ggcacatgtt ctatgagctc 12060

tcctggatgg tgggacaggc gctacgtctc aaccagtttc atttctcgcg acgaattaca 12120

12180

caacggggac tttactacgg ccgatataat gtttaatgtt aaagtggcat gtgccttttc 12240

aaagactgga atagaagatg atacattatg caaaccctttt catttctttg ccaatgcaac 12300

attgcacaat ttaaccatga ttagatcggt aactcttcga gcgcacgaaa gccatttaaa 12360

ggaatgggtg gcacggagag gtggtaacgt ccctgcagtg ctacttgagt ctaccatgta 12420

tcatgcatcc aatctgccta gaaatttcag ggatttctac ataaagtctc cagatgatta 12480

taagtataat cacctagatg ggccatctgt aatgctcatc actgacagac ctagtgaaga 12540

tttggatggg aggctcgttc accaaagtga catttttact actacaagtc ctataaaaca 12600

ggtccggtat gaagagcatc agtcacatac aaagcagtat cctgtaaaca aaatacaagc 12660

tataattttt ttgatagggt taggctcgtt cattggaagc atattcgtag ttttggtagt 12720

12780

ccggtatgtg tataccaggc tatgatcacg tgtgaaactt gggcggacct gtatcatatg 12840

tacaccgtcc ctattcgttt atagccagta cgtgttatct gcacatagag gaacatgtgt 12900

catactggga tcgcatgcat ggtatgtgtg actctaatat tattctgtat cataataaaa 12960

acacagtgca tggtatatag aggatcgctg gtaagcacta cggtagacca atcggctcag 13020

attgcattct ttggcatcga taccgttgtt aatttatatg gcaaagtctt gttcatggga 13080

13140

aaccgctgct attctattgc gcatgcaaca tattacgccg actgtcctat aatcagttct 13200

acggtattca gaggatgccg ggacgccgtt gtttatacta ggccccacag cag 13253

<210> 29

<211> 12248

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 29

ggcgcgccac tggagaacgg catgaccgca aaaggcgttg tagagatcga tcccacgaac 60

tctcaggcga tcgtgtcagt cgccataaac agcgacgatc gtctccagga tctgaacggt 120

tttcttctca acgatcatca gtatatgagg aactgaacct gatatttagc cgagggaaac 180

gcaggttaaa aaccctatca agcgattgcg attttcgcgt atctagtaaa aatagatggg 240

cttcggtact agccttcgcc gccaactctg aatatgccct tcgtggacct catataacat 300

ggcattgttt gttggatgcg gggccggaat taagaagaac attcgaaata cgagcaaaaa 360

tttcggccct ggcatgtgct gcgcgagaat cggtacttcg gggagaaagt tttatcggag 420

ctttgggtag tgcagaggaa actctatctt ggttgaaaat gcatgcgacc ctgcacttga 480

ttctggttaa ccacgatcca atttttaaga cggctggcgc ggtcctagat aacctccgct 540

taaaactagc cccaatattg atgtgcagat ataacacaga aaaacgatca atggaagaca 600

tgctacggcg gtcatctccc gaagacatca ccgattccct aacaatgtgc ctgattatgt 660

tatcgcgcat tcgtcgtacc atgcgcaccg caggaaataa atatagctat atgatagatc 720

caatgaatcg tatgtctaat tacactccag gcgaatgtat gacaggtata ttgcgatata 780

ttgacgaaca tgctagaagg tgtcctgatc acatatgtaa tttgtatatc acatgtacac 840

ttatgccgat gtatgtgcac gggcgatatt tctattgtaa ttcatttttt tgttagtaaa 900

ctaccacagg ctgtccggaa atctaagtta atgaataaag tagatggtta atactcattg 960

cttagaattg gactactttt aattctcttt aatgttcgta ttaaataaaa acatctttaa 1020

1080

tcttcggaga gtgtactcgc cacggtggtt ggaacatcac tatgtctaca cgtcaaattt 1140

aagcacgtca ggtctgtcga ggacaagaaa tggttaacta gtgtttcaat tattcttata 1200

aacgttaagc attgtaagcc ccccggccgt ccgcagcaac aatttactag tatgccgtgg 1260

gctccgggac tatcacggat gtccaattcg cacatgcata taatttttct agggtctctc 1320

1380

aaatgaaggt gctccaacac ggtcttcaaa gcaaccggca taccagcaaa cacagactgc 1440

aactccccgc tgcaatgatt ggttataaac agtaatctgt cttctggaag tatatttcgc 1500

ccgacaatcc acggcgcccc caaagttaaa aaccatccat gtgtatttgc gtcttctctg 1560

ttaaaagaat attgactggc attttcccgt tgaccgccag atatccaaag tacagcacga 1620

tgttgcacgg acgactttgc agtcaccagc cttcctttcc acccccccac caacaaaatg 1680

tttatcgtag gacccatatc cgtaataagg atgggtctgg cagcaacccc ataggcgcct 1740

cggcgtggta gttctcgagg ccttaattaa gtcgacggca gagtcgcaga cgcccctatt 1800

ggacgtcaaa attgtagagg tgaagttttc aaacgatggc gaagtaacgg cgacttgcgt 1860

ttccaccgtc aaatctccct atagggtaga aactaattgg aaagtagacc tcgtagatgt 1920

aatggatgaa atttctggga acagtcccgc cggggttttt aacagtaatg agaaatggca 1980

gaaacagctg tactacagag taaccgatgg aagaacatcg gtccagctaa tgtgcctgtc 2040

gtgcacgagc cattctccgg aaccttactg tcttttcgac acgtctctta tagcgaggga 2100

aaaagatatc gcgccagagt tatactttac ctctgatccg caaacggcat actgcacaat 2160

aactctgccg tccggcgttg ttccgagatt cgaatggagc cttaataatg tttcactgcc 2220

ggaatatttg acggccacga ccgttgtttc gcataccgct ggccaaagta cagtgtggaa 2280

gagcagcgcg agagcaggcg aggcgtggat ttctggccgg ggaggcaata tatacgaatg 2340

caccgtcctc atctcagacg gcactcgcgt tactacgcga aaggagaggt gcttaacaaa 2400

cacatggatt gcggtggaaa acggtgctgc tcaggcgcag ctgtattcac tcttttctgg 2460

acttgtgtca ggattatgcg ggagcatatc tgctttgtac gcaacgctat ggaccgccat 2520

ttatttttga ggaatgcttt ttggactatc gtactgcttt cttccttcgc tagccagagc 2580

accgccgccg tcacgtacga ctacatttta ggccgtcgcg cgctcgacgc gctaaccata 2640

ccggcggttg gcccgtataa cagatacctc actagggtat caagaggctg cgacgttgtc 2700

gagctcaacc cgatttctaa cgtggacgac atgatatcgg cggccaaaga aaaagagaag 2760

gggggccctt tcgaggcctc cgtcgtctgg ttctacgtga ttaagggcga cgacggcgag 2820

gacaagtact gtccaatcta tagaaaagag tacagggaat gtggcgacgt acaactgcta 2880

tctgaatgcg ccgttcaatc tgcacagatg tgggcagtgg actatgttcc tagcaccctt 2940

gtatcgcgaa atggcgcggg actgactata ttctccccca ctgctgcgct ctctggccaa 3000

3060

aacgatcgct gtttaaagat cgggtcgcag cttaactttt taccgtcgaa atgctggaca 3120

acagaacagt atcagactgg atttcaaggc gaacaccttt atccgatcgc agacaccaat 3180

acacgacacg cggacgacgt atatcgggga tacgaagata ttctgcagcg ctggaataat 3240

ttgctgagga aaaagaatcc tagcgcgcca gaccctcgtc cagatagcgt cccgcaagaa 3300

3360

aaggcccctc cagaagactc ggaggacgac atgcaggcag aggcttctgg agaaaatcct 3420

gccgccctcc ccgaagacga cgaagtcccc gaggacaccg agcacgatga tccaaactcg 3480

gatcctgact attacaatga catgcccgcc gtgatcccgg tggaggagac tactaaaagt 3540

tctaatgccg tctccatgcc catattcgcg gcgttcgtag cctgcgcggt cgcgctcgtg 3600

gggctactgg tttggagcat cgtaaaatgc gcgcgtagct aatcgagcct agaataggtg 3660

gtttcttcct acatgccacg cctcacgctc ataatataaa tcacatggaa tagcatacca 3720

atgcctattc attgggacgt tcgaaaagca tggcatcgct acttggaact ctggctctcc 3780

ttgccgcgac gctcgcaccc ttcggcgcga tgggaatcgt gatcactgga aatcacgtct 3840

ccgccaggat tgacgacgat cacatcgtga tcgtcgcgcc tcgccccgaa gctacaattc 3900

aactgcagct atttttcatg cctggccaga gaccccacaa accctactca ggaaccgtcc 3960

gcgtcgcgtt tcggtctgat ataacaaacc agtgctacca ggaacttagc gaggagcgct 4020

ttgaaaattg accgagtaca 4080

cgttctccgc ctcgaacaga ctaaccggac ctccacaccc gtttaagctc actatacgaa 4140

atcctcgtcc gaacgacagc gggatgttct acgtaattgt tcggctagac gacaccaaag 4200

aacccattga cgtcttcgcg atccaactat cggtgtatca attcgcgaac accgccgcga 4260

ctcgcggact ctattccaag gcttcgtgtc gcaccttcgg attacctacc gtccaacttg 4320

aggcctatct caggaccgag gaaagttggc gcaactggca agcgtacgtt gccacgggagg 4380

ccacgacgac cagcgccgag gcgacaaccc cgacgcccgt cactgcaacc agcgcctccg 4440

aacttgaagc ggaacacttt acctttccct ggctagaaaa tggcgtggat cattacgaac 4500

cgacacccgc aaacgaaaat tcaaacgtta ctgtccgtct cgggacaatg agccctacgc 4560

taattggggt aaccgtggct gccgtcgtga gcgcaacgat cggcctcgtc attgtaattt 4620

ccatcgtcac cagaaacatg tgcaccccgc accgaaaatt agacacggtc tcgcaagacg 4680

acgaagaacg ttcccaaact agaagggaat cgcgaaaatt tggacccatg gttgcgtgcg 4740

aaataaacaa gggggctgac caggatagtg aacttgtgga actggttgcg attgttaacc 4800

cgtctgcgct aagctcgcc gactcaataa aaatgtgatt aagtctgaat gtggctctcc 4860

aatcatttcg attctctaat ctcccaatcc tctcaaaagg ggcagtatcg gacacggact 4920

gggaggggcg tacacgatag ttatatggta cagcagaggc ctctgaacac ttaggaggag 4980

aattcagccg gggagagccc ctgttgagta ggcttgggag catattgcag gatgaacatg 5040

ttagtgatag ttctcgcctc ttgtcttgcg cgcctaactt ttgcgacgcg acacgtcctc 5100

tttttggaag gcactcaggc tgtcctcggg gaagatgatc ccagaaacgt tccggaaggg 5160

actgtaatca aatggacaaa agtcctgcgg aacgcgtgca agatgaaggc ggccgatgtc 5220

tgctcttcgc ctaactattg ctttcatgat ttaatttacg acggaggaaa gaaagactgc 5280

ccgcccgcgg gacccctgtc tgcaaacctg gtaattttac taaagcgcgg cgaaagcttc 5340

ccgggttaat taaggccctc gaggatacat ccaaagaggt tgagtattct ctctacactt 5400

cttgttaaat ggaaagtgca tttgcttgtt cttacaatcg gcccgagtct cgttcacagc 5460

gcctcgttca cacttaaacc acaaatagtc tacaggctat atgggagcca gactgaaact 5520

cacatatgac taatattcgg gggtgttagt cacgtgtagc ccattgtgtg catataacga 5580

tgttggacgc gtcctttattc gcggtgtact tgatactatg gcagcgagca tgggatattc 5640

atcctcgtca tcgttaacat ctctacgggt tcagaatgtt tggcatgtcg tcgatccttt 5700

gcccatcgtt gcaaattaca agtccgatcg ccatgaccgc gataagcctg taccatgtgg 5760

cattagggtg acatctcgat catacattat aagaccaacg tgcgagtctt ccaaagacct 5820

gcacgccttc ttcttcggat tgtcaacggg ttcttcagaa tctatgccca tatctggcgt 5880

tgagaccatt gtgcgtttaa tgaacaataa agcggcatgc catggaaagg agggctgcag 5940

atctccattt tctcacgcca ctatcctgga cgctgtagac gataattata ccatgaatat 6000

agagggggta tgtttccact gccactgtga tgataagttt tctccagatt gttggatatc 6060

tgcattttct gctgccgaac aaacttcatc gctatgcaaa gagatgcgtg tgtacacgcg 6120

6180

aaaataattg accagtgaac aatttgttta atgttagttt attcaatgca ttggttgcaa 6240

atattcatta cttctccaat cccaggtcat tctttagcga gatgatgtta tgacattgct 6300

gtgaaaatta ctacaggata tatttttaag atgcaggagt aacaatgtgc atagtaggcg 6360

tagttatcgc agacgtgcaa cgcttcgcat ttgagttacc gaagtgccca acagtgctgc 6420

ggttatggtt tatgcgcaca gaatccatgc atgtcctaat tgaaccatcc gatttttctt 6480

ttaatcgcga tcgttgtttg ggcaactgcg ttatttcaga tctaaaaaat ttacccttta 6540

tgaccatcac atctctctgg ctcatacccc gcttggataa gatatcatgt agattccgcc 6600

ctaagaaatg caaactaaca ttattgtcgg ttccatatac acttccatct tgtccttcga 6660

aaataacaaa ctcgcgcaat agaccgtccg tacatgcatg gccgatgtgt gtcaacatca 6720

ttggtctgct agatcccgat gggacgaatc gtacagtcgt cgctccagca ttggcaaaaa 6780

tccccagata ccctccatgc ggcaaatcta aattgcgacc ccgaagagac tgcaccaaag 6840

tcttatcgac gcacgctgat ttttttgaac agcgggagcc cattatcttc agtggagcgt 6900

agacgggcga ggctaattat gtgacatagc aacactgcat gtatgttttt ataaatcaat 6960

aagagtacat aatttattac gtatcatttc cgtttgtaat atactgtata catcatccac 7020

actattagtc agcactagcg cgcgggcgca cgttacaata gcagcgtgcc cgttatctat 7080

attgtccgat atttacacat aacatttcat cgacatgatt aaatacctaa gtactgcaca 7140

cagatgttta atgtatatcg tcatataaat tatatcgcta ggacagaccc aaacgacctt 7200

tatcccaaac agtcagatcc tcttctcaag tgtcgatttc tgttatggaa tatgcatacc 7260

ctggcccaga aattgcacgc acgagcgtag tgaatgcgtc attggtttta catttaaagg 7320

ctaaatgcac aaattcttta gacgacagca catcgttaaa tagcatctct agcgttctta 7380

tgaatgctaa gcattggagt cctcctggtc ggccacaata acagctgagt atcataccct 7440

gagctccggg gttgtcgcac atagcggatt cgtataaaaca taggattttc cgcgaatcca 7500

tcagttgcaa aaatctgtta ggctccatca acaacgctgg atttacttca gatccacgcg 7560

taaagtaatg gtgctcgaat accgttttta gagttgtcgg catttcaagg aacaaagaat 7620

tcatttcttc attgcaacga cgcgccagaa atcccaagac ctctttgggt agtatgttct 7680

tgcctataaa acacggcgtt ccaagtgcca ggaaccacgc atgtgttact gttggggcgt 7740

attcagaaat aaagcggggt ttatgcggct tttgaagctc ggatatccaa agtatcgctt 7800

gctgatgaac gagcgatgta gctgttacaa aacctccttt ccatcctcca gtcaacataa 7860

tatttatcgg cctacctatg tccgtaataa gtattggtcg ggcaattatt ccgtatgagg 7920

tcttgcagga ataagctctt agggacagcc agcttggata tggtgcgaaa cagaccttct 7980

cggcttcaga atgtcgctcc gcagtctctt cgtgtcggtg catcttagat ccaccatcaa 8040

tgtgtgcagc attgactccc gcccgtcgaa tattcctttt gttacgatgc agtaatgagc 8100

acgatcatgg gcggggcgat gacgttctat ttgcatgtct gcgaacaatt tgcgtcagtc 8160

atacagctat ggagtgggcc atttctggcc gtcaacttaa aaacgcgaac cgcagacata 8220

tgtatttgca tgcaaagacg tatcttcgta tttctgggca tcttcaaatg ctctggccaa 8280

tatggcaatg aatttggatt cgtttgacgc cgatggtatg cagtgcaaat gtgccaatag 8340

cccacatccg aaaaagttat ttgtcataca agcaggtgtt aagtagcaat cacataaagg 8400

caccagacgc ctcatggcat cataatgaat agctccttct ccccactggga accactgaca 8460

aaatctgcga gtatattccg caaaccacat tttatttctc atagaaacta ccctaaatcc 8520

ttttaacggg aagaagaatc ctagatagtg cttgaagtca tgactgttac tgctgcaata 8580

acactgtata ttatttataa attccgtttg tctaggtatc tgatgtaggc attccgatcc 8640

ctttactatt gcgtcttcac gaccaaatgg gaatgcgcca aaatccccac acctcatcac 8700

cctggaggca gattgtgtat tattaatatc cgccgattga agcacaaaac ggtacggtac 8760

tgttcctaat tctggtatag attctatggt caaaagtctg catatccccg acattgccat 8820

gagatcacac agtccaagta gcatgtttat tgagtcactc agactgtcaa cgtccctcgc 8880

cgcaccacca atcgaaaata aagtatctac gcaagttata gctccgcatt ttctatcgct 8940

agcagcaatc gcgacgcaaa acataaaggc catgttggga tttgaactct ctggggggct 9000

tgttatcttc tgcaccgtcg cagtcgcagt tttccgaaat ttatgtctaa tatattttcc 9060

ggccgtgctc caatcggccg aaaagaatct gcgtattacc agactcattg acgggccgat 9120

aaagaccata aaacaaaatt cctgtgcact ccctcctcca gttttgccat cgtccaagtc 9180

ccgtaacttt ttttgcgttt cgaggagcaa gcgttcgtta tccctaccca cacttgtttt 9240

ccaccgtttt cttattataa gcggttgtat cgccaacgcg tcaccgcagg ttgtcacata 9300

cagtgatggc atacttgaac gtgcaacaac gcgctcgctt tgcaaatcta agtcattgac 9360

catcaaatcg cgttgagagg atagccaggc atcttttttc ctagtatggt gacggtgcag 9420

ccaccccaac tcagttcttg taaaaaaagc tattggcggg aatttatgtt ctgaggtgca 9480

ttctatattt atgagtccat caaatgccat taaccagatt cgtatttttt cgctcgaccc 9540

ggcatcacta tggatacaat acctttctat ggcccatttc agctctcgaa ccaaccacac 9600

9660

atggcatccg agcgctctta ctgtacggtc ggatacaccc atggtttttc ctttatatag 9720

tcgggttata gtctgtcggg tttggcggta gcacggagta gtttgatttt taagaatcga 9780

aaaccggctt ggagagacca ctgtcgaata tttgtccgta tactctacac gtgagtgttg 9840

tccattccta ggtatattca tctgttcgga taccttcaat tgctgttcag gcataacctt 9900

aaagcatatg ttatgttgta catcaaaact tggtgagtta tgttcgattg ccgcgcataa 9960

agaatcgtac atgagcgttt ctgctaacat actatctata ttctcacacg cccctgcata 10020

tactgttcct attccaaatt cacgttttgc cccatcggct atctgctccc aaaaagttgt 10080

aatataggtg ccgctggggtg cgaaattttc atcagttgta ttcctgataa actgaatcac 10140

tttacataat ttttgccaca tatctgcgtg cagccatagt atcgaacccg tgggctcgga 10200

gacgacagtg cgtacaatgg gtattttacc tttccccaac aaaataatgg tatacaagtt 10260

aggtccgtac ctagacctta atgtttccaa ttcttctgaa tcactgcact ctcgtaggggg 10320

agtaacggta ataatttcgt ctctgagccc cgttttgcgt tgaaaactaa tcacattaga 10380

taatgtgcaa tcggtttctt ttatccggat acatctaagt attatgacat cggtggtcat 10440

tgtttccatc aacgaccatc ttttacgatc gcccatacta ctcatggacg ttgtcggtgt 10500

tgaaaaatca ccagaattgc aacggatctc tgggtaccat gctgctgatg gaattggcgg 10560

ttttaattgt tgtttcagtc tattattgct atctttggcg gggttgaata atgtgggggg 10620

agagtgattg caggaatccg aatgggtcaa taaaacgacc gtgctccgtt ctgccggcgc 10680

cgatccgatt gaagctatat acttcgcttc tctccccact tttccaattt gatccggaaa 10740

taaaacggcc ccggacaaca gtatcgtacg atccggatcc ggatcctgct tgcctacaga 10800

agaatcaaca tctcgcccca atattctggt caaaactggc tcgctcatgg caacgcggac 10860

gtttcccccg gtggccagtc ttaatggtta atgttctttt cggcaatctt atacatcagc 10920

gggttgcgtg aatactggtc acagttcagt catttactac acaccagcaa tacgacgacg 10980

11040

atgtcgtacg ggcggtgcgg ggttggatcc tcggcaaaga gatcctcgta attcggcggt 11100

gggagcggag ggtaaagacg cgggtggggga tctccctccg gaccgcgcgc cgggcgcggt 11160

tcgaaaatgc tttccgcctc gctcagtgtc aacgccaagt attcgggcgg gctgggggcc 11220

ggaatatctc ccgcgacttc ttctatcggc gcggaattgg agtcgcggtc gtggcgcgct 11280

tctagcgtcg tcaacggaag tccattttcg gggtctcccg gtgggcgttc agcgtccatc 11340

gtcgtatatg ctctaacaca cgtctcgcta tattaaaaaa aagaagagta tcggtcagtg 11400

11460

11520

11580

11640

ccacatctcc cgaattagta gaggaaagcc cgcgctgtcg gtttggggac atatcgatct 11700

tttttgtgtt tttcctagga cccttttgcc agatcgtaca aagtcgcgtc ttatgagcgg 11760

acgttcttac tgcagctcgg taggagtggg gcagggttag atttcgtcgg cgtttcggcc 11820

11880

11940

ttagcgccat catgtggttc gtggcggtgg aaaggctatt atgtgttagg ggagagacca 12000

cgtgatcggc atgcaaatga gcaaggcgaa cgcgtcagcg ttcgcactgc gaaccaataa 12060

tatatatatt atactattgg ctttaggtgc gaacgtccgg ctagtccaat agcggggtcg 12120

cgtttcgtac cacgtgttat agaccgccct aaactcgcac tcgggggtcc ggccgcgccc 12180

agacagggcg gagacgtgcc acaggggctt taaaacaccg cttcgggcac cgttcatctc 12240

ggcgcgcc 12248

<210> 30

<211> 13068

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 30

aattccagac taaatgcccc ggcccaattt gtcaagtgtg cagtcacgga ggcgtcgacc 60

gtgtccccgg cattaaacag gaaagcgtta aagtttttga atgttaggtc acaggtacaa 120

acataaatgt ttgtacaaac aggtaacagg tacaaacata aatgccccgg cataaatgtc 180

ccttacggcg gatcgaaacg acattaggca tactcgggta ccattttgca ttccgatcag 240

cacggatgaa attaggcagg aatgcggttt atattatgcg gcattggaca aacgatatgg 300

cattgattgg cagtttatga atgtcttcat gtggggcgta aacggattcc tattggttca 360

gaagacaacg acgatatatt tagagagaaa aagctaccca gcataggata aacacacatt 420

gagcattgag agacataggt atcggtatgg atgggaaaac tacacacgtg aacaccaaac 480

gacttatata ctcgagcggt gatactactg agcaagaatg cactgcatct gagccactga 540

atgaagactg tgatgaaaat gtgaccatcg atggaattgg agaagaatat gcgcagttct 600

tcatgtcccc gcaatgggtc ccaaatctac atcgcttgag cgaggatacc aaaaaggtat 660

accgatgtat ggtttccaac agactcaatt attttcccta ttatgaggcg ttcaggcggt 720

ctttgtttga tatgtatatg ctaggtcggt tggggcgtcg acttaagcga tctgactgggg 780

agactattat gcatctgtca ccaacgcaaa gtcggcgtct acatagaact ttaagatttg 840

tggagcgtag aattatccca tctaacagtt atatacgcac atcgggccac gttccgcctt 900

cgagggcact tccgacagat acgaatttaa agatggatga ataattaaat tggaaagagt 960

aactacatta atcgagcgtc atgacggcgt cccgtgaaaa tgggaatttt ctactcgaaa 1020

1080

ggcaacatac ataatgtgca tgcgaaacca ctttttcagt gtacgctgac attgtgcaac 1140

acggaggggt agcatctaca tacaatatat gttgattaat gattggagaa aaaactatgc 1200

agctcgccga tcatatggct aactcgcctt cgtctatatg gcggaccccg cgggaaaaat 1260

cgacgtacca tctgatttac aacaccagta atgaacatgt cgcatccctg cccagatctg 1320

tgcgcccatt ggcgcggatc gttgtgaatg ccgccgaaac acttcaggtc ggtatgagag 1380

ccgggaggcc gccatcagca ggagtttggc gagaggtgtt tgatagaatg atgacagcct 1440

tccgtgacca cgagcctact gcgacattta atgctgcaaa tcccattaga aaaatggtcg 1500

agacagttct acagaataat gaagagcccc cgcggacgca tgctgaaatg ggtaatcgcc 1560

ttatgaacat tatgtactgg tgttgcttgg gacacgcagg acaatgctcg atatggcagt 1620

tgtacgagac gaatcaggcc attttaagtt tattagatga agtggttatc ggcacaacaa 1680

atcccttttg caccctcgag caatactgga agccattatg caccgcaatc gccaacaagg 1740

ggacctcatc gcttgttgag gatgccaaag tggccgagta cctggttagc atgcgcaaat 1800

tgatataaca taggcacgct ctgatgttac agaccacaat accgcataca tttattgtaa 1860

ggttgttaat aaaggtttat tctatgtaag actacaatac tttcgacatt gcttgtatac 1920

atattaaata ctttctcaag ttcctattac ataaaatggg atctatcatt acattcgtta 1980

agagtctgga taattttact gtttgccagc ttcgatcttg gaacgtactg tggatagtgc 2040

cttacttgga atcgtgaaaa tttgaaacgt ccattatttg gatatcttcc ggttgtccca 2100

2160

aatcggggac caacaacgcg tgggtccaca ctcattcgga aattttccga tgattctgaa 2220

tatttattgc cgctcgttac gagtcgttgg acatatctgt aatacatttc ttcttctgaa 2280

ggatcgctgc acatttgatc tatacattgg ccaggatgtt caagtctcag atgttgcatt 2340

ctggcacagc acaactttat ggcatttccg atgtaatcgt ccggcagccc tgggggagtt 2400

ctatattcgc atattgggat ggtaaggaca atagcagatc tcgcaacctc cagggaggct 2460

2520

tatcctttac tagcgccgat tgagagcatc gtcgtccaat tttctaaatg gaaagaaaac 2580

aaggcgggca agagtgttcc aaacattttc attttcggcg aatctctcaa atcccatggc 2640

gtgcaattga ttgcaaaatt ggcacttccg ttcacgtttg tatctccaaa ctctaagaca 2700

cttttaattg aaaaactacg ttctagtgtg gaaagaaacc tataggcaga ccatagaact 2760

atttgacacc acatatcttt ttgtatgtca aactgaccat gatcgtatgt tgctgaatgc 2820

actagggcaa ttcgctcgcg cgactccata cattgaataa ttccacacgt cagctcatcg 2880

gttagcaagg tccagtagtt gaagtcattt atttttcccc gcggctggcc aaatctacct 2940

ctgggaatat ccaagttgtc gaatatgatc gcaccggctc tggtcatggt gaaggaactg 3000

tagcataaag acgcaggtat cataggggta atattttttt attcactcac atactaaaag 3060

taacgcatat tagcaccatg tatgggctat caattgacat ttgcgtagca ctacatcacg 3120

3180

gggtttatct actattgaat ttttcccctat ctgtgataca cttgggagcc tctacaagca 3240

tattgccatc atgtacgttt ttatctactg tcttaacgcc catgggaacg gaggcgtcgt 3300

3360

tggactcgat accaagcccc tgcagctggg gaacgtctgg tggagagccg ataatttgat 3420

atacgcacgc catattactg tcgttgaagt acgccttatc ttctatgttt tcaaatttag 3480

gttcccaagt ggacgtgaga agtgtttgta tctcacatgg aatggcccaa ggcattccag 3540

cccaggtgcc tggtacttta atggcaaaca aacgttttgg tagaggtatt gattctattg 3600

3660

caagctgcgg ccgctctaga actagtggat cccccgggct gcagcccaat gtggaattcg 3720

cccttgcaca ttgttactcc tgcatcttaa aaatatatcc tgtagtaatt ttcacagcaa 3780

tgtcataaca tcatctcgct aaagaatgac ctgggattgg agaagtaatg aatatttgca 3840

accaatgcat tgaataaact aacattaaac gaattcacta gtggatcccc caactccgcc 3900

cgttttatga ctagaaccaa tagtttttaa tgccaaatgc actgaaatcc cctaatttgc 3960

aaagccaaac gccccctatg tgagtaatac ggggactttt tacccaattt cccaagcgga 4020

aagcccccta atacactcat atggcatatg aatcagcacg gtcatgcact ctaatggcgg 4080

cccatagggga ctttccacat agggggcgtt caccatttcc cagcataggg gtggtgactc 4140

aatggccttt acccaagtac attgggtcaa tgggaggtaa gccaatgggt ttttcccatt 4200

actggcaagc acactgagtc aaatgggact ttccactggg ttttgcccaa gtacattggg 4260

tcaatgggag gtgagccaat gggaaaaacc cattgctgcc aagtacactg actcaatagg 4320

gactttccaa tgggtttttc cattgttggc aagcatataa ggtcaatgtg ggtgagtcaa 4380

tagggacttt ccattgtatt ctgcccagta cataaggtca atagggggtg aatcaacagg 4440

aaagtcccat tggagccaag tacactgcgt caatagggac tttccattgg gttttgccca 4500

gtacataagg tcaatagggg atgagtcaat gggaaaaacc cattggagcc aagtacactg 4560

actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag ggggtgagtc 4620

aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc caatgggttt 4680

tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt actggcacgt 4740

atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt caataggggt 4800

gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga ctttccattg 4860

ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc ccattattgg 4920

cacgtacata aggtcaatag gggtgagtca ttgggttttt ccagccaatt taattaaaac 4980

gccatgtact ttcccaccat tgacgtcaat gggctattga aactaatgca acgtgacctt 5040

taaacggtac tttcccatag ctgattaatg ggaaagtacc gttctcgagc caatacacgt 5100

caatgggaag tgaaagggca gccaaaacgt aacaccgccc cggttttccc ctggaaattc 5160

catattggca cgcattctat tggctgagct gcgttctacg tgggtataag aggcgcgacc 5220

agcgtcggta ccgtcgcagt cttcggtctg accaccgtag aacgcagagc tcctcgctgc 5280

aggcggccgc tctagaactc gtcgatcgca gcgatgacaa acctgcaaga tcaaacccaa 5340

cagattgttc cgttcatacg gagccttctg atgccaacaa ccggaccggc gtccattccg 5400

gacgacaccc tggagaagca cactctcagg tcagagacct cgacctacaa tttgactgtg 5460

ggggacacag ggtcagggct aattgtcttt ttccctggat tccctggctc aattgtgggt 5520

gctcactaca cactgcagag caatgggaac tacaagttcg atcagatgct cctgactgcc 5580

cagaacctac cggccagcta caactactgc agactagtga gtcggagtct cacagtgagg 5640

tcaagcacac tccctggtgg cgtttatgca ctaaacggca ccataaacgc cgtgaccttc 5700

caaggaagcc tgagtgaact gacagatgtt agctacaatg ggttgatgtc tgcaacagcc 5760

aacatcaacg acaaaattgg gaatgtcctg gtaggggaag gggtcactgt cctcagccta 5820

cccacatcat atgatcttgg gtatgtgagg cttggtgacc ccattcccgc tatagggctt 5880

gacccaaaaa tggtagctac atgcgacagc agtgacaggc ccagagtcta caccataact 5940

gcagccgatg attaccaatt ctcatcacag taccaaccag gtggggtaac aatcacactg 6000

ttctcagcca acattgatgc tatcacaagc ctcagcattg ggggagagct cgtgtttcaa 6060

acaagcgtcc aaggccttgt actgggcgcc accatctacc ttataggctt tgatgggact 6120

6180

atgccattca atcttgtcat tccaaccaat gagataaccc agccaatcac atccatcaaa 6240

ctggagatag tgacctccaa aagtggtggt caggcagggg atcagatgtc atggtcggca 6300

agtgggagcc tagcagtgac gatccatggt ggcaactatc caggggccct ccgtcccgtc 6360

acactagtag cctacgaaag agtggcaaca ggatccgtcg ttacggtcgc tggggtgagt 6420

aacttcgagc tgattccaaa tcctgaacta gcaaagaacc tggttacaga atacggccga 6480

tttgacccag gagccatgaa ctacacaaaa ttgatactga gtgagaggga ccgtcttggc 6540

atcaagaccg tctggccaac aagggagtac actgattttc gtgagtactt catggaggtg 6600

gccgacctca actctcccct gaagattgca ggagcatttg gcttcaaaga cataatccgg 6660

gctataagga ggtaagcttg atctagagcg gccgcggggga tccagacatg ataagataca 6720

aaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 6780

6840

acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttcggatc 6900

ctctagagtc gacaattatt tcatttaata acatatagcc caaagacctc tatgaacatt 6960

tagtttcccg tatactcaac ggcgcgtgta cacacaaggg cgaattccac agtggatatc 7020

aagcttaatt aagtaccgag ctcgaattgg cgcgccaggt caattccctg gcattatgcc 7080

cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 7140

attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 7200

cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 7260

caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 7320

cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 7380

agacgccatc cacgctgttt tgacctccat agaagacacc gggcgcgccg gatccatggg 7440

ccccagacct tctaccaaga acccagtacc tatgatgctg actgtccgag tcgcgctggt 7500

actgagttgc atctgtccgg caaactccat tgatggcagg cctcttgcgg ctgcaggaat 7560

tgtggttaca ggagacaaag ccgtcaacat atacacctca tcccagacag gatcaatcat 7620

agttaagctc ctcccgaatc tgcccaagga taaggaggca tgtgcgaaag cccccttggga 7680

tgcatacaac aggacattga ccactttgct cacccccctt ggtgactcta tccgtaggat 7740

acaagagtct gtgactacat ctggaggggg gagacagggg cgccttatag gcgccattat 7800

tggcggtgtg gctcttgggg ttgcaactgc cgcacaaata acagcggccg cagctctgat 7860

acaagccaaa caaaatgctg ccaacatcct ccgacttaaa gagagcattg ccgcaaccaa 7920

tgaggctgtg catgaggtca ctgacggatt atcgcaacta gcagtggcag ttgggaagat 7980

gcagcagttt gttaatgacc aatttaataa aacagctcag gaattagact gcatcaaaat 8040

tgcacagcaa gttggtgtag agctcaacct gtacctaacc gaattgacta cagtattcgg 8100

accacaaatc acttcacctg ctttaaacaa gctgactatt caggcacttt acaatctagc 8160

tggtggaaat atggattact tattgactaa gttaggtgta gggaacaatc aactcagctc 8220

attaatcggt agcggcttaa tcaccggtaa ccctattcta tacgactcac agactcaact 8280

cttgggtata caggtaactc taccttcagt cgggaaccta aataatatgc gtgccaccta 8340

cttggaaacc ttatccgtaa gcacaaccag gggatttgcc tcggcacttg tcccaaaagt 8400

ggtgacacag gtcggttctg tgatagaaga acttgacacc tcatactgta tagaaactga 8460

cttagattta tattgtacaa gaatagtaac gttccctatg tcccctggta tttattcctg 8520

cttgagcggc aatacgtcgg cctgtatgta ctcaaagacc gaaggcgcac ttactacacc 8580

atacatgact atcaaaggtt cagtcatcgc caactgcaag atgacaacat gtagatgtgt 8640

aaaccccccg ggtatcatat cgcaaaacta tggagaagcc gtgtctctaa tagataaaca 8700

atcatgcaat gttttatcct taggcgggat aactttaagg ctcagtgggg aattcgatgt 8760

aacttatcag aagaatatctct caatacaaga ttctcaagta ataataacag gcaatcttga 8820

tatctcaact gagcttggga atgtcaacaa ctcgatcagt aatgctttga ataagttaga 8880

ggaaagcaac agaaaactag acaaagtcaa tgtcaaactg actagcacat ctgctctcat 8940

tacctatatc gttttgacta tcatatctct tgttttttggt atacttagcc cgattctagc 9000

atgctaccta atgtacaagc aaaaggcgca acaaaagacc ttattatggc ttgggaataa 9060

tactctagat cagatgagg ccactacaaa aatgtgagga tctctcgagg aattctagat 9120

cccacgtcac tattgtatac tctatattat actctatgtt atactctgta atcctactca 9180

ataaacgtgt cacgcctgtg aaaccgtact aagtctcccg tgtcttctta tcaccatcag 9240

gtgacatcct cgcccaggct gtcaatcatg ccggtatcga ttccagtagc accggcccca 9300

cgctgacaac ccactcttgc agcgttagca gcgcccctct taacaagccg acccccacca 9360

gcgtcgcggt tactaacact cctctccccg acctgcaact agtgcggccg cagcttgcct 9420

ccgattctag cattacatag ccggtcagta gatcctgcca ttcggtagcg caaccggcta 9480

catcttcaaa cagtctcacg ataaatgcat ctctcgttcc tgccaatccg gaaccgggca 9540

taccactccc gcctgccgat ttaattctca caattgggcg atgccggcgg ggcaaaacga 9600

atgtggattt ggcaaaccga cacaggtctg ctgtacggac taatatgggc acacccacat 9660

cattcttcag atgctccatg cattgttcta tgagaaagat ccatagggtg gaggcagcgt 9720

cacgagatcg cccaggcaat cgatcgcatt cgtctagtaa agtgacgaga gttatcatgc 9780

acacacccat gccccgcct tccgaataac tggagctgtg gaagatcgga aacgtctttt 9840

tgactgccgg tctcgtacta ctttcgcaca ggtgtatacc cggacgcgta ctatatattt 9900

tatatcatcc aacgtcccga aattacatac gtggcggcga tggaagtaga tgttgagtct 9960

tcgaaagtaa gtgcctcgaa tatgggtatt gtctgtgaaa atatcgaaag cggtacgacg 10020

gttgcagaac cgtcgatgtc gccagatact agtaacaata gcttcgataa cgaagacttc 10080

10140

gaatcttcgt gccgtgaaca acgagcggcg tgcgaacttc gaaagtgttc gtgtcctacg 10200

tctgccgtgc gcatgcaata cagtattctt tcatctctcg ctccgggttc agagggtcat 10260

gtatatatat gtactagata cggggacgcg gaccaaaaaa aatgcatagt gaaggcagtc 10320

gttggaggaa agaatcccgg gagggaagtg gatattttaa aaaccatctc acataaatca 10380

attataaaat taatccatgc ctataaatgg aaaaatgttg tgtgtatggc aatgcgtgta 10440

tatcgttatg atcttttcac atatattgac ggagtcggcc ctatgcccct tcaacagatg 10500

atctatattc aacgtggact actagaggcg ctagcataca tacatgaaag gggcatcatt 10560

10620

gacttcggtg ctgcatgcca actaggagat tgtatagata cgccccaatg ttacggttgg 10680

10740

acagatattt ggagtgccgg attggttcta tatgagatgg caattaaaaa tgtaccattg 10800

tttagtaagc aggtgaaaag ttcgggatct cagctgagat ccataatacg gtgcatgcaa 10860

10920

tatgcggttc gtgtacgacc gccttatacc attcctcgag ttataagaaa tggggggatg 10980

ccaatggatg ttgaatatgt catttctaaa atgcttacgt ttgaccagga gttcagacct 11040

tctgctaagg aaatattgaa tatgccccta tttactaagg cgccgattaa cctgcttaat 11100

atcacaccct ctgacagtgt ctaacggtat acaggcggga gcgggtcgtg gcgtcatcat 11160

11220

actgaaacgc catattggtt tcttggatat gtctacaaca attagttaaa ttgctatgtt 11280

11340

atataaagta cataacgtgt ttagaatacc cagaatccga atagtccgcg ggggcgtctt 11400

taaatctgtg acactctttg 11460

tgtgatgatt attgtcacca cttcgaagat ggcttcgaca ttcatgatgt tctggtgttt 11520

11580

ttatcacg gataagtcaa ccaatattag aatacccacg ccattatttg tatcaacgga 11640

aaactcttat cccacaaaac atgtaatcta cgatgaaaac tgtggcttcg ctgtactcaa 11700

11760

tgatgcgacg gtcgcgtggt ttgttctcgg taaaatgtgt gccagattaa tatatttgcg 11820

cgaattttat aactgctcga caaatgagcc ttttggcaca tgttctatga gctctcctgg 11880

atggtgggac aggcgctacg tctcaaccag tttcatttct cgcgacgaat tacagctggt 11940

ttttgcagcg ccgtcccgag aattagatgg tttatatacg cgcgtagtag ttgtcaacgg 12000

ggactttact acggccgata taatgtttaa tgttaaagtg gcatgtgcct tttcaaagac 12060

tggaatagaa gatgatacat tatgcaaacc ctttcatttc tttgccaatg caacattgca 12120

12180

ggtggcacgg agaggtggta acgtccctgc agtgctactt gagtctacca tgtatcatgc 12240

12300

taatcaccta gatgggccat ctgtaatgct catcactgac agacctagtg aagatttgga 12360

tgggaggctc gttcaccaaa gtgacatttt tactactaca agtcctataa aacaggtccg 12420

gtatgaagag catcagtcac atacaaagca gtatcctgta aacaaaatac aagctataat 12480

ttttttgata gggttaggct cgttcattgg aagcatattc gtagttttgg tagtatggat 12540

tatacgcaga tattgcaatg gagcgcggag tgggggaacg ccccccagtc ctcgccggta 12600

tgtgtatacc aggctatgat cacgtgtgaa acttgggcgg acctgtatca tatgtacacc 12660

gtccctattc gtttatagcc agtacgtgtt atctgcacat agaggaacat gtgtcatact 12720

gggatcgcat gcatggtatg tgtgactcta atattattct gtatcataat aaaaacacag 12780

tgcatggtat atagaggatc gctggtaagc actacggtag accaatcggc tcagattgca 12840

ttctttggca tcgataccgt tgttaattta tatggcaaag tcttgttcat gggagatcag 12900

tatttggagg aaatatactc tggaacgatg gaaatactca aatggaatca agctaaccgc 12960

tgctattcta ttgcgcatgc aacatattac gccgactgtc ctataatcag ttctacggta 13020

ttcagaggat gccgggacgc cgttgtttat actaggcccc acagcaga 13068

<210> 31

<211> 14598

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 31

cgcgccactg gagaacggca tgaccgcaaa aggcgttgta gagatcgatc ccacgaactc 60

tcaggcgatc gtgtcagtcg ccataaacag cgacgatcgt ctccaggatc tgaacggttt 120

tcttctcaac gatcatcagt atatgaggaa ctgaacctga tatttagccg agggaaacgc 180

aggttaaaaa ccctatcaag cgattgcgat tttcgcgtat ctagtaaaaa tagatgggct 240

tcggtactag ccttcgccgc caactctgaa tatgcccttc gtggacctca tataacatgg 300

360

tcggccctgg catgtgctgc gcgagaatcg gtacttcggg gagaaagttt tatcggagct 420

ttgggtagtg cagaggaaac tctatcttgg ttgaaaatgc atgcgaccct gcacttgatt 480

ctggttaacc acgatccaat ttttaagacg gctggcgcgg tcctagataa cctccgctta 540

aaactagccc caatattgat gtgcagatat aacacagaaa aacgatcaat ggaagacatg 600

ctacggcggt catctcccga agacatcacc gattccctaa caatgtgcct gattatgtta 660

tcgcgcattc gtcgtaccat gcgcaccgca ggaaataaat atagctatat gatagatcca 720

atgaatcgta tgtctaatta cactccaggc gaatgtatga caggtatatt gcgatatatt 780

gacgaacatg ctagaaggtg tcctgatcac atatgtaatt tgtatatcac atgtacactt 840

atgccgatgt atgtgcacgg gcgatatttc tattgtaatt catttttttg ttagtaaact 900

accacaggct gtccggaaat ctaagttaat gaataaagta gatggttaat actcattgct 960

tagaattgga ctacttttaa ttctctttaa tgttcgtatt aaataaaaac atctttaata 1020

aacttcagcc tcttcgctta ttgtagaaat tgagtattca aaatcatgtt caaagccgtc 1080

ttcggagagt gtactcgcca cggtggttgg aacatcacta tgtctacacg tcaaatttaa 1140

gcacgtcagg tctgtcgagg acaagaaatg gttaactagt gtttcaatta ttcttataaa 1200

1260

tccgggacta tcacggatgt ccaattcgca catgcatata atttttctag ggtctctcat 1320

ttcgagaaat cttcggggat ccatcagcaa tgcgggctgt agtcccgatt cccgtttcaa 1380

atgaaggtgc tccaacacgg tcttcaaagc aaccggcata ccagcaaaca cagactgcaa 1440

ctccccgctg caatgattgg ttataaacag taatctgtct tctggaagta tatttcgccc 1500

gacaatccac ggcgccccca aagttaaaaa ccatccatgt gtatttgcgt cttctctgtt 1560

aaaagaatat tgactggcat tttcccgttg accgccagat atccaaagta cagcacgatg 1620

ttgcacggac gactttgcag tcaccagcct tcctttccac ccccccacca acaaaatgtt 1680

tatcgtagga cccatatccg taataaggat gggtctggca gcaaccccat aggcgcctcg 1740

gcgtggtagt tctcgaggcc ttaattaagt cgacggcaga gtcgcagacg cccctattgg 1800

acgtcaaaat tgtagaggtg aagttttcaa acgatggcga agtaacggcg acttgcgttt 1860

ccaccgtcaa atctccctat agggtagaaa ctaattggaa agtagacctc gtagatgtaa 1920

tggatgaaat ttctgggaac agtcccgccg gggtttttaa cagtaatgag aaatggcaga 1980

aacagctgta ctacagagta accgatggaa gaacatcggt ccagctaatg tgcctgtcgt 2040

gcacgagcca ttctccggaa ccttactgtc ttttcgacac gtctcttata gcgagggaaa 2100

aagatatcgc gccagagtta tactttacct ctgatccgca aacggcatac tgcacaataa 2160

ctctgccgtc cggcgttgtt ccgagattcg aatggagcct taataatgtt tcactgccgg 2220

aatatttgac ggccacgacc gttgtttcgc ataccgctgg ccaaagtaca gtgtggaaga 2280

gcagcgcgag agcaggcgag gcgtggattt ctggccgggg aggcaatata tacgaatgca 2340

ccgtcctcat ctcagacggc actcgcgtta ctacgcgaaa ggagaggtgc ttaacaaaca 2400

catggattgc ggtggaaaac ggtgctgctc aggcgcagct gtattcactc ttttctggac 2460

ttgtgtcagg attatgcggg agcatatctg ctttgtacgc aacgctatgg accgccattt 2520

atttttgagg aatgcttttt ggactatcgt actgctttct tccttcgcta gccagagcac 2580

cgccgccgtc acgtacgact acattttagg ccgtcgcgcg ctcgacgcgc taaccatacc 2640

ggcggttggc ccgtataaca gatacctcac tagggtatca agaggctgcg acgttgtcga 2700

gctcaacccg atttctaacg tggacgacat gatatcggcg gccaaagaaa aagagaaggg 2760

gggccctttc gaggcctccg tcgtctggtt ctacgtgatt aagggcgacg acggcgagga 2820

caagtactgt ccaatctata gaaaagagta cagggaatgt ggcgacgtac aactgctatc 2880

tgaatgcgcc gttcaatctg cacagatgtg ggcagtggac tatgttccta gcacccttgt 2940

atcgcgaaat ggcgcgggac tgactatatt ctcccccact gctgcgctct ctggccaata 3000

3060 cttgctgacc ctgaaaatcg ggagatttgc gcaaacagct

3120 cgatcgctgt ttaaagatcg ggtcgcagct

agaacagtat cagactggat ttcaaggcga acacctttat ccgatcgcag acaccaatac 3180

acgacacgcg gacgacgtat atcggggata cgaagatatt ctgcagcgct ggaataattt 3240

gctgaggaaa aagaatccta gcgcgccaga ccctcgtcca gatagcgtcc cgcaagaaat 3300

3360

ggcccctcca gaagactcgg aggacgacat gcaggcagag gcttctggag aaaatcctgc 3420

cgccctcccc gaagacgacg aagtccccga ggacaccgag cacgatgatc caaactcgga 3480

tcctgactat tacaatgaca tgcccgccgt gatcccggtg gaggagacta ctaaaagttc 3540

taatgccgtc tccatgccca tattcgcggc gttcgtagcc tgcgcggtcg cgctcgtggg 3600

gctactggtt tggagcatcg taaaatgcgc gcgtagctaa tcgagcctag aataggtggt 3660

ttcttcctac atgccacgcc tcacgctcat aatataaatc acatggaata gcataccaat 3720

gcctattcat tgggacgttc gaaaagcatg gcatcgctac ttggaactct ggctctcctt 3780

gccgcgacgc tcgcaccctt cggcgcgatg ggaatcgtga tcactggaaa tcacgtctcc 3840

gccaggattg acgacgatca catcgtgatc gtcgcgcctc gccccgaagc tacaattcaa 3900

ctgcagctat ttttcatgcc tggccagaga ccccacaaac cctactcagg aaccgtccgc 3960

gtcgcgtttc ggtctgatat aacaaaccag tgctaccagg aacttagcga ggagcgcttt 4020

gaaaattgca ctcatcgatc gtcttctgtt tttgtcggct gtaaagtgac cgagtacacg 4080

ttctccgcct cgaacagact aaccggacct ccacacccgt ttaagctcac tatacgaaat 4140

cctcgtccga acgacagcgg gatgttctac gtaattgttc ggctagacga caccaaagaa 4200

cccattgacg tcttcgcgat ccaactatcg gtgtatcaat tcgcgaacac cgccgcgact 4260

cgcggactct attccaaggc ttcgtgtcgc accttcggat tacctaccgt ccaacttgag 4320

gcctatctca ggaccgagga aagttggcgc aactggcaag cgtacgttgc cacggaggcc 4380

acgacgacca gcgccgaggc gacaaccccg acgcccgtca ctgcaaccag cgcctccgaa 4440

cttgaagcgg aacactttac ctttccctgg ctagaaaatg gcgtggatca ttacgaaccg 4500

acacccgcaa acgaaaattc aaacgttact gtccgtctcg ggacaatgag ccctacgcta 4560

attggggtaa ccgtggctgc cgtcgtgagc gcaacgatcg gcctcgtcat tgtaatttcc 4620

atcgtcacca gaaacatgtg caccccgcac cgaaaattag acacggtctc gcaagacgac 4680

gaagaacgtt cccaaactag aagggaatcg cgaaaatttg gacccatggt tgcgtgcgaa 4740

ataaacaagg gggctgacca ggatagtgaa cttgtggaac tggttgcgat tgttaacccg 4800

tctgcgctaa gctcgcccga ctcaataaaa atgtgattaa gtctgaatgt ggctctccaa 4860

tcatttcgat tctctaatct cccaatcctc tcaaaagggg cagtatcgga cacggactgg 4920

gaggggcgta cacgatagtt atatggtaca gcagaggcct ctgaacactt aggaggagaa 4980

ttcagccggg gagagcccct gttgagtagg cttgggagca tattgcagga tgaacatgtt 5040

agtgatagtt ctcgcctctt gtcttgcgcg cctaactttt gcgacgcgac acgtcctctt 5100

tttggaaggc actcaggctg tcctcgggga agatgatccc agaaacgttc cggaagggac 5160

tgtaatcaaa tggacaaaag tcctgcggaa cgcgtgcaag atgaaggcgg ccgatgtctg 5220

ctcttcgcct aactattgct ttcatgattt aatttacgac ggaggaaaga aagactgccc 5280

gcccgcggga cccctgtctg caaacctggt aattttacta aagcgcggcg aaagcttcgc 5340

gccaggtcaa ttccctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 5400

cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 5460

aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc 5520

aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 5580

gccccattga cgcaaatggg cggtagcgtg tacggtggga ggtctatata agcagagctc 5640

gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa 5700

gacaccggtt gcgccgccac catgggcccc agaccttcta ccaagaaccc agtacctatg 5760

atgctgactg tccgagtcgc gctggtactg agttgcatct gtccggcaaa ctccattgat 5820

ggcaggcctc ttgcggctgc aggaattgtg gttacaggag acaaagccgt caacatatac 5880

acctcatccc agacaggatc aatcatagtt aagctcctcc cgaatctgcc caaggataag 5940

6000

ccccttggtg actctatccg taggatacaa gagtctgtga ctacatctgg aggggggaga 6060

caggggcgcc ttataggcgc cattattggc ggtgtggctc ttggggttgc aactgccgca 6120

caaataacag cggccgcagc tctgatacaa gccaaacaaa atgctgccaa catcctccga 6180

cttaaagaga gcattgccgc aaccaatgag gctgtgcatg aggtcactga cggattatcg 6240

6300

gctcaggaat tagactgcat caaaattgca cagcaagttg gtgtagagct caacctgtac 6360

ctaaccgaat tgactacagt attcggacca caaatcactt cacctgcttt aaacaagctg 6420

actattcagg cactttacaa tctagctggt ggaaatatgg attacttatt gactaagtta 6480

ggtgtaggga acaatcaact cagctcatta atcggtagcg gcttaatcac cggtaaccct 6540

attctatacg actcacagac tcaactcttg ggtatacagg taactctacc ttcagtcggg 6600

aagctaaata atatgcgtgc cacctacttg gaaaccttat ccgtaagcac aaccagggga 6660

6720

gacacctcat actgtataga aactgactta catttatatt gtacaagaat agtaacgttc 6780

6840

aagaccgaag gcgcacttac tacaccatac atgactatca aaggttcagt catcgccaac 6900

tgcaagatga caacatgtag atgtgtaaac cccccgggta tcatatcgca aaactatgga 6960

gaagccgtgt ctctaataga taaacaatca tgcaatgttt tatccttagg cgggataact 7020

ttaaggctca gtggggaatt cgatgtaact tatcagaaga atatctcaat acaagattct 7080

caagtaataa taacaggcaa tcttgatatc tcaactgagc ttgggaatgt caacaactcg 7140

atcagtaatg ctttgaataa gttagaggaa agcaacagaa aactagacaa agtcaatgtc 7200

aaactgacta gcacatctgc tctcattacc tatatcgtgt tgactatcat atctcttgtt 7260

tttggtatac ttagcctgat tctagcatgc tacctaatgt acaagcaaaa ggcgcaacaa 7320

aagaccttat tatggcttgg gaataatact ctagatcaga tgagagccac tacaaaaatg 7380

tgaggatctc tcgaggaatt ctagatccca cgtcactatt gtatactcta tattatactc 7440

tatgttatac tctgtaatcc tactcaataa acgtgtcacg cctgtgaaac cgtactaagt 7500

ctcccgtgtc ttcttatcac catcaggtga catcctcgcc caggctgtca atcatgccgg 7560

tatcgattcc agtagcaccg gccccacgct gacaacccac tcttgcagcg ttagcagcgc 7620

ccctcttaac aagccgaccc ccaccagcgt cgcggttact aacactcctc tccccgacct 7680

gcaactagta agcttcccgg gttaattaag gccctcgagg atacatccaa agaggttgag 7740

tattctctct acacttcttg ttaaatggaa agtgcatttg cttgttctta caatcggccc 7800

gagtctcgtt cacagcgcct cgttcacact taaaccacaa atagtctaca ggctatatgg 7860

gagccagact gaaactcaca tatgactaat attcgggggt gttagtcacg tgtagcccat 7920

tgtgtgcata taacgatgtt ggacgcgtcc ttattcgcgg tgtacttgat actatggcag 7980

cgagcatggg atattcatcc tcgtcatcgt taacatctct acgggttcag aatgtttggc 8040

atgtcgtcga tcctttgccc atcgttgcaa attacaagtc cgatcgccat gaccgcgata 8100

agcctgtacc atgtggcatt agggtgacat ctcgatcata cattataaga ccaacgtgcg 8160

agtcttccaa agacctgcac gccttcttct tcggattgtc aacgggttct tcagaatcta 8220

tgcccatatc tggcgttgag accattgtgc gtttaatgaa caataaagcg gcatgccatg 8280

gaaaggaggg gtagacgata 8340

attataccat gaatatagag ggggtatgtt tccactgcca ctgtgatgat aagttttctc 8400

cagattgttg gatatctgca ttttctgctg ccgaacaaac ttcatcgcta tgcaaagaga 8460

tgcgtgtgta cacgcgccgt tgagtatacg ggaaactaaa tgttcataga ggtctttggg 8520

ctatatgtta ttaaataaaa taattgacca gtgaacaatt tgtttaatgt tagtttattc 8580

aatgcattgg ttgcaaatat tcattacttc tccaatccca ggtcattctt tagcgagatg 8640

atgttatgac attgctgtga aaattactac aggatatatt tttaagatgc aggagtaaca 8700

atgtgcatag taggcgtagt tatcgcagac gtgcaacgct tcgcatttga gttaccgaag 8760

tgcccaacag tgctgcggtt atggtttatg cgcacagaat ccatgcatgt cctaattgaa 8820

8880

aaaaatttac ccttttatgac catcacatct ctctggctca taccccgctt ggataagata 8940

tcatgtagat tccgccctaa gaaatgcaaa ctaacattat tgtcggttcc atatacactt 9000

ccatcttgtc cttcgaaaat aacaaactcg cgcaatagac cgtccgtaca tgcatggccg 9060

atgtgtgtca acatcattgg tctgctagat cccgatggga cgaatcgtac agtcgtcgct 9120

ccagcattgg caaaaatccc cagataccct ccatgcggca aatctaaatt gcgaccccga 9180

agagactgca ccaaagtctt atcgacgcac gctgattttt ttgaacagcg ggagcccatt 9240

atcttcagtg gagcgtagac gggcgaggct aattatgtga catagcaaca ctgcatgtat 9300

gtttttataa atcaataaga gtacataatt tattacgtat catttccgtt tgtaatatac 9360

tgtatacatc atccacacta ttagtcagca ctagcgcgcg ggcgcacgtt acaatagcag 9420

9480

acctaagtac tgcacacaga tgtttaatgt atatcgtcat ataaattata tcgctaggac 9540

agacccaaac gacctttatc ccaaacagtc agatcctctt ctcaagtgtc gatttctgtt 9600

atggaatatg cataccctgg cccagaaatt gcacgcacga gcgtagtgaa tgcgtcattg 9660

gttttacatt taaaggctaa atgcacaaat tctttagacg acagcacatc gttaaaatagc 9720

atctctagcg ttcttatgaa tgctaagcat tggagtcctc ctggtcggcc acaataacag 9780

ctgagtatca taccctgagc tccggggttg tcgcacatag cggattcgta taaacatagg 9840

attttccgcg aatccatcag ttgcaaaaat ctgttaggct ccatcaacaa cgctggattt 9900

acttcagatc cacgcgtaaa gtaatggtgc tcgaataccg tttttagagt tgtcggcatt 9960

tcaaggaaca aagaattcat ttcttcattg caacgacgcg ccagaaatcc caagacctct 10020

ttgggtagta tgttcttgcc tataaaacac ggcgttccaa gtgccaggaa ccacgcatgt 10080

gttactgttg gggcgtattc agaaataaag cggggtttat gcggcttttg aagctcggat 10140

atccaaagta tcgcttgctg atgaacgagc gatgtagctg ttacaaaacc tcctttccat 10200

10260

attattccgt atgaggtctt gcaggaataa gctcttaggg acagccagct tggatatggt 10320

10380

ttagatccac catcaatgtg tgcagcattg actcccgccc gtcgaatatt ccttttgtta 10440

cgatgcagta atgagcacga tcatgggcgg ggcgatgacg ttctatttgc atgtctgcga 10500

acaatttgcg tcagtcatac agctatggag tgggccattt ctggccgtca acttaaaaac 10560

10620

caaatgctct ggccaatatg gcaatgaatt tggattcgtt tgacgccgat ggtatgcagt 10680

gcaaatgtgc caatagccca catccgaaaa agttatttgt catacaagca ggtgttaagt 10740

agcaatcaca taaaggcacc agacgcctca tggcatcata atgaatagct ccttctcccc 10800

actggaacca ctgacaaaat ctgcgagtat attccgcaaa ccacatttta tttctcatag 10860

aaactaccct aaatcctttt aacgggaaga agaatcctag atagtgcttg aagtcatgac 10920

10980

gtaggcattc cgatcccttt actattgcgt cttcacgacc aaatgggaat gcgccaaaat 11040

ccccacacct catcaccctg gaggcagatt gtgtattatt aatatccgcc gattgaagca 11100

caaaacggta cggtactgtt cctaattctg gtatagattc tatggtcaaa agtctgcata 11160

tccccgacat tgccatgaga tcacacagtc caagtagcat

tgtcaacgtc cctcgccgca ccaccaatcg aaaataaagt atctacgcaa gttatagctc 11280

11340

aactctctgg ggggcttgtt atcttctgca ccgtcgcagt cgcagttttc cgaaatttat 11400

gtctaatata ttttccggcc gtgctccaat cggccgaaaa gaatctgcgt attaccagac 11460

tcattgacgg gccgataaag accataaaac aaaattcctg tgcactccct cctccagttt 11520

tgccatcgtc caagtcccgt aacttttttt gcgtttcgag gagcaagcgt tcgttatccc 11580

tacccacact tgttttccac cgttttctta ttataagcgg ttgtatcgcc aacgcgtcac 11640

cgcaggttgt cacatacagt gatggcatac ttgaacgtgc aacaacgcgc tcgctttgca 11700

aatctaagtc attgaccatc aaatcgcgtt

tatggtgacg gtgcagccac cccaactcag ttcttgtaaa aaaagctatt ggcgggaatt 11820

tgccattaac cagattcgta 11880

ttttttcgct cgacccggca tcactatgga tacaatacct ttctatggcc catttcagct 11940

ctcgaaccaa ccacacggac aattgactaa cataagtatg atctttatca cagtcgcacc 12000

catctgagtt atatttatgg catccgagcg ctcttactgt acggtcggat acacccatgg 12060

ttttccttt atatagtcgg gttatagtct gtcgggtttg gcggtagcac ggagtagttt 12120

12180

ctacacgtga gtgttgtcca ttcctaggta tattcatctg ttcggatacc ttcaattgct 12240

gttcaggcat aaccttaaag catatgttat gttgtacatc aaaacttggt gagttatgtt 12300

12360

cacacgcccc tgcatatact gttcctattc caaattcacg ttttgcccca tcggctatct 12420

gctcccaaaa agttgtaata taggtgccgc tgggtgcgaa attttcatca gttgtattcc 12480

tgataaactg aatcacttta cataattttt gccacatatc tgcgtgcagc catagtatcg 12540

12600

taatggtata caagttaggt ccgtacctag accttaatgt ttccaattct tctgaatcac 12660

tgcactctcg taggggagta acggtaataa tttcgtctct gagccccgtt ttgcgttgaa 12720

12780

tgacatcggt ggtcattgtt tccatcaacg accatctttt acgatcgccc atactactca 12840

tggacgttgt cggtgttgaa aaatcaccag aattgcaacg gatctctgggg taccatgctg 12900

ctgatggaat tggcggtttt aattgttgtt tcagtctatt attgctatct ttggcggggt 12960

tgaataatgt ggggggagag tgattgcagg aatccgaatg ggtcaataaa acgaccgtgc 13020

tccgttctgc cggcgccgat ccgattgaag ctatatactt cgcttctctc cccacttttc 13080

13140

cctgcttgcc tacagaagaa tcaacatctc gccccaatat tctggtcaaa actggctcgc 13200

tcatggcaac gcggacgttt cccccggtgg ccagtcttaa tggttaatgt tcttttcggc 13260

aatcttatac atcagcgggt tgcgtgaata ctggtcacag ttcagtcatt tactacacac 13320

13380

gaccgcgtcc gaggcgatgt cgtacgggcg gtgcggggtt ggatcctcgg caaagagatc 13440

ctcgtaattc ggcggtggga gcggagggta aagacgcggg tggggatctc cctccggacc 13500

13560

gggcgggctg ggggccggaa tatctcccgc gacttcttct atcggcgcgg aattggagtc 13620

13680

gcgttcagcg tccatcgtcg tatatgctct aacacacgtc tcgctatatt aaaaaaaaga 13740

agagtatcgg tcagtgtcga gtgtcgccga caatgtcgcg agttctcggc gatttaattt 13800

ttggaactgc tccctatgaa tcccgtaact gtagcgcccg cgcagaaagc cgccatcaga 13860

ccaactacgt gtctgttcga tgtttgcccg ccgatcgctt taccgattaa ggttccggcg 13920

13980

gtgcgagatg gagaaaccac atctcccgaa ttagtagagg aaagcccgcg ctgtcggttt 14040

14100

cgcgtcttat gagcggacgt tcttactgca gctcggtagg agtggggcag ggttagattt 14160

14220

ggtgggggtg agcgcttccg gagttgcgat ctccgatctc gagccgcagc ccggcggtgt 14280

ctctttcagt ggagcgttag cgccatcatg tggttcgtgg cggtggaaag gctattatgt 14340

gttaggggag agaccacgtg atcggcatgc aaatgagcaa ggcgaacgcg tcagcgttcg 14400

cactgcgaac caataatata tatattatac tattggcttt aggtgcgaac gtccggctag 14460

tccaatagcg gggtcgcgtt tcgtaccacg tgttatagac cgccctaaac tcgcactcgg 14520

gggtccggcc gcgcccagac agggcggaga cgtgccacag gggctttaaa acaccgcttc 14580

catctcgg 14598

<210> 32

<211> 10681

<212> DNA

<213> Artificial sequence

<220>

<223> Expression cassette

<400> 32

aattccagac taaatgcccc ggcccaattt gtcaagtgtg cagtcacgga ggcgtcgacc 60

gtgtccccgg cattaaacag gaaagcgtta aagtttttga atgttaggtc acaggtacaa 120

acataaatgt ttgtacaaac aggtaacagg tacaaacata aatgccccgg cataaatgtc 180

ccttacggcg gatcgaaacg acattaggca tactcgggta ccattttgca ttccgatcag 240

cacggatgaa attaggcagg aatgcggttt atattatgcg gcattggaca aacgatatgg 300

cattgattgg cagtttatga atgtcttcat gtggggcgta aacggattcc tattggttca 360

gaagacaacg acgatatatt tagagagaaa aagctaccca gcataggata aacacacatt 420

gagcattgag agacataggt atcggtatgg atgggaaaac tacacacgtg aacaccaaac 480

gacttatata ctcgagcggt gatactactg agcaagaatg cactgcatct gagccactga 540

atgaagactg tgatgaaaat gtgaccatcg atggaattgg agaagaatat gcgcagttct 600

tcatgtcccc gcaatgggtc ccaaatctac atcgcttgag cgaggatacc aaaaaggtat 660

accgatgtat ggtttccaac agactcaatt attttcccta ttatgaggcg ttcaggcggt 720

ctttgtttga tatgtatatg ctaggtcggt tggggcgtcg acttaagcga tctgactgggg 780

agactattat gcatctgtca ccaacgcaaa gtcggcgtct acatagaact ttaagatttg 840

tggagcgtag aattatccca tctaacagtt atatacgcac atcgggccac gttccgcctt 900

cgagggcact tccgacagat acgaatttaa agatggatga ataattaaat tggaaagagt 960

aactacatta atcgagcgtc atgacggcgt cccgtgaaaa tgggaatttt ctactcgaaa 1020

1080

ggcaacatac ataatgtgca tgcgaaacca ctttttcagt gtacgctgac attgtgcaac 1140

acggaggggt agcatctaca tacaatatat gttgattaat gattggagaa aaaactatgc 1200

agctcgccga tcatatggct aactcgcctt cgtctatatg gcggaccccg cgggaaaaat 1260

cgacgtacca tctgatttac aacaccagta atgaacatgt cgcatccctg cccagatctg 1320

tgcgcccatt ggcgcggatc gttgtgaatg ccgccgaaac acttcaggtc ggtatgagag 1380

ccgggaggcc gccatcagca ggagtttggc gagaggtgtt tgatagaatg atgacagcct 1440

tccgtgacca cgagcctact gcgacattta atgctgcaaa tcccattaga aaaatggtcg 1500

agacagttct acagaataat gaagagcccc cgcggacgca tgctgaaatg ggtaatcgcc 1560

ttatgaacat tatgtactgg tgttgcttgg gacacgcagg acaatgctcg atatggcagt 1620

tgtacgagac gaatcaggcc attttaagtt tattagatga agtggttatc ggcacaacaa 1680

atcccttttg caccctcgag caatactgga agccattatg caccgcaatc gccaacaagg 1740

ggacctcatc gcttgttgag gatgccaaag tggccgagta cctggttagc atgcgcaaat 1800

tgatataaca taggcacgct ctgatgttac agaccacaat accgcataca tttattgtaa 1860

ggttgttaat aaaggtttat tctatgtaag actacaatac tttcgacatt gcttgtatac 1920

atattaaata ctttctcaag ttcctattac ataaaatggg atctatcatt acattcgtta 1980

agagtctgga taattttact gtttgccagc ttcgatcttg gaacgtactg tggatagtgc 2040

cttacttgga atcgtgaaaa tttgaaacgt ccattatttg gatatcttcc ggttgtccca 2100

2160

aatcggggac caacaacgcg tgggtccaca ctcattcgga aattttccga tgattctgaa 2220

tatttattgc cgctcgttac gagtcgttgg acatatctgt aatacatttc ttcttctgaa 2280

ggatcgctgc acatttgatc tatacattgg ccaggatgtt caagtctcag atgttgcatt 2340

ctggcacagc acaactttat ggcatttccg atgtaatcgt ccggcagccc tgggggagtt 2400

ctatattcgc atattgggat ggtaaggaca atagcagatc tcgcaacctc cagggaggct 2460

2520

tatcctttac tagcgccgat tgagagcatc gtcgtccaat tttctaaatg gaaagaaaac 2580

aaggcgggca agagtgttcc aaacattttc attttcggcg aatctctcaa atcccatggc 2640

gtgcaattga ttgcaaaatt ggcacttccg ttcacgtttg tatctccaaa ctctaagaca 2700

cttttaattg aaaaactacg ttctagtgtg gaaagaaacc tataggcaga ccatagaact 2760

atttgacacc acatatcttt ttgtatgtca aactgaccat gatcgtatgt tgctgaatgc 2820

actagggcaa ttcgctcgcg cgactccata cattgaataa ttccacacgt cagctcatcg 2880

gttagcaagg tccagtagtt gaagtcattt atttttcccc gcggctggcc aaatctacct 2940

ctgggaatat ccaagttgtc gaatatgatc gcaccggctc tggtcatggt gaaggaactg 3000

tagcataaag acgcaggtat cataggggta atattttttt attcactcac atactaaaag 3060

taacgcatat tagcaccatg tatgggctat caattgacat ttgcgtagca ctacatcacg 3120

3180

gggtttatct actattgaat ttttcccctat ctgtgataca cttgggagcc tctacaagca 3240

tattgccatc atgtacgttt ttatctactg tcttaacgcc catgggaacg gaggcgtcgt 3300

3360

tggactcgat accaagcccc tgcagctggg gaacgtctgg tggagagccg ataatttgat 3420

atacgcacgc catattactg tcgttgaagt acgccttatc ttctatgttt tcaaatttag 3480

gttcccaagt ggacgtgaga agtgtttgta tctcacatgg aatggcccaa ggcattccag 3540

cccaggtgcc tggtacttta atggcaaaca aacgttttgg tagaggtatt gattctattg 3600

3660

caagctgcgg ccgctctaga actagtggat cccccgggct gcagcccaat gtggaattcg 3720

cccttgcaca ttgttactcc tgcatcttaa aaatatatcc tgtagtaatt ttcacagcaa 3780

tgtcataaca tcatctcgct aaagaatgac ctgggattgg agaagtaatg aatatttgca 3840

accaatgcat tgaataaact aacattaaac gaattcacta gtggatcccc caactccgcc 3900

cgttttatga ctagaaccaa tagtttttaa tgccaaatgc actgaaatcc cctaatttgc 3960

aaagccaaac gccccctatg tgagtaatac ggggactttt tacccaattt cccaagcgga 4020

aagcccccta atacactcat atggcatatg aatcagcacg gtcatgcact ctaatggcgg 4080

cccatagggga ctttccacat agggggcgtt caccatttcc cagcataggg gtggtgactc 4140

aatggccttt acccaagtac attgggtcaa tgggaggtaa gccaatgggt ttttcccatt 4200

actggcaagc acactgagtc aaatgggact ttccactggg ttttgcccaa gtacattggg 4260

tcaatgggag gtgagccaat gggaaaaacc cattgctgcc aagtacactg actcaatagg 4320

gactttccaa tgggtttttc cattgttggc aagcatataa ggtcaatgtg ggtgagtcaa 4380

tagggacttt ccattgtatt ctgcccagta cataaggtca atagggggtg aatcaacagg 4440

aaagtcccat tggagccaag tacactgcgt caatagggac tttccattgg gttttgccca 4500

gtacataagg tcaatagggg atgagtcaat gggaaaaacc cattggagcc aagtacactg 4560

actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag ggggtgagtc 4620

aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc caatgggttt 4680

tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt actggcacgt 4740

atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt caataggggt 4800

gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga ctttccattg 4860

ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc ccattattgg 4920

cacgtacata aggtcaatag gggtgagtca ttgggttttt ccagccaatt taattaaaac 4980

gccatgtact ttcccaccat tgacgtcaat gggctattga aactaatgca acgtgacctt 5040

taaacggtac tttcccatag ctgattaatg ggaaagtacc gttctcgagc caatacacgt 5100

caatgggaag tgaaagggca gccaaaacgt aacaccgccc cggttttccc ctggaaattc 5160

catattggca cgcattctat tggctgagct gcgttctacg tgggtataag aggcgcgacc 5220

agcgtcggta ccgtcgcagt cttcggtctg accaccgtag aacgcagagc tcctcgctgc 5280

aggcggccgc tctagaactc gtcgatcgca gcgatgacaa acctgcaaga tcaaacccaa 5340

cagattgttc cgttcatacg gagccttctg atgccaacaa ccggaccggc gtccattccg 5400

gacgacaccc tggagaagca cactctcagg tcagagacct cgacctacaa tttgactgtg 5460

ggggacacag ggtcagggct aattgtcttt ttccctggat tccctggctc aattgtgggt 5520

gctcactaca cactgcagag caatgggaac tacaagttcg atcagatgct cctgactgcc 5580

cagaacctac cggccagcta caactactgc agactagtga gtcggagtct cacagtgagg 5640

tcaagcacac tccctggtgg cgtttatgca ctaaacggca ccataaacgc cgtgaccttc 5700

caaggaagcc tgagtgaact gacagatgtt agctacaatg ggttgatgtc tgcaacagcc 5760

aacatcaacg acaaaattgg gaatgtcctg gtaggggaag gggtcactgt cctcagccta 5820

cccacatcat atgatcttgg gtatgtgagg cttggtgacc ccattcccgc tatagggctt 5880

gacccaaaaa tggtagctac atgcgacagc agtgacaggc ccagagtcta caccataact 5940

gcagccgatg attaccaatt ctcatcacag taccaaccag gtggggtaac aatcacactg 6000

ttctcagcca acattgatgc tatcacaagc ctcagcattg ggggagagct cgtgtttcaa 6060

acaagcgtcc aaggccttgt actgggcgcc accatctacc ttataggctt tgatgggact 6120

6180

atgccattca atcttgtcat tccaaccaat gagataaccc agccaatcac atccatcaaa 6240

ctggagatag tgacctccaa aagtggtggt caggcagggg atcagatgtc atggtcggca 6300

agtgggagcc tagcagtgac gatccatggt ggcaactatc caggggccct ccgtcccgtc 6360

acactagtag cctacgaaag agtggcaaca ggatccgtcg ttacggtcgc tggggtgagt 6420

aacttcgagc tgattccaaa tcctgaacta gcaaagaacc tggttacaga atacggccga 6480

tttgacccag gagccatgaa ctacacaaaa ttgatactga gtgagaggga ccgtcttggc 6540

atcaagaccg tctggccaac aagggagtac actgattttc gtgagtactt catggaggtg 6600

gccgacctca actctcccct gaagattgca ggagcatttg gcttcaaaga cataatccgg 6660

gctataagga ggtaagcttg atctagagcg gccgcggggga tccagacatg ataagataca 6720

aaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 6780

6840

acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttcggatc 6900

6960

tagtttcccg tatactcaac ggcgcgtgta cacacaaggg cgaattccac agtggatatc 7020

aagcttagct tgcctccgat tctagcatta catagccggt cagtagatcc tgccattcgg 7080

tagcgcaacc ggctacatct tcaaacagtc tcacgataaa tgcatctctc gttcctgcca 7140

atcgggaacc gggcatacca ctcccgcctg ccgatttaat tctcacaatt gggcgatgcc 7200

ggcggggcaa aacgaatgtg gatttggcaa accgacacag gtctgctgta cggactaata 7260

tgggcacacc cacatcattc ttcagatgct ccatgcattg ttctatgaga aagatccata 7320

gggtggaggc agcgtcacga gatcgcccag gcaatcgatc gcattcgtct agtaaagtga 7380

cgagagttat catgcacaca cccatgccca cgccttccga ataactggag ctgtggaaga 7440

tcggaaacgt ctttttgact gccggtctcg tactactttc gcacaggtgt atacccggac 7500

gcgtactata tattttatat catccaacgt ccgaaattac atacgtggcg gcgatggaag 7560

tagatgttga gtcttcgaaa gtaagtgcct cgaatatggg tattgtctgt gaaaatatcg 7620

aaagcggtac gacggttgca gaaccgtcga tgtcgccaga tactagtaac aatagcttcg 7680

ataacgaaga cttccgtggg cctgaatacg atgtggagat aaataccaga aaatctgcta 7740

atcttgatcg tatggaatct tcgtgccgtg aacaacgagc ggcgtgcgaa cttcgaaagt 7800

gttcgtgtcc tacgtctgcc gtgcgcatgc aatacagtat tctttcatct ctcgctccgg 7860

gttcagaggg tcatgtatat atatgtacta gatacgggga cgcggaccaa aaaaaatgca 7920

tagtgaaggc agtcgttgga ggaaagaatc ccgggaggga agtggatatt ttaaaaacca 7980

tctcacataa atcaattata aaattaatcc atgcctataa atggaaaaat gttgtgtgta 8040

tggcaatgcg tgtatatcgt tatgatcttt tcacatatat tgacggagtc ggccctatgc 8100

cccttcaaca gatgatctat attcaacgtg gactactaga ggcgctagca tacatacatg 8160

aaaggggcat cattcaccga gacgtaaaga cggagaatat attcttggat aatcacgaaa 8220

atgcagtttt gggtgacttc ggtgctgcat gccaactagg agattgtata gatacgcccc 8280

aatgttacgg ttggagcgga actgtggaaa caaattcgcc ggaattatct gcacttgatc 8340

cgtattgcac aaaaacagat atttggagtg ccggattggt tctatatgag atggcaatta 8400

aaaatgtacc attgtttagt aagcaggtga aaagttcggg atctcagctg agatccataa 8460

tacggtgcat gcaagtgcat gaactggagt ttccccgcaa cgattctacc aacctctgta 8520

aacatttcaa acaatatgcg gttcgtgtac gaccgcctta taccattcct cgagttataa 8580

gaaatggggg gatgccaatg gatgttgaat atgtcatttc taaaatgctt acgtttgacc 8640

aggagttcag accttctgct aaggaaatat tgaatatgcc cctatttact aaggcgccga 8700

ttaacctgct taatatcaca ccctctgaca gtgtctaacg gtatacaggc gggagcgggt 8760

8820

ctgattcatt gagaactgaa acgccatatt ggtttcttgg atatgtctac aacaattagt 8880

taaattgcta tgttctactg cgagtaacat ttgataagtt gtaagagacg ggcgactcat 8940

gtcgaagttg acgaatataa agtacataac gtgtttagaa tacccagaat ccgaatagtc 9000

cgcggggggcg tcttctcgcg tgagtaccaa atactgagtt gaacttgaaa atgctaaatc 9060

tgtgacactc tttgtgtgat gattattgtc accacttcga agatggcttc gacattcatg 9120

atgttctggt gtttgtttgg aatcgtaata gcgcttgttt cgtccaagtc tgacaacaaa 9180

gaaaatctga agaattatat cacggataag tcaaccaata ttagaatacc cacgccatta 9240

tttgtatcaa cggaaaactc ttatcccaca aaacatgtaa tctacgatga aaactgtggc 9300

ttcgctgtac tcaatcctat aagtgacccc aaatatgtcc ttttgagcca gcttctaatg 9360

ggaaggcgca aatatgatgc gacggtcgcg tggtttgttc tcggtaaaat gtgtgccaga 9420

ttaatatatt tgcgcgaatt ttataactgc tcgacaaatg agccttttgg cacatgttct 9480

atgagctctc ctggatggtg ggacaggcgc tacgtctcaa ccagtttcat ttctcgcgac 9540

gaattacagc tggtttttgc agcgccgtcc cgagaattag atggtttata tacgcgcgta 9600

gtagttgtca acggggactt tactacggcc gatataatgt ttaatgttaa agtggcatgt 9660

gccttttcaa agactggaat agaagatgat acattatgca aaccctttca tttctttgcc 9720

aatgcaacat tgcacaattt aaccatgatt agatcggtaa ctcttcgagc gcacgaaagc 9780

catttaaagg aatgggtggc acggagaggt ggtaacgtcc ctgcagtgct acttgagtct 9840

accatgtatc atgcatccaa tctgcctaga aatttcaggg atttctacat aaagtctcca 9900

gatgattata agtataatca cctagatggg ccatctgtaa tgctcatcac tgacagacct 9960

agtgaagatt tggatgggag gctcgttcac caaagtgaca tttttactac tacaagtcct 10020

10080

atacaagcta taattttttt gatagggtta ggctcgttca ttggaagcat attcgtagtt 10140

ttggtagtat ggattatacg cagatattgc aatggagcgc ggagtggggg aacgcccccc 10200

agtcctcgcc ggtatgtgta taccaggcta tgatcacgtg tgaaacttgg gcggacctgt 10260

atcatatgta caccgtccct attcgtttat agccagtacg tgttatctgc acatagagga 10320

acatgtgtca tactgggatc gcatgcatgg tatgtgtgac tctaatatta ttctgtatca 10380

taataaaaac acagtgcatg gtatatagag gatcgctggt aagcactacg gtagaccaat 10440

cggctcagat tgcattcttt ggcatcgata ccgttgttaa tttatatggc aaagtcttgt 10500

10560

10620

tcagttctac ggtattcaga ggatgccggg acgccgttgt ttatactagg ccccacagca 10680

g 10681

<---

Claims (22)

1. Recombinant non-pathogenic Marek's disease virus (rMDV _np ) to help protect chickens from a virus selected from the group consisting of Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), infectious bursitis virus (IBDV), Marek's disease virus (MDV) ) and any combination thereof that contains at least three heterologous nucleotide sequences; where the first heterologous nucleotide sequence encodes one or more antigens from the first avian pathogen, the second heterologous nucleotide sequence encodes one or more antigens from the second avian pathogen, and the third heterologous nucleotide sequence encodes one or more antigens from the third avian pathogen; where the first heterologous nucleotide sequence, the second heterologous nucleotide sequence and the third heterologous nucleotide sequence, each located in a non-essential site of the rMDV genome _np ; and where the first chicken pathogen is infectious bursitis virus (IBDV), the second chicken pathogen is infectious laryngotracheitis virus (ILTV) and the third chicken pathogen is Newcastle disease virus (NDV). 2. rMDV _np according to claim 1, wherein the first heterologous nucleotide sequence contains the coding sequence for infectious bursitis virus viral protein 2 (IBDV VP2), the second heterologous nucleotide sequence contains the coding sequence for infectious laryngotracheitis virus (gD ILTV) glycoprotein D and the coding sequence infectious laryngotracheitis virus glycoprotein I (gI ILTV) and the third heterologous nucleotide sequence contains the coding sequence for a Newcastle disease virus (F NDV) fusion protein; and where the first heterologous nucleotide sequence consists of the first heterologous nucleic acid or the second heterologous nucleic acid, the second heterologous nucleotide sequence consists of the first heterologous nucleic acid or the second heterologous nucleic acid, and the third heterologous nucleotide sequence consists of the first heterologous nucleic acid or the second heterologous nucleic acid; where the first heterologous nucleic acid is located in the first non-essential site in the rMDV _np genome, and the second heterologous nucleic acid is located in the second non-essential site in the rMDV _np genome; and wherein the first non-essential site and the second non-essential site are the same or different. 3. rMDV _np according to claim 2, wherein the first heterologous nucleic acid comprises a coding sequence for ILTV gD, a coding sequence for ILTV gI, and a coding sequence for NDV F, and wherein the second heterologous nucleic acid comprises a coding sequence for IBDV VP2. 4. rMDV _np according to claim 3, in which the first heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 28, and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 27 or the first heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 31 , and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 32. 5. rMDV _np according to claim 2, wherein the first heterologous nucleic acid comprises a coding sequence for IBDV VP2 and a coding sequence for NDV F, and wherein the second heterologous nucleic acid comprises a coding sequence for ILTV gD and a coding sequence for ILTV gI. 6. rMDV _np according to claim 5, wherein the first heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 30, and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 29. 7. rMDV _np according to claim 2, wherein the first heterologous nucleic acid comprises a coding sequence for IBDV VP2, a coding sequence for ILTV gD, and a coding sequence for ILTV gI, and wherein the second heterologous nucleic acid comprises a coding sequence for F NDV. 8. rMDV _np according to claim 3, in which the first heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 23, and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 26 or the first heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 21 , and the second heterologous nucleic acid contains the nucleotide sequence of SEQ ID NO: 22. 9. rMDV _np according to claim 2, 3, 4, 5, 6, 7 or 8, wherein the first non-essential site and the second non-essential site are selected from the group consisting of unique short site 2 (US2), unique long site 54.5 (UL54 .5), unique long site 7/8 (UL7/8), unique long site 40 (UL40), unique long site 43 (UL43), unique long site 45/46 (UL45/46), unique long site 55 (UL55 ), unique short site 10 (US10), region between US10 and unique short open reading frame 3 (SORF3), region between US2 and SORF3, intergenic region 1 (IG1) site, intergenic region 2 (IG2) site, and intergenic region 3 site (IG3). 10. rMDV _np according to claim 2, 3, 5, 7 or 9, wherein the IBDV VP2 protein coding sequence of the first heterologous nucleotide sequence is under functional control of the mCMV mouse cytomegalovirus (IE1) immediate early gene 1 promoter; the ILTV gD protein coding sequence of the second heterologous nucleotide sequence is under the functional control of the endogenous ILTV gD promoter; the ILTV gI protein coding sequence of the second heterologous nucleotide sequence is under the functional control of the endogenous ILTV gI promoter; and the NDV F coding sequence is under functional control of the human cytomegalovirus immediate early gene 1 (hCMV IE1) promoter. 11. rMDV _np according to claim 2, 3, 4, 5, 6, 7, 8, 9 or 10, wherein the first non-essential site is a US2 site and the second non-essential site is a UL54.5 site. 12. rMDV _np according to claim 2, 3, 4, 5, 6, 7, 8, 9 or 10, wherein the first non-essential site is the UL54.5 site and the second non-essential site is the US2 site. 13. rMDVnp according to any one of paragraphs. 1-12, wherein rMDVnp is a recombinant turkey herpesvirus (rHVT). 14. Immunogenic composition for chickens to a virus selected from the group consisting of Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), infectious bursitis virus (IBDV), Marek's disease virus (MDV) and any combination thereof, containing rMDV _np according to any one of paragraphs. 1-13. 15. Chicken vaccine against a virus selected from the group consisting of Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), infectious bursitis virus (IBDV), Marek's disease virus (MDV) and any combination thereof, containing an immunogenic composition according to claim 14 with a pharmaceutically acceptable carrier. 16. A method of assisting in protecting chickens from a virus selected from the group consisting of Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), infectious bursitis virus (IBDV), Marek's disease virus (MDV), and any combination thereof, comprising administering vaccines according to item 15.

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