U.S. patent application number 16/594436 was filed with the patent office on 2020-05-28 for her2 antigenic polypeptide compositions, and methods for their use in treatment and prevention of carcinomas.
This patent application is currently assigned to Wayne State University. The applicant listed for this patent is Wayne State University. Invention is credited to Heather GIBSON, Richard JONES, Joyce REYES, Wei-Zen WEI.
Application Number | 20200164052 16/594436 |
Document ID | / |
Family ID | 53878941 |
Filed Date | 2020-05-28 |
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United States Patent
Application |
20200164052 |
Kind Code |
A1 |
WEI; Wei-Zen ; et
al. |
May 28, 2020 |
HER2 ANTIGENIC POLYPEPTIDE COMPOSITIONS, AND METHODS FOR THEIR USE
IN TREATMENT AND PREVENTION OF CARCINOMAS
Abstract
Antigenic polypeptides of the growth factor receptor HER2, for
breaking the tolerance of a host against self HER2. The antigenic
polypeptides include HER2 polypeptides with single amino acid
substitutions of lysine for glutamine, arginine for glutamine, or
aspartic acid for asparagine. Gene expression constructs, vaccine
compositions, and immunization methods including the substituted
HER2 polypeptides. Methods for immunizing mammalian subjects with
heterologous unsubstituted HER2 antigenic polypeptides, including
polypeptides of feline and bear HER2. A diagnostic method of
determining whether a mammalian subject is sufficiently
immunocompetent to respond to immunotherapies directed at breaking
tolerance to self HER2.
Inventors: |
WEI; Wei-Zen; (Grosse Pointe
Farms, MI) ; GIBSON; Heather; (Madison Heights,
MI) ; JONES; Richard; (Fayetteville, NY) ;
REYES; Joyce; (Rochester Hills, MI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Wayne State University |
Detroit |
MI |
US |
|
|
Assignee: |
Wayne State University
Detroit
MI
|
Family ID: |
53878941 |
Appl. No.: |
16/594436 |
Filed: |
October 7, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15120621 |
Aug 22, 2016 |
10434152 |
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PCT/US2015/016542 |
Feb 19, 2015 |
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16594436 |
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61942071 |
Feb 20, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 2039/53 20130101;
G01N 2800/52 20130101; A61K 2039/552 20130101; A61K 39/39 20130101;
A61K 2039/572 20130101; C07K 16/3015 20130101; C07K 14/71 20130101;
A61K 39/001106 20180801; A61K 2039/55522 20130101; A61K 39/0011
20130101; A61K 2039/575 20130101; G01N 33/574 20130101; C07K 16/32
20130101; C07K 16/2863 20130101 |
International
Class: |
A61K 39/00 20060101
A61K039/00; G01N 33/574 20060101 G01N033/574; C07K 16/32 20060101
C07K016/32; C07K 16/30 20060101 C07K016/30; C07K 16/28 20060101
C07K016/28; C07K 14/71 20060101 C07K014/71; A61K 39/39 20060101
A61K039/39 |
Goverment Interests
GRANT INFORMATION
[0001] This invention was made with government support under grant
no. CA076340 awarded by the National Institutes of Health and grant
no. W81XWH-11-1-0050 awarded by the US ARMY/Medical Research and
Materiel Command. The government has certain rights in the
invention.
Claims
1. Antigenic polypeptides of HER2, for breaking tolerance to the
self HER2 of an animal subject, said antigenic polypeptides
including at least one point mutation in the extracellular domain
of HER2.
2. The antigenic peptides according to claim 1, wherein said point
mutation induces the substitution of glutamine with lysine with
(Q-K) in the amino acid sequence QLRSLTEILKGGVLI (SEQ ID NO: 109)
of HER2 domain I, rendering said amino acid sequence
KLRSLTEILKGGVLI (SEQ ID NO: 110).
3. Isolated antigenic polypeptides for inducing immune response
against HER2 in a subject of a mammalian species, said polypeptides
comprising at least the extracellular domain of the HER2 of an
animal species, said extracellular domain including an amino acid
substitution of glutamine with lysine (Q-K), or with a conservative
amino acid of lysine, said substitution being at position 119 of
mature feline HER2, or at a homologous position of the mature HER2
of another animal species.
4. The antigenic polypeptides according to claim 3, selected from
the group consisting of mature feline HER2 having a Q-K
substitution at position 119 (mfeHER2-Q119K); mature bear HER2
having a Q-K substitution at position 119 (mbearHER2-Q119K); mature
human HER2 having a Q-K substitution at position 119
(mhumHER2-Q119K); mature mouse HER-2 having a Q-K substitution at
position 120 (mmouseHER2-Q120K); mature rat HER2 having a Q-K
substitution at position 120 (mratHER2-Q120K); and mature human rat
chimeric HER2 having a Q-K substitution at position 119
(mE2Neu-Q119K).
5. The antigenic polypeptides according to claim 3 wherein said
antigenic polypeptides comprise the signal peptide and
extracellular and transmembrane domains of HER2 (precursor HER2)
and said amino acid substitution of glutamine with lysine (Q-K) or
a conservative amino acid of lysine, is at position 141 of
precursor feline HER2, or at a homologous position of the precursor
HER2 of another animal species.
6. The antigenic polypeptides according to claim 5, selected from
the group consisting of precursor feline HER-2 having a Q-K
substitution at position 141 (prefeHER2-Q141K); precursor bear
HER-2 having a Q-K substitution at position 141
(prebearHER2-Q141K); precursor human HER-2 having a Q-K
substitution at position 141 (prehumHER2-Q141K); precursor mouse
HER-2 having a Q-K substitution at position 142
(premouseHER2-Q142K); precursor rat HER-2 having a Q-K substitution
at position 145 (preratHER2-Q145K); and precursor human rat
chimeric HER2 having a Q-K substitution at position 141
(preE2Neu-Q141K).
7. The antigenic polypeptides according to claim 4, wherein said
mfeHER2-Q119K includes SEQ ID NO: 1, said mbearHER2-Q119K includes
SEQ ID NO: 2, said mhumHER2-Q119K includes SEQ ID NO: 3, said
mmouseHER2-Q120K includes SEQ ID NO: 4, said mratHER2-Q120K
includes SEQ ID NO: 5, and said mE2Neu-Q119K includes SEQ ID NO:
6.
8. The antigenic polypeptides according to claim 6, wherein said
prefeHER2-Q141K includes SEQ ID NO: 7, said prebearHER2-Q141K
includes SEQ ID NO: 8, said prehumHER2-Q141K includes SEQ ID NO: 9,
said premouseHER2-Q142K includes SEQ ID NO: 10, said
preratHER2-Q145K includes SEQ ID NO: 11, and said preE2Neu-Q141K)
includes SEQ ID NO: 12.
9. A method for inducing immune response to HER2 in a mammalian
subject, comprising the steps of: administering an effective amount
of the vaccine composition including an effective amount of a gene
expression construct including a nucleic acid sequence encoding an
antigenic polypeptide of the HER2 of an animal species, said
antigenic polypeptide comprising at least the extracellular domain
of HER2, including an amino acid substitution of glutamine for
lysine (Q-K) or for a conservative amino acid of K, said gene
construct additionally including at least one promoter operatively
linked to said nucleic acid sequence encoding a HER2 polypeptide,
for expression of said antigenic peptide in a living cell, wherein
said nucleic acid sequence additionally encodes the signal peptide
of HER2 (precursor HER2), and said amino acid substitution of
glutamine for lysine (Q-K) or for a conservative amino acid of K,
is encoded at amino acid 141 of precursor feline HER2, wherein said
nucleic acid sequence encoding an antigenic polypeptide is selected
from the group consisting of nucleic acid sequences encoding
precursor feline HER2, and wherein said amino acid substitution is
encoded at amino acid 141 (prefeHER2-Q141K); precursor bear HER2,
wherein said substitution is encoded at amino acid 141
(prebearHER2-Q141K); precursor human HER2, wherein said
substitution is encoded at amino acid 141 (prehumHER2-Q141K);
precursor mouse HER2, wherein said substitution is encoded at amino
acid 142 (premouseHER2-Q142K); precursor rat HER2, wherein said
substitution is encoded at amino acid 145 (preratHER2-Q145K); and
precursor human rat chimeric HER2, wherein said substitution is
encoded at amino acid 141 (mE2Neu-Q141K), wherein said nucleotide
sequence encoding prefeHER2-Q141K includes SEQ ID NO: 19; said
nucleotide sequence encoding prebearHER2-Q141K includes SEQ ID NO:
20; said nucleotide sequence encoding prehumHER2-Q141K includes SEQ
ID NO: 21; said nucleotide sequence encoding premouseHER2-Q142K
includes SEQ ID NO: 22; said nucleotide sequence encoding
preratHER2-Q145K includes SEQ ID NO: 23; and said nucleotide
sequence encoding mE2Neu-Q141K includes SEQ ID NO: 24, and an
effective amount of an adjuvant; and inducing an immune response to
HER2.
10. A method for inducing immune response to HER2 in a mammalian
subject, including the steps of: administering, to a mammalian
subject, an effective amount of a gene construct comprising a
nucleic acid sequence encoding a heterologous antigenic polypeptide
selected from the group consisting of precursor unsubstituted bear
HER2 (prebearHER2); mature unsubstituted bear HER2 (mbearHER2);
precursor unsubstituted feline HER2 (prefeHER2); and mature
unsubstituted feline HER2 (mfeHER2); the gene construct
additionally including at least one promoter for expression of said
antigenic peptide in a living cell; administering an effective
amount of an immunological adjuvant; expressing said gene construct
in cells of the mammalian subject; and inducing an immune response
against HER2 in the mammalian subject.
11. The method according to claim 10, wherein the prebearHER2
includes SEQ ID NO: 37; the mbearHER2 includes SEQ ID NO: 38; the
prefeHER2 includes SEQ ID NO: 39; and the mfeHER2 includes SEQ ID
NO: 40.
12. The method according to claim 10, wherein the nucleic acid
sequence encoding prebearHER2 includes SEQ ID NO: 41; the nucleic
acid sequence encoding mbearHER2 includes SEQ ID NO: 42; the
nucleic acid sequence encoding prefeHER2 includes SEQ ID NO: 43;
and the nucleic acid sequence encoding mfeHER2 includes SEQ ID NO:
44.
13. The method according to claim 10, wherein the step of
administering an effective amount of an immunological adjuvant is
further defined as administering an effective amount of GM-CSF.
14. The method according to claim 10, wherein the mammalian subject
is further defined as a subject hosting a population of
HER2-expressing pathological cells, additionally including the step
of eliminating or reducing the population of HER2-expressing
pathological cells.
15. The method according to claim 14, wherein the HER2 expressing
pathological cells are further defined as mammary carcinoma
cells.
16. A method for inducing immune response to HER2 in a cat,
including the steps of: administering, to a cat, an effective
amount of a gene expression construct encoding an antigenic
polypeptide selected from the group consisting of: precursor
unsubstituted bear HER2 (prebearHER2); mature unsubstituted bear
HER2 (mbearHER2); precursor unsubstituted feline HER2 (prefeHER2);
mature unsubstituted feline HER2 (mfeHER2); precursor unsubstituted
human HER2 (prehumHER2); mature unsubstituted human HER2
(mhumHER2); precursor unsubstituted mouse HER2 (premouseHER2);
mature unsubstituted mouse HER2 (mmouseHER2); precursor
unsubstituted rat HER2 (preratHER2); mature unsubstituted rat HER2
(mratHER2); precursor human rat chimeric HER2 (preE2Neu); and
mature human rat chimeric HER2 (mE2Neu); the gene expression
construct additionally including at least one promoter for
expression of the antigenic peptide in a living cell; administering
an effective amount of an immunological adjuvant; expressing the
gene construct in cells of the cat; and inducing an immune response
against HER2 in the cat.
17. The method according to claim 16, wherein the prebearHER2
includes SEQ ID NO: 37; the mbearHER2 includes SEQ ID NO: 38; the
prefeHER2 includes SEQ ID NO: 39; the mfeHER2 includes SEQ ID NO:
40; the prehumHER includes SEQ ID NO: 45; the mhumHER2 includes SEQ
ID NO: 46; the premouseHER2 includes SEQ ID NO: 47; the mmouseHER2
includes SEQ ID NO: 48; the preratHER2 includes SEQ ID NO: 49; the
mratHER2 includes SEQ ID NO: 50; the preE2Neu includes SEQ ID NO:
51; and the mE2Neu includes SEQ ID NO: 52.
18. The method according to claim 16, wherein the nucleic acid
sequence encoding prebearHER2 includes SEQ ID NO: 41; the nucleic
acid sequence encoding mbearHER2 includes SEQ ID NO: 42; the
nucleic acid sequence encoding prefeHER2 includes SEQ ID NO: 43;
the nucleic acid sequence encoding mfeHER2 includes SEQ ID NO: 44;
the nucleic acid sequence encoding prehumHER includes SEQ ID NO:
53; the nucleic acid sequence encoding mhumHER2 includes SEQ ID NO:
54; said nucleic acid sequence encoding premouseHER2 includes SEQ
ID NO: 55; the nucleic acid sequence encoding mmouseHER2 includes
SEQ ID NO: 56; the nucleic acid sequence encoding preratHER2
includes SEQ ID NO: 57; the nucleic acid sequence encoding mratHER2
includes SEQ ID NO: 58; the nucleic acid sequence encoding preE2Neu
includes SEQ ID NO: 59; and the nucleic acid sequence encoding
mE2Neu includes SEQ ID NO: 60.
19. The method according to claim 16, wherein the cat is further
defined as a cat hosting a population of HER2-expressing
pathological cells, additionally including the step of eliminating
or reducing the population of HER2-expressing pathological
cells.
20. The method according to claim 19, wherein the HER2 expressing
pathological cells are further defined as mammary carcinoma
cells.
21. Antigenic polypeptides for inducing immune response against
HER2 in a mammalian subject, said polypeptides comprising at least
the extracellular and transmembrane domains of human HER2, said
extracellular domain including an amino acid substitution selected
from the following amino acid substitutions: glutamine with lysine
(Q-K) or a conservative amino acid of lysine, at position 141 of
precursor humanHER2 (prehumHER2-Q141K); glutamine with lysine (Q-K)
or a conservative amino acid of lysine, at position 119 of mature
human HER2 (mhumHER2-Q119K); glutamine with lysine (Q-K) or a
conservative amino acid of lysine, at position 329 of precursor
humanHER2 (prehumHER2-Q329K); glutamine with lysine (Q-K) or a
conservative amino acid of lysine, at position 307 of mature
humanHER2 (mhumHER2-Q307K); glutamine with arginine (Q-R) or a
conservative amino acid of arginine, said substitution being at
position 429 of precursor human HER2 (prehumHER2-Q429R); glutamine
with arginine (Q-R) or a conservative amino acid of R, at position
407 of mature human HER2 mhumHER2-Q407R; asparagine with aspartic
acid (N-D) or a conservative amino acid of aspartic acid, at
position 438 of precursor human HER2 (prehumHER2-N438D); and
asparagine with aspartic acid (N-D), or a conservative amino acid
of aspartic acid, at position 416 of mature human HER2
mhumHER2-N416D.
22. The antigenic polypeptides according to claim 21, wherein said
prehumHER2-Q141K includes SEQ ID NO: 9; said mhumHER2-Q119K
includes SEQ ID NO: 3; said prehumHER2-Q329K includes SEQ ID NO:
28; said mhumHER2-Q307K includes SEQ ID NO: 25; said
prehumHER2-Q429R includes SEQ ID NO: 29; said mhumHER2-Q407R
includes SEQ ID NO: 26; said prehumHER2-N438D includes SEQ ID NO:
30; and said mhumHER2-N416D includes SEQ ID NO: 27.
23. A gene expression construct comprising a nucleic acid sequence
encoding an antigenic polypeptide of HER2, said nucleic acid
sequence encoding a substituted HER2 polypeptide selected from the
group including: prehumHER2-Q141K, mhumHER2-Q119K, prehumHER2-Q329K
mhumHER2-Q307K, prehumHER2-Q429R, mhumHER2-Q407R, prehumHER2-N438D,
and mhumHER2-N416D, said gene construct additionally including at
least one promoter operatively linked to said nucleic acid sequence
encoding a HER2 polypeptide, for expression of said antigenic
polypeptide in a living cell.
24. The gene expression construct according to claim 23, wherein
said nucleic acid sequence is selected from the group consisting
of: SEQ ID NO: 21, encoding said prehumHER2-Q141K; SEQ ID NO: 34,
encoding said prehumHER2-Q329K; SEQ ID NO: 35, encoding
prehumHER2-Q429R; SEQ ID NO: 36, encoding prehumHER2-N438D; SEQ ID
NO: 15, encoding said mhumHER2-Q119K SEQ ID NO: 31, encoding said
mhumHER2-Q307K; SEQ ID NO: 32, encoding mhumHER2-Q407R; and SEQ ID
NO: 33, encoding mhumHER2-N416D.
25. A vaccine composition for inducing immunity to HER2 in a
mammalian subject, comprising an effective amount of the gene
expression construct according to claim 23, and an effective amount
of an adjuvant.
26. The vaccine composition according to claim 25, wherein said
adjuvant is granulocyte macrophage colony stimulating factor
(GM-CSF)
27. The vaccine composition according to claim 26, wherein said
GM-CSF is delivered to the mammalian subject as an expression
vector comprising a polynucleotide encoding GM-CSF, for expression
of said polynucleotide in the mammalian subject.
28. The vaccine composition according to claim 26, wherein said
GM-CSF is delivered to the mammalian subject as GM-CSF protein.
29. Monoclonal antibodies selective for substituted HER2
polypeptides, the substituted HER2 polypeptides being selected from
the group consisting of: prefeHER2-Q141K, mfeHER2-Q119K,
prebearHER2-Q141K, mbearHER2-Q119K, prehumHER2-Q141K,
mhumHER2-Q119K, premouseHER2-Q142K, mmouseHER2-Q120K,
preratHER2-Q145K, mratHER2-Q120K, preE2Neu-Q141K, mE2Neu-Q119K,
prehumHER2-Q329K, mhumHER2-Q307K, prehumHER2-Q429R, mhumHER2-Q407R;
prehumHER2-N438D, and mhumHER2-N416D.
30. A diagnostic method of determining whether a mammalian subject
is sufficiently immunocompetent to respond to immunotherapy
directed at self HER2, including the steps of: administering to the
mammalian subject an effective amount of a vaccine known to induce
immune response to self HER2 of the species of the mammalian
subject; determining that an immune response to the HER2 antigen is
induced in the mammalian subject by the vaccine; and recognizing
the mammalian subject as being sufficiently immunocompetent to
respond to immunotherapy directed at self HER2.
31. The diagnostic method of claim 30, wherein the vaccine is
selected from the group consisting of the vaccine according to
claim 14 and the vaccine according to claim 34.
32. The diagnostic method according to claim 30, wherein the step
of determining that an immune response is induced is further
defined as the step of determining whether there a T cell response
has been induced, a B cell response has been induced, or a mixed T
and B cell response has been induced.
Description
TECHNICAL FIELD
[0002] The invention relates to the field of antigens and vaccines
for inducing a host to initiate immune response to the growth
factor receptor HER2, and particularly for breaking tolerance to
self HER2. The invention related particularly to the induction of
anti-HER2 immunity for the treatment and prevention of mammary
carcinomas and other HER2 expressing tumors in humans and other
mammalian species.
BACKGROUND OF THE INVENTION
[0003] One of the foremost barriers to cancer immunotherapy and
immunoprevention is the phenomenon of tolerance, the immune
system's safeguard against autoimmune disease. Most tumor antigens
are self antigens showing little or no difference from their normal
counterparts in amino acid sequence and three dimensional
structure.
[0004] The immune system generally becomes tolerant to self
antigens early in life. T lymphocyte clones specifically reactive
to self antigens are either deleted or anergized during thymic
development, or are kept in check at the periphery, mainly by
diverse populations of regulatory T cells (Treg). Especially
important are natural Treg which develop in the thymus upon high
affinity recognition of antigens in the thymic stroma (Colombo and
Piconese, 2007). It is often impossible to predict an antigen and
immunization protocol that will break tolerance to a self antigen
to achieve effective vaccination. This problem has defeated the
development of many vaccines intended to induce immune response
against tumor antigens (Wei et al, 2004).
[0005] A most promising tumor antigen in breast and other
carcinomas is HER2 (ErbB-2, neu). HER2 is amplified in .about.30%
of all breast cancers and is over-expressed in several other
epithelial-derived neoplasms including ovarian cancer, small cell
lung cancer, and cancers of the head and neck (Slamon, et al.,
1989, Yu and Hung, 2000; Tzahar and Yarden. 1998).
[0006] HER2 receptors include an extracellular domain (ECD) of
about 630 amino acids, a single membrane-spanning transmembrane
region (TM), and an intracellular domain (ICD) including a
cytoplasmic tyrosine kinase. The ECD contains four domains arranged
as a tandem repeat of a two-domain unit consisting of a
.about.190-amino acid L domain (domains I and III) followed by a
.about.120-amino acid cysteine-rich domain (domains II and IV)
(Witton, 2003, Roskoski, 2014).
[0007] Other members of the HER family of receptors, HER1, HER3,
and HER4, bind extracellular growth factor (EGF) family ligands,
but HER2 itself does not. Instead, it acts as a co-receptor, the
preferred binding partner of the other HER family receptors. Ligand
binding brings about heterodimerization of HER family receptors
with HER2, leading to tyrosine kinase activation, and the
activation of downstream signaling pathways. Overexpression of
HER2, commonly seen in carcinomas, promotes spontaneous receptor
dimerization and the activation of signaling pathways, in the
absence of a ligand (Olayioye, 2001).
[0008] The presence of HER2 specific T cells and antibodies in
breast and ovarian cancer patients indicate this molecule as a
target of immunoprevention and therapy (Disis, et al., 1994;
Peoples, et al., 1995; Fisk, et al., 1997; Kobayashi, et al.,
2000). Passive immunotherapy, by administration of the anti-HER2
moAb (monoclonal antibody), Herceptin.RTM., is used to treat
patients with advanced breast cancer (Cobleigh, et al., 1999).
Unfortunately, since ErbB-2 is a self antigen, and its sequence is
typically unmodified in cancer, tumor hosts show strong immune
tolerance against immune tolerance to HER2.
[0009] A HER2 tolerance breaking strategy that has shown some
promise is to immunize a host with xenogeneic (heterologous) HER2,
that is, HER2 from a different species than that of the immunized
host. The strategy depends on the development of antigens that are
sufficiently foreign to the HER2 of the host species to break
tolerance to HER2, but sufficiently similar to elicit T cells and
antibodies that cross react with the host HER2.
[0010] Some success has been attained with this strategy. It was
found, for example, that heterologous vaccination with rat HER2
(rat neu) produced a degree of T cell response in
human-HER2-tolerant transgenic mice. More complete responses were
produced by vaccinating the transgenic mice with a hybrid antigen
combining components of rat and human HER2 (Jacob, et al, 2006;
Jacob, et al., 2010).
[0011] There is a need for more effective tolerance breaking
antigens, for use in therapeutic and preventative vaccination
against mammary carcinoma and other HER2-expressing cancers. There
is also a need for monoclonal antibodies to such antigens, because
such antibodies are themselves potential cross reacting reagents
that can target HER2 expressing tumor cells.
[0012] There is also a need for antigens and methods useful for
breaking tolerance to self HER2 in cats, for the treatment and
prevention of mammary carcinoma in domestic feline populations.
Feline mammary cancer is an important veterinary problem. The
domestic cat population is estimated at 1 billion worldwide
(Mullikin, et al., 2010) with approximately 95 million residing in
US households
(www.humanesociety.org/issues/pet_overpopulation/facts/pet_ownership_stat-
istics.html) About 15% of unsprayed domestic cats spontaneously
develop mammary tumors, 90% of which are malignant. Most of the
malignancies are adenocarcinomas, with progression and
histopathology similar to that of human breast cancer. HER2
expression has been reported in these tumors (Hayden, et al., 1971;
Munson and Moresco, 2007; Gimenez, et al., 2010; Soares, et al.,
2013; DeMaria, et al., 2005). Furthermore, successful HER2-targeted
immunotherapies in outbred cat populations can lead directly to
improved immunotherapies for human patients, which is not the case
for immunotherapies developed with inbred rodent model populations.
That is because the amino acid sequences of human and feline HER2
are more similar than those of human and mouse or rat neu (see,
e.g., FIG. 6B), and because outbred cat populations exhibit a
genetic diversity similar to that of human populations. There is
therefore a need for antigens, vaccines, and methods for breaking
tolerance to self HER2 for the therapeutic and preventative
vaccination of mammary carcinomas of domestic cats.
[0013] Even with improved antigens and vaccines, a roadblock to
breaking HER2 tolerance is the immunocompromised status of many
cancer patients at the time of presentation for treatment. A
competent immune system is required to meet the challenge of
mounting a response to a self antigen. The induction of regulatory
T cells, and the effects chemotherapy and radiation treatments can
all contribute to a compromised immune system. There is a need for
a diagnostic method for screening human and animal candidates for
immunocompetence before the start of extended courses of
tolerance-breaking immunotherapies.
SUMMARY OF THE INVENTION
[0014] The present invention provides antigenic HER2 polypeptides
for breaking tolerance to self HER2 of an animal subject, the HER2
polypeptides including at least one point mutation in the
extracellular domain of HER2.
[0015] The present invention also provides isolated HER2 antigenic
polypeptides for inducing immune response against HER2 in a subject
of a mammalian species. The HER2 polypeptides include an amino
substitution of glutamine with lysine at position 141 of precursor
feline HER2, or at position 119 of mature feline HER2, or at
homologous positions of the HER2 of other species.
[0016] The present invention further provides HER2 gene expression
constructs for the expression of these substituted antigenic HER2
polypeptides in living cells.
[0017] The present invention still further provides HER2 vaccine
compositions for inducing immunity to HER2 in a mammalian subject,
including an effective amount of one of the substituted HER2 gene
expression constructs, and an effective amount of an adjuvant.
[0018] The present invention also provides methods for inducing an
immune response to HER2 in a mammalian subject, including the steps
of administering a substituted HER2 vaccine composition, and
inducing an immune response to HER2.
[0019] The present invention further provides a method for inducing
immune response to HER2 in a mammalian subject, using heterologous
HER2 polypeptides. The method includes the steps of administering
an effective amount of a gene expression construct encoding a
heterologous unsubstituted (i.e. wild type) feline or bear HER2
polypeptide; administering an effective amount of an immunological
adjuvant; expressing the gene construct in cells of the mammalian
subject; and inducing an immune response against HER2 in the
mammalian subject.
[0020] The present invention still further provides a method for
inducing immune response to HER2 in a cat, including the steps of
administering, to a cat, an effective amount of a gene expression
construct encoding an antigenic polypeptide of human HER2, bear
HER2, mouse HER2, rat neu, or human-rat chimeric HER2neu;
administering an effective amount of an immunological adjuvant;
expressing the gene construct in the cells of the cat, and inducing
an immune response against HER2 in the cat.
[0021] The present invention also provides antigenic polypeptides
for inducing immune response against HER2 in a mammalian subject,
the antigenic polypeptides including either a substitution of
glutamine with lysine at position 329 of precursor human HER2; a
substitution of glutamine with lysine at position 307 of mature
human HER2; a substitution of glutamine with arginine at position
429 of precursor human HER2; a substitution of glutamine with
arginine at position 407 of mature human HER2; a substitution of
asparagine with arginine at position 438 of precursor human HER2;
and a substitution of asparagine with arginine at position 416 of
mature human HER2. The invention provides these antigenic peptides
as isolated peptides, gene expression vectors, and vaccine
compositions.
[0022] The present invention further provides monoclonal antibodies
selective for all of the previously mentioned substituted antigenic
HER2 polypeptides.
[0023] The present invention still further provides a diagnostic
test to determine whether a mammalian subject is sufficiently
immunocompetent to respond to immunotherapy directed at self
HER2.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawing(s) will be provided by the Office
upon request and payment of the necessary fee.
[0025] Other advantages of the present invention are readily
appreciated as the same becomes better understood by reference to
the following detailed description when considered in connection
with the accompanying drawings wherein:
[0026] FIG. 1 shows a table of primer pairs used for the
authentication of feline mammary tumor lines;
[0027] FIG. 2 shows the authentication of the feline mammary tumor
lines K248 and K12 via short tandem repeat (STR) analysis at 4
loci: FCA733 (Felis catus Failed Axon Connector, human homolog),
FCA 749 (Thyroglobulin), MACROD2 (MACRO Domain Containing 2), and
feline mtCR (Mitochondrial Control Region/d-Loop); PCR
amplification was conducted using primer pairs listed in the table
above; template DNAs were prepared with a DNeasy kit (Qiagen) from
cultures of cat mammary tumor lines K248 and K12 or the human
ovarian cancer line SKOV3 as a negative control; the left-hand
panel shows predicted product sizes of STR regions from FCA733
(lanes 1-3), FCA749 (lanes 4-6), or MACROD2 (lanes 8-10); ladder (1
kb+, Invitrogen) is in lane 7; the right-hand panel shows product
sizes of the feline mtCR PCR amplified in duplicate, with ladder in
lanes 3 and 6; Amplified products matched expected sizes based on
the current cat genome sequence assembly (c.f.
www.ncbi.nlm.nih.gov/genome/guide/cat/) and the Felis catus
mitochondrion (NC_001700.1); PCR products were verified by
sequencing;
[0028] FIG. 3A shows expression of HER2 in feline mammary carcinoma
by flow cytometric analysis of cell surface EGFR (HER1), HER2, HER3
and MHCI expression (open histograms) in FMC cell lines K12 and
K248; human SKOV3 cells are included as controls; shaded histograms
are unstained controls;
[0029] FIG. 3B shows expression of HER2 in feline mammary carcinoma
by immunohistochemical analysis of three primary FMC samples and
FMC line K248 outgrowth in SCID mice using polyclonal Ab to huHER2;
H&E stains are shown in parallel;
[0030] FIG. 4A shows RTK (receptor tyrosine kinase) analysis of
protein lysates from primary FMC tumor with paired uninvolved
stromal tissue; phospho-Akt (S473) detection is indicated with a
box;
[0031] FIG. 4B shows RTK analysis of FMC cell lines K12 and K248
were analyzed by PathScan RTK signaling array (Cell Signaling
Technology). Phospho-Akt (S473) detection is indicated with a box;
human SKOV3 was the control;
[0032] FIG. 5A shows RTK activity in FMC, as determined by Western
blot analysis of total HER2, pHER2 (Y1248), total Akt and pAkt
(S473) in primary FMC samples; feline PBL (peripheral blood
leukocytes) was the negative control.
[0033] FIG. 5B shows the sensitivity of FMC cell lines to receptor
tyrosine kinase inhibitors; cells were cultured in gefitinib or
lapatinib for 48 h and the percentage of proliferative cells was
measured with Alamar Blue by comparison to untreated controls;
*p<0.001 two-way ANOVA with Dunnet's posttest;
[0034] FIG. 6A shows the amino acid sequences of human, cat, bear,
rat, and mouse HER2, compared by clustal alignment ([*], identical
a.a., [:], strongly similar a.a., [.], weakly similar a.a.); the
signal peptide and transmembrane region are indicated with
lines;
[0035] FIG. 6B summarizes the results of BLASTP analysis of
full-length HER2 a.a. sequence identity;
[0036] FIG. 6C (top panel) shows a schematic of HER2 domains, and
(bottom panel) flow cytometric analyses of the binding of four moAb
to human HER2 (TA-1, N29, N12, Trastuzumab) and one moAb to rat neu
(7.16.4); 3T3 cells transfected to express the indicated HER2 ECTM
were stained with 1 .mu.g/mL of the indicated moAb (open histogram)
with detection by PE-conjugated secondary antibody; secondary
antibody alone (shaded histogram) was the negative control;
[0037] FIG. 6D shows an annotated amino acid sequence of precursor
feline HER2, highlighting the site of the 141 substitution;
[0038] FIG. 7 shows a construct for the expression of
prefeHer2-Q141K ("feHER2-K") in a host animal;
[0039] FIG. 8A shows the sequence of the feHER2ecd-hFc fusion
protein;
[0040] FIG. 8B shows a schematic of the feHER2ecd-hFc fusion
protein (top panel) and a Western blot verifying its composition
(bottom panel);
[0041] FIG. 9A shows a flow cytometric analysis of the binding of
3T3/prefeHER2-Q141K by moAb to HER2/neu, with staining and analysis
conducted as in FIG. 6C;
[0042] FIG. 9B shows antibody response to pprefeHER2,
pprefeHER2-Q141K and pprehuHER2 in BALB/c mice before (Pre) and two
weeks after 1.times. and 2.times. vaccination given in 2 wk
intervals; horizontal lines depict the mean for each group
(*p<0.05, **p<0.005 one-way ANOVA with Tukey's posttest);
[0043] FIG. 9C shows T cell response to feline and human HER2 in
2.times. vaccinated mouse splenocytes (n=4) or naive mouse
splenocytes (n=3) as measured by ELISPOT in triplicates using 10
.mu.g recombinant antigen (*p<0.05, **p<0.01 two-way ANOVA
with Dunnet's posttest);
[0044] FIG. 10A shows induction of anti-HER2 antibody in cats
including feline anti-huHER2 (left panel), rat neu (rat HER2)
(middle panel) or feHER2 (right panel) IgG titer induced by pE2Neu
and pfeGM-CSF vaccination; serum samples were collected just before
and 2 weeks after each of 4 vaccinations given in 3 week intervals.
Antigen binding was analyzed by flow cytometry using transfected
3T3 cells;
[0045] FIG. 10B shows induction of anti-HER2 antibody in cats,
including feline anti-huHER2 (left), feHER2-K (middle) or feHER2
(right) IgG titer induced by the indicated vaccines; serum samples
were collected just before and 2 weeks after each of 4 vaccinations
given in 3 week intervals; antigen binding was analyzed by flow
cytometry using transfected 3T3 cells;
[0046] FIG. 10C shows the binding of immune sera to 3T3/feHER2
cells (top left panel), K12 tumor cells (top right panel) or K248
tumor cells (bottom panel), either prior to (shaded histogram) and
two weeks after, the 4th vaccination (open histogram) with the
indicated vaccines; each overlayed histogram represents an
individual cat;
[0047] FIG. 11A shows a specificity control of mouse immune serum:
binding of moAb TA-1 at indicated concentrations to 3T3 cells
expressing prefeHER2, prefeHER2-Q141K, or prehuHER2; un-transfected
3T3 cells were the control;
[0048] FIG. 11B shows a specificity control of mouse immune serum:
binding of immune serum from vaccinated mice to un-transfected 3T3
cells; each panel depicts the binding of pre-, post 1.times. and
post 2.times. vaccination serum from an individual mouse;
[0049] FIG. 11C shows a specificity control of feline immune serum,
based on an experiment with 3 cats; pre-vaccination (gray shaded)
and two weeks post 4.times. vaccination (open histogram); serum was
incubated at a 1:50 dilution with untransfected 3T3 cells followed
by PE-anti-feline IgG; each panel represents an individual cat;
[0050] FIG. 11D shows a specificity control of feline immune serum,
based on an experiment with 15 cats; pre-vaccination (gray shaded)
and two weeks post 4.times. vaccination (open histogram); serum was
incubated at a 1:50 dilution with untransfected 3T3 cells followed
by PE-anti-feline IgG. Each panel represents an individual cat;
[0051] FIG. 12A shows a cartoon model of the effect of a Q141K
substitution in feline HER2;
[0052] FIG. 12B shows a space filling model of the effect of a
Q141K substitution in feline HER2;
[0053] FIG. 12C shows an surface electrostatic model of the effect
of a Q141K substitution in feline HER2;
[0054] FIG. 13A shows the induction of human HER2-specific T cells
in cats; PBMC were harvested after 4.times. vaccination and
stimulated with 10 .mu.g/mL rhuHER2-huFc or an IgG control; results
are presented as IFN.gamma. spot forming units (SFU)/10.sup.6 PBMC;
*p<0.05, **p<0.01 Mann-Whitney test;
[0055] FIG. 13B shows the induction of feline HER2-specific T cells
in cats; PBMC were harvested after 4.times. vaccination and
stimulated with 10 .mu.g/mL rfeHER2-huFc or human IgG control;
results are presented as IFN.gamma. spot forming units
(SFU)/10.sup.6 PBMC; *p<0.05, **p<0.01 Mann-Whitney test;
[0056] FIG. 13C shows induction of feline HER2-specific T cells in
cats; HER2 specific T cell response to feline HER2 is shown after a
total of 7 vaccinations with pfeHER2-K with or without pE2Neu;
*p<0.05, **p<0.01 Mann-Whitney test;
[0057] FIG. 13D shows human HER2 specific T cell response induced
by pE2Neu or pbearHER2 followed by pfeHER2; *p<0.05, **p<0.01
Mann-Whitney test;
[0058] FIG. 13E shows feline HER2-specific T cell response induced
by pE2Neu or pbearHER2 followed by pfeHER2; *p<0.05, **p<0.01
Mann-Whitney test;
[0059] FIG. 14A shows the binding of anti-human HER2 antibodies to
3T3 cells transiently transfected by lipofection with human HER2
mutant constructs encoding amino acid substitutions, with binding
of moAb to HER2, Ab5 and N12 was measured by flow cytometry; Ab4
specific to rat neu is the negative control;
[0060] FIG. 14B shows the location of the amino acid substitutions
shown in FIG. 14A on a schematic diagram of human HER2;
[0061] FIG. 15 shows a summary of results of a flow cytometric
analysis of humoral immune response of wild-type BALB/c mice to
human HER2 including amino acid substitutions; wild type BALB/c
mice were electro-vaccinated i.m. with 60 ug HER2 construct+60 ug
pmGMCSF divided in two sites; serum was collected 3 weeks after
immunization and binding to Her2-expressing SKOV3 cells was
assessed by flow cytometry;
[0062] FIG. 16 shows binding of HER2 transgenic (HER2Tg) mouse
immune serum to 3T3 EKB cells transfected to express WT human
HER2;
[0063] FIG. 17 shows a characterization of HER2 and EGFR expression
in the human cancer cell lines used to detect the presence of HER2
and EGFR antibodies in the sera of immunized mice;
[0064] FIG. 18 shows an analysis of the binding of anti-human HER2
antibodies, induced in BALB Her2Tg mice by two rounds of
vaccination with HER2 mutant DNA encoding amino acid substitutions,
to the human cancer cell lines characterized in FIG. 17; *
p<0.05, **p<0.005, by 2-tail student's t test; and
[0065] FIG. 19 shows an analysis of humoral immune response in BALB
Her2Tg mice to human Her2 ECTM including amino acid substitutions;
female BALB Her2Tg mice were electro-vaccinated twice at a 2-week
interval; serum was collected 2 weeks after immunizations, and
binding to HER2-expressing SKOV3 and TNBC SUM159 cells was assessed
by flow cytometry; HER2 Ab levels elicited by each mutant construct
were compared to the wild type pE2TM; * p<0.05, **p<0.005, by
2-tail student's t test.
DETAILED DESCRIPTION OF THE INVENTION
[0066] The compositions and methods according to the present
invention represent solutions to the problem of immunological
tolerance of tumor hosts to the HER2 antigens of their tumors. They
represent the first reported examples of deliberately introduced
point mutations that convert HER2 polypeptides into antigens
capable of breaking tolerance to self HER2, and inducing an immune
response reactive to self HER2. In an exemplary embodiment, a point
mutation brings about the substitution of glutamine with lysine
(Q-K) in the amino acid sequence QLRSLTEILKGGVLI (SEQ ID NO: 109)
of HER2 domain I, rendering the sequence KLRSLTEILKGGVLI (SEQ ID
NO: 110).
[0067] In a related embodiment, the present invention includes
isolated antigenic polypeptides for breaking tolerance and inducing
immune response against HER2 in a mammalian subject. The
polypeptides include a substitution of glutamine with lysine with
(Q-K) or a with a conservative amino acid of lysine, at position
141 (Q141K) of precursor feline HER2, or at a homologous position
of the precursor HER2 of another animal species. Precursor HER2
(preHER2) is defined as HER2 including a signal peptide, an
extracellular domain (ECD) and a transmembrane domain (TM). For
brevity, the term Q-K substitution will refer to both a
substitution of Q with K, and to a substitution of Q with a
conservative amino acid, such as arginine. Also included in this
embodiment are antigenic polypeptides of mature HER2 (mHER2), that
is, HER2 lacking a signal peptide, and including the ECD and TM. In
mHER2, the Q-K substitution is at position 119 of mfeHER2, or at a
homologous position of the mature HER2 of another species.
[0068] The specification of the Q141K or Q119K substitution at a
"homologous position of the HER2 of another animal species"
indicates that the substitution is made to the Q that is in the
lead position in a highly conserved 15 aa sequence in domain I of
ErbB2, the conserved sequence being QLRSLTEILKGGVLI (see, for
example, the underlined feature of SEQ ID NO: 39 as shown in FIG.
6D for precursor feline HER2. Exemplary amino acid and nucleotide
sequences of HER2 polypeptides are listed in Table 4.
[0069] A standardized notation system will be used to denote
specific forms of HER2 polypeptides. The notation will refer both
to a polypeptide gene product, and to the gene construct employed
to induce expression of the gene product in an organism. In the
notation system, the animal species from which the HER2 is derived
will be abbreviated and italicized and placed before HER2. The
identity and location of an amino acid substitution if any, will be
hyphenated after HER2. A wild-type HER2 will lack a designated
substitution. The designation of a precursor or mature form will be
abbreviated "pre" or "m", respectively, and placed to the left of
the species. If the construct is included in a vector, the vector
abbreviation will appear as the left-most term. For example:
##STR00001##
[0070] An annotated amino acid sequence of unsubstituted prefeHER2
(SEQ ID NO: 39) and prefeHER2-Q141K (SEQ ID NO: 7) is shown in FIG.
6D. Other exemplary embodiments of the antigenic HER2 polypeptides
having a Q141K or homologous substitution are described in Example
2. They include precursor feline HER-2 having a Q-K substitution at
position 141 (prefeHER2-Q141K), precursor bear HER-2 having a Q-K
mutation at position 141 (prebearHER2-Q141K), precursor human HER-2
having a Q-K mutation at position 141 (prehumHER2-Q141K), precursor
mouse HER-2 having a Q-K mutation at position 142
(premouseHER2-Q142K), precursor rat HER-2 having a Q-K mutation at
position 145 (preratHER2-Q145K), and precursor human rat chimeric
HER2 having a Q-K mutation at position 141 (preE2Neu-Q141K). They
also include mature feline HER2 having a Q-K substitution at
position 119 (mfeHER2-Q119K); mature bear HER2 having a Q-K
substitution at position 119 (mbearHER2-Q119K); mature human HER2
having a Q-K substitution at position 119 (mhumHER2-Q119K); mature
mouse HER-2 having a Q-K substitution at position 120
(mmouseHER2-Q120K); mature rat HER2 (rat neu) having a Q-K
substitution at position 120 (mratHER2-Q120K); and mature human rat
chimeric HER2 having a Q-K substitution at position 119
(mE2Neu-Q119K)
[0071] Exemplary amino acid sequences for each of the substituted
antigens as follows: prefeHER2-Q141K, SEQ ID NO: 7;
prebearHER2-Q141K, SEQ ID NO: 8; prehumHER2-Q141K, SEQ ID NO: 9;
premouseHER2-Q142K, SEQ ID NO: 10; preratHER2-Q145K, SEQ ID NO: 11;
preE2Neu-Q141K, SEQ ID NO: 12; mfeHER2-Q119K, SEQ ID NO: 1;
mbearHER2-Q119K, SEQ ID NO: 2; mhumHER2-Q119K, SEQ ID NO: 3;
mmouseHER2-Q120K, SEQ ID NO: 4; mratHER2-Q120K, SEQ ID NO: 5; and
mE2Neu-Q119K, SEQ ID NO: 6. It will be understood that the
disclosed amino acid sequences are exemplary, and that the present
invention encompasses all immunologically equivalent sequences.
[0072] The development of the antigenic HER2 polypeptides of the
present invention was initiated on the basis of experiments in
which it was found that heterologous electrovaccination with rat
neu (rat HER2) overcame T cell tolerance in human HER2 transgenic
(Tg) mice. The term "heterologous", when used to refer to HER2, or
another antigen, indicates that the antigen is derived from an
animal species or species hybrid that is different from the species
of the animal being vaccinated. Unfortunately, heterologous
vaccination of human HER2 Tg mice with rat neu did not produce an
effective humoral (B cell) response. The resulting immune sera did
not cross react with human HER2 (Jacob, et al., 2006).
[0073] A hybrid antigen was next developed, which included portions
of human HER2 and rat HER2 (rat neu), which will be referred to as
E2Neu. This hybrid antigen included human HER2 extracellular
domains (ECD) 1/2, rat neu ECD 3/4 and the rat neu transmembrane
domain. E2Neu was incorporated into a gene construct for expression
in animals, specifically the plasmid vector pE2Neu. The vector was
delivered, as a component of a vaccine, to human HER2 Tg mice. The
pE2Neu vaccine was found to induce both humoral and cellular (T
cell) immunity against human HER2 in human HER2 Tg mice (Jacob, et
al., 2010).
[0074] Because a chimeric human/rat form of HER2 broke tolerance as
well as the pure heterologous rat form, and gave a more complete
immune response than the pure rat form, it was hypothesized that
results can be further improved by immunization with forms of HER2
that are more minimally altered from the HER2 of a human host or
experimental animal.
[0075] In one test of this hypotheses, animal hosts were immunized
with self HER2 containing point mutations, to afford immunogenicity
while preserving HER2 epitopes. In experiments disclosed in
Examples 3 and 4, certain point mutations in HER2 were found to
confer enhanced immunogenicity in vaccination experiments. Each
point mutation produced a single amino acid substitution in
HER2.
[0076] Vaccination experiments with wild type and substituted forms
of HER2 are described in Example 3. These vaccination experiments
were performed not only in mice but also in a novel and highly
realistic outbred cat tumor immunity model, which was utilized in
experiments disclosed in Examples 1-4. Feline HER2 is more closely
related to human HER2 than are the HER2s of mice or rats (FIG. 6B).
Outbred cats show a range of genetic diversity, including MHC
molecule diversity, comparable to that of humans, and not found in
inbred rodent models.
[0077] In the experiments of Example 3, cats were immunized with a
genetic vaccine including an expression vector which induced
expression of the prefeHER2Q141K as well as wild type controls and
heterologous HER2 forms. It was found that prefeHer2-Q141K was
sufficiently foreign to break tolerance to self HER2 (i.e. feHER2)
in outbred cats, inducing both antibodies and T cells reactive with
feHer2. The antibodies and T cells were also reactive with Her2
molecules of humans and other species. Because these findings were
obtained in the highly realistic outbred cat vaccine test system,
it is reasonably predictable that they will also be applicable to
other animal species. That is, it is predictable that HER2-K mutant
antigens of cats and other animal species, in which Q141 or a
homologous Q is substituted with K, will produce anti HER2 immunity
in those animal species, when included in an appropriate vaccine
composition. Indeed, it found, in experiments disclosed in Example
4, that the homologous substituted human HER2 polypeptide,
prehumHER2-Q141K, broke tolerance to human HER2 in human HER2
transgenic mice.
[0078] Extrapolating from these findings, it is also predictable
that the corresponding mature forms of HER2-Q141K, will also be
effective in inducing immunity. The lack of a signal peptide is
expected to have no effect on the reactivity of the Q-K
substitution epitope, which is over a hundred residues distant, in
the ECD. When expressed in a host cell, the mature forms of HER2
would not be processed into the secretory pathway and inserted into
the cell membrane, but they would nonetheless be available to
antigen presenting cells upon apoptosis or necrosis of the
expressing cells. Thus, the previously enumerated mature forms of
HER2, including a Q-K substitution at position 119 of feline HER2,
or at a homologous position in other species, are also encompassed
by the present invention.
[0079] In a related embodiment, the present invention includes gene
expression constructs such as those utilized in Examples 2-4. The
expression constructs include a nucleic acid sequence encoding an
antigenic polypeptide of the HER2 of an animal species, and
specifically encoding the Q141K substitution at position 141 of
precursor feline HER2 (prefeHER2-Q141K), or at position 119 of
mature feline HER2 (mfeHER2-Q119K), and at homologous positions of
the precursor and mature forms of HER2 of other animal species. The
expression construct additionally includes at least one promoter
operatively linked to said nucleic acid sequence encoding a HER2
polypeptide, for expression of said antigenic peptide in a living
cell.
[0080] The encoded constructs, and exemplary nucleic acid sequences
encoding them, include: mfeHER2-Q119K, SEQ ID NO: 13;
mbearHER2-Q119K, SEQ ID NO: 14; mhumHER2-Q119K, SEQ ID NO: 15;
mmouseHER2-Q120K, SEQ ID NO: 16; mratHER2-Q120K, SEQ ID NO: 17;
mE2Neu-Q119K, SEQ ID NO: 18; prefeHER2-Q141K; SEQ ID NO: 19;
prebearHER2-Q141K, SEQ ID NO: 20; prehumHER2-Q141K, SEQ ID NO: 21;
premouseHER2-Q142K, SEQ ID NO: 22; preratHER2-Q145K, SEQ ID NO: 23;
and mE2Neu-Q141K, SEQ ID NO: 24. It will be understood that the
recited nucleic acid sequences are only exemplary, and that each
specified polypeptide can be encoded by one or more synonymous
nucleic acid sequences without departing from the scope of the
present invention. The nucleic acid sequences are preferably DNA
sequences, but may alternatively comprise at least one RNA
molecule.
[0081] The gene construct also includes at least one promoter
operatively linked to the nucleic acid sequence encoding a HER2 Q-K
substituted polypeptide, to promote expression of the gene product
in a mammal or other organism. The promoter or other regulatory
element is selected to ensure that the nucleic acid sequence is
transcribed and translated into the antigenic polypeptide upon
introduction into a living cell. An exemplary promoter is the
cytomegalovirus (CMV) promoter, but any suitable promoter known in
the art can be utilized, including, but not limited to, the
cytomegalovirus (CMV) promoter, the Rous sarcoma virus (RSV)
promoter; the SV40 virus promoter, and the mammalian housekeeping
promoter EF1 (elongation factor 1).
[0082] A preferred expression vector is the naked DNA plasmid
vector pVAX1 (Life Technologies, Grand Island, N.Y.), but any
suitable vector system known in the art can be employed with
routine modifications, depending on the cell type in which
expression is to be obtained. In the Examples, gene constructs are
expressed in mammalian hosts and in cultured mammalian cells, but
with suitable expression vectors they can also be expressed in
bacteria, yeasts, insect cells, and any other desired host.
Appropriate techniques or references thereto can be found in Green
and Sambrook (2012).
[0083] In a related embodiment, the present invention includes a
vaccine composition for inducing immunity to HER2 in a mammalian
subject. The vaccine composition includes an effective amount of at
least one of the previously enumerated isolated HER2 polypeptide
antigens or, more preferably, an effective amount of at least one
of the previously mentioned gene expression constructs. The gene
expression construct is expressible the living cells of the
mammalian subject. The vaccine composition preferably includes an
adjuvant to amplify immune response to the antigen. The present
invention also includes methods for inducing immune response to
HER2 in a mammalian subject, including the steps of administering
an effective amount of the vaccine composition, and inducing an
immune response to HER2.
[0084] An effective amount of vaccine composition is defined as one
which produces an observable antigen-specific humoral and/or
cellular immune response, and if administered as a therapy, a
reduction in a population HER2-expressing target cells. The
effective amount of a particular vaccine composition can be
determined by one skilled in the art on the basis of preliminary
trials in which increasing doses are given, and, as warranted,
multiple courses of administration are tested. The extent of T or B
cell response is measured by, for example, ELISA, cytotoxicity or
growth suppression assays, and ELISPOT or other cytokine release
assays. The effective amount can be adjusted to account for
differences in host weight, species, or physical condition.
[0085] Exemplary gene constructs for vaccination techniques,
according to the present invention, include HER2 antigens encoded
into the naked DNA plasmid expression vector pVax, as described in
detail in Examples 3 and 4. The preferred adjuvant is GM-CSF,
administered either in soluble form or as a nucleic acid expression
vector, which results in GM-CSF expression at a vaccination site.
Preferably, the GM-CSF is preferably delivered as an expression
plasmid.
[0086] Alternatively, the HER2 antigen gene constructs of the
present invention can be cloned into any plasmid, or bacterial, or
viral vector that can serve as a vaccine vector, to transfect or
transduce mammalian cells. Examples include a retrovirus vector, an
adenovirus vector, a lentivirus vector, a vaccinia virus vector, a
pox virus vector, an adenovirus-associated vector, a virus-like
particle, a Salmonella vector, a Shigella vector, a Listeria
vector, a Yersinia vector, and an Escherichia vector. Techniques
for expression of proteins using viral vectors can be found in
Adolph, K. ed. "Viral Genome Methods" CRC Press, Florida (1996) and
in Harrop, et al., 2006. Techniques for the use of attenuated
bacterial vectors such as Salmonella, Shigella, Listeria, Yersinia,
and Escherichia species are found for example in Vassaux et al.,
2006. Vaccination is preferably accompanied by a cytokine adjuvant,
such as -1, -2, -3, -6, -12, gamma-interferon, tumor necrosis
factor, GM-CSF, or flt-3 ligand, delivered either as an expression
construct, or as a cytokine protein.
[0087] Although electrovaccination is the preferred delivery mode,
the gene expression constructs of the present invention can
alternatively be packaged into liposomes or coated onto colloidal
gold particles prior to administration. The gene expression
constructs can then be administered intradermally, subcutaneously
or intramuscularly by injection or by gas driven particle
bombardment. Alternatively, the gene expression constructs can be
administered to host cells ex vivo. Host cells, such as bone-marrow
derived cells, can be induced to express the HER2 polypeptide
antigens, and then reintroduced to the host to effect immunization.
Appropriate techniques can be found in Sudowe, S. and Reske-Kunz,
A. B. eds. "Biolistic DNA Delivery: Methods and Protocols", Humana
Press, New York City (2012), and Raz, E., ed. "Gene Vaccination:
Theory and Practice, Springer, New York City (1998).
[0088] The antigenic HER2 polypeptides of the present invention can
also be administered as isolated polypeptides, as a component of a
vaccine composition that can also include at least one adjuvant,
including but not limited to incomplete or complete Freund's
adjuvant, alum, QS21, TITERMAX; cytokines, and cytokines such as,
interleukins-1, -2, -3, -6, -12, gamma-interferon, tumor necrosis
factor, GM-CSF, or flt-3 ligand. The vaccine composition, with or
without adjuvant, can be administered in a pure preparation, or
admixed with a pharmaceutically acceptable carrier, diluent, or
excipient, as a sterile suspension, emulsion, or in a lipid carrier
such as a liposome. Delivery can be by subcutaneous, intradermal,
intramuscular, intranasal, or intravenous routes.
[0089] In another embodiment, the present invention includes
antibodies that selectively bind each of the substituted HER2
antigenic polypeptides disclosed herein. The term "selectively
binds", when applied to an antibody of the present invention,
indicates that the antibody (a) binds at a level above background
to a particular substituted HER2 antigenic polypeptide, with
background taken as the level of binding of a nonspecific reference
agent such as a matched immunoglobulin isotype control; and (b)
binds at a level at or below background to the corresponding wild
type HER2 peptide.
[0090] With the amino acid sequences disclsosed herein, one skilled
in the art can readily generate antibodies selective for each of
the substituted HER2 polypeptides. Thus, the present invention
includes monoclonal antibodies selective for substituted HER2
polypeptides, the substituted HER2 polypeptides being selected from
the group consisting of: prefeHER2-Q141K mfeHER2-Q119K,
prebearHER2-Q141K, mbearHER2-Q119K, prehumHER2-Q141K,
mhumHER2-Q119K, premouseHER2-Q142K, mmouseHER2-Q120K,
preratHER2-Q145K, mratHER2-Q120K, preE2Neu-Q141K, mE2Neu-Q119K,
prehumHER2-Q329K, mhumHER2-Q307K, prehumHER2-Q429R, mhumHER2-Q407R;
prehumHER2-N438D, and mhumHER2-N416D. Monoclonal antibodies
according to the present invention can be obtained by using the
polypeptides or their antigenic fragments as the antigens. For
example, an antibody can be obtained by preparing hybridomas by
fusion of myelomas, or other mammalian cells capable of infinite
proliferation, with antibody-producing cells collected from mammals
immunized with one of the antigens. Hybridoma clones capable of
producing monoclonal antibody are cultured in vivo or in vitro. The
preferred method of immunizing a mammal is by electrovaccination
with expression vectors encoding mutant HER2, as described in
Examples 3 and 4.
[0091] Alternatively, immunizations can be performed with purified
or partially purified antigenic polypeptides. For example, the
antigens can be in the form of live or killed cells expressing
mutant HER2, or the partially purified culture supernatants or
homogenates of such cells. In another alternative, the antigens are
prepared chemically by peptide synthesis based on the amino acid
sequences disclosed herein for the substituted polypeptides.
[0092] Immunization with antigenic polypeptides is performed by
techniques well known in the art. For example, antigens, preferably
in combination appropriate adjuvants, are injected into mammals
intradermally, subcutaneously, intramuscularly, intraperitoneally,
or intravenously. Rodents such as rats, mice and hamsters can be
used. Depending upon species, the total dose of the antigens is
generally in the range of about 5-500 .mu.g of purified antigen or
equivalent per animal.
[0093] Immunizations are performed 2-5 times at an interval of 1-2
Weeks. After the course of immunization, the animal's spleen is
extracted and dispersed into a suspension of spleen cells. The
antibody-producing cells and the myeloma cells obtained in the
above are fused into a cell fusion mixture containing the objective
hybridomas.
[0094] Suitable myeloma cells include mouse myeloma lines, such as
P3-NS1-Ag4-1 cells (ATCC TIB18), P3-X63-Ag8 cells (ATCC TIB9),
SP2/O--Ag14 cells (ATCC CRL1581), and other mutants which lack the
HGPRT (hypoxanthine-guanine phosphoribosyltransferase) gene.
[0095] Cell fusion is carried out by techniques well known in the
art, such as incubation with polyethylene glycol or Sendai virus,
or by electric pulse. For example, fusion partners are suspended in
fusion media containing fusion accelerators, and incubated at about
30-40.degree. C. for about 1-5 min. Conventional serum-free media
such as minimum essential medium (MEM), RPMI 1640 medium, and
Iscove's Modified Dulbecco's Medium (IMDM) are preferred fusion
media. To select hybridomas, the resultant cell fusion mixture is
transferred to selection media such as HAT medium, and incubated at
about 30-40.degree. C. for from 3 days to 3 weeks, after which
point only hybridomas are expected to survive. Detailed fusion and
selection protocols are found in Harlow and Lane 1988 and Pandey,
2010.
[0096] Hybridomas are cultured as monoclonal populations, and
antibodies secreted into culture are screened for reactivity with
the immunizing antigen, preferably expressed on cells. Screening is
preferably by immunofluorescence assays such as those described in
Example 1. Alternatively, well known assays such as enzyme linked
immunoassay and radioimmunoassay are used. See, for example Harlow
and Lane, 1988
[0097] For selection of antibodies specifically reactive to the
substituted HER2 antigens of the present invention, immunoassay
screening techniques are employed to determine which of the
monoclonal antibodies are reactive with the substituted antigen but
not to the corresponding non-substituted antigen. Preferably, the
immunoassay is an immunofluorescence assay, and the targets for
immunofluorescence staining are cells transfected with either the
substituted or non-substituted form of the HER2 antigen. Exemplary
techniques for transfecting 3T3 cells with HER2 antigen constructs,
and for assaying specific antibody binding to the transfected
cells, are described in Piechocki, et al., 2001. Hybridoma clones
secreting antibodies that bind specifically to the substituted form
of the HER2 antigen, but not to the unsubstituted form, are
producing the selective antibodies of the present invention. These
clones are cultured or stored according to standard techniques. The
monoclonal antibodies produced by these clones are harvested from
culture in vivo, for example from animal ascites culture, or in
vitro, from culture medium. The antibodies are purified by
techniques such as protein A or Protein G affinity chromatography,
salting out, dialysis, ultrafiltration, ion-exchange
chromatography, affinity chromatography, high performance liquid
chromatography (HPLC), gel electrophoresis, and isoelectrophoresis,
or an appropriate combination of techniques. Once purified, the
monoclonal antibodies are refrigerated, frozen, or lyophilized for
storage.
[0098] Alternatively, one skilled in the art can utilize the HER2
antigens of the present invention to generate monoclonal antibodies
by antibody phage display techniques. For example, cats are
vaccinated with feHER2 or feHER2-Q141K, and RNA is extracted from
bone marrow cells or PBLs. The RNA is used for the preparation of
oligo dT-primed cDNA libraries, as described by Konthur and Walter
(2002). Variable heavy (VH) and variable light (VL) chain orfs are
PCR amplified based on reported sequences
(www.ncbi.nlm.nih.gov/genome?term=felis%20catus), then Ig cDNA
libraries are generated following the protocol described by Hammers
and Stanley (2014). For screening purposes, phagemids such as
pCombo3X are used to express these two chains as an scFv fused to
the pill minor capsid protein of an engineered filamentous
bacteriophage (originally derived from M13) (Hammers and Stanley
2014). The cognate antigen used for screening is recombinant fe
HER2ecd-Fc (SEQ ID NO: 69), which is immobilized on sterile dishes
via Fc binding. Screening is carried out with the
pCombo3x/feHER2scFv library. A round of screening consists of
binding of the feHER2scFv library to immobilized feHER2ecd-Fc to
capture clones with high-affinity binding ("biopanning"), washing
to remove background non-binding phage, elution of bound
phage/scFv, infection of competent E. coli, expression and recovery
via addition of helper phage, and preparation of recovered
phage/scFv library for another round of screening. Several rounds
of such screening are required to achieve sufficient purity of the
aHER2 fe-scFv for functional and genetic analysis. Affinities of
.about.1 nMolar are readily achievable (Hammers and Stanley 2014;
Carmen and Jemutus 2002). The cloned aHER2-fe-scFv can be 1) used
directly as a recombinant scFv, 2) stabilized by fusion to an Fc
domain (as in SEQ ID NO: 69) for use in vivo, or 3) reconstructed
into full-length IgG light and heavy chains for production of aHER2
monoclonal Ab's of interest.
[0099] Human anti-HER2 mAb's are engineered by the same protocol
using human PBLs for generating the initial Ig cDNA libraries.
[0100] In another embodiment, the present invention includes
methods for immunizing a mammalian subject against HER2, using
heterologous, unsubstituted ("wild type") antigenic polypeptides of
bear HER2 or feline HER2. While these HER2 polypeptides are known,
naturally occurring polypeptides, their inclusion in a method to
induce immunity against HER2 represents a novel use. The methods
include vaccination with an effective amount of expression
construct, as previously described for the methods involving Q to K
mutants of HER2. The unsubstituted bear and feline HER2
polypeptides were found to be effective antigens in experiments
testing the hypothesis that anti-HER2 response can be achieved by
immunization of a host with HER2 that is heterologous, but
relatively closely matched to self HER2. Cat and bear HER2 are more
closely related to human HER2 than is, for example, rodent HER2
(FIG. 6B). As disclosed in Example 3, bear HER2 broke the HER2
tolerance of cats, and induced both antibodies and T cells that
reacted against feline HER2.
[0101] Therefore, the present invention includes a method for
inducing immune response to HER2 in a mammalian subject, beginning
with the step of administering, to a mammalian subject, an
effective amount of a gene construct comprising a nucleic acid
sequence encoding a heterologous antigenic polypeptide selected
from the group consisting of precursor unsubstituted bear HER2
(prebearHER2); mature unsubstituted bear HER2 (mbearHER2);
precursor unsubstituted feline HER2 (prefeHER2); and mature
unsubstituted feline HER2 (mfeHER2). The gene construct
additionally includes at least one promoter for expression of said
antigenic peptide in a living cell. The step of administering the
gene construct is followed by the steps of administering an
effective amount of an immunological adjuvant, expressing the gene
construct in cells of the mammalian subject, and inducing an immune
response against HER2 in the mammalian subject.
[0102] In another embodiment, the present invention includes
methods for immunizing cats against self HER2 with vaccines
including antigenic polypeptides including the ECD and TM domains
of wild type human, mouse, rat, or bear, HER2, and the human-rat
hybrid E2neu. Both the precursor and mature forms are included. Use
of these antigens for the immunization in experimental rodent
models is known, but their administration to cats as an effective
anti-HER2 vaccine in cats is a novel use. These antigens are known
but their use in methods of inducing immunity to self HER2 in cats
is novel. Again, the methods include vaccination with an effective
amount of expression construct, as previously described. Exemplary
antigenic polypeptides included in the HER2 antigens include:
prebearHER2, SEQ ID NO: 37; mbearHER2, SEQ ID NO: 38; prefeHER2.
SEQ ID NO: 39; mfeHER2, SEQ ID NO: 40; prehumHER, SEQ ID NO: 45;
mhumHER2, SEQ ID NO: 46; premouseHER2, SEQ ID NO: 47; mmouseHER2,
SEQ ID NO: 48; preratHER2, SEQ ID NO: 49; mratHER2, SEQ ID NO: 50;
preE2Neu, SEQ ID NO: 51; and mE2Neu, SEQ ID NO: 52. Where the
antigenic polypeptides are administered in the form of expression
constructs, they can be encoded in the following exemplary
polynucleotide sequences, or in synonymous sequences thereof:
prebearHER2, SEQ ID NO: 41; mbearHER2, SEQ ID NO: 42; prefeHER2,
SEQ ID NO: 43; mfeHER2, SEQ ID NO: 44; prehumHER, SEQ ID NO: 53;
mhumHER2, SEQ ID NO: 54; premouseHER2, SEQ ID NO: 55; mmouseHER2,
SEQ ID NO: 56; preratHER2, SEQ ID NO: 57; mratHER2, SEQ ID NO: 58;
preE2Neu, SEQ ID NO: 59; and mE2Neu, SEQ ID NO: 60.
[0103] An additional panel of substituted human HER2 polypeptides
was also found to break tolerance to human HER2 in mice
transgenically expressing human HER2 (human Tg mice), in
experiments described in Example 4. Human HER2 is self HER2 in
human HER2 Tg mice, and these mice are well known to exhibit strong
tolerance to human HER2 (Piechocki, et al., 2003). The additional
HER2 polypeptides found to be effective tolerance-breakers included
precursor human HER2 having a Q-K substitution at amino acid 329,
or a substitution of arginine for glutamine (Q-R) at amino acid
429, or a substitution of aspartic acid for asparagine (N-D) at
position 438. The mature forms of these substituted HER2 antigens
are reasonably predicted to be equivalently immunogenic to the
precursor forms, for reasons previously stated.
[0104] Therefore, the present invention provides antigenic
polypeptides for inducing immune response against HER2 in a
mammalian subject, The polypeptides include at least the
extracellular and transmembrane domains of human HER2, the
extracellular domain including at least one of the following amino
acid substitutions: glutamine with lysine (Q-K) or a conservative
amino acid of lysine, at position 141 of precursor humanHER2
(prehumHER2-Q141K); glutamine with lysine (Q-K) or a conservative
amino acid of lysine, at position 119 of mature humanHER2
(mhumHER2-Q119K); glutamine with lysine (Q-K) or a conservative
amino acid of lysine, at position 329 of precursor humanHER2
(prehumHER2-Q329K); glutamine with lysine (Q-K) or a conservative
amino acid of lysine, at position 307 of mature humanHER2
(mhumHER2-Q307K); glutamine with arginine (Q-R), or a conservative
amino acid of arginine, at position 429 of precursor human HER2
(prehumHER2-Q429R); glutamine with arginine (Q-R), or a
conservative amino acid of arginine, at position 407 of mature
human HER2 (mhumHER2-Q407R); asparagine with aspartic acid (N-D),
or a conservative amino acid of aspartic acid, at position 438 of
precursor human HER2 (prehumHER2-N438D); and asparagine with
aspartic acid (N-D), or a conservative amino acid of aspartic acid,
at position 416 of mature human HER2 (mhumHER2-N416D).
[0105] Exemplary amino acid sequences for each of the substituted
antigens as follows: prehumHER2-Q141K (SEQ ID NO: 9);
mhumHER2-Q119K (SEQ ID NO: 3;); prehumHER2-Q329K (SEQ ID NO: 28);
mhumHER2-Q307K (SEQ ID NO: 25); prehumHER2-Q429R (SEQ ID NO: 29);
mhumHER2-Q407R (SEQ ID NO: 26); prehumHER2-N438D (SEQ ID NO: 30);
and mhumHER2-N416D (SEQ ID NO: 27). It will be understood that the
disclosed amino acid sequences are exemplary, and that the present
invention encompasses all immunologically equivalent sequences.
[0106] The antigenic peptides are provided as isolated
polypeptides, and also as polypeptides encoded as nucleic acid
sequences in gene constructs. Each gene construct additionally
includes at least one promoter operatively linked to the nucleic
acid sequence, with the promoter inducing the expression of the
encoded antigenic polypeptide in a living cell.
[0107] The encoded constructs, and exemplary nucleic acid sequences
encoding them, include: prehumHER2-Q141K, SEQ ID NO: 21;
mhumHER2-Q119K, SEQ ID NO: 15; prehumHER2-Q329K, SEQ ID NO: 34;
mhumHER2-Q307K, SEQ ID NO: 231; prehumHER2-Q429R, SEQ ID NO: 35;
mhumHER2-Q407R, SEQ ID NO: 32; prehumHER2-N438D, SEQ ID NO: 36; and
mhumHER2-N416D, SEQ ID NO: 33. It will be understood that the
recited nucleic acid sequences are only exemplary, and that each
specified polypeptide can be encoded by one or more synonymous
nucleic acid sequences without departing from the scope of the
present invention. The nucleic acid sequences are preferably DNA
sequences, but may alternatively comprise at least one RNA
molecule.
[0108] In a related embodiment, the present invention provides the
additional panel of HER2 antigen constructs in a vaccine
composition for inducing immunity to HER2 in a mammalian subject,
the vaccine composition including an effective amount of at least
one of the gene constructs and an effective amount of an adjuvant.
An exemplary use of these vaccine compositions is disclosed in
Example 4.
[0109] In another related embodiment, the present invention
includes monoclonal antibodies selective for each member of the
additional panel of substituted human HER2 polypeptides. As
previously stated, with the amino acid sequences disclosed herein,
one skilled in the art can readily generate antibodies selective
for each of the substituted HER2 polypeptides.
[0110] In order to respond to vaccines against self HER2, a host
must possess an immune system sufficiently competent to overcome
tolerance in response to vaccination. The immunodepression
characteristic of mammary carcinomas and other cancers can prevent
the success of even the most potent vaccine. In one embodiment, the
present invention includes a diagnostic method for determining
whether a host is capable of making a response to self HER2.
[0111] In the diagnostic method, a sufficient amount of vaccine
known to break tolerance to the HER2 of host species is
administered to a candidate host for anti-HER2 immunotherapy. A
vaccine "known to induce immune response" is a vaccine which has
been shown to produce a measurable benefit to at least a subset of
similarly disposed mammalian hosts, for example in a clinical or
preclinical trial. The vaccines include those that incorporate the
substituted HER2 polypeptide antigens of the present invention,
which are shown to induce immunity in Examples 3 and 4. After the
vaccine has been administered, the subject is monitored for the
development of cellular and/or humoral immunity to self HER2, for
example by means of the immunoassays described in Example 3. If
detectable immune response is detected, then the candidate host is
recognized as being sufficiently immunocompetent to respond to
immunotherapy directed at self HER2. Ideally, the diagnostic test
represents a minimal, initial course of vaccination, with the
results determining whether a more extensive course of vaccination
will next be administered.
[0112] The invention is further described in detail in reference to
the following examples, which are provided for the purpose of
illustration only, and are not intended to be limiting. Thus, the
present invention should in no way be construed as being limited to
the following examples, but rather, be construed to encompass any
and all variations which become evident as a result of the teaching
provided herein.
Example 1
[0113] Expression and Function of HER-2 in Feline Mammary Carcinoma
Cell Lines and Explants.
[0114] Materials and Methods
[0115] Animals and Tissues
[0116] BALB/c mice were purchased from Charles River Laboratory.
Pathogen free (SPF) purpose bred domestic shorthair cats aged 6
months-2 years were obtained from Liberty Research, Inc (Liberty,
N.Y.). Animals were housed and maintained in the Department of
Laboratory Animal Resource (DLAR) facility at the Wayne State
University School of Medicine in accordance with Institutional
Animal Care and Use Committee guidelines. The experimental cats
were adopted as domestic pets by the care taker community after
completion of the study. A black bear legally harvested in Ontario,
Canada was the donor of the liver tissue.
[0117] Feline mammary carcinoma (FMC) samples were obtained from
mastectomy tissues of two feline patients treated at Oakland
Veterinary Referral Services (OVRS) in Michigan with consent from
the cat owners (Table 1).
TABLE-US-00001 TABLE 1 Feline Mammary Carcinoma Patients Cat Breed
Sex Spayed Age OVRS-1A Rex F Y 8 OVRS-1B 9 OVRS-2 DSH F Y 13
CSU-133 Persian F N 12 CSU-418 DLH F Y 5 CSU-1646 DSH F Y 5
[0118] OVRS-1A and OVRS-1B are two independent primary tumors from
the same cat. Three additional mammary tumor samples with paired,
uninvolved stromal tissues were purchased from Colorado State
University (CSU-133, 418, and 1646).
[0119] Cell Lines
[0120] K248 established from a pulmonary metastasis of a Siamese
cat mammary carcinoma was provided by Dr. John Hilkens and the late
Dr. Wim Misdorp at the Netherland Cancer Institute (Minke, et al.,
2010). Mammary carcinoma line K12 from a 14 year old cat was
established by Dr. William Hardy, Jr. and provided by Dr. Jaime
Modiano of the University of Pennsylvania, PA (Modiano, et al.,
1991). SKOV3 cells were purchased from the American Type Culture
Collection. MCF7 cells were obtained from Lisa Polin of the
Karmanos Cancer Institute. All cells were maintained in Dulbecco's
Modified Eagle's Medium supplemented with fetal bovine sera,
penicillin and streptomycin. The feline origin of K248 and K12
cells was authenticated by short tandem repeat (STR) analysis of
four loci (FIGS. 1 and 2.).
[0121] Immunohistochemical Analysis
[0122] Pathological diagnoses were performed according to the WHO
classification for tumors in domestic animals. For feline HER2
detection, epitopes were retrieved with sodium citrate buffer (pH
6.0) and histological grade primary antibodies were applied
according to manufacturer's recommendation (HER2, clone Z4881,
Invitrogen) followed by broad-spectrum HRP polymer conjugate
(SuperPicTure.TM. Polymer Detection Kit, Zymed) and DAB substrate
(Pierce Biotech). Feline mammary tumor cells, K248, were injected
subcutaneously in SCID mice. Tumor explants were used as
controls.
[0123] Cell Proliferation Assay
[0124] Cells were plated at 2-5,000/well in 96-well plates and
treated with gefitinib or lapatinib in quintuplicate for 48 h.
Alamar Blue reagent (Life Technologies) was added and fluorescence
measured after 3-4 h. The % proliferative activity was determined
relative to the average of untreated samples.
[0125] Western Blot Analysis
[0126] Cells or tissues were lysed in a non-ionic detergent lysis
buffer (Gibson, et al., 2013) with protease inhibitor cocktail
(Roche Diagnostics) immediately after the addition of phosphatase
inhibitors (NEB). Total protein was quantified by BCA assay (Pierce
Biotech). Ten .mu.g protein was boiled in Laemmli buffer, separated
with 8% SDS-polyacrylamide (PAGE) gel and transferred onto PVDF
membrane for overnight incubation with antibody to HER2
(42/c-erbB-2, BD Biosciences), phospho-HER2 Y1248 (polyclonal, Cell
Signaling Technology), Akt (polyclonal, Cell Signaling Technology),
phospho-Akt S473 (587F11, Cell Signaling Technology) or b-Actin
(I-19, Santa Cruz Biotech). After washing in TBS-Tween, membranes
were incubated with horseradish peroxidase (HRP)-conjugated
secondary antibody before washing and development using enhanced
chemiluminescent reagents (Thermo Scientific).
[0127] Flow Cytometric Analysis
[0128] HER2/neu epitopes were detected by moAb TA-1 (Calbiochem),
Trastuzumab (Genentech), 7.16.4 (Calbiochem), N12, and N29
(hybridoma lines were generous gifts of Dr. Yosef Yarden, Weissman
Institute, Isreal). moAb to human EGFR (528, Santa Cruz Biotech),
HER3 (SGP1, eBioscience) and HLA-ABC (W6/32, eBioscience) were used
as indicated. Phycoerythrin-conjugated goat anti-mouse or
anti-human IgG was the secondary antibody (Jackson ImmunoResearch).
Flow cytometric analysis was performed using FACS Canto II and data
analyzed with FlowJo (Tree Star).
[0129] To measure antibody level in immune sera, mouse or feline
sera were incubated with 3T3 cells transfected to express the
designated antigen and detected by PE-conjugated anti-mouse or
feline IgG secondary antibody (SantaCruz). Mouse antibody
concentrations were extrapolated from a standard curve of HER2 moAb
TA-1. Feline antibody titers were determined by serial dilution
until binding was no longer detected above isotype control.
[0130] Results
[0131] In a step toward establishing an outbred cat model of HER2
immunity, the expression and signaling functions of HER2 in feline
mammary carcinoma cell lines was characterized. These HER2
properties were also compared those of spontaneous feline mammary
carcinomas and known HER2-expressing cell lines.
[0132] Expression of HER2 in Feline Mammary Carcinoma (FMC)
[0133] Expression of ERBB family receptor tyrosine kinase (RTK) in
FMC was measured by flow cytometry. Surface expression of HER1
(EGFR), HER2, and HER3 was detected in K12 and K248 cells, using
moAbs to their human homologs (FIG. 3A) (FIG. 3A). MHC I expression
was also detected with moAb W6/32 to a constant region of human MHC
I. Control human ovarian cancer cell line SKOV3 showed elevated
HER2, while lacking HER3 expression (Gostring, et al., 2012).
Feline HER2 expression in primary FMC clinical samples (Table 1)
was detected by IHC staining (FIG. 3B). Membrane staining of HER2
was detected in all three primary FMC samples and in K248 explant,
consistent with membrane staining of K248 cells by flow cytometry
(FIG. 3A). Membranous expression of HER2 in FMC would be expected
to permit recognition by effectors of both humoral and cellular
immunity. Cytoplasmic staining was also detected in OVRS-1A and the
K248 explant and may indicate accumulation of incompletely or
incorrectly processed HER2. The clinical significance of
cytoplasmic HER2 remains unclear but T cells would be expected to
recognize cytoplasmically derived peptides in the context of MHC I
and II molecules.
[0134] Receptor Tyrosine Kinase (RTK) Activity in FMC
[0135] Activation of RTK signaling in FMC was tested using a human
RTK array (R&D Systems), FIGS. 4A and 4B. Although
cross-reactivity with feline antigens by all antibodies in this
array was not verified, AKT phosphorylation (S473) was elevated in
3/3 FMC tissue samples CSU-133, 418, and 1646, compared to their
paired, uninvolved stromal tissue. AKT phosphorylation was also
observed in K12 and K248 cells consistent with RTK pathway
activation in FMC. HER2 (Y1248) and downstream AKT (S473)
phosphorylation in primary FMC tissue was further tested by Western
blotting (FIG. 5A). Total and phosphorylated HER2 and AKT were
detected in FMC OVRS-2, CSU-133, CSU-418 and CSU-1646,
demonstrating activation of HER2 and downstream RTK signaling
events.
[0136] To further test if ERRB RTK signaling is required for FMC
cell proliferation, FMC cell lines K12 and K248 were cultured with
or without ERRB family tyrosine kinase inhibitors gefitinib or
lapatinib. Both K12 and K248 exhibited dose-dependent inhibition of
cell proliferation. SKOV3 and MCF7 cells were the positive and
negative control, respectively (FIG. 5B). Therefore, FMC express
functional tyrosine kinase receptors which trigger downstream
signaling and cell proliferation.
[0137] The results show that the FMC cell lines are useful models
for the induction and effects of anti-HER2 immune response.
Example 2
[0138] Cloning and Characterization of Substituted Feline Her2, and
Her2 of Other Species
[0139] Materials and Methods
[0140] DNA Cloning and Construction
[0141] Cloning primer sequences are shown in Table 2.
TABLE-US-00002 TABLE 2 CLONING PRIMERS cDNA product Primer Sequence
(5' to 3') SEQ ID NO Feline HER2 ECTM Forward ATTAC TACAA GCTTG
AGACC ATGGA GCTGG SEQ ID NO: 70 (prefeHER2) CGGCC TGGT Feline HER2
ECTM Reverse TACTA ATCTA GATCA CATCG TGTAC TTCCG SEQ ID NO: 71
(prefeHER2) GATCT TCTG Feline HER2 ECD- Forward CACCA AGCTT GAGAC
CATGG AGCTG G SEQ ID NO: 72 Fc (HER2 ECD) Feline HER2 ECD- Reverse
GATTT GGGCT CGGAC GTCAC AGGGC TGG SEQ ID NO: 73 Fc (HER2 ECD)
Feline HER2 ECD- Forward CTGTG ACGTC CGAGC CCAAA TCTTG TGAC SEQ ID
NO: 74 Fc (Fc tag) Feline HER2 ECD- Reverse TCTAG ATTAT TTACC CGGAG
ACAGG GAGAG Fc (Fc tag) GCTC SEQ ID NO: 75 Black bear HER2 Forward
TAAGC TTGAG ACCAT GGAGC TGGCG GCCTG ECTM GTG SEQ ID NO: 76
(prebearHER2) Black bear HER2 Reverse CTCTA GATTC ACATC GTGTA CTTCC
GGATC ECTM TTC SEQ ID NO: 77 (prebearHER2) Feline GM-CSF Forward
CACCA TGTGG CTGCA GAACC TGCTT TTCCT G SEQ ID NO: 78 Feline GM-CSF
Reverse TTACT TCTGG TCTGG TCCCC AGCAG TC SEQ ID NO: 79
prefeHER2-Q141-K Forward GCGGGAGCTGaAGCTCCGAAG SEQ ID NO: 103 and
mfeHER2- Q119-K prefeHER2-Q141-K Reverse AGCCCTCCTAGGGCAGCCCCTGTAG
SEQ ID NO: 104 and mfeHER2- Q119-K prebearher2-Q141K Forward
AAGCCTCACAGAGATCCTGAAG SEQ ID NO: 111 and mbearher2- Q119K
prebearher2-Q141K Reverse CGAAGCTTCAGCTCCCGCAGCCCTC SEQ ID NO: 112
and mbearher2- Q119K pre mouseHER2- Forward AGTCTCACAGAGATCTTGAAGG
SEQ ID NO: 113 Q142K and mmouseHER2- Q120K pre mouseHER2- Reverse
TCGAAGCTTCAGCTCCCGCAGCCC SEQ ID NO: 114 Q142K and mmouseHER2- Q120K
preratHER2-Q145K Forward GCTTCGAAGTCTCACAGAGATC SEQ ID NO: 115 and
mratHER2- Q120K preratHER2-Q145K Reverse TTCAGCTCCCGCAGCCCCTCTG SEQ
ID NO: 116 and mratHER2- Q120K pre humHER2- Forward
GCTGAAGCTTCGAAGCCTCACAG SEQ ID NO: 117 Q141K, preE2Neu- Q141K,
mhumHER2- Q119K, and mE2Neu-Q119K pre humHER2- Reverse
TCCCGCAGGCCTCCTGGGGAGG SEQ ID NO: 118 Q141K, preE2Neu- Q141K,
mhumHER2- Q119K, and mE2Neu-Q119K
[0142] Feline HER2 (ERBB2) cDNA was cloned from cell line K248
(39), K12 (40), and the ovary of a domestic shorthair cat using a
Protoscript kit (New England Biolabs) which showed identical
sequences for all three sources. The confirmed full-length
precursor feline ERBB2 (prefeHER2) cDNA sequence has been submitted
to Genbank (# JN990983). For vaccination, a stop codon was
introduced after codon 687 to delete the oncogenic intracellular
domain, then subcloned into pVax1, giving pprefeHER2 which contains
the signal peptide, extracellular and transmembrane domains of
feline HER2 (FIG. 7). The pprefeHER2-K nucleotide substitution in
codon 141 of extracellular domain I was cag->aag, based on our
reported sequence of feline ERBB2 (Genbank JN990983) and was
generated by PCR-based methods and verified by DNA sequencing.
Homologous Q-K substituted antigens for bear, mouse, and other
species of HER2 are similiarly generated, using the primers given
in Table 2.
[0143] Black bear HER2 cDNA was similarly cloned from the liver
tissue of a black bear. The cDNA sequence was submitted to Genbank
(# JQ040508). DNA vaccine pprebearHER2 encoding the signal peptide,
extracellular and transmembrane domains was constructed by PCR
similar to pprefeHER2.
[0144] Results
[0145] Feline HER2 (ERBB2) cDNA cloned from K12, K248 and normal
feline ovary showed identical sequences (GenBank Accession
JN990983). The amino acid. translate of full-length feline HER2
shared 93% sequence identity with human HER2 (FIGS. 6A and 6B).
Black bear HER2 (GenBank Accession JQ040508) had 96% and 92% a.a.
sequence identity with feline and human HER2, respectively. The
comparative data in FIG. 6B also show that human HER2 is more
closely related to the HER2 of cats and black bears than they are
to the HER2 of experimental rodents.
[0146] The feline, black bear, human and rat HER2 (rat neu)
extracellular and transmembrane regions (ECTM) were individually
transfected into 3T3 cells. Epitope expression was compared by
staining with moAbs to human HER2 (TA-1, N12, N29 and trastuzumab)
or rat neu (7.16.4) (McKenzie, et al., 1989; Stancovski, et al.,
1991; Hudziak, et al., 1989; Drebin, et al., 1984) (FIG. 6C).
Feline HER2 was recognized by all five moAbs. The closely related
black bear HER2 was recognized by four moAbs, except trastuzumab,
signifying structural disparity at this epitope. moAb 7.16.4
identifies a rat neu epitope which is present in feline and black
bear, but not human HER2. Overall, there are high levels of epitope
sharing among HER2 molecules from these 4 species, with cat HER2
expressing all 5 epitopes recognized by the panel of moAbs.
Example 3
[0147] Immunogenicity of Substituted and Heterologous her Antigenic
Polypeptides
[0148] Materials and Methods
[0149] Generation of Recombinant Feline HER2 and Human Fc Fusion
Protein-feHER2ecd-Fc
[0150] The secreted fusion protein feHER2ecd-hFc (SEQ ID NO: 69)
was generated to serve as a stimulator for T cells in vitro.
feHER2ecd-hFc was synthesized by fusing the 3' end of the signal
peptide-extracellular domain region of feHER2 (codons 1-653) to the
hinge-CH2-CH3 region of human IGHG1. This codon 1-653 region of
feHER2 (Genbank JN990983) was PCR amplified with forward primer 5'
CACCA AGCTT GAGAC CATGG AGCTG G (SEQ ID NO: 72) and reverse primer
5'-GATTT GGGCT CGGAC GTCAC AGGGC TGG (SEQ ID NO: 73), giving a 1986
bp product. IGHG1 cDNA (BC080557; Openbiosystems) was PCR amplified
with primers 5'-CTGTG ACGTC CGAGC CCAAA TCTTG TGAC (SEQ ID NO: 74)
and 5'-TCTAG ATTAT TTACC CGGAG ACAGG GAGAG GCTC (SEQ ID NO: 75),
giving a 716 bp product consisting of codons 248-479. These two
DNAs, which overlap by 22 bases, were fused by overlap
extension-primed DNA synthesis giving 2676 bp product, which was
then cloned into the HindIII and XbaI sites of the mammalian
expression vector pVax1. The sequence coded by this feHER2ecd-hFc
fusion cDNA, confirmed by DNA sequence analysis, is shown in FIG.
8A (feHER2ecd in uppercase, hFc in lowercase).
[0151] A schematic of feHER2ecd-hFc is shown in the upper panel of
FIG. 8B. A Western blot verifying the fusion protein is shown in
the lower panel of FIG. 8B. For production of feHER2ecd-hFc, murine
3T3 cells were transfected with the pVax/feHER2-hFc vector, and
recombinant feHER2-Fc in culture supernatant was quantified by
ELISA using mouse anti-human HER2 capture moAb (clone TA-1,
Calbiochem), which cross-reacts with feline HER2. Rabbit anti-human
IgG was the detection antibody (Jackson Immunoresearch). huHER2- Fc
was purchased from Sino Biologicals.
[0152] Stimulation of T Cells In Vitro with Recombinant
HER2ecd-Fc.
[0153] Feline PBMC were isolated by ficoll separation (GE
Healthcare). Cells were plated at 2.times.105/well in round bottom
96-well plates and cultured with 10 .mu.g/mL feHER2Fc (3T3
supernatant equivalent as described above), huHER2Fc, human IgG
control or control 3T3 conditioned medium for 72 h. Total well
contents were then transferred to feline IFN.gamma. ELISPOT plates
(R&D Systems) and incubated for an additional 48 h prior to
enumeration.
[0154] Analysis of T Cell Response by ELISPOT
[0155] Mouse splenocytes or feline PBMC isolated by Ficoll
separation (GE Healthcare) were maintained in Roswell Park Memorial
Institute Medium supplemented with fetal bovine sera,
penicillin/streptomycin. Feline PBMC were supplemented with 0.5
ng/mL feline IL-2 (R&D Systems). Cells were plated at
2.times.10.sup.5/well in round bottom 96-well plates and cultured
with 10 .mu.g/mL feHER2Fc (3T3 supernatant equivalent as described
above), huHER2Fc, human IgG control (Jackson Immunolabs) or control
3T3 conditioned medium for 48 (mouse) or 72 (feline) hours. Total
well contents were then transferred to mouse or or feline (R&D
Systems) IFN.gamma. ELISPOT plates and incubated for an additional
48 hours prior to detection and enumeration as per manufacturer
protocol. Visualized cytokine spots were enumerated using the
ImmunoSpot analyzer (CTL, Shaker Heights, Ohio) and expressed as
the number of cytokine-producing cells per 10.sup.6 splenocytes or
PBMC.
[0156] Feline GM-CSF
[0157] (CSF2) cDNA was amplified from a randomly-primed cDNA
library (Protoscript kit from New England Biolabs) prepared from
ConA-stimulated feline peripheral blood mononuclear cells (PBMC).
Codons 1 through 67 were PCR amplified with forward primer 5'-ATGTG
GCTGC AGAAC CTGCT TTTCC TG (SEQ ID NO: 80) and reverse primer
5'-CTCAG GGTCA AACAT TTCAG AGAC (SEQ ID NO: 81). Codons 60 through
145 were amplified with primers 5'-GTCTC TGAAA TGTTT GACCC TGAGG
(SEQ ID NO: 82) and 5'-TTACT TCTGG TCTGG TCCCC AGCAG TC (SEQ ID NO:
83). These two PCR products with fused by overlap extension priming
PCR, giving a 435 bp full-length CSF2 orf, which was cloned into
expression vector pcDNA3.1 blunt Topo (Invitrogen). The orf
sequence from a clone in the correct orientation was in accord with
the consensus of feline CSF2 cDNAs in Genbank (AY878357,
NM001009840, AF053007 and AF138140).
[0158] Electrovaccination of Mice and Cats
[0159] Mice were injected with an admix of 50 .mu.g each of vaccine
plasmid and plasmid encoding murine GM-CSF (pmuGM-CSF) in 50 .mu.l
PBS in the gastrocnemius muscle (Jacob, et al., 2006). Conductive
gel was applied on the skin over the injection sites.
Electroporation was conducted with NEPA21 electroporator (Napagene)
using a tweezer electrode. Three 50 msec degenerating bipolar
pulses of 100 V were administered at each site. Cats were injected
with 1.5 mg each of HER2 vaccine plasmid and pfeGM-CSF in 1.5 mL
PBS, divided equally over three injection sites in the biceps
femoris or quadriceps. Two rounds of electroporation were applied
to each site as described using a 1.5 cm2 caliper electrode
(BTX).
[0160] Results
[0161] Note that in FIGS. 9-13, an abbreviated notation is used for
the HER2 polypeptides. The term Q141K is shortened to "K". The
prefix "pre" is omitted. For example, the plasmid
"pprefeHER2-Q141K" is given as pfeHER2.
[0162] Immunogenicity of Substituted and Heterologous Forms of
HER2.
[0163] Recombinant prefeHER2-Q141K was expressed in 3T3 cells and
characterized by flow cytometry (FIG. 9A). Positive staining by the
panel of five moAbs indicated preservation of 5 HER2/neu epitopes
after Q.fwdarw.K substitution. Improved binding of feHER2-K by neu
specific moAb 7.16.4 compared to WT prefeHER2 (compare to FIG. 6C)
suggests a possible structural alteration of this epitope.
[0164] The immunogenicity of prefeHER2 and prefeHER2-Q141K was
initially characterized in BALB/c mice by electrovaccination with
pprefeHER2, prefeHER2-Q141K, or control pprehuHER2 (encoding human
HER2 ECTM), each of which were admixed with pmuGM-CSF encoding
murine GM-CSF (Jacob, et al., 2010; Radkevich-Brown, et al., 2009;
Jacob, et al., 2006; Jacob, et al., 2007) Antisera of vaccinated
mice were tested for reactivity against 3T3 cells transfected with
pprefeHER2 ("3T3/HER2"), pprefeHer2-Q141K ("3T3/HER2-K"), or
control pprehuHER2 ("3T3/huHER2"). Expression of with pprefeHER2,
pprefeHer2-Q141K, or control pprehuHER2. on individually
transfected 3T3 cells was comparable, as verified with moAb TA-1
binding at 2 different concentrations (FIG. 11A).
[0165] After 2.times. immunization, mice produced 59.+-.19,
49.+-.13 and 39.+-.20 .mu.g/mL IgG to their cognate antigens,
respectively, as measured with 3T3 cells transfected with
individual test antigens (FIG. 9B). The results indicate that
antibodies induced by vaccination with pprefeHER2 or
pprefeHER2-Q141K are highly cross-reactive, with both antibodies
recognizing dominant foreign epitopes on feline HER2 (FIG. 9B left
and middle panels). Therefore, the Q141K substitution appears to
create only subtle changes which did not alter the immune response
to the dominant foreign epitopes. Modest cross-reactivity between
feline and human HER2 immune sera was also observed. Neither pre-,
nor post-vaccination mouse serum bound un-transfected 3T3 cells
(FIG. 11B), supporting HER2 specificity of the immune sera.
[0166] To measure T cell response after vaccination, a feline HER2
extracellular domain (ECD) and human Ig Fc fusion protein,
feHER2-Fc, was generated as the test antigen, as previously
described and shown in FIG. 8B. Splenocytes from immunized BALB/c
mice were stimulated by incubation with feHER2-Fc or control
huHER2-Fc, and IFN.gamma.-producing T cells were enumerated by
ELISPOT.
[0167] The results (FIG. 9C) show that T cell responses to feline
HER2 were induced by vaccination with pprefeHER2 ("pfeHER2"),
pprefeHER2-Q141K ("pfeHER2-K") or pprehuHER2 ("phuHER2"). This
finding indicates cross-reactivity between feline and human HER2
antigen. This finding also indicates that the Q141K substitution
does not alter mouse T cell response to the WT HER2 protein. The
greater responses produced by in vitro stimulation with huHER2-Fc,
relative to feHER2-Fc, probably reflects the different forms of the
stimulating preparations, with purified huHER2-Fc protein
stimulating more effectively than secreted feHER2-Fc in culture
supernatant.
[0168] Anti-HER2 Vaccinations in the Feline Model System
[0169] The feasibility of DNA electrovaccination in cats was
initially tested with ppreE2Neu encoding a fusion protein of human
HER2 and rat neu. This construct was previously found to be
effective at inducing both humoral and cellular immunity in HER2 Tg
mice (Jacob, et al., 2010).
[0170] Three healthy purpose-bred, pathogen-free domestic shorthair
cats 12-24 months of age (Liberty Research Inc. Liberty, N.Y.) were
injected with ppreE2Neu and pfeGM-CSF in three legs in the biceps
femoralis or quadriceps. Each injection site was subjected to 2
rounds of electroporation. Vaccination was administered 4.times. at
3 week intervals. Blood was collected through the jugular vein 2
weeks after each vaccination.
[0171] Humoral Response of Cats to ppreE2Neu
[0172] Antibodies to huHER2, rat neu, and feHER2 were quantitated
by flow cytometric analysis of feline antibody binding to 3T3 cells
transfected with prehuHER2, rat neu, or prefeHER2, as previously
described.
[0173] Human HER2 binding IgG reached a titer of 1:400,000 in two
of three cats and 1:100,000 in the third (FIG. 10A, left panel).
Rat neu binding Ab ranged from 1:25,000 to 1:100,000 (FIG. 10A,
middle panel). The robust response validated the effectiveness of
DNA electrovaccination in cats.
[0174] Vaccination with ppreE2Neu induced antibody that cross
reacted with wild type feline HER2 as determined by binding to 3T3
cells expressing prefeHER2 ("3T3/feHER2" in FIG. 10A, right panel).
Neither pre-, nor post 4th-vaccination feline serum stained
non-transfected 3T3 cells, illustrating the HER2/neu-specific
reactivity of the immune sera (FIG. 11C).
[0175] Humoral Responses of Cats to Vaccines Including Bear and
Substituted Feline HER2
[0176] A panel of HER2 vaccines were tested in fifteen additional
healthy cats between 5-8 months of age. Cats were electrovaccinated
four times with pprefeHER2, pprefeHER2-K, pprebearHER2, ppreE2Neu,
or an admixture of pprefeHER2-K and ppreE2Neu. Results are shown in
FIG. 10B. In each panel of FIG. 10B, the antibody target-binding
cell type is listed at the top of the panel. The graphs show IgG
titers obtained with each of five vaccines, which are listed in the
FIG. 10B legend.
[0177] Consistent with the results shown previously in FIG. 10A,
the six cats that received ppreE2Neu either alone or in combination
with pprefeHER2-K developed high levels of IgG antibody to human
HER2 (FIG. 10B, left panel). Peak titers were achieved after three
vaccinations. Immune sera induced by pprebearHER2 cross-reacted
with human HER2, and the titers increased after each booster
immunization. pprefeHER2 or pprefeHER2-K immunization did not
induce significant antibody response to human HER2.
[0178] Recognition of prefeHER2-Q141K by immune sera was measured
by their binding to 3T3/prefeHER2-Q141K (FIG. 10B, middle panel,
"3T3/feHER2-K"). A prominent response (.about.1:120,000) was
induced by pfeHER2-K+pE2Neu. Individually, pfeHER2-K, pE2Neu or
pbearHER2 induced moderate titers averaging .about.1:6,000. The
enhancement of immune response to pprefeHER2-K and to pbearHER2 by
admixture with pE2Neu suggests a synergistic or adjuvant effect of
the heterologous pE2Neu. Vaccination with pprefeHER2 did not
generate a significant antibody response to prefeHER2-Q141K.
[0179] Antibodies binding to 3T3 cells expressing wild type
prefeHER2 ("3T3/feHER2") were detected at a dilution of between
1:1,600-1:3,200 of immune sera from pE2Neu, pfeHER2-K+pE2Neu, or
pbearHER2 vaccinated cats (FIG. 10B, right panel). These results
show that these heterologous vaccines break tolerance to self
feHER2, and induce antibodies which cross react with self feHER2.
The specificity of HER2 antigen recognition was validated by the
absence of binding to un-transfected 3T3 cells after 4.times.
vaccination (FIG. 11D).
[0180] Immune sera from mice immunized only with pprefeHER2 or
pprefeHER2-K showed negligible antibody binding to 3T3/feHER2 (FIG.
10B, right panel). The pprefeHER2-K vaccine showed much greater
effect, however, when the immune sera were tested against feline
mammary carcinoma cells. These cells constitute a much more
realistic system, for the carcinoma cells express not only HER2 but
also HER1 and HER3, which are capable of heterodimerizing with HER2
(Olayioye, 2001).
[0181] FIG. 10C shows the results of antibody binding experiments,
in which immune sera were tested at 1:50-1:100 dilution, not only
against 3T3 expressing only prefeHER2 (left panel), but also
against feline mammary carcinoma cells of cell lines K12 (middle
panel) and K248 (right panel), which express HER1, HER2, and HER3.
Sera from cats immunized with pprefeHer2 ("pfeHER2") showed little
binding to any of the test cells. Sera from cats immunized with
pprefeHer2-Q141K ("pfeHER2-K") also showed little or low level
binding to 3T3/feHER2, but they showed significant binding to K12
and K248 cells (FIG. 10C, middle and right panel, second line of
each panel). Although other immune sera recognized 3T3/feHER2, they
reacted minimally with K12 or K248 cells.
[0182] These findings indicate that vaccines including feline HER2
with a Q to K mutation at position 141 is sufficiently foreign to
break tolerance to normal self HER2 in the cat model. The findings
also suggest that vaccines including feline HER2 with a Q to K
mutation at position 141 induce antibodies specific for epitopes
whose expression is related to some combination of HER2, HER1
and/or HER3, a combination that is exposed naturally on feline
mammary carcinoma cells.
[0183] The findings also indicate that prebearHER2, when
administered as a heterologous vaccination to cats, is sufficiently
foreign from feline HER2 to break tolerance, but sufficiently
similar to induce antibodies that cross react with feline HER2.
[0184] FIGS. 12A-12C show the predicted 3D structure of
feHER2-Q141K domains I-Ill using cartoon and space-filling models.
The predicted effect of substituting Q with K at a.a. 141 is
portrayed by an electrostatic surface model (SYBYL-X 2.1.1
software; Tripos), where the electropositive side chain of K is
indicated by red and Q is shown in blue (FIG. 12C). It is
hypothesized that feHER2-Q141K recapitulates a cryptic HER2 epitope
naturally displayed when wild type feHER2 is heterodimerized or
associated with other ERBB family members on a cell surface. Cats
immunized with heterologous ppreE2Neu or pprebearHER2 produced
antibodies that recognize dominant epitopes on wild type feHER2 as
well as feHER2-Q141K expressed on 3T3 cells, but not the cryptic
epitope exhibited by K12 or K248 cells. This seemingly subtle, but
potentially critical difference in HER2 epitope recognition could
only have been detected in vaccination experiments with the outbred
cat system, wherein genetically un-manipulated cats naturally
express HER1, 2, and 3.
[0185] T Cell Responses of Cats to Substituted and Heterologous
HER2 Vaccines
[0186] Cats were vaccinated with plasmids encoding prefeHER2-Q141K
("pfeHER2-K" in FIG. 13) or the heterologous non-substituted
antigens prebearHER2 ("pbearHER2") or preE2neu ("pE2neu"), with
combinations of these plasmids, as admixtures or as initial and
booster vaccinations. The reactivity of the induced T cells to
human HER2 was measured first. Immune PBMC were cultured with
recombinant huHER2-Fc for three days before IFN.gamma. ELISPOT
analysis (FIG. 13A). The 6 cats receiving pE2Neu, with or without
pfeHER2-K, responded to human HER2 at 130-750 SFU/10.sup.6 PBMC,
validating the effectiveness of DNA electrovaccination (FIG. 13A).
pbearHER2 immune T cells also cross-reacted with human HER2,
producing an average of .about.160 SFU/10.sup.6 PBMC. Vaccination
with pfeHER2-K alone did not induce T cells that recognized human
HER2.
[0187] IFN-.gamma. T cell response to feHER2-Fc was measured to
evaluate reactivity to self HER2. Of the 10 evaluated cats, three
produced significant feHER2 specific T cell responses, with one cat
each from the pfeHER2-K (.about.100 SFU per million cells), pE2Neu
(.about.270 IFN.gamma. spots) and pbearHER2 (.about.280 IFN.gamma.
spots) groups (FIG. 13B). The observed T cell responses indicate
the immunogenic nature of both the heterologous bear and E2Neu and
the Q-K substituted HER2 vaccines. The 30% response rate may
reflect the heterogeneous genetic background of outbred cats, as in
humans.
[0188] Three bi-weekly booster vaccinations were given to five cats
that received pfeHER2-K or pfeHER2-K+pE2Neu (FIG. 13C). Only 1 of
the 3 cats receiving the admixed vaccine converted from a
non-responder to a responder after three boosters (.about.135
SFU/10.sup.6 PBMC). Therefore, a total of 40% T cell response rate
to self HER2 was achieved in healthy cats. Those cats initially
receiving pE2Neu or pbearHER2 were boosted 3.times. with pfeHER2-K
(FIG. 13D). Responses to recombinant human or feline HER2 did not
increase, suggesting that cross-reactive T cells, not common
epitopes in HER2 from pE2Neu or bear HER2, contributed to feline
HER2 reactivity.
[0189] The cats tolerated the vaccination procedure without signs
of pain or discomfort after they recovery from anesthesia. No
adverse side effects were detected 6-12 months after the final
vaccination and the cats continue to thrive.
Conclusion
[0190] Taken together, the experimental results disclosed in
Example 3 validate a new vaccine design strategy of including a
single residue substitution in a tumor self antigen. The results
show that, in the feline mammary cancer test system, a feline HER2
antigenic polypeptide with a Q to K substitution at position 141 is
sufficiently foreign to break tolerance, yet induces antibodies
cross reactive with normal feline HER2, and with the HER2 molecules
of humans and other species. Because the outbred cat system is a
realistic system which reflects antigen and MHC diversity of
natural human and animal populations, it is reasonably predictable
that other species of HER2 including a Q to K substitution at
position 141, or at an analogous position, will also be effective
at immunizing against mammary carcinomas and other HER2 expressing
cancers in other animal species, including humans.
[0191] The results of immunizations of cats with wild type bear
HER2 validate the strategy of immunization of a host with HER2 that
is heterologous, but relatively closely matched to self HER2. This
indicates that bear HER2 is an effective antigen for the
immunization and treatment of feline cancer hosts. Because of the
similarity of bear, cat, and human HER2 (FIG. 6B), it is reasonably
predictable that bear HER2 will be an effective antigen for
immunizing humans, and possibly other mammals, against
HER2-expressing tumor cells. This conclusion is further supported
by the generalizability of results from the highly realistic,
genetically diverse, outbred cat model.
Example 4
[0192] Additional Antigenic Her2 Polypeptides Including Amino Acid
Substitutions
[0193] To further test the hypothesis that minimally altered
variants of self HER2 can break tolerance and induce immunity to
self HER2, an additional panel of substituted HER2 was generated.
All of the substituted forms were based on the precursor form of
human HER2 (prehumHER2).
[0194] Materials and Methods
[0195] Generation of Substituted Variants of Human HER2.
[0196] Seven vaccine expression plasmids were constructed. Each
encoded either a single point-mutated human precursor HER2 ECTM
construct (pprehumHER2-Q141K, SEQ ID NO: 9; pprehumHER2-Q213K, SEQ
ID NO: 84; pprehumHER2-Q239K, SEQ ID NO: 85; pprehumHER2-Q329K SEQ
ID NO: 28; pprehumHER2-Q429R SEQ ID NO: 29; and pprehumHER2-N438D
SEQ ID NO: 30); or a human precursor HER2 ECTM construct with 3
a.a. substitutions (pprehumHER2-NNT 124-126 DSG, SEQ ID NO: 85).
The primer sequences for all constructs are listed in Table 3
below. For brevity, the constructs, and the polypeptides they
encode, will be referred to in the following disclosure only by
their substitutions, that is, respectively, Q141K, Q213K, Q239K,
Q329K Q429R N438D, and NNT124DSG. In FIGS. 16 and 18, the terms are
further abbreviated to, respectively, 141, 213, 239, 329, 429, 438,
and 124.
TABLE-US-00003 TABLE 3 PCR primers for generating HER2 point
variants AA Primer sequence (5'.fwdarw.3'; reverse primer, ital.;
Species substitution mutant, lowercase) Human q141k
GCTGaAGCTTCGAAGCCTCACAGAG (SEQ ID NO: 87) '' ''
TCCCGCAGGCCTCCTGGGGAGGC (SEQ ID NO: 88) '' q213k
TGAGGATTGTaAGAGCCTGAC (SEQ ID NO: 89) '' '' GAACTCTCTCCCCAGCAG (SEQ
ID NO: 90) '' q239k CTGCCATGAGaAGTGTGCTGC (SEQ ID NO: 91) '' ''
CAGTCAGTGGGCAGTGGC (SEQ ID NO: 92) '' q320k
AAGAGGTGACAGCAGAGGATGGAAC (SEQ ID NO: 93) '' ''
tGTTGTGCAGGGGGCAGACGAG (SEQ ID NO: 94) '' q329k
GGATGGAACAaAGCGGTGTGA (SEQ ID NO: 95) '' '' TCTGCTGTCACCTCTTGG (SEQ
ID NO: 96) '' q429r CCAGAACCTGagAGTAATCCGGG (SEQ ID NO: 97) '' ''
AAGACGCTGAGGTCAGGC (SEQ ID NO: 98) '' n438d AATTCTGCACgATGGCGCCTA
(SEQ ID NO: 99) '' '' CGTCCCCGGATTACTTGC (SEQ ID NO: 100) ''
nnt124dsg ggcACACCTGTCACAGGGGCCTCCCCAG (SEQ ID NO: 101)
actgtcCAGCGGGTCTCCATTGTCTAGCAC (SEQ ID NO: 102) Cat q141k
GCGGGAGCTGaAGCTCCGAAG (SEQ ID NO: 103) '' ''
AGCCCTCCTAGGGCAGCCCCTGTAG (SEQ ID NO: 104) '' r398q
TGAGCAGCTCcaAGTGTTTGAGGCTCTGGAG (SEQ ID NO: 105) '' ''
GGCTGCAGGGGGGCAGTG (SEQ ID NO: 106) '' n421d CAGCTTGCCTgACCTCAGTGTC
(SEQ ID NO: 107) '' '' TCTGGCCACGCTGAGATG (SEQ ID NO: 108)
[0197] New England Biolab's Q5 Site-Directed Mutagenesis Kit was
used with these primers to generate the variant vaccines, which
were confirmed by DNA sequencing.
[0198] Stable HER2 expression of six gene constructs was confirmed
in transiently transfected 3T3 cells by flow cytometry using
anti-human HER2 moAbs Ab5 and N12 (FIGS. 14A and 14B). One
construct, Q213K, was not expressed, and was removed from the test
panel.
[0199] Electrovaccination of Mice and Cats
[0200] Mice were injected with an admix of 50 .mu.g each of vaccine
plasmid and plasmid encoding murine GM-CSF (pmuGM-CSF) in 50 .mu.l
PBS in the gastrocnemius muscle. Conductive gel was applied on the
skin over the injection sites. Electroporation was conducted with
NEPA21 electroporator (Napagene) using a tweezer electrode. Three
50 msec degenerating bipolar pulses of 100 V were administered at
each site. Cats were injected with 1.5 mg each of HER2 vaccine
plasmid and pfeGM-CSF in 1.5 mL PBS, divided equally over three
injection sites in the biceps femoris or quadriceps. Two rounds of
electroporation were applied to each site as described using a 1.5
cm2 caliper electrode (BTX).
[0201] Results
[0202] In Vivo Expression of Substituted HER2 Polypeptides and
Immune Activation in Wild Type (WT) BALB/c Mice
[0203] The six verified constructs were advanced to vaccination
tests as plasmid vaccines in wild type BALB/c mice. The plasmids
pE2TM (prehumHER2) and pE2Neu (human HER2-rat neu hybrid ECTM) were
employed as controls. There were 3 mice in each group. It was
previously reported that pE2Neu induced significantly higher levels
of anti-human HER2 Ab and T cell response in HER2 Tg mice than does
pE2TM, showing the efficacy to overcome immune tolerance by
incorporating heterologous neu sequence in ECD domains 3 and 4
(Jacob, et al., 2010). Because human HER2 with or without mutations
are foreign proteins in WT mice, anti-HER2 antibody response is
expected as long as the vaccine construct is expressed in vivo. Of
the 6 mutants, five constructs except prehumHER2-Q239K induced
anti-human HER2 Ab in at least one animal to show their successful
expression in vivo and antibody induction after a single
electrovaccination (FIG. 15).
[0204] Wild type BALB/c mice were electro-vaccinated i.m. with 60
ug HER2 construct+60 ug pmGMCSF divided in two sites. Serum was
collected 3 weeks after immunization and binding to Her2-expressing
SKOV3 cells was assessed by flow cytometry.
[0205] Of the 6 substituted polypeptides, all induced anti-human
HER2 Ab in at least one animal (FIG. 15). This shows that the
substituted polypeptides, prehumHER2-Q141K, prehumHER2-Q329K,
prehumHER2-Q329K, prehumHER2-Q429R, prehumHER2-N438D, and
prehumHER2-NNT124-126DSG were successfully expressed in vivo, and
induced the expression of anti human-HER2 antibody after a single
electrovaccination (FIG. 15).
[0206] Immune Activation in Human HER2 Transgenic (Tg) Mice
[0207] The five positive mutant constructs identified in WT BALB/c
mice were advanced to human HER2 transgenic (Tg) mice (in BALB/c
background). Mice received electrovaccinations twice, 2 wks apart,
and immune sera were collected 2 wks after the final vaccination.
Binding of immune sera to 3T3 cells that express human HER2, but
not other human ERBB members, showed induction of anti-human HER2
antibodies by all 5 test vaccines as well as by control pE2TM or
pE2Neu (FIG. 16).
[0208] The finer specificity of the immune sera was further tested
with 5 human cancer cell lines. Breast cancer line SKBR3 and
ovarian cancer line SKOV3 have amplified HER2 as shown by the
binding to moAb Ab5, N12, N29 and Herceptin (FIG. 17). They also
express HER1 (EGFR). Three triple negative breast cancer cell lines
SUM149, SUM 159 and MDA MB231 show lower levels of HER2 expression.
MDAMB231 has elevated HER1 (EGFR) expression.
[0209] All test immune sera except those from mice immunized with
prehumHER2-NNT124DSG ("124") showed significant binding to the 5
human cancer cell lines (FIG. 18), indicating their immunogenic
nature. To determine whether the substituted HER2 vaccines were
more effective in overcoming immune tolerance than WT pE2TM, the
change in their antibody level relative to pE2TM immune serum was
calculated (FIG. 19). Of interest is that Q329K immune serum that
showed comparable binding to SKOV3 as pE2TM immune serum. The same
immune serum appears to bind MDA MB231 more effectively than pE2TM
immune serum (p<0.05). Note that SKOV3 cells express high levels
of HER2, moderate EGFR and no HER3 (see FIG. 3A). MDA MB231 cells
express moderate levels of HER2, elevated HER1. It is hypothesized
that HER2 epitopes on SKOV 3 and MDA-MB231 differ, and some unique
HER2 epitopes on MDA MB231 are mimicked by HER2 Q329K. Perhaps MDA
MB231 and K12/K248 cat mammary tumor cells share common HER2
structural features. Feline HER2 Q141K immune sera preferentially
bind K12/K248, but minimally to 3T3 cells expressing feline
HER2.
Discussion
[0210] It is proposed that an increased immune response to
substituted HER2 vaccines is due to alteration of the amino acid
charges or position of charges of HER2. The aa substitutions can be
visualized by space-filling modeling (Protein Data Bank ID #2a91;
www.rcsb.org/;) 3D view of "The crystal structure of a truncated
ErbB2 ectodomain reveals an active conformation, poised to interact
with other ErbB receptors" (Garrett, et al., 2003). Amino acid
substitutions on the outer surface of HER2 3D structure would
presumably be directly accessible to B-cell receptors, and
predispose to anti-HER2 antibody production. In contrast, the Q329K
substitution which also increases the charge is beneath the
surface, but may still trigger subtle changes in antigenicity.
Conclusion
[0211] The results indicate that prehumHER2-Q141K prehumHER2-Q329K,
prehumHER2-Q429R, and prehumHER2-N438D, are effective antigens for
breaking tolerance to self human HER2, as determined in the
transgenic human HER2 mouse model. The finding that
prehumHER2-Q141K breaks tolerance to self human HER2 reinforces the
previously disclosed finding from the cat system, that
prefeHER2-Q141K breaks tolerance to self feline HER2 (Example 3).
The concordance of these two findings supports the generalizability
of results from the outbred cat model to human HER2
immunobiology.
[0212] In addition, as previously stated, it is predictable that
the mature forms of these substituted antigenic polypeptides,
mhumHER2-Q119K, mhumHER2-Q407K, mhumHER2-Q429R, and mhumHER2-N438D
are also effective as tolerance-breaking antigens. The lack of a
signal peptide is expected to have no effect on the reactivity of
the Q-K substitution epitope, which is over a hundred residues
distant, in the ECD. Even if not processed onto the surface of a
host cell, the mature forms of HER2 are nonetheless be available to
antigen presenting cells upon apoptosis or necrosis of the
expressing cells. Exemplary amino acid sequences of these
substituted HER2 antigens are: mhumHER2-Q119K, SEQ ID NO: 3;
mhumHER2-Q407K, SEQ ID NO: 26; mhumHER2-Q429, SEQ ID NO: 29; and
mhumHER2-N438D, SEQ ID NO: 30.
[0213] Finally, it is predictable that conservatively substituted
variants of the substituted peptides of the present invention will
also be effective antigens. For example, arginine can be
substituted for lysine 141 in prehumHER2-Q141K and lysine 329 in
prehumHER2-Q329K; lysine can be substituted for arginine 429 in
prehumHER2-Q429R, and glutamic acid can be substituted for aspartic
acid 438 prehumHER2-N438D.
[0214] The invention has been described in an illustrative manner,
and it is to be understood that the terminology that has been used
is intended to be in the nature of words of description rather than
of limitation.
[0215] Obviously, many modifications and variations of the present
invention are possible in light of the above teachings. It is,
therefore, to be understood that within the scope of the appended
claims, the invention can be practiced otherwise than as
specifically described.
TABLE-US-00004 TABLE 4 Amino acid and nucleotide sequences for
substituted and wild type HER2 polypeptides mfeHER2-Q119K, SEQ ID
NO: 1 1 tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tylhanasls
lqdiqevqg 61 yvliahnqvk qvplqrlriv rgtqlfedny alavldngdp ldsgtpatga
lgglrelkl 121 rslteilkgg vliqrnpqlc hqdtilwkdi fhknnqlalm
lidtnrsrac pcspackds 181 hcwgassgdc qsltrtvcag gcarckgpqp
tdccheqcaa gctgpkhsdc aclhfnhsg 241 icelhcpalv tyntdtfesm
pnpegrytfg ascvtacpyn ylstdvgsct vcplnnqev 301 taedgtqrce
kcskpcarvc yglgmehlre aravtsaniq efvgckkifg laflpesfe 361
gdpasntapl qpeqlrvfea leeitgylyi sawpdslpnl svfqnlrvir rvlhdgays
421 ltlqglgisw lglrslrelg sgialihrns rlcfvhtvpw dqlfrnphqa
lhsanrped 481 ecageglacy plcahghcwg pgptqcvncs qflrgqecve
ecrviqglpr yvkdrfclp 541 chpecqpqng svtclgsead qcvacahykd
ppfcvarcps gvkpdlsfmp wkfadeegt 601 cqpcpincth scadldekgc
paeqraspvt siiaavvgil lvvvvglvlg likrrrqki 661 rkytm
mbearHER2-Q119K, SEQ ID NO: 2 1 tqvctgtdmk lrlpaspeth ldmlrhlyqa
cqvvqgnlel tylpanasls lqdiqevqg 61 yvliahsqvr qvplqrlriv rgtqlfedny
alavldngep pkgdtsvaga pgglrelkl 121 rslteilkgg vliqrnpqlc
hqdtliwkdi fhknnqlalt lidtnrsrac pcspackdp 181 hcwgassgdc
qsltrtvcag gcarckgpkp tdccheqcaa gctgpkhsdc aclhfnhsg 241
icelhcpalv tyntdtfesm pnpegrytfg ascvtacpyn ylstdvgsct vcplnnqev
301 taedgtqrce kcsrpcarvc yglgmehlre aravtsaniq efagckkifg
laflpesfe 361 gdpasntapl qpeqlrvfea leeitgylyi sawpdslpnl
svfqnlrvir rvlhdgays 421 ltlqglgisw lglrslrelg sglalihrna
rlcfihtvpw eqlfrnphqa lhsanrpea 481 ecvgeglacy plcahghcwg
pgptqcvncs qflrgqecve ecrelhglpr yvkdryclp 541 chpecrpqng
svtcfgsead qcvacahykd ppscvarcps gvkpdlsfmp wkfadeegt 601
cqpcpincth scgdldergc paeqraspvt siiaavvgil lavvmglvlg likrrrqki
661 rkytm mhumHER2-Q119K, SEQ ID NO: 3 1 tqvctgtdmk lrlpaspeth
ldmlrhlyqg cqvvqgnlel tylptnasls lqdiqevqg 61 yvliahnqvr qvplqrlriv
rgtqlfedny alavldngdp lnnttpvtga pgglrelKl 121 rslteilkgg
vliqrnpqlc yqdtilwkdi fhknnqlalt lidtnrsrac pcspmckgs 181
rcwgessedc qsltrtvcag gcarckgplp tdccheqcaa gctgpkhsdc aclhfnhsg
241 icelhcpalv tyntdtfesm pnpegrytfg ascvtacpyn ylstdvgsct
vcplhnqev 301 taedgtqrce kcskpcarvc yglgmehlre vravtsaniq
efagckkifg laflpesfd 361 gdpasntapl qpeqlqvfet leeitgylyi
sawpdslpdl svfqnlqvir rilhngays 421 ltlqglgisw lglrslrelg
sglalihhnt hlcfvhtvpw dqlfrnphqa lhtanrped 481 ecvgeglach
qlcarghcwg pgptqcvncs qflrgqecve ecrvlqglpr yvnarhclp 541
chpecqpqng svtcfgpead qcvacahykd ppfcvarcps gvkpdlsymp wkfpdeega
601 cqpcpincth scvdlddkgc paeqrasplt siisavvgil lvvvlgvvfg
likrrqqki 661 rkytm mmouseHER2-Q120K , SEQ ID NO: 4 1 tqvctgtdmk
lrlpaspeth ldmlrhlyqg cqvvqgnlel tylpanasls flqdiqevqg 61
ymliahnrvk hvplqrlriv rgtqlfedky alavldnrdp ldnvttaapg rtpeglrelk
121 lrslteilkg gvlirgnpql cyqdmvlwkd vlrknnqlap vdmdtnrsra
cppcaptckd 181 nhcwgesped cqiltgtict sgcarckgrl ptdccheqca
agctgpkhsd claclhfnhs 241 gicelhcpal ityntdtfes mlnpegrytf
gascvttcpy nylstevgsc tlvcppnnqe 301 vtaedgtqrc ekcskpcagv
cyglgmehlr garaitsdni qefagokkif gslaflpesf 361 dgnpssgvap
lkpehlqvfe tleeitgyly isawpesfqd lsvfqnlrvi rgrilhdgay 421
sltlqglgih slglrslrel gsglalihrn thlcfvntvp wdqlfrnphq allhsgnrpe
481 eacgleglvc nslcarghcw gpgptqcvnc sqflrgqecv eecrvwkglp
reyvrgkhcl 541 pchpecqpqn ssetcygsea dqceacahyk dssscvarcp
sgvkpdlsym piwkypdeeg 601 icqpcpinct hscvdlderg cpaeqraspv
tfiiatvvgv llfliivvvi gilikrrrqk 661 irkytm mratHER2-Q120K, SEQ ID
NO: 5 1 tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tyvpanasls
flqdiqevqg 61 ymliahnqvk rvplqrlriv rgtqlfedky alavldnrdp
qdnvaastpg rtpeglrelk 121 lrslteilkg gvlirgnpql cyqdmv1wkd
vfrknnqlap vdidtnrsra cppcapackd 181 nhcwgesped cqiltgtict
sgcarckgrl ptdccheqca agctgpkhsd claclhfnhs 241 gicelhcpal
vtyntdtfes mhnpegrytf gascvttcpy nylstevgsc tlvcppnnqe 301
vtaedgtqrc ekcskpcary cyglgmehlr garaitsdnv qefdgckkif gslaflpesf
361 dgdpssgiap lrpeqlqvfe tleeitgyly isawpdslrd lsvfqnlrii
rgrilhdgay 421 sltlqglgih slglrslrel gsglalihrn ahlcfvhtvp
wdqlfrnphq allhsgnrpe 481 edcgleglvc nslcahghcw gpgptqcvnc
shflrgqecv eecrvwkglp reyvsdkrcl 541 pchpecqpqn ssetcfgsea
dqcaacahyk dssscvarcp sgvkpdlsym piwkypdeeg 601 icqpcpinct
hscvdlderg cpaeqraspv tfiiatvvgv llflilvvvv gilikrrrqk 661 irkytm
mE2Neu-Q119K, SEQ ID NO: 6 1 tqvctgtdmk lrlpaspeth ldmlrhlyqg
cqvvqgnlel tylptnasls flqdiqevqg 61 yvliahnqvr qvplqrlriv
rgtqlfedny alavldngdp lnnttpvtga spgglrelkl 121 rslteilkgg
vliqrnpqlc yqdtilwkdi fhknnqlalt lidtnrsrac hpcspmckgs 181
rcwgessedc qsltrtvcag gcarckgplp tdccheqcaa gctgpkhsdc laclhfnhsg
241 icelhcpalv tyntdtfesm pnpegrytfg ascvtacpyn ylstdvgsct
lvcplhnqev 301 taedgtqrce kcskpcarvc yglgmehlre vravtsaniq
efagckkifg slaflpesfd 361 gdpasntaef aplrpeqlqv fetleeitgy
lyisawpdsl rdlsvfqnlr iirgrilhdg 421 aysltlqglg ihslglrslr
elgsglalih rnahlcfvht vpwdqlfrnp hqallhsgnr 481 peedcglegl
vcnslcahgh cwgpgptqcv ncshflrgqe cveecrvwkg lpreyvsdkr 541
clpchpecqp qnssetcfgs eadqcaacah ykdssscvar cpsgvkpdls ympiwkypde
601 egicqpcpin cthscvdlde rgcpaeqras pvtfiiatvv gvllflilvv
vvgilikrrr 661 qkirkytm prefeHER2-Q141K, SEQ ID NO: 7 1 melaawcrwg
lllallpsga tgtqvctgtd mklrlpaspe thldmlrhly qgcqvvqgnl 61
eltylhanas lsflqdiqev qgyvliahnq vkqvplqrlr ivrgtqlfed nyalavldng
121 dpldsgtpat gaalgglrel klrslteilk ggvliqrnpq lchqdtilwk
difhknnqla 181 lmlidtnrsr acqpcspack dshcwgassg dcqsltrtvc
aggcarckgp qptdccheqc 241 aagctgpkhs dclaclhfnh sgicelhcpa
lvtyntdtfe smpnpegryt fgascvtacp 301 ynylstdvgs ctivcpinnq
evtaedgtqr cekcskpcar vcyglgmehl rearavtsan 361 iqefvgckki
fgslaflpes fegdpasnta plqpeqlrvf ealeeitgyl yisawpdslp 421
nlsvfqnlrv irgrvlhdga ysltlqglgi swlglrslre lgsglalihr nsrlcfvhtv
481 pwdqlfrnph qallhsanrp edecagegla cyplcahghc wgpgptqcvn
csqflrgqec 541 veecrvlqgl preyvkdrfc lpchpecqpq ngsvtclgse
adqcvacahy kdppfcvarc 601 psgvkpdlsf mpiwkfadee gtcqpcpinc
thscadldek gcpaeqrasp vtsiiaavvg 661 illvvvvglv lgilikrrrq kirkytm
prebearHER2-Q141K SEQ ID NO: 8 1 melaawcrwg lllallpsga agtqvctgtd
mklrlpaspe thldmlrhly qacqvvqgnl 61 eltylpanas lsflqdiqev
qgyvliahsq vrqvplqrlr ivrgtqlfed nyalavldng 121 eppkgdtsva
gatpgglrel klrslteilk ggvliqrnpq lchqdtilwk difhknnqla 181
ltlidtnrsr acqpcspack dphcwgassg dcqsltrtvc aggcarckgp kptdccheqc
241 aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt
fgascvtacp 301 ynylstdvgs ctivcpinnq evtaedgtqr cekcsrpcar
vcyglgmehl rearavtsan 361 iqefagckki fgslaflpes fegdpasnta
plqpeqlrvf ealeeitgyl yisawpdslp 421 nlsvfqnlrv irgrvlhdga
ysltlqglgi swlglrslre lgsglalihr narlcfihtv 481 pweqlfrnph
qallhsanrp eaecvgegla cyplcahghc wgpgptqcvn csqflrgqec 541
veecrelhgl preyvkdryc lpchpecrpq ngsvtcfgse adqcvacahy kdppscvarc
601 psgvkpdlsf mpiwkfadee gtcqpcpinc thscgdlder gcpaeqrasp
vtsiiaavvg 661 illavvmglv lgilikrrrq kirkytm prehumHER-Q141K SEQ ID
NO: 9 1 melaalcrwg lllallppga astqvctgtd mklrlpaspe thldmlrhly
qgcqvvqgnl 61 eltylptnas lsflgdigev qgyvliahnq vrqvplqrlr
ivrgtqlfed nyalavldng 121 dpinnttpvt gaspgglrel Klrslteilk
ggvliqrnpq lcyqdtilwk difhknnqla 181 ltlidtnrsr achpcspmck
gsrcwgesse dcqsltrtvc aggcarckgp 1ptdccheqc 241 aagctgpkhs
dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp 301
ynylstdvgs ctivcplhnq evtaedgtqr cekcskpcar vcyglgmehl revravtsan
361 iqefagckki fgslaflpes fdgdpasnta plqpeqlqvf etleeitgyl
yisawpdslp 421 dlsvfqnlqv irgrilhnga ysltlqglgi swlglrslre
lgsglalihh nthlcfvhtv 481 pwdqlfrnph qallhtanrp edecvgegla
chqlcarghc wgpgptqcvn csqflrggec 541 veecrvlqgl preyvnarhc
lpchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc 601 psgvkpdlsy
mpiwkfpdee gacqpcpinc thscvdlddk gcpaegrasp ltsiisavvg 661
illvvvlgvv fgilikrrqq kirkytm premouseHER-Q142K, SEQ ID NO: 10 1
melaawcrwg fllallspga agtqvctgtd mklrlpaspe thldmlrhly qgcqvvqgnl
61 eltylpanas lsflgdigev qgymliahnr vkhvplqrlr ivrgtqlfed
kyalavldnr 121 dpldnvttaa pgrtpeglre lklrslteil kggvlirgnp
qlcygdmvlw kdvlrknnql 181 apvdmdtnrs racppcaptc kdnhcwgesp
edcgiltgti ctsgcarckg rlptdccheq 241 caagctgpkh sdclaclhfn
hsgicelhcp alityntdtf esmlnpegry tfgascvttc 301 pynylstevg
sctivcppnn gevtaedgtq rcekcskpca gvcyglgmeh lrgaraitsd 361
nigefagckk ifgslaflpe sfdgnpssgv aplkpehlqv fetleeitgy lyisawpesf
421 qdlsvfqnlr virgrilhdg aysltlqglg ihslglrslr elgsglalih
rnthlcfvnt 481 vpwdqlfrnp hgallhsgnr peeacglegl vcnslcargh
cwgpgptqcv ncsqflrgge
541 cveecrvwkg lpreyvrgkh clpchpecqp qnssetcygs eadqceacah
ykdssscvar 601 cpsgvkpdls ympiwkypde egicqpcpin cthscvdlde
rgcpaegras pvtfiiatvv 661 gvllfliivv vigilikrrr qkirkytm
preratHER2-Q145K, SEQ ID NO: 11 1 miimelaawc rwgfllallp pgiagtqvct
gtdmklrlpa spethldmlr hlyggcqvvg 61 gnleltyvpa naslsflqdi
gevggymlia hnqvkrvplq rlrivrgtql fedkyalavl 121 dnrdpqdnva
astpgrtpeg lrelklrslt eilkggvlir gnpq1cygdm vlwkdvfrkn 181
nqlapvdidt nrsracppca packdnhcwg espedcgilt gtictsgcar ckgrlptdcc
241 heqcaagctg pkhsdclacl hfnhsgicel hcpalvtynt dtfesmhnpe
grytfgascv 301 ttcpynylst evgsctivcp pnnqevtaed gtqrcekcsk
pcarvcyglg mehlrgarai 361 tsdnvqefdg ckkifgslaf lpesfdgdps
sgiaplrpeq lqvfetleei tgylyisawp 421 dslrdlsvfq nlriirgril
hdgaysltlq glgihslglr slrelgsgla lihrnahlcf 481 vhtvpwdqlf
rnphgallhs gnrpeedcgl eglvcnslca hghcwgpgpt qcvncshflr 541
ggecveecry wkglpreyvs dkrclpchpe cqpqnssetc fgseadqcaa cahykdsssc
601 varcpsgvkp dlsympiwky pdeegicqpc pincthscvd ldergcpaeq
raspvtfiia 661 tvvgvllfli lvvvvgilik rrrqkirkyt m preE2Neu-Q141K
SEQ ID NO: 12 1 melaalcrwg lllallppga astqvctgtd mklrlpaspe
thldmlrhly qgcqvvqgnl 61 eltylptnas lsflqdiqev qgyvliahnq
vrqvplqrlr ivrgtqlfed nyalavldng 121 dplnnttpvt gaspgglrel
klrslteilk ggvliqrnpq lcyqdtilwk difhknnqla 181 ltlidtnrsr
achpcspmck gsrcwgesse dcqsltrtvc aggcarckgp 1ptdccheqc 241
aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp
301 ynylstdvgs ctivcplhnq evtaedgtqr cekcskpcar vcyglgmehl
revravtsan 361 igefagckki fgslaflpes fdgdpasnta efaplrpeql
qvfetleeit gylyisawpd 421 slrdlsvfqn lriirgrilh dgaysltlqg
lgihslglrs lrelgsglal ihrnahlcfv 481 htvpwdqlfr nphqallhsg
nrpeedcgle glvcnslcah ghcwgpgptq cvncshflrg 541 qecveecrvw
kglpreyvsd krclpchpec qpqnssetcf gseadqcaac ahykdssscv 601
arcpsgvkpd lsympiwkyp deegicqpcp incthscvdl dergcpaeqr aspvtfiiat
661 vvgvllflil vvvvgilikr rrqkirkytm mfeHER2-Q119K, SEQ ID NO: 13 1
acccaagtgt gcaccggcac agacatgaag ctgcggctcc cagccagtcc cgagacccac
61 ctggacatgc tccgccacct ctaccagggc tgtcaagtgg tacagggcaa
cctggagctc 121 acctacctgc atgccaatgc cagcctctcc ttcctgcagg
atatccagga ggtgcaaggc 181 tatgtgctca ttgcccacaa ccaagtgaaa
caggtcccac tgcagaggct acgaatcgtg 241 cgaggcaccc agctctttga
ggacaactac gccctggccg tgctggacaa cggagaccca 301 ctggacagtg
gcacccctgc tacaggggct gccctaggag ggctgcggga gctgaagctc 361
cgaagcctca cagagatcct gaagggaggg gtcctcattc agcggaaccc gcagctctgc
421 caccaggaca cgattctgtg gaaggacatc ttccacaaga acaaccagct
ggccctcatg 481 ctgatagaca ccaaccgctc tcgggcctgc caaccctgtt
ctccagcttg taaagactcc 541 cactgctggg gagcaagttc cggggactgt
cagagcttga ctcgaactgt ctgtgctggc 601 ggctgtgccc gctgcaaggg
cccgcagccc accgactgct gccacgagca atgtgctgct 661 ggctgcacgg
gccccaagca ttctgactgc ctggcctgcc tccacttcaa ccacagtggc 721
atctgtgagc tgcactgccc agccctggtc acctacaaca cggacacctt cgaatccatg
781 cccaaccctg agggccgtta taccttcggt gccagctgtg tgactgcctg
tccctacaac 841 tacctgtcta cggacgtggg atcctgcacc ctggtctgtc
ccctgaacaa ccaagaggtg 901 acagctgagg atggaacaca gcggtgtgag
aaatgcagca agccctgtgc ccgagtgtgc 961 tacggcctag gcatggagca
cctgcgggag gcgagggcag tcaccagtgc caacatccaa 1021 gaatttgtcg
gctgcaagaa gatctttggg agcctggcgt ttctgccaga gagctttgag 1081
ggggacccag cctccaacac tgcccccctg cagcctgagc agctcagagt gtttgaggct
1141 ctggaggaga ttacaggtta cctgtacatc tcagcgtggc cagacagctt
gcctaacctc 1201 agtgtcttcc agaacctcag agtgatccgg ggccgagttc
tgcatgacgg tgcttactcg 1261 ctgacccttc aagggctggg catcagctgg
ctggggctgc gctcgctgcg ggagctgggc 1321 agtgggctgg ccctcatcca
ccgcaactcc cgcctctgct tcgtacacac ggtgccctgg 1381 gaccagctct
tccggaaccc ccaccaggcc ctgctccaca gcgccaaccg gccagaggac 1441
gagtgcgcgg gtgagggcct ggcctgctat ccgctgtgtg cccacgggca ctgctggggt
1501 ccgggaccca cccagtgtgt caactgcagc cagttccttc ggggccagga
gtgcgtggag 1561 gaatgccgag tattgcaggg gcttccccgg gagtatgtga
aggataggtt ctgtctgcca 1621 tgccacccgg agtgtcagcc ccagaatggc
tcagtgacct gcttgggctc ggaagctgac 1681 cagtgtgtgg cctgtgccca
ctacaaggac cctcctttct gtgtggctcg ctgccccagt 1741 ggggtgaaac
ctgacctctc cttcatgccc atctggaagt tcgcagatga ggagggcacg 1801
tgccagccat gccccatcaa ctgcacccac tcctgtgcgg acctggacga gaagggctgc
1861 cccgccgagc agagagccag ccctgtgacg tccatcattg ctgctgtggt
gggcattctg 1921 ctggtcgtgg ttgtggggct ggtccttggc atcctaatca
agcgaaggcg gcagaagatc 1981 cggaagtaca cgatg mbearHER2-Q119K, SEQ ID
NO: 14 1 acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc
cgagacccac 61 ctggatatgc tccgccacct ctaccaggcc tgtcaagtgg
tacagggtaa cctggagctc 121 acctacctgc ccgccaatgc cagcctgtcc
ttcctgcagg atatccagga ggtacagggc 181 tatgtgctca ttgctcacag
ccaagtgaga caggtcccgc tgcagaggct ccgaatcgtg 241 cgaggcaccc
agctctttga ggacaactac gccctggccg tgctggacaa tggagagccg 301
cccaaggggg acacctctgt ggcaggggct accccaggag ggctgcggga gctgaagctt
361 cgaagcctca cagagatcct gaagggaggg gtcttgattc agcggaaccc
acagctctgc 421 caccaggaca cgattttgtg gaaggacatc ttccacaaga
acaaccagct ggccctcacg 481 ctgatagaca ccaaccgctc tcgggcctgc
caaccctgtt ctccagcctg taaagacccc 541 cactgctggg gagcaagttc
cggggactgt cagagcttga cacgaaccgt ctgtgccggc 601 ggctgtgccc
gctgcaaggg cccaaaaccc actgactgct gccatgagca gtgcgcggct 661
ggctgcacgg gccccaagca ctcggactgc ctggcctgcc ttcacttcaa ccacagtggc
721 atctgtgagc tgcactgccc agccctggtc acctacaaca cggacacgtt
cgaatccatg 781 cccaaccctg agggccgata caccttcggt gccagctgtg
tgaccgcctg tccctacaac 841 tacctgtcca cggacgtggg atcctgcacc
ctggtctgtc ccctgaacaa ccaagaggtg 901 acggctgagg atggcaccca
gcggtgtgag aaatgcagca gaccctgtgc ccgagtgtgc 961 tatggtctgg
gcatggagca cctgcgggag gcgagggcgg tcaccagcgc caacatccaa 1021
gagttcgccg gctgcaagaa gatctttggg agcctggcgt ttctgccaga gagcttcgag
1081 ggagacccag cctccaacac tgcccccctg cagcctgaac agctcagagt
gttcgaggcc 1141 ctggaggaga tcacaggtta cctgtatatc tcagcgtggc
cggacagctt gcctaacctc 1201 agtgtcttcc agaacctgcg agtaatccgg
ggacgagttc tgcatgatgg cgcctactcg 1261 ctgaccctgc aagggctggg
catcagctgg ctggggctgc gctcgctgcg ggaactgggc 1321 agcgggctgg
ccctcatcca ccgcaacgcc cgcctctgct tcatccacac ggtgccctgg 1381
gagcagctct tccggaaccc ccaccaagcc ctgctgcaca gtgccaaccg gccggaggcc
1441 gagtgcgtgg gcgagggcct ggcctgctac ccgctgtgcg cccatgggca
ctgctggggt 1501 ccggggccca cccagtgcgt caactgcagc caattccttc
ggggccagga gtgcgtggag 1561 gaatgccgag aactgcacgg gctaccccgg
gaatatgtga aggacagata ctgtctgcca 1621 tgccaccccg agtgtcggcc
ccagaatggc tcagtgacct gctttgggtc ggaggctgac 1681 cagtgtgtgg
cctgcgccca ctacaaggac cctccctcct gcgtggctcg ctgccccagt 1741
ggtgtgaaac ccgacctctc tttcatgccc atttggaagt ttgcagatga ggagggcaca
1801 tgccagccgt gccccatcaa ctgcacccac tcctgtgggg acctggacga
gaggggctgc 1861 cccgccgaac agagagccag ccctgtgaca tccatcattg
ccgctgtggt gggcattctg 1921 ctggccgtgg tcatggggct ggtcctcggc
atcctgatca agcgaaggcg acagaagatc 1981 cggaagtaca cgatg
mhumHER2-Q119K, SEQ ID NO: 15 1 acccaagtgt gcaccggcac agacatgaag
ctgcggctcc ctgccagtcc cgagacccac 61 ctggacatgc tccgccacct
ctaccagggc tgccaggtgg tgcagggaaa cctggaactc 121 acctacctgc
ccaccaatgc cagcctgtcc ttcctgcagg atatccagga ggtgcagggc 181
tacgtgctca tcgctcacaa ccaagtgagg caggtcccac tgcagaggct gcggattgtg
241 cgaggcaccc agctctttga ggacaactat gccctggccg tgctagacaa
tggagacccg 301 ctgaacaata ccacccctgt cacaggggcc tccccaggag
gcctgcggga gctgAagctt 361 cgaagcctca cagagatctt gaaaggaggg
gtcttgatcc agcggaaccc ccagctctgc 421 taccaggaca cgattttgtg
gaaggacatc ttccacaaga acaaccagct ggctctcaca 481 ctgatagaca
ccaaccgctc tcgggcctgc cacccctgtt ctccgatgtg taagggctcc 541
cgctgctggg gagagagttc tgaggattgt cagagcctga cgcgcactgt ctgtgccggt
601 ggctgtgccc gctgcaaggg gccactgccc actgactgct gccatgagca
gtgtgctgcc 661 ggctgcacgg gccccaagca ctctgactgc ctggcctgcc
tccacttcaa ccacagtggc 721 atctgtgagc tgcactgccc agccctggtc
acctacaaca cagacacgtt tgagtccatg 781 cccaatcccg agggccggta
tacattcggc gccagctgtg tgactgcctg tccctacaac 841 tacctttcta
cggacgtggg atcctgcacc ctcgtctgcc ccctgcacaa ccaagaggtg 901
acagcagagg atggaacaca gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc
961 tatggtctgg gcatggagca cttgcgagag gtgagggcag ttaccagtgc
caatatccag 1021 gagtttgctg gctgcaagaa gatctttggg agcctggcat
ttctgccgga gagctttgat 1081 ggggacccag cctccaacac tgccccgctc
cagccagagc agctccaagt gtttgagact 1141 ctggaagaga tcacaggtta
cctatacatc tcagcatggc cggacagcct gcctgacctc 1201 agcgtcttcc
agaacctgca agtaatccgg ggacgaattc tgcacaatgg cgcctactcg 1261
ctgaccctgc aagggctggg catcagctgg ctggggctgc gctcactgag ggaactgggc
1321 agtggactgg ccctcatcca ccataacacc cacctctgct tcgtgcacac
ggtgccctgg 1381 gaccagctct ttcggaaccc gcaccaagct ctgctccaca
ctgccaaccg gccagaggac 1441 gagtgtgtgg gcgagggcct ggcctgccac
cagctgtgcg cccgagggca ctgctggggt 1501 ccagggccca cccagtgtgt
caactgcagc cagttccttc ggggccagga gtgcgtggag 1561 gaatgccgag
tactgcaggg gctccccagg gagtatgtga atgccaggca ctgtttgccg 1621
tgccaccctg agtgtcagcc ccagaatggc tcagtgacct gttttggacc
ggaggctgac
1681 cagtgtgtgg cctgtgccca ctataaggac cctcccttct gcgtggcccg
ctgccccagc 1741 ggtgtgaaac ctgacctctc ctacatgccc atctggaagt
ttccagatga ggagggcgca 1801 tgccagcctt gccccatcaa ctgcacccac
tcctgtgtgg acctggatga caagggctgc 1861 cccgccgagc agagagccag
ccctctgacg tccatcatct ctgcggtggt tggcattctg 1921 ctggtcgtgg
tcttgggggt ggtctttggg atcctcatca agcgacggca gcagaagatc 1981
cggaagtaca cgatgtag mmouseHER2-Q120K, SEQ ID NO: 16 1 acccaagtgt
gtaccggtac cgacatgaag ttgcgactcc ctgccagtcc tgagacccac 61
ctggacatgc ttcgccacct ctaccagggc tgtcaggtgg tgcagggcaa tttggagctt
121 acctacctgc ccgccaatgc cagcctctca ttcctgcagg acatccagga
agtccaggga 181 tacatgctca tcgctcacaa ccgagtgaaa cacgtcccac
tgcagaggtt gcgcatcgtg 241 agagggactc agctctttga ggacaagtat
gccctggctg tgctagacaa ccgagaccct 301 ttggacaacg tcaccaccgc
cgccccaggc agaaccccag aagggctgcg ggagctgaag 361 cttcgaagtc
tcacagagat cttgaaggga ggagttttga tccgtgggaa ccctcagctc 421
tgctaccagg acatggtttt gtggaaggat gtcctccgta agaataacca gctggctcct
481 gtcgacatgg acaccaatcg ttcccgggcc tgtccacctt gtgccccaac
ctgcaaagac 541 aatcactgtt ggggtgagag tcctgaagac tgtcagatct
tgactggcac catctgtact 601 agtggctgtg cccggtgcaa gggccggctg
cccactgact gttgccatga gcagtgtgct 661 gcaggctgca cgggtcccaa
gcattctgac tgcctggcct gcctccactt caatcatagt 721 ggtatctgtg
agctgcactg cccggccctc atcacctaca acacagacac cttcgagtcc 781
atgctcaacc ctgagggtcg ctacaccttt ggtgccagct gtgtgaccac ctgcccctac
841 aactacctct ccacggaagt gggatcctgc actctggtct gtcccccgaa
caaccaagag 901 gtcacagctg aggacggaac acagcggtgt gagaaatgca
gcaagccctg tgctggagta 961 tgctatggtc tgggcatgga gcacctccga
ggggcgaggg ccatcaccag tgacaatatc 1021 caggagtttg ctggctgcaa
gaagatcttt gggagcctgg catttttgcc ggagagcttt 1081 gatgggaacc
cctcctccgg cgttgcccca ctgaagccag agcatctcca agtgttcgaa 1141
accctggagg agatcacagg ttacctatac atttcagcat ggccagagag cttccaagac
1201 ctcagtgtct tccagaacct tcgggtcatt cggggacgga ttctccatga
tggtgcttac 1261 tcattgacgt tgcaaggcct ggggattcac tcactggggc
tacgctcact gcgggagctg 1321 ggcagtggat tggctctcat tcaccgcaac
acccatctct gctttgtaaa cactgtacct 1381 tgggaccagc tcttccggaa
cccgcaccag gccctactcc acagtgggaa ccggccagaa 1441 gaggcatgtg
gtcttgaggg cttggtctgt aactcactgt gtgcccgtgg gcactgctgg 1501
gggccagggc ccacccagtg tgtcaactgc agtcagttcc tccggggcca ggagtgtgtg
1561 gaggagtgcc gagtatggaa ggggctcccc agggagtatg tgaggggcaa
gcactgtctg 1621 ccatgccacc ccgagtgtca gcctcaaaac agctcggaga
cctgctatgg atcggaggct 1681 gaccagtgtg aggcttgtgc ccactacaag
gactcatctt cctgtgtggc tcgctgcccc 1741 agtggtgtga agccagacct
ctcctacatg cctatctgga agtacccgga tgaggagggc 1801 atatgtcagc
catgccccat caactgcacc cactcatgtg tggacctgga cgaacgaggc 1861
tgcccagcag agcagagagc cagcccagtg acattcatca ttgcaactgt ggtgggcgtc
1921 ctgttgttcc tgatcatagt ggtggtcatt ggaatcctaa tcaaacgaag
gcgacagaag 1981 atccggaagt ataccatg mratHER2-Q120K, SEQ ID NO: 17 1
acccaagtgt gtaccggcac agacatgaag ttgcggctcc ctgccagtcc tgagacccac
61 ctggacatgc tccgccacct gtaccagggc tgtcaggtag tgcagggcaa
cttggagctt 121 acctacgtgc ctgccaatgc cagcctctca ttcctgcagg
acatccagga agttcagggt 181 tacatgctca tcgctcacaa ccaggtgaag
cgcgtcccac tgcaaaggct gcgcatcgtg 241 agagggaccc agctctttga
ggacaagtat gccctggctg tgctagacaa ccgagatcct 301 caggacaatg
tcgccgcctc caccccaggc agaaccccag aggggctgcg ggagctgaag 361
cttcgaagtc tcacagagat cctgaaggga ggagttttga tccgtgggaa ccctcagctc
421 tgctaccagg acatggtttt gtggaaggac gtcttccgca agaataacca
actggctcct 481 gtcgatatag acaccaatcg ttcccgggcc tgtccacctt
gtgcccccgc ctgcaaagac 541 aatcactgtt ggggtgagag tccggaagac
tgtcagatct tgactggcac catctgtacc 601 agtggttgtg cccggtgcaa
gggccggctg cccactgact gctgccatga gcagtgtgcc 661 gcaggctgca
cgggccccaa gcattctgac tgcctggcct gcctccactt caatcatagt 721
ggtatctgtg agctgcactg cccagccctc gtcacctaca acacagacac ctttgagtcc
781 atgcacaacc ctgagggtcg ctacaccttt ggtgccagct gcgtgaccac
ctgcccctac 841 aactacctgt ctacggaagt gggatcctgc actctggtgt
gtcccccgaa taaccaagag 901 gtcacagctg aggacggaac acagcgttgt
gagaaatgca gcaagccctg tgctcgagtg 961 tgctatggtc tgggcatgga
gcaccttcga ggggcgaggg ccatcaccag tgacaatgtc 1021 caggagtttg
atggctgcaa gaagatcttt gggagcctgg catttttgcc ggagagcttt 1081
gatggggacc cctcctccgg cattgctccg ctgaggcctg agcagctcca agtgttcgaa
1141 accctggagg agatcacagg ttacctgtac atctcagcat ggccagacag
tctccgtgac 1201 ctcagtgtct tccagaacct tcgaatcatt cggggacgga
ttctccacga tggcgcgtac 1261 tcattgacac tgcaaggcct ggggatccac
tcgctggggc tgcgctcact gcgggagctg 1321 ggcagtggat tggctctgat
tcaccgcaac gcccatctct gctttgtaca cactgtacct 1381 tgggaccagc
tcttccggaa cccacatcag gccctgctcc acagtgggaa ccggccggaa 1441
gaggattgtg gtctcgaggg cttggtctgt aactcactgt gtgcccacgg gcactgctgg
1501 gggccagggc ccacccagtg tgtcaactgc agtcatttcc ttcggggcca
ggagtgtgtg 1561 gaggagtgcc gagtatggaa ggggctcccc cgggagtatg
tgagtgacaa gcgctgtctg 1621 ccgtgtcacc ccgagtgtca gcctcaaaac
agctcagaga cctgctttgg atcggaggct 1681 gatcagtgtg cagcctgcgc
ccactacaag gactcgtcct cctgtgtggc tcgctgcccc 1741 agtggtgtga
aaccggacct ctcctacatg cccatctgga agtacccgga tgaggagggc 1801
atatgccagc cgtgccccat caactgcacc cactcctgtg tggatctgga tgaacgaggc
1861 tgcccagcag agcagagagc cagcccggtg acattcatca ttgcaactgt
agtgggcgtc 1921 ctgctgttcc tgatcttagt ggtggtcgtt ggaatcctaa
tcaaacgaag gagacagaag 1981 atccggaagt atacgatg mE2Neu-Q119K, SEQ ID
NO: 18 1 acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc
cgagacccac 61 ctggacatgc tccgccacct ctaccagggc tgccaggtgg
tgcagggaaa cctggaactc 121 acctacctgc ccaccaatgc cagcctgtcc
ttcctgcagg atatccagga ggtgcagggc 181 tacgtgctca tcgctcacaa
ccaagtgagg caggtcccac tgcagaggct gcggattgtg 241 cgaggcaccc
agctctttga ggacaactat gccctggccg tgctagacaa tggagacccg 301
ctgaacaata ccacccctgt cacaggggcc tccccaggag gcctgcggga gctgaagctt
361 cgaagcctca cagagatctt gaaaggaggg gtcttgatcc agcggaaccc
ccagctctgc 421 taccaggaca cgattttgtg gaaggacatc ttccacaaga
acaaccagct ggctctcaca 481 ctgatagaca ccaaccgctc tcgggcctgc
cacccctgtt ctccgatgtg taagggctcc 541 cgctgctggg gagagagttc
tgaggattgt cagagcctga cgcgcactgt ctgtgccggt 601 ggctgtgccc
gctgcaaggg gccactgccc actgactgct gccatgagca gtgtgctgcc 661
ggctgcacgg gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc
721 atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt
tgagtccatg 781 cccaatcccg agggccggta tacattcggc gccagctgtg
tgactgcctg tccctacaac 841 tacctttcta cggacgtggg atcctgcacc
ctcgtctgcc ccctgcacaa ccaagaggtg 901 acagcagagg atggaacaca
gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc 961 tatggtctgg
gcatggagca cttgcgagag gtgagggcag ttaccagtgc caatatccag 1021
gagtttgctg gctgcaagaa gatctttggg agcctggcat ttctgccgga gagctttgat
1081 ggggacccag cctccaacac tgccgaattc gctccgctga ggcctgagca
gctccaagtg 1141 ttcgaaaccc tggaggagat cacaggttac ctgtacatct
cagcatggcc agacagtctc 1201 cgtgacctca gtgtcttcca gaaccttcga
atcattcggg gacggattct ccacgatggc 1261 gcgtactcat tgacactgca
aggcctgggg atccactcgc tggggctgcg ctcactgcgg 1321 gagctgggca
gtggattggc tctgattcac cgcaacgccc atctctgctt tgtacacact 1381
gtaccttggg accagctctt ccggaaccca catcaggccc tgctccacag tgggaaccgg
1441 ccggaagagg attgtggtct cgagggcttg gtctgtaact cactgtgtgc
ccacgggcac 1501 tgctgggggc cagggcccac ccagtgtgtc aactgcagtc
atttccttcg gggccaggag 1561 tgtgtggagg agtgccgagt atggaagggg
ctcccccggg agtatgtgag tgacaagcgc 1621 tgtctgccgt gtcaccccga
gtgtcagcct caaaacagct cagagacctg ctttggatcg 1681 gaggctgatc
agtgtgcagc ctgcgcccac tacaaggact cgtcctcctg tgtggctcgc 1741
tgccccagtg gtgtgaaacc ggacctctcc tacatgccca tctggaagta cccggatgag
1801 gagggcatat gccagccgtg ccccatcaac tgcacccact cctgtgtgga
tctggatgaa 1861 cgaggctgcc cagcagagca gagagccagc ccggtgacat
tcatcattgc aactgtagtg 1921 ggcgtcctgc tgttcctgat cttagtggtg
gtcgttggaa tcctaatcaa acgaaggaga 1981 cagaagatcc ggaagtatac gatg
prefeHER2-Q141, SEQ ID NO: 19 1 atggagctgg cggcctggtg ccgctggggg
ctcctcctcg ccctcctgcc ctccggagcc 61 acgggcaccc aagtgtgcac
cggcacagac atgaagctgc ggctcccagc cagtcccgag 121 acccacctgg
acatgctccg ccacctctac cagggctgtc aagtggtaca gggcaacctg 181
gagctcacct acctgcatgc caatgccagc ctctccttcc tgcaggatat ccaggaggtg
241 caaggctatg tgctcattgc ccacaaccaa gtgaaacagg tcccactgca
gaggctacga 301 atcgtgcgag gcacccagct ctttgaggac aactacgccc
tggccgtgct ggacaacgga 361 gacccactgg acagtggcac ccctgctaca
ggggctgccc taggagggct gcgggagctg 421 aagctccgaa gcctcacaga
gatcctgaag ggaggggtcc tcattcagcg gaacccgcag 481 ctctgccacc
aggacacgat tctgtggaag gacatcttcc acaagaacaa ccagctggcc 541
ctcatgctga tagacaccaa ccgctctcgg gcctgccaac cctgttctcc agcttgtaaa
601 gactcccact gctggggagc aagttccggg gactgtcaga gcttgactcg
aactgtctgt 661 gctggcggct gtgcccgctg caagggcccg cagcccaccg
actgctgcca cgagcaatgt 721 gctgctggct gcacgggccc caagcattct
gactgcctgg cctgcctcca cttcaaccac 781 agtggcatct gtgagctgca
ctgcccagcc ctggtcacct acaacacgga caccttcgaa 841 tccatgccca
accctgaggg ccgttatacc ttcggtgcca gctgtgtgac tgcctgtccc 901
tacaactacc tgtctacgga cgtgggatcc tgcaccctgg tctgtcccct
gaacaaccaa
961 gaggtgacag ctgaggatgg aacacagcgg tgtgagaaat gcagcaagcc
ctgtgcccga 1021 gtgtgctacg gcctaggcat ggagcacctg cgggaggcga
gggcagtcac cagtgccaac 1081 atccaagaat ttgtcggctg caagaagatc
tttgggagcc tggcgtttct gccagagagc 1141 tttgaggggg acccagcctc
caacactgcc cccctgcagc ctgagcagct cagagtgttt 1201 gaggctctgg
aggagattac aggttacctg tacatctcag cgtggccaga cagcttgcct 1261
aacctcagtg tcttccagaa cctcagagtg atccggggcc gagttctgca tgacggtgct
1321 tactcgctga cccttcaagg gctgggcatc agctggctgg ggctgcgctc
gctgcgggag 1381 ctgggcagtg ggctggccct catccaccgc aactcccgcc
tctgcttcgt acacacggtg 1441 ccctgggacc agctcttccg gaacccccac
caggccctgc tccacagcgc caaccggcca 1501 gaggacgagt gcgcgggtga
gggcctggcc tgctatccgc tgtgtgccca cgggcactgc 1561 tggggtccgg
gacccaccca gtgtgtcaac tgcagccagt tccttcgggg ccaggagtgc 1621
gtggaggaat gccgagtatt gcaggggctt ccccgggagt atgtgaagga taggttctgt
1681 ctgccatgcc acccggagtg tcagccccag aatggctcag tgacctgctt
gggctcggaa 1741 gctgaccagt gtgtggcctg tgcccactac aaggaccctc
ctttctgtgt ggctcgctgc 1801 cccagtgggg tgaaacctga cctctccttc
atgcccatct ggaagttcgc agatgaggag 1861 ggcacgtgcc agccatgccc
catcaactgc acccactcct gtgcggacct ggacgagaag 1921 ggctgccccg
ccgagcagag agccagccct gtgacgtcca tcattgctgc tgtggtgggc 1981
attctgctgg tcgtggttgt ggggctggtc cttggcatcc taatcaagcg aaggcggcag
2041 aagatccgga agtacacgat g prebearHER2-Q141K, SEQ ID NO: 20 1
atggagctgg cggcctggtg ccgctggggg ctcctcctcg ccctcctgcc ctccggagcc
61 gcgggcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc
cagtcccgag 121 acccacctgg atatgctccg ccacctctac caggcctgtc
aagtggtaca gggtaacctg 181 gagctcacct acctgcccgc caatgccagc
ctgtccttcc tgcaggatat ccaggaggta 241 cagggctatg tgctcattgc
tcacagccaa gtgagacagg tcccgctgca gaggctccga 301 atcgtgcgag
gcacccagct ctttgaggac aactacgccc tggccgtgct ggacaatgga 361
gagccgccca agggggacac ctctgtggca ggggctaccc caggagggct gcgggagctg
421 aagcttcgaa gcctcacaga gatcctgaag ggaggggtct tgattcagcg
gaacccacag 481 ctctgccacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggcc 541 ctcacgctga tagacaccaa ccgctctcgg
gcctgccaac cctgttctcc agcctgtaaa 601 gacccccact gctggggagc
aagttccggg gactgtcaga gcttgacacg aaccgtctgt 661 gccggcggct
gtgcccgctg caagggccca aaacccactg actgctgcca tgagcagtgc 721
gcggctggct gcacgggccc caagcactcg gactgcctgg cctgccttca cttcaaccac
781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct acaacacgga
cacgttcgaa 841 tccatgccca accctgaggg ccgatacacc ttcggtgcca
gctgtgtgac cgcctgtccc 901 tacaactacc tgtccacgga cgtgggatcc
tgcaccctgg tctgtcccct gaacaaccaa 961 gaggtgacgg ctgaggatgg
cacccagcgg tgtgagaaat gcagcagacc ctgtgcccga 1021 gtgtgctatg
gtctgggcat ggagcacctg cgggaggcga gggcggtcac cagcgccaac 1081
atccaagagt tcgccggctg caagaagatc tttgggagcc tggcgtttct gccagagagc
1141 ttcgagggag acccagcctc caacactgcc cccctgcagc ctgaacagct
cagagtgttc 1201 gaggccctgg aggagatcac aggttacctg tatatctcag
cgtggccgga cagcttgcct 1261 aacctcagtg tcttccagaa cctgcgagta
atccggggac gagttctgca tgatggcgcc 1321 tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc gctgcgggaa 1381 ctgggcagcg
ggctggccct catccaccgc aacgcccgcc tctgcttcat ccacacggtg 1441
ccctgggagc agctcttccg gaacccccac caagccctgc tgcacagtgc caaccggccg
1501 gaggccgagt gcgtgggcga gggcctggcc tgctacccgc tgtgcgccca
tgggcactgc 1561 tggggtccgg ggcccaccca gtgcgtcaac tgcagccaat
tccttcgggg ccaggagtgc 1621 gtggaggaat gccgagaact gcacgggcta
ccccgggaat atgtgaagga cagatactgt 1681 ctgccatgcc accccgagtg
tcggccccag aatggctcag tgacctgctt tgggtcggag 1741 gctgaccagt
gtgtggcctg cgcccactac aaggaccctc cctcctgcgt ggctcgctgc 1801
cccagtggtg tgaaacccga cctctctttc atgcccattt ggaagtttgc agatgaggag
1861 ggcacatgcc agccgtgccc catcaactgc acccactcct gtggggacct
ggacgagagg 1921 ggctgccccg ccgaacagag agccagccct gtgacatcca
tcattgccgc tgtggtgggc 1981 attctgctgg ccgtggtcat ggggctggtc
ctcggcatcc tgatcaagcg aaggcgacag 2041 aagatccgga agtacacgat g
prehumHER2-Q141K, SEQ ID NO: 21 1 atggagctgg cggccttgtg ccgctggggg
ctcctcctcg ccctcttgcc ccccggagcc 61 gcgagcaccc aagtgtgcac
cggcacagac atgaagctgc ggctccctgc cagtcccgag 121 acccacctgg
acatgctccg ccacctctac cagggctgcc aggtggtgca gggaaacctg 181
gaactcacct acctgcccac caatgccagc ctgtccttcc tgcaggatat ccaggaggtg
241 cagggctacg tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca
gaggctgcgg 301 attgtgcgag gcacccagct ctttgaggac aactatgccc
tggccgtgct agacaatgga 361 gacccgctga acaataccac ccctgtcaca
ggggcctccc caggaggcct gcgggagctg 421 Aagcttcgaa gcctcacaga
gatcttgaaa ggaggggtct tgatccagcg gaacccccag 481 ctctgctacc
aggacacgat tttgtggaag gacatcttcc acaagaacaa ccagctggct 541
ctcacactga tagacaccaa ccgctctcgg gcctgccacc cctgttctcc gatgtgtaag
601 ggctcccgct gctggggaga gagttctgag gattgtcaga gcctgacgcg
cactgtctgt 661 gccggtggct gtgcccgctg caaggggcca ctgcccactg
actgctgcca tgagcagtgt 721 gctgccggct gcacgggccc caagcactct
gactgcctgg cctgcctcca cttcaaccac 781 agtggcatct gtgagctgca
ctgcccagcc ctggtcacct acaacacaga cacgtttgag 841 tccatgccca
atcccgaggg ccggtataca ttcggcgcca gctgtgtgac tgcctgtccc 901
tacaactacc tttctacgga cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa
961 gaggtgacag cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc
ctgtgcccga 1021 gtgtgctatg gtctgggcat ggagcacttg cgagaggtga
gggcagttac cagtgccaat 1081 atccaggagt ttgctggctg caagaagatc
tttgggagcc tggcatttct gccggagagc 1141 tttgatgggg acccagcctc
caacactgcc ccgctccagc cagagcagct ccaagtgttt 1201 gagactctgg
aagagatcac aggttaccta tacatctcag catggccgga cagcctgcct 1261
gacctcagcg tcttccagaa cctgcaagta atccggggac gaattctgca caatggcgcc
1321 tactcgctga ccctgcaagg gctgggcatc agctggctgg ggctgcgctc
actgagggaa 1381 ctgggcagtg gactggccct catccaccat aacacccacc
tctgcttcgt gcacacggtg 1441 ccctgggacc agctctttcg gaacccgcac
caagctctgc tccacactgc caaccggcca 1501 gaggacgagt gtgtgggcga
gggcctggcc tgccaccagc tgtgcgcccg agggcactgc 1561 tggggtccag
ggcccaccca gtgtgtcaac tgcagccagt tccttcgggg ccaggagtgc 1621
gtggaggaat gccgagtact gcaggggctc cccagggagt atgtgaatgc caggcactgt
1681 ttgccgtgcc accctgagtg tcagccccag aatggctcag tgacctgttt
tggaccggag 1741 gctgaccagt gtgtggcctg tgcccactat aaggaccctc
ccttctgcgt ggcccgctgc 1801 cccagcggtg tgaaacctga cctctcctac
atgcccatct ggaagtttcc agatgaggag 1861 ggcgcatgcc agccttgccc
catcaactgc acccactcct gtgtggacct ggatgacaag 1921 ggctgccccg
ccgagcagag agccagccct ctgacgtcca tcatctctgc ggtggttggc 1981
attctgctgg tcgtggtctt gggggtggtc tttgggatcc tcatcaagcg acggcagcag
2041 aagatccgga agtacacgat gtag premouseHER2-Q142K, SEQ ID NO: 22 1
atggagctgg cggcctggtg ccgttggggg ttcctcctcg ccctcctgtc ccccggagcc
61 gcgggtaccc aagtgtgtac cggtaccgac atgaagttgc gactccctgc
cagtcctgag 121 acccacctgg acatgcttcg ccacctctac cagggctgtc
aggtggtgca gggcaatttg 181 gagcttacct acctgcccgc caatgccagc
ctctcattcc tgcaggacat ccaggaagtc 241 cagggataca tgctcatcgc
tcacaaccga gtgaaacacg tcccactgca gaggttgcgc 301 atcgtgagag
ggactcagct ctttgaggac aagtatgccc tggctgtgct agacaaccga 361
gaccctttgg acaacgtcac caccgccgcc ccaggcagaa ccccagaagg gctgcgggag
421 ctgaagcttc gaagtctcac agagatcttg aagggaggag ttttgatccg
tgggaaccct 481 cagctctgct accaggacat ggttttgtgg aaggatgtcc
tccgtaagaa taaccagctg 541 gctcctgtcg acatggacac caatcgttcc
cgggcctgtc caccttgtgc cccaacctgc 601 aaagacaatc actgttgggg
tgagagtcct gaagactgtc agatcttgac tggcaccatc 661 tgtactagtg
gctgtgcccg gtgcaagggc cggctgccca ctgactgttg ccatgagcag 721
tgtgctgcag gctgcacggg tcccaagcat tctgactgcc tggcctgcct ccacttcaat
781 catagtggta tctgtgagct gcactgcccg gccctcatca cctacaacac
agacaccttc 841 gagtccatgc tcaaccctga gggtcgctac acctttggtg
ccagctgtgt gaccacctgc 901 ccctacaact acctctccac ggaagtggga
tcctgcactc tggtctgtcc cccgaacaac 961 caagaggtca cagctgagga
cggaacacag cggtgtgaga aatgcagcaa gccctgtgct 1021 ggagtatgct
atggtctggg catggagcac ctccgagggg cgagggccat caccagtgac 1081
aatatccagg agtttgctgg ctgcaagaag atctttggga gcctggcatt tttgccggag
1141 agctttgatg ggaacccctc ctccggcgtt gccccactga agccagagca
tctccaagtg 1201 ttcgaaaccc tggaggagat cacaggttac ctatacattt
cagcatggcc agagagcttc 1261 caagacctca gtgtcttcca gaaccttcgg
gtcattcggg gacggattct ccatgatggt 1321 gcttactcat tgacgttgca
aggcctgggg attcactcac tggggctacg ctcactgcgg 1381 gagctgggca
gtggattggc tctcattcac cgcaacaccc atctctgctt tgtaaacact 1441
gtaccttggg accagctctt ccggaacccg caccaggccc tactccacag tgggaaccgg
1501 ccagaagagg catgtggtct tgagggcttg gtctgtaact cactgtgtgc
ccgtgggcac 1561 tgctgggggc cagggcccac ccagtgtgtc aactgcagtc
agttcctccg gggccaggag 1621 tgtgtggagg agtgccgagt atggaagggg
ctccccaggg agtatgtgag gggcaagcac 1681 tgtctgccat gccaccccga
gtgtcagcct caaaacagct cggagacctg ctatggatcg 1741 gaggctgacc
agtgtgaggc ttgtgcccac tacaaggact catcttcctg tgtggctcgc 1801
tgccccagtg gtgtgaagcc agacctctcc tacatgccta tctggaagta cccggatgag
1861 gagggcatat gtcagccatg ccccatcaac tgcacccact catgtgtgga
cctggacgaa 1921 cgaggctgcc cagcagagca gagagccagc ccagtgacat
tcatcattgc aactgtggtg 1981 ggcgtcctgt tgttcctgat catagtggtg
gtcattggaa tcctaatcaa acgaaggcga 2041 cagaagatcc ggaagtatac
catg
preratHER2-Q145K, SEQ ID NO: 23 1 atgatcatca tggagctggc ggcctggtgc
cgctgggggt tcctcctcgc cctcctgccc 61 cccggaatcg cgggcaccca
agtgtgtacc ggcacagaca tgaagttgcg gctccctgcc 121 agtcctgaga
cccacctgga catgctccgc cacctgtacc agggctgtca ggtagtgcag 181
ggcaacttgg agcttaccta cgtgcctgcc aatgccagcc tctcattcct gcaggacatc
241 caggaagttc agggttacat gctcatcgct cacaaccagg tgaagcgcgt
cccactgcaa 301 aggctgcgca tcgtgagagg gacccagctc tttgaggaca
agtatgccct ggctgtgcta 361 gacaaccgag atcctcagga caatgtcgcc
gcctccaccc caggcagaac cccagagggg 421 ctgcgggagc tgaagcttcg
aagtctcaca gagatcctga agggaggagt tttgatccgt 481 gggaaccctc
agctctgcta ccaggacatg gttttgtgga aggacgtctt ccgcaagaat 541
aaccaactgg ctcctgtcga tatagacacc aatcgttccc gggcctgtcc accttgtgcc
601 cccgcctgca aagacaatca ctgttggggt gagagtccgg aagactgtca
gatcttgact 661 ggcaccatct gtaccagtgg ttgtgcccgg tgcaagggcc
ggctgcccac tgactgctgc 721 catgagcagt gtgccgcagg ctgcacgggc
cccaagcatt ctgactgcct ggcctgcctc 781 cacttcaatc atagtggtat
ctgtgagctg cactgcccag ccctcgtcac ctacaacaca 841 gacacctttg
agtccatgca caaccctgag ggtcgctaca cctttggtgc cagctgcgtg 901
accacctgcc cctacaacta cctgtctacg gaagtgggat cctgcactct ggtgtgtccc
961 ccgaataacc aagaggtcac agctgaggac ggaacacagc gttgtgagaa
atgcagcaag 1021 ccctgtgctc gagtgtgcta tggtctgggc atggagcacc
ttcgaggggc gagggccatc 1081 accagtgaca atgtccagga gtttgatggc
tgcaagaaga tctttgggag cctggcattt 1141 ttgccggaga gctttgatgg
ggacccctcc tccggcattg ctccgctgag gcctgagcag 1201 ctccaagtgt
tcgaaaccct ggaggagatc acaggttacc tgtacatctc agcatggcca 1261
gacagtctcc gtgacctcag tgtcttccag aaccttcgaa tcattcgggg acggattctc
1321 cacgatggcg cgtactcatt gacactgcaa ggcctgggga tccactcgct
ggggctgcgc 1381 tcactgcggg agctgggcag tggattggct ctgattcacc
gcaacgccca tctctgcttt 1441 gtacacactg taccttggga ccagctcttc
cggaacccac atcaggccct gctccacagt 1501 gggaaccggc cggaagagga
ttgtggtctc gagggcttgg tctgtaactc actgtgtgcc 1561 cacgggcact
gctgggggcc agggcccacc cagtgtgtca actgcagtca tttccttcgg 1621
ggccaggagt gtgtggagga gtgccgagta tggaaggggc tcccccggga gtatgtgagt
1681 gacaagcgct gtctgccgtg tcaccccgag tgtcagcctc aaaacagctc
agagacctgc 1741 tttggatcgg aggctgatca gtgtgcagcc tgcgcccact
acaaggactc gtcctcctgt 1801 gtggctcgct gccccagtgg tgtgaaaccg
gacctctcct acatgcccat ctggaagtac 1861 ccggatgagg agggcatatg
ccagccgtgc cccatcaact gcacccactc ctgtgtggat 1921 ctggatgaac
gaggctgccc agcagagcag agagccagcc cggtgacatt catcattgca 1981
actgtagtgg gcgtcctgct gttcctgatc ttagtggtgg tcgttggaat cctaatcaaa
2041 cgaaggagac agaagatccg gaagtatacg atg preE2Neu-Q141K, SEQ ID
NO: 24 1 atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc
ccccggagcc 61 gcgagcaccc aagtgtgcac cggcacagac atgaagctgc
ggctccctgc cagtcccgag 121 acccacctgg acatgctccg ccacctctac
cagggctgcc aggtggtgca gggaaacctg 181 gaactcacct acctgcccac
caatgccagc ctgtccttcc tgcaggatat ccaggaggtg 241 cagggctacg
tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca gaggctgcgg 301
attgtgcgag gcacccagct ctttgaggac aactatgccc tggccgtgct agacaatgga
361 gacccgctga acaataccac ccctgtcaca ggggcctccc caggaggcct
gcgggagctg 421 aagcttcgaa gcctcacaga gatcttgaaa ggaggggtct
tgatccagcg gaacccccag 481 ctctgctacc aggacacgat tttgtggaag
gacatcttcc acaagaacaa ccagctggct 541 ctcacactga tagacaccaa
ccgctctcgg gcctgccacc cctgttctcc gatgtgtaag 601 ggctcccgct
gctggggaga gagttctgag gattgtcaga gcctgacgcg cactgtctgt 661
gccggtggct gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt
721 gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca
cttcaaccac 781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacaga cacgtttgag 841 tccatgccca atcccgaggg ccggtataca
ttcggcgcca gctgtgtgac tgcctgtccc 901 tacaactacc tttctacgga
cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa 961 gaggtgacag
cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga 1021
gtgtgctatg gtctgggcat ggagcacttg cgagaggtga gggcagttac cagtgccaat
1081 atccaggagt ttgctggctg caagaagatc tttgggagcc tggcatttct
gccggagagc 1141 tttgatgggg acccagcctc caacactgcc gaattcgctc
cgctgaggcc tgagcagctc 1201 caagtgttcg aaaccctgga ggagatcaca
ggttacctgt acatctcagc atggccagac 1261 agtctccgtg acctcagtgt
cttccagaac cttcgaatca ttcggggacg gattctccac 1321 gatggcgcgt
actcattgac actgcaaggc ctggggatcc actcgctggg gctgcgctca 1381
ctgcgggagc tgggcagtgg attggctctg attcaccgca acgcccatct ctgctttgta
1441 cacactgtac cttgggacca gctcttccgg aacccacatc aggccctgct
ccacagtggg 1501 aaccggccgg aagaggattg tggtctcgag ggcttggtct
gtaactcact gtgtgcccac 1561 gggcactgct gggggccagg gcccacccag
tgtgtcaact gcagtcattt ccttcggggc 1621 caggagtgtg tggaggagtg
ccgagtatgg aaggggctcc cccgggagta tgtgagtgac 1681 aagcgctgtc
tgccgtgtca ccccgagtgt cagcctcaaa acagctcaga gacctgcttt 1741
ggatcggagg ctgatcagtg tgcagcctgc gcccactaca aggactcgtc ctcctgtgtg
1801 gctcgctgcc ccagtggtgt gaaaccggac ctctcctaca tgcccatctg
gaagtacccg 1861 gatgaggagg gcatatgcca gccgtgcccc atcaactgca
cccactcctg tgtggatctg 1921 gatgaacgag gctgcccagc agagcagaga
gccagcccgg tgacattcat cattgcaact 1981 gtagtgggcg tcctgctgtt
cctgatctta gtggtggtcg ttggaatcct aatcaaacga 2041 aggagacaga
agatccggaa gtatacgatg mhumHER2-Q307K, SEQ ID NO: 25 1 tqvctgtdmk
lrlpaspeth ldmlrhlyqg cqvvqgnlel tylptnasls flqdiqevqg 61
yvliahnqvr qvplqrlriv rgtqlfedny alavldngdp lnnttpvtga spgglrelql
121 rslteilkgg vliqrnpqlc yqdtilwkdi fhknnqlalt lidtnrsrac
hpcspmckgs 181 rcwgessedc qsltrtvcag gcarckgplp tdccheqcaa
gctgpkhsdc laclhfnhsg 241 icelhcpalv tyntdtfesm pnpegrytfg
ascvtacpyn ylstdvgsct lvcplhnqev 301 taedgtKrce kcskpcarvc
yglgmehlre vravtsaniq efagckkifg slaflpesfd 361 gdpasntapl
qpeqlqvfet leeitgylyi sawpdslpdl svfqnlqvir grilhngays 421
ltlqglgisw lglrslrelg sglalihhnt hlcfvhtvpw dqlfrnphqa llhtanrped
481 ecvgeglach qlcarghcwg pgptqcvncs qflrgqecve ecrvlqglpr
eyvnarhclp 541 chpecqpqng svtcfgpead qcvacahykd ppfcvarcps
gvkpdlsymp iwkfpdeega 601 cqpcpincth scvdlddkgc paeqrasplt
siisavvgil lvvvlgvvfg ilikrrqqki 661 rkytm mhumHER2-Q407R, SEQ ID
NO: 26 1 tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tylptnasls
flqdiqevqg 61 yvliahnqvr qvplqrlriv rgtqlfedny alavldngdp
lnnttpvtga spgglrelql 121 rslteilkgg vliqrnpqlc yqdtilwkdi
fhknnqlalt lidtnrsrac hpcspmckgs 181 rcwgessedc qsltrtvcag
gcarckgplp tdccheqcaa gctgpkhsdc laclhfnhsg 241 icelhcpalv
tyntdtfesm pnpegrytfg ascvtacpyn ylstdvgsct lvcplhnqev 301
taedgtqrce kcskpcarvc yglgmehlre vravtsaniq efagckkifg slaflpesfd
361 gdpasntapl qpeqlqvfet leeitgylyi sawpdslpdl svfqn1Rvir
grilhngays 421 ltlqglgisw lglrslrelg sglalihhnt hlcfvhtvpw
dqlfrnphqa llhtanrped 481 ecvgeglach qlcarghcwg pgptqcvncs
qflrgqecve ecrvlqglpr eyvnarhclp 541 chpecqpqng svtcfgpead
qcvacahykd ppfcvarcps gvkpdlsymp iwkfpdeega 601 cqpcpincth
scvdlddkgc paeqrasplt siisavvgil lvvvlgvvfg ilikrrqqki 661 rkytm
mhumHER2-N416D SEQ ID NO: 27 1 tqvctgtdmk lrlpaspeth ldmlrhlyqg
cqvvqgnlel tylptnasls flqdiqevqg 61 yvliahnqvr qvplqrlriv
rgtqlfedny alavldngdp lnnttpvtga spgglrelql 121 rslteilkgg
vliqrnpqlc yqdtilwkdi fhknnqlalt lidtnrsrac hpcspmckgs 181
rcwgessedc qsltrtvcag gcarckgplp tdccheqcaa gctgpkhsdc laclhfnhsg
241 icelhcpalv tyntdtfesm pnpegrytfg ascvtacpyn ylstdvgsct
lvcplhnqev 301 taedgtqrce kcskpcarvc yglgmehlre vravtsaniq
efagckkifg slaflpesfd 361 gdpasntapl qpeqlqvfet leeitgylyi
sawpdslpdl svfqnlqvir grilhDgays 421 ltlqglgisw lglrslrelg
sglalihhnt hlcfvhtvpw dqlfrnphqa llhtanrped 481 ecvgeglach
qlcarghcwg pgptqcvncs qflrgqecve ecrvlqglpr eyvnarhclp 541
chpecqpqng svtcfgpead qcvacahykd ppfcvarcps gvkpdlsymp iwkfpdeega
601 cqpcpincth scvdlddkgc paeqrasplt siisavvgil lvvvlgvvfg
ilikrrqqki 661 rkytm prehumHER2-Q329K, SEQ ID NO: 28 1 melaalcrwg
lllallppga astqvctgtd mklrlpaspe thldmlrhly qgcqvvqgnl 61
eltylptnas lsflgdigev qgyvliahnq vrqvplqrlr ivrgtqlfed nyalavldng
121 dpinnttpvt gaspgglrel qlrslteilk ggvliqrnpq lcyqdtilwk
difhknnqla 181 ltlidtnrsr achpcspmck gsrcwgesse dcqsltrtvc
aggcarckgp 1ptdccheqc 241 aagctgpkhs dclaclhfnh sgicelhcpa
lvtyntdtfe smpnpegryt fgascvtacp 301 ynylstdvgs ctlvcplhnq
evtaedgtKr cekcskpcar vcyglgmehl revravtsan 361 igefagokki
fgslaflpes fdgdpasnta plqpeqlqvf etleeitgyl yisawpdslp 421
dlsvfqnlqv irgrilhnga ysltlqglgi swlglrslre lgsglalihh nthlcfvhtv
481 pwdqlfrnph qallhtanrp edecvgegla chqlcarghc wgpgptqcvn
csqflrggec 541 veecrvlqgl preyvnarhc lpchpecqpq ngsvtcfgpe
adqcvacahy kdppfcvarc 601 psgvkpdlsy mpiwkfpdee gacqpcpinc
thscvdlddk gcpaegrasp ltsiisavvg 661 illvvvlgvv fgilikrrqq kirkytm
prehumHER2-Q429R, SEQ ID NO: 29 1 melaalcrwg lllallppga astqvctgtd
mklrlpaspe thldmlrhly qgcqvvqgnl 61 eltylptnas lsflgdigev
qgyvliahnq vrqvplqrlr ivrgtqlfed nyalavldng 121 dpinnttpvt
gaspgglrel qlrslteilk ggvliqrnpq lcyqdtilwk difhknnqla 181
ltlidtnrsr achpcspmck gsrcwgesse dcqsltrtvc aggcarckgp
1ptdccheqc
241 aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt
fgascvtacp 301 ynylstdvgs ctlvcplhnq evtaedgtqr cekcskpcar
vcyglgmehl revravtsan 361 igefagokki fgslaflpes fdgdpasnta
plqpeqlqvf etleeitgyl yisawpdslp 421 dlsvfqn1Rv irgrilhnga
ysltlqglgi swlglrslre lgsglalihh nthlcfvhtv 481 pwdqlfrnph
qallhtanrp edecvgegla chqlcarghc wgpgptqcvn csqflrggec 541
veecrvlqgl preyvnarhc lpchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc
601 psgvkpdlsy mpiwkfpdee gacqpcpinc thscvdlddk gcpaegrasp
ltsiisavvg 661 illvvvlgvv fgilikrrqq kirkytm prehumHER2-N438D, SEQ
ID NO: 30 1 melaalcrwg lllallppga astqvctgtd mklrlpaspe thldmlrhly
qgcqvvqgnl 61 eltylptnas lsflgdigev qgyvliahnq vrqvplqrlr
ivrgtqlfed nyalavldng 121 dpinnttpvt gaspgglrel qlrslteilk
ggvliqrnpq lcyqdtilwk difhknnqla 181 ltlidtnrsr achpcspmck
gsrcwgesse dcqsltrtvc aggcarckgp 1ptdccheqc 241 aagctgpkhs
dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp 301
ynylstdvgs ctlvcplhnq evtaedgtqr cekcskpcar vcyglgmehl revravtsan
361 igefagokki fgslaflpes fdgdpasnta plqpeqlqvf etleeitgyl
yisawpdslp 421 dlsvfqnlqv irgrilhDga ysltlqglgi swlglrslre
lgsglalihh nthlcfvhtv 481 pwdqlfrnph qallhtanrp edecvgegla
chqlcarghc wgpgptqcvn csqflrggec 541 veecrvlqgl preyvnarhc
lpchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc 601 psgvkpdlsy
mpiwkfpdee gacqpcpinc thscvdlddk gcpaegrasp ltsiisavvg 661
illvvvlgvv fgilikrrqq kirkytm mhumHER2-Q307K, SEQ ID NO: 31 1
acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac
61 ctggacatgc tccgccacct ctaccagggc tgccaggtgg tgcagggaaa
cctggaactc 121 acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg
atatccagga ggtgcagggc 181 tacgtgctca tcgctcacaa ccaagtgagg
caggtcccac tgcagaggct gcggattgtg 241 cgaggcaccc agctctttga
ggacaactat gccctggccg tgctagacaa tggagacccg 301 ctgaacaata
ccacccctgt cacaggggcc tccccaggag gcctgcggga gctgcagctt 361
cgaagcctca cagagatctt gaaaggaggg gtcttgatcc agcggaaccc ccagctctgc
421 taccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggctctcaca 481 ctgatagaca ccaaccgctc tcgggcctgc cacccctgtt
ctccgatgtg taagggctcc 541 cgctgctggg gagagagttc tgaggattgt
cagagcctga cgcgcactgt ctgtgccggt 601 ggctgtgccc gctgcaaggg
gccactgccc actgactgct gccatgagca gtgtgctgcc 661 ggctgcacgg
gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc 721
atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt tgagtccatg
781 cccaatcccg agggccggta tacattcggc gccagctgtg tgactgcctg
tccctacaac 841 tacctttcta cggacgtggg atcctgcacc ctcgtctgcc
ccctgcacaa ccaagaggtg 901 acagcagagg atggaacaAa gcggtgtgag
aagtgcagca agccctgtgc ccgagtgtgc 961 tatggtctgg gcatggagca
cttgcgagag gtgagggcag ttaccagtgc caatatccag 1021 gagtttgctg
gctgcaagaa gatctttggg agcctggcat ttctgccgga gagctttgat 1081
ggggacccag cctccaacac tgccccgctc cagccagagc agctccaagt gtttgagact
1141 ctggaagaga tcacaggtta cctatacatc tcagcatggc cggacagcct
gcctgacctc 1201 agcgtcttcc agaacctgca agtaatccgg ggacgaattc
tgcacaatgg cgcctactcg 1261 ctgaccctgc aagggctggg catcagctgg
ctggggctgc gctcactgag ggaactgggc 1321 agtggactgg ccctcatcca
ccataacacc cacctctgct tcgtgcacac ggtgccctgg 1381 gaccagctct
ttcggaaccc gcaccaagct ctgctccaca ctgccaaccg gccagaggac 1441
gagtgtgtgg gcgagggcct ggcctgccac cagctgtgcg cccgagggca ctgctggggt
1501 ccagggccca cccagtgtgt caactgcagc cagttccttc ggggccagga
gtgcgtggag 1561 gaatgccgag tactgcaggg gctccccagg gagtatgtga
atgccaggca ctgtttgccg 1621 tgccaccctg agtgtcagcc ccagaatggc
tcagtgacct gttttggacc ggaggctgac 1681 cagtgtgtgg cctgtgccca
ctataaggac cctcccttct gcgtggcccg ctgccccagc 1741 ggtgtgaaac
ctgacctctc ctacatgccc atctggaagt ttccagatga ggagggcgca 1801
tgccagcctt gccccatcaa ctgcacccac tcctgtgtgg acctggatga caagggctgc
1861 cccgccgagc agagagccag ccctctgacg tccatcatct ctgcggtggt
tggcattctg 1921 ctggtcgtgg tcttgggggt ggtctttggg atcctcatca
agcgacggca gcagaagatc 1981 cggaagtaca cgatgtga mhumHER2-Q407R, SEQ
ID NO: 32 1 acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc
cgagacccac 61 ctggacatgc tccgccacct ctaccagggc tgccaggtgg
tgcagggaaa cctggaactc 121 acctacctgc ccaccaatgc cagcctgtcc
ttcctgcagg atatccagga ggtgcagggc 181 tacgtgctca tcgctcacaa
ccaagtgagg caggtcccac tgcagaggct gcggattgtg 241 cgaggcaccc
agctctttga ggacaactat gccctggccg tgctagacaa tggagacccg 301
ctgaacaata ccacccctgt cacaggggcc tccccaggag gcctgcggga gctgcagctt
361 cgaagcctca cagagatctt gaaaggaggg gtcttgatcc agcggaaccc
ccagctctgc 421 taccaggaca cgattttgtg gaaggacatc ttccacaaga
acaaccagct ggctctcaca 481 ctgatagaca ccaaccgctc tcgggcctgc
cacccctgtt ctccgatgtg taagggctcc 541 cgctgctggg gagagagttc
tgaggattgt cagagcctga cgcgcactgt ctgtgccggt 601 ggctgtgccc
gctgcaaggg gccactgccc actgactgct gccatgagca gtgtgctgcc 661
ggctgcacgg gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc
721 atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt
tgagtccatg 781 cccaatcccg agggccggta tacattcggc gccagctgtg
tgactgcctg tccctacaac 841 tacctttcta cggacgtggg atcctgcacc
ctcgtctgcc ccctgcacaa ccaagaggtg 901 acagcagagg atggaacaca
gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc 961 tatggtctgg
gcatggagca cttgcgagag gtgagggcag ttaccagtgc caatatccag 1021
gagtttgctg gctgcaagaa gatctttggg agcctggcat ttctgccgga gagctttgat
1081 ggggacccag cctccaacac tgccccgctc cagccagagc agctccaagt
gtttgagact 1141 ctggaagaga tcacaggtta cctatacatc tcagcatggc
cggacagcct gcctgacctc 1201 agcgtcttcc agaacctgAG agtaatccgg
ggacgaattc tgcacaatgg cgcctactcg 1261 ctgaccctgc aagggctggg
catcagctgg ctggggctgc gctcactgag ggaactgggc 1321 agtggactgg
ccctcatcca ccataacacc cacctctgct tcgtgcacac ggtgccctgg 1381
gaccagctct ttcggaaccc gcaccaagct ctgctccaca ctgccaaccg gccagaggac
1441 gagtgtgtgg gcgagggcct ggcctgccac cagctgtgcg cccgagggca
ctgctggggt 1501 ccagggccca cccagtgtgt caactgcagc cagttccttc
ggggccagga gtgcgtggag 1561 gaatgccgag tactgcaggg gctccccagg
gagtatgtga atgccaggca ctgtttgccg 1621 tgccaccctg agtgtcagcc
ccagaatggc tcagtgacct gttttggacc ggaggctgac 1681 cagtgtgtgg
cctgtgccca ctataaggac cctcccttct gcgtggcccg ctgccccagc 1741
ggtgtgaaac ctgacctctc ctacatgccc atctggaagt ttccagatga ggagggcgca
1801 tgccagcctt gccccatcaa ctgcacccac tcctgtgtgg acctggatga
caagggctgc 1861 cccgccgagc agagagccag ccctctgacg tccatcatct
ctgcggtggt tggcattctg 1921 ctggtcgtgg tcttgggggt ggtctttggg
atcctcatca agcgacggca gcagaagatc 1981 cggaagtaca cgatgtga
mhumHER2-N416D, SEQ ID NO: 33 1 acccaagtgt gcaccggcac agacatgaag
ctgcggctcc ctgccagtcc cgagacccac 61 ctggacatgc tccgccacct
ctaccagggc tgccaggtgg tgcagggaaa cctggaactc 121 acctacctgc
ccaccaatgc cagcctgtcc ttcctgcagg atatccagga ggtgcagggc 181
tacgtgctca tcgctcacaa ccaagtgagg caggtcccac tgcagaggct gcggattgtg
241 cgaggcaccc agctctttga ggacaactat gccctggccg tgctagacaa
tggagacccg 301 ctgaacaata ccacccctgt cacaggggcc tccccaggag
gcctgcggga gctgcagctt 361 cgaagcctca cagagatctt gaaaggaggg
gtcttgatcc agcggaaccc ccagctctgc 421 taccaggaca cgattttgtg
gaaggacatc ttccacaaga acaaccagct ggctctcaca 481 ctgatagaca
ccaaccgctc tcgggcctgc cacccctgtt ctccgatgtg taagggctcc 541
cgctgctggg gagagagttc tgaggattgt cagagcctga cgcgcactgt ctgtgccggt
601 ggctgtgccc gctgcaaggg gccactgccc actgactgct gccatgagca
gtgtgctgcc 661 ggctgcacgg gccccaagca ctctgactgc ctggcctgcc
tccacttcaa ccacagtggc 721 atctgtgagc tgcactgccc agccctggtc
acctacaaca cagacacgtt tgagtccatg 781 cccaatcccg agggccggta
tacattcggc gccagctgtg tgactgcctg tccctacaac 841 tacctttcta
cggacgtggg atcctgcacc ctcgtctgcc ccctgcacaa ccaagaggtg 901
acagcagagg atggaacaca gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc
961 tatggtctgg gcatggagca cttgcgagag gtgagggcag ttaccagtgc
caatatccag 1021 gagtttgctg gctgcaagaa gatctttggg agcctggcat
ttctgccgga gagctttgat 1081 ggggacccag cctccaacac tgccccgctc
cagccagagc agctccaagt gtttgagact 1141 ctggaagaga tcacaggtta
cctatacatc tcagcatggc cggacagcct gcctgacctc 1201 agcgtcttcc
agaacctgca agtaatccgg ggacgaattc tgcacGatgg cgcctactcg 1261
ctgaccctgc aagggctggg catcagctgg ctggggctgc gctcactgag ggaactgggc
1321 agtggactgg ccctcatcca ccataacacc cacctctgct tcgtgcacac
ggtgccctgg 1381 gaccagctct ttcggaaccc gcaccaagct ctgctccaca
ctgccaaccg gccagaggac 1441 gagtgtgtgg gcgagggcct ggcctgccac
cagctgtgcg cccgagggca ctgctggggt 1501 ccagggccca cccagtgtgt
caactgcagc cagttccttc ggggccagga gtgcgtggag 1561 gaatgccgag
tactgcaggg gctccccagg gagtatgtga atgccaggca ctgtttgccg 1621
tgccaccctg agtgtcagcc ccagaatggc tcagtgacct gttttggacc ggaggctgac
1681 cagtgtgtgg cctgtgccca ctataaggac cctcccttct gcgtggcccg
ctgccccagc 1741 ggtgtgaaac ctgacctctc ctacatgccc atctggaagt
ttccagatga ggagggcgca 1801 tgccagcctt gccccatcaa ctgcacccac
tcctgtgtgg acctggatga caagggctgc 1861 cccgccgagc agagagccag
ccctctgacg tccatcatct ctgcggtggt tggcattctg 1921 ctggtcgtgg
tcttgggggt ggtctttggg atcctcatca agcgacggca gcagaagatc 1981
cggaagtaca cgatgtga prehumHER2-Q329K, SEQ ID NO: 34 1 atggagctgg
cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc ccccggagcc
61 gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc
cagtcccgag 121 acccacctgg acatgctccg ccacctctac cagggctgcc
aggtggtgca gggaaacctg 181 gaactcacct acctgcccac caatgccagc
ctgtccttcc tgcaggatat ccaggaggtg 241 cagggctacg tgctcatcgc
tcacaaccaa gtgaggcagg tcccactgca gaggctgcgg 301 attgtgcgag
gcacccagct ctttgaggac aactatgccc tggccgtgct agacaatgga 361
gacccgctga acaataccac ccctgtcaca ggggcctccc caggaggcct gcgggagctg
421 cagcttcgaa gcctcacaga gatcttgaaa ggaggggtct tgatccagcg
gaacccccag 481 ctctgctacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggct 541 ctcacactga tagacaccaa ccgctctcgg
gcctgccacc cctgttctcc gatgtgtaag 601 ggctcccgct gctggggaga
gagttctgag gattgtcaga gcctgacgcg cactgtctgt 661 gccggtggct
gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt 721
gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca cttcaaccac
781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct acaacacaga
cacgtttgag 841 tccatgccca atcccgaggg ccggtataca ttcggcgcca
gctgtgtgac tgcctgtccc 901 tacaactacc tttctacgga cgtgggatcc
tgcaccctcg tctgccccct gcacaaccaa 961 gaggtgacag cagaggatgg
aacaAagcgg tgtgagaagt gcagcaagcc ctgtgcccga 1021 gtgtgctatg
gtctgggcat ggagcacttg cgagaggtga gggcagttac cagtgccaat 1081
atccaggagt ttgctggctg caagaagatc tttgggagcc tggcatttct gccggagagc
1141 tttgatgggg acccagcctc caacactgcc ccgctccagc cagagcagct
ccaagtgttt 1201 gagactctgg aagagatcac aggttaccta tacatctcag
catggccgga cagcctgcct 1261 gacctcagcg tcttccagaa cctgcaagta
atccggggac gaattctgca caatggcgcc 1321 tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc actgagggaa 1381 ctgggcagtg
gactggccct catccaccat aacacccacc tctgcttcgt gcacacggtg 1441
ccctgggacc agctctttcg gaacccgcac caagctctgc tccacactgc caaccggcca
1501 gaggacgagt gtgtgggcga gggcctggcc tgccaccagc tgtgcgcccg
agggcactgc 1561 tggggtccag ggcccaccca gtgtgtcaac tgcagccagt
tccttcgggg ccaggagtgc 1621 gtggaggaat gccgagtact gcaggggctc
cccagggagt atgtgaatgc caggcactgt 1681 ttgccgtgcc accctgagtg
tcagccccag aatggctcag tgacctgttt tggaccggag 1741 gctgaccagt
gtgtggcctg tgcccactat aaggaccctc ccttctgcgt ggcccgctgc 1801
cccagcggtg tgaaacctga cctctcctac atgcccatct ggaagtttcc agatgaggag
1861 ggcgcatgcc agccttgccc catcaactgc acccactcct gtgtggacct
ggatgacaag 1921 ggctgccccg ccgagcagag agccagccct ctgacgtcca
tcatctctgc ggtggttggc 1981 attctgctgg tcgtggtctt gggggtggtc
tttgggatcc tcatcaagcg acggcagcag 2041 aagatccgga agtacacgat gtga
prehumHER2-Q429R, SEQ ID NO: 35 1 atggagctgg cggccttgtg ccgctggggg
ctcctcctcg ccctcttgcc ccccggagcc 61 gcgagcaccc aagtgtgcac
cggcacagac atgaagctgc ggctccctgc cagtcccgag 121 acccacctgg
acatgctccg ccacctctac cagggctgcc aggtggtgca gggaaacctg 181
gaactcacct acctgcccac caatgccagc ctgtccttcc tgcaggatat ccaggaggtg
241 cagggctacg tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca
gaggctgcgg 301 attgtgcgag gcacccagct ctttgaggac aactatgccc
tggccgtgct agacaatgga 361 gacccgctga acaataccac ccctgtcaca
ggggcctccc caggaggcct gcgggagctg 421 cagcttcgaa gcctcacaga
gatcttgaaa ggaggggtct tgatccagcg gaacccccag 481 ctctgctacc
aggacacgat tttgtggaag gacatcttcc acaagaacaa ccagctggct 541
ctcacactga tagacaccaa ccgctctcgg gcctgccacc cctgttctcc gatgtgtaag
601 ggctcccgct gctggggaga gagttctgag gattgtcaga gcctgacgcg
cactgtctgt 661 gccggtggct gtgcccgctg caaggggcca ctgcccactg
actgctgcca tgagcagtgt 721 gctgccggct gcacgggccc caagcactct
gactgcctgg cctgcctcca cttcaaccac 781 agtggcatct gtgagctgca
ctgcccagcc ctggtcacct acaacacaga cacgtttgag 841 tccatgccca
atcccgaggg ccggtataca ttcggcgcca gctgtgtgac tgcctgtccc 901
tacaactacc tttctacgga cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa
961 gaggtgacag cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc
ctgtgcccga 1021 gtgtgctatg gtctgggcat ggagcacttg cgagaggtga
gggcagttac cagtgccaat 1081 atccaggagt ttgctggctg caagaagatc
tttgggagcc tggcatttct gccggagagc 1141 tttgatgggg acccagcctc
caacactgcc ccgctccagc cagagcagct ccaagtgttt 1201 gagactctgg
aagagatcac aggttaccta tacatctcag catggccgga cagcctgcct 1261
gacctcagcg tcttccagaa cctgAGagta atccggggac gaattctgca caatggcgcc
1321 tactcgctga ccctgcaagg gctgggcatc agctggctgg ggctgcgctc
actgagggaa 1381 ctgggcagtg gactggccct catccaccat aacacccacc
tctgcttcgt gcacacggtg 1441 ccctgggacc agctctttcg gaacccgcac
caagctctgc tccacactgc caaccggcca 1501 gaggacgagt gtgtgggcga
gggcctggcc tgccaccagc tgtgcgcccg agggcactgc 1561 tggggtccag
ggcccaccca gtgtgtcaac tgcagccagt tccttcgggg ccaggagtgc 1621
gtggaggaat gccgagtact gcaggggctc cccagggagt atgtgaatgc caggcactgt
1681 ttgccgtgcc accctgagtg tcagccccag aatggctcag tgacctgttt
tggaccggag 1741 gctgaccagt gtgtggcctg tgcccactat aaggaccctc
ccttctgcgt ggcccgctgc 1801 cccagcggtg tgaaacctga cctctcctac
atgcccatct ggaagtttcc agatgaggag 1861 ggcgcatgcc agccttgccc
catcaactgc acccactcct gtgtggacct ggatgacaag 1921 ggctgccccg
ccgagcagag agccagccct ctgacgtcca tcatctctgc ggtggttggc 1981
attctgctgg tcgtggtctt gggggtggtc tttgggatcc tcatcaagcg acggcagcag
2041 aagatccgga agtacacgat gtga prehumHER2-N438D SEQ ID NO: 36 1
atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc ccccggagcc
61 gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc
cagtcccgag 121 acccacctgg acatgctccg ccacctctac cagggctgcc
aggtggtgca gggaaacctg 181 gaactcacct acctgcccac caatgccagc
ctgtccttcc tgcaggatat ccaggaggtg 241 cagggctacg tgctcatcgc
tcacaaccaa gtgaggcagg tcccactgca gaggctgcgg 301 attgtgcgag
gcacccagct ctttgaggac aactatgccc tggccgtgct agacaatgga 361
gacccgctga acaataccac ccctgtcaca ggggcctccc caggaggcct gcgggagctg
421 cagcttcgaa gcctcacaga gatcttgaaa ggaggggtct tgatccagcg
gaacccccag 481 ctctgctacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggct 541 ctcacactga tagacaccaa ccgctctcgg
gcctgccacc cctgttctcc gatgtgtaag 601 ggctcccgct gctggggaga
gagttctgag gattgtcaga gcctgacgcg cactgtctgt 661 gccggtggct
gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt 721
gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca cttcaaccac
781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct acaacacaga
cacgtttgag 841 tccatgccca atcccgaggg ccggtataca ttcggcgcca
gctgtgtgac tgcctgtccc 901 tacaactacc tttctacgga cgtgggatcc
tgcaccctcg tctgccccct gcacaaccaa 961 gaggtgacag cagaggatgg
aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga 1021 gtgtgctatg
gtctgggcat ggagcacttg cgagaggtga gggcagttac cagtgccaat 1081
atccaggagt ttgctggctg caagaagatc tttgggagcc tggcatttct gccggagagc
1141 tttgatgggg acccagcctc caacactgcc ccgctccagc cagagcagct
ccaagtgttt 1201 gagactctgg aagagatcac aggttaccta tacatctcag
catggccgga cagcctgcct 1261 gacctcagcg tcttccagaa cctgcaagta
atccggggac gaattctgca cGatggcgcc 1321 tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc actgagggaa 1381 ctgggcagtg
gactggccct catccaccat aacacccacc tctgcttcgt gcacacggtg 1441
ccctgggacc agctctttcg gaacccgcac caagctctgc tccacactgc caaccggcca
1501 gaggacgagt gtgtgggcga gggcctggcc tgccaccagc tgtgcgcccg
agggcactgc 1561 tggggtccag ggcccaccca gtgtgtcaac tgcagccagt
tccttcgggg ccaggagtgc 1621 gtggaggaat gccgagtact gcaggggctc
cccagggagt atgtgaatgc caggcactgt 1681 ttgccgtgcc accctgagtg
tcagccccag aatggctcag tgacctgttt tggaccggag 1741 gctgaccagt
gtgtggcctg tgcccactat aaggaccctc ccttctgcgt ggcccgctgc 1801
cccagcggtg tgaaacctga cctctcctac atgcccatct ggaagtttcc agatgaggag
1861 ggcgcatgcc agccttgccc catcaactgc acccactcct gtgtggacct
ggatgacaag 1921 ggctgccccg ccgagcagag agccagccct ctgacgtcca
tcatctctgc ggtggttggc 1981 attctgctgg tcgtggtctt gggggtggtc
tttgggatcc tcatcaagcg acggcagcag 2041 aagatccgga agtacacgat gtga
prebearHER2, SEQ ID NO: 37 1 melaawcrwg lllallpsga agtqvctgtd
mklrlpaspe thldmlrhly qacqvvqgnl 61 eltylpanas lsflqdiqev
qgyvliahsq vrqvplqrlr ivrgtqlfed nyalavldng 121 eppkgdtsva
gatpgglrel qlrslteilk ggvliqrnpq lchqdtilwk difhknnqla 181
ltlidtnrsr acqpcspack dphcwgassg dcqsltrtvc aggcarckgp kptdccheqc
241 aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt
fgascvtacp 301 ynylstdvgs ctlvcpinnq evtaedgtqr cekcsrpcar
vcyglgmehl rearavtsan 361 iqefagckki fgslaflpes fegdpasnta
plq8peqlrvf ealeeitgyl yisawpdslp 421 nlsvfqnlrv irgrvlhdga
ysltlqglgi swlglrslre lgsglalihr narlcfihtv 481 pweqlfrnph
qallhsanrp eaecvgegla cyplcahghc wgpgptqcvn csqflrgqec 541
veecrelhgl preyvkdryc lpchpecrpq ngsvtcfgse adqcvacahy kdppscvarc
601 psgvkpdlsf mpiwkfadee gtcqpcpinc thscgdlder gcpaeqrasp
vtsiiaavvg 661 illavvmglv lgilikrrrq kirkytm mbearHER2, SEQ ID NO:
38 1 tqvctgtdmk lrlpaspeth ldmlrhlyqa cqvvqgnlel tylpanasls
flqdiqevqg 61 yvliahsqvr qvplqrlriv rgtqlfedny alavldngep
pkgdtsvaga tpgglrelql 121 rslteilkgg vliqrnpqlc hqdtilwkdi
fhknnqlalt lidtnrsrac qpcspackdp 181 hcwgassgdc qsltrtvcag
gcarckgpkp tdccheqcaa gctgpkhsdc laclhfnhsg 241 icelhcpalv
tyntdtfesm pnpegrytfg ascvtacpyn ylstdvgsct lvcpinnqev 301
taedgtqrce kcsrpcarvc yglgmehlre aravtsaniq efagckkifg slaflpesfe
361 gdpasntapl qpeqlrvfea leeitgylyi sawpdslpnl svfqnlrvir
grvlhdgays
421 ltlqglgisw lglrslrelg sglalihrna rlcfihtvpw eqlfrnphqa
llhsanrpea 481 ecvgeglacy plcahghcwg pgptqcvncs qflrgqecve
ecrelhglpr eyvkdryclp 541 chpecrpqng svtcfgsead qcvacahykd
ppscvarcps gvkpdlsfmp iwkfadeegt 601 cqpcpincth scgdldergc
paeqraspvt siiaavvgil lavvmglvlg ilikrrrqki 661 rkytm prefeHER2,
SEQ ID NO: 39 1 melaawcrwg lllallpsga tgtqvctgtd mklrlpaspe
thldmlrhly qgcqvvqgnl 61 eltylhanas lsflqdiqev qgyvliahnq
vkqvplqrlr ivrgtqlfed nyalavldng 121 dpldsgtpat gaalgglrel
qlrslteilk ggvliqrnpq lchqdtilwk difhknnqla 181 lmlidtnrsr
acqpcspack dshcwgassg dcqsltrtvc aggcarckgp qptdccheqc 241
aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp
301 ynylstdvgs ctlvcpinnq evtaedgtqr cekcskpcar vcyglgmehl
rearavtsan 361 iqefvgckki fgslaflpes fegdpasnta plqpeqlrvf
ealeeitgyl yisawpdslp 421 nlsvfqnlrv irgrvlhdga ysltlqglgi
swlglrslre lgsglalihr nsrlcfvhtv 481 pwdqlfrnph qallhsanrp
edecagegla cyplcahghc wgpgptqcvn csqflrgqec 541 veecrvlqgl
preyvkdrfc lpchpecqpq ngsvtclgse adqcvacahy kdppfcvarc 601
psgvkpdlsf mpiwkfadee gtcqpcpinc thscadldek gcpaeqrasp vtsiiaavvg
661 illvvvvglv lgilikrrrq kirkytm mfeHER2 SEQ ID NO: 40 1
tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tylhanasls flqdiqevqg
61 yvliahnqvk qvplqrlriv rgtqlfedny alavldngdp ldsgtpatga
algglrelql 121 rslteilkgg vliqrnpqlc hqdtilwkdi fhknnqlalm
lidtnrsrac qpcspackds 181 hcwgassgdc qsltrtvcag gcarckgpqp
tdccheqcaa gctgpkhsdc laclhfnhsg 241 icelhcpalv tyntdtfesm
pnpegrytfg ascvtacpyn ylstdvgsct lvcpinnqev 301 taedgtqrce
kcskpcarvc yglgmehlre aravtsaniq efvgckkifg slaflpesfe 361
gdpasntapl qpeqlrvfea leeitgylyi sawpdslpnl svfqnlrvir grvlhdgays
421 ltlqglgisw lglrslrelg sglalihrns rlcfvhtvpw dqlfrnphqa
llhsanrped 481 ecageglacy plcahghcwg pgptqcvncs qflrgqecve
ecrvlqglpr eyvkdrfclp 541 chpecqpqng svtclgsead qcvacahykd
ppfcvarcps gvkpdlsfmp iwkfadeegt 601 cqpcpincth scadldekgc
paeqraspvt siiaavvgil lvvvvglvlg ilikrrrqki 661 rkytm prebearHER2,
SEQ ID NO: 41 1 atggagctgg cggcctggtg ccgctggggg ctcctcctcg
ccctcctgcc ctccggagcc 61 gcgggcaccc aagtgtgcac cggcacagac
atgaagctgc ggctccctgc cagtcccgag 121 acccacctgg atatgctccg
ccacctctac caggcctgtc aagtggtaca gggtaacctg 181 gagctcacct
acctgcccgc caatgccagc ctgtccttcc tgcaggatat ccaggaggta 241
cagggctatg tgctcattgc tcacagccaa gtgagacagg tcccgctgca gaggctccga
301 atcgtgcgag gcacccagct ctttgaggac aactacgccc tggccgtgct
ggacaatgga 361 gagccgccca agggggacac ctctgtggca ggggctaccc
caggagggct gcgggagctg 421 cagcttcgaa gcctcacaga gatcctgaag
ggaggggtct tgattcagcg gaacccacag 481 ctctgccacc aggacacgat
tttgtggaag gacatcttcc acaagaacaa ccagctggcc 541 ctcacgctga
tagacaccaa ccgctctcgg gcctgccaac cctgttctcc agcctgtaaa 601
gacccccact gctggggagc aagttccggg gactgtcaga gcttgacacg aaccgtctgt
661 gccggcggct gtgcccgctg caagggccca aaacccactg actgctgcca
tgagcagtgc 721 gcggctggct gcacgggccc caagcactcg gactgcctgg
cctgccttca cttcaaccac 781 agtggcatct gtgagctgca ctgcccagcc
ctggtcacct acaacacgga cacgttcgaa 841 tccatgccca accctgaggg
ccgatacacc ttcggtgcca gctgtgtgac cgcctgtccc 901 tacaactacc
tgtccacgga cgtgggatcc tgcaccctgg tctgtcccct gaacaaccaa 961
gaggtgacgg ctgaggatgg cacccagcgg tgtgagaaat gcagcagacc ctgtgcccga
1021 gtgtgctatg gtctgggcat ggagcacctg cgggaggcga gggcggtcac
cagcgccaac 1081 atccaagagt tcgccggctg caagaagatc tttgggagcc
tggcgtttct gccagagagc 1141 ttcgagggag acccagcctc caacactgcc
cccctgcagc ctgaacagct cagagtgttc 1201 gaggccctgg aggagatcac
aggttacctg tatatctcag cgtggccgga cagcttgcct 1261 aacctcagtg
tcttccagaa cctgcgagta atccggggac gagttctgca tgatggcgcc 1321
tactcgctga ccctgcaagg gctgggcatc agctggctgg ggctgcgctc gctgcgggaa
1381 ctgggcagcg ggctggccct catccaccgc aacgcccgcc tctgcttcat
ccacacggtg 1441 ccctgggagc agctcttccg gaacccccac caagccctgc
tgcacagtgc caaccggccg 1501 gaggccgagt gcgtgggcga gggcctggcc
tgctacccgc tgtgcgccca tgggcactgc 1561 tggggtccgg ggcccaccca
gtgcgtcaac tgcagccaat tccttcgggg ccaggagtgc 1621 gtggaggaat
gccgagaact gcacgggcta ccccgggaat atgtgaagga cagatactgt 1681
ctgccatgcc accccgagtg tcggccccag aatggctcag tgacctgctt tgggtcggag
1741 gctgaccagt gtgtggcctg cgcccactac aaggaccctc cctcctgcgt
ggctcgctgc 1801 cccagtggtg tgaaacccga cctctctttc atgcccattt
ggaagtttgc agatgaggag 1861 ggcacatgcc agccgtgccc catcaactgc
acccactcct gtggggacct ggacgagagg 1921 ggctgccccg ccgaacagag
agccagccct gtgacatcca tcattgccgc tgtggtgggc 1981 attctgctgg
ccgtggtcat ggggctggtc ctcggcatcc tgatcaagcg aaggcgacag 2041
aagatccgga agtacacgat g mbearHER2. SEQ ID NO: 42 1 acccaagtgt
gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac 61
ctggatatgc tccgccacct ctaccaggcc tgtcaagtgg tacagggtaa cctggagctc
121 acctacctgc ccgccaatgc cagcctgtcc ttcctgcagg atatccagga
ggtacagggc 181 tatgtgctca ttgctcacag ccaagtgaga caggtcccgc
tgcagaggct ccgaatcgtg 241 cgaggcaccc agctctttga ggacaactac
gccctggccg tgctggacaa tggagagccg 301 cccaaggggg acacctctgt
ggcaggggct accccaggag ggctgcggga gctgcagctt 361 cgaagcctca
cagagatcct gaagggaggg gtcttgattc agcggaaccc acagctctgc 421
caccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct ggccctcacg
481 ctgatagaca ccaaccgctc tcgggcctgc caaccctgtt ctccagcctg
taaagacccc 541 cactgctggg gagcaagttc cggggactgt cagagcttga
cacgaaccgt ctgtgccggc 601 ggctgtgccc gctgcaaggg cccaaaaccc
actgactgct gccatgagca gtgcgcggct 661 ggctgcacgg gccccaagca
ctcggactgc ctggcctgcc ttcacttcaa ccacagtggc 721 atctgtgagc
tgcactgccc agccctggtc acctacaaca cggacacgtt cgaatccatg 781
cccaaccctg agggccgata caccttcggt gccagctgtg tgaccgcctg tccctacaac
841 tacctgtcca cggacgtggg atcctgcacc ctggtctgtc ccctgaacaa
ccaagaggtg 901 acggctgagg atggcaccca gcggtgtgag aaatgcagca
gaccctgtgc ccgagtgtgc 961 tatggtctgg gcatggagca cctgcgggag
gcgagggcgg tcaccagcgc caacatccaa 1021 gagttcgccg gctgcaagaa
gatctttggg agcctggcgt ttctgccaga gagcttcgag 1081 ggagacccag
cctccaacac tgcccccctg cagcctgaac agctcagagt gttcgaggcc 1141
ctggaggaga tcacaggtta cctgtatatc tcagcgtggc cggacagctt gcctaacctc
1201 agtgtcttcc agaacctgcg agtaatccgg ggacgagttc tgcatgatgg
cgcctactcg 1261 ctgaccctgc aagggctggg catcagctgg ctggggctgc
gctcgctgcg ggaactgggc 1321 agcgggctgg ccctcatcca ccgcaacgcc
cgcctctgct tcatccacac ggtgccctgg 1381 gagcagctct tccggaaccc
ccaccaagcc ctgctgcaca gtgccaaccg gccggaggcc 1441 gagtgcgtgg
gcgagggcct ggcctgctac ccgctgtgcg cccatgggca ctgctggggt 1501
ccggggccca cccagtgcgt caactgcagc caattccttc ggggccagga gtgcgtggag
1561 gaatgccgag aactgcacgg gctaccccgg gaatatgtga aggacagata
ctgtctgcca 1621 tgccaccccg agtgtcggcc ccagaatggc tcagtgacct
gctttgggtc ggaggctgac 1681 cagtgtgtgg cctgcgccca ctacaaggac
cctccctcct gcgtggctcg ctgccccagt 1741 ggtgtgaaac ccgacctctc
tttcatgccc atttggaagt ttgcagatga ggagggcaca 1801 tgccagccgt
gccccatcaa ctgcacccac tcctgtgggg acctggacga gaggggctgc 1861
cccgccgaac agagagccag ccctgtgaca tccatcattg ccgctgtggt gggcattctg
1921 ctggccgtgg tcatggggct ggtcctcggc atcctgatca agcgaaggcg
acagaagatc 1981 cggaagtaca cgatg prefeHER2, SEQ ID NO: 43 1
atggagctgg cggcctggtg ccgctggggg ctcctcctcg ccctcctgcc ctccggagcc
61 acgggcaccc aagtgtgcac cggcacagac atgaagctgc ggctcccagc
cagtcccgag 121 acccacctgg acatgctccg ccacctctac cagggctgtc
aagtggtaca gggcaacctg 181 gagctcacct acctgcatgc caatgccagc
ctctccttcc tgcaggatat ccaggaggtg 241 caaggctatg tgctcattgc
ccacaaccaa gtgaaacagg tcccactgca gaggctacga 301 atcgtgcgag
gcacccagct ctttgaggac aactacgccc tggccgtgct ggacaacgga 361
gacccactgg acagtggcac ccctgctaca ggggctgccc taggagggct gcgggagctg
421 cagctccgaa gcctcacaga gatcctgaag ggaggggtcc tcattcagcg
gaacccgcag 481 ctctgccacc aggacacgat tctgtggaag gacatcttcc
acaagaacaa ccagctggcc 541 ctcatgctga tagacaccaa ccgctctcgg
gcctgccaac cctgttctcc agcttgtaaa 601 gactcccact gctggggagc
aagttccggg gactgtcaga gcttgactcg aactgtctgt 661 gctggcggct
gtgcccgctg caagggcccg cagcccaccg actgctgcca cgagcaatgt 721
gctgctggct gcacgggccc caagcattct gactgcctgg cctgcctcca cttcaaccac
781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct acaacacgga
caccttcgaa 841 tccatgccca accctgaggg ccgttatacc ttcggtgcca
gctgtgtgac tgcctgtccc 901 tacaactacc tgtctacgga cgtgggatcc
tgcaccctgg tctgtcccct gaacaaccaa 961 gaggtgacag ctgaggatgg
aacacagcgg tgtgagaaat gcagcaagcc ctgtgcccga 1021 gtgtgctacg
gcctaggcat ggagcacctg cgggaggcga gggcagtcac cagtgccaac 1081
atccaagaat ttgtcggctg caagaagatc tttgggagcc tggcgtttct gccagagagc
1141 tttgaggggg acccagcctc caacactgcc cccctgcagc ctgagcagct
cagagtgttt 1201 gaggctctgg aggagattac aggttacctg tacatctcag
cgtggccaga cagcttgcct 1261 aacctcagtg tcttccagaa cctcagagtg
atccggggcc gagttctgca tgacggtgct 1321 tactcgctga cccttcaagg
gctgggcatc agctggctgg ggctgcgctc gctgcgggag 1381 ctgggcagtg
ggctggccct catccaccgc aactcccgcc tctgcttcgt acacacggtg 1441
ccctgggacc agctcttccg gaacccccac caggccctgc tccacagcgc caaccggcca
1501 gaggacgagt gcgcgggtga gggcctggcc tgctatccgc tgtgtgccca
cgggcactgc
1561 tggggtccgg gacccaccca gtgtgtcaac tgcagccagt tccttcgggg
ccaggagtgc 1621 gtggaggaat gccgagtatt gcaggggctt ccccgggagt
atgtgaagga taggttctgt 1681 ctgccatgcc acccggagtg tcagccccag
aatggctcag tgacctgctt gggctcggaa 1741 gctgaccagt gtgtggcctg
tgcccactac aaggaccctc ctttctgtgt ggctcgctgc 1801 cccagtgggg
tgaaacctga cctctccttc atgcccatct ggaagttcgc agatgaggag 1861
ggcacgtgcc agccatgccc catcaactgc acccactcct gtgcggacct ggacgagaag
1921 ggctgccccg ccgagcagag agccagccct gtgacgtcca tcattgctgc
tgtggtgggc 1981 attctgctgg tcgtggttgt ggggctggtc cttggcatcc
taatcaagcg aaggcggcag 2041 aagatccgga agtacacgat g mfeHER2, SEQ ID
NO: 44 1 acccaagtgt gcaccggcac agacatgaag ctgcggctcc cagccagtcc
cgagacccac 61 ctggacatgc tccgccacct ctaccagggc tgtcaagtgg
tacagggcaa cctggagctc 121 acctacctgc atgccaatgc cagcctctcc
ttcctgcagg atatccagga ggtgcaaggc 181 tatgtgctca ttgcccacaa
ccaagtgaaa caggtcccac tgcagaggct acgaatcgtg 241 cgaggcaccc
agctctttga ggacaactac gccctggccg tgctggacaa cggagaccca 301
ctggacagtg gcacccctgc tacaggggct gccctaggag ggctgcggga gctgcagctc
361 cgaagcctca cagagatcct gaagggaggg gtcctcattc agcggaaccc
gcagctctgc 421 caccaggaca cgattctgtg gaaggacatc ttccacaaga
acaaccagct ggccctcatg 481 ctgatagaca ccaaccgctc tcgggcctgc
caaccctgtt ctccagcttg taaagactcc 541 cactgctggg gagcaagttc
cggggactgt cagagcttga ctcgaactgt ctgtgctggc 601 ggctgtgccc
gctgcaaggg cccgcagccc accgactgct gccacgagca atgtgctgct 661
ggctgcacgg gccccaagca ttctgactgc ctggcctgcc tccacttcaa ccacagtggc
721 atctgtgagc tgcactgccc agccctggtc acctacaaca cggacacctt
cgaatccatg 781 cccaaccctg agggccgtta taccttcggt gccagctgtg
tgactgcctg tccctacaac 841 tacctgtcta cggacgtggg atcctgcacc
ctggtctgtc ccctgaacaa ccaagaggtg 901 acagctgagg atggaacaca
gcggtgtgag aaatgcagca agccctgtgc ccgagtgtgc 961 tacggcctag
gcatggagca cctgcgggag gcgagggcag tcaccagtgc caacatccaa 1021
gaatttgtcg gctgcaagaa gatctttggg agcctggcgt ttctgccaga gagctttgag
1081 ggggacccag cctccaacac tgcccccctg cagcctgagc agctcagagt
gtttgaggct 1141 ctggaggaga ttacaggtta cctgtacatc tcagcgtggc
cagacagctt gcctaacctc 1201 agtgtcttcc agaacctcag agtgatccgg
ggccgagttc tgcatgacgg tgcttactcg 1261 ctgacccttc aagggctggg
catcagctgg ctggggctgc gctcgctgcg ggagctgggc 1321 agtgggctgg
ccctcatcca ccgcaactcc cgcctctgct tcgtacacac ggtgccctgg 1381
gaccagctct tccggaaccc ccaccaggcc ctgctccaca gcgccaaccg gccagaggac
1441 gagtgcgcgg gtgagggcct ggcctgctat ccgctgtgtg cccacgggca
ctgctggggt 1501 ccgggaccca cccagtgtgt caactgcagc cagttccttc
ggggccagga gtgcgtggag 1561 gaatgccgag tattgcaggg gcttccccgg
gagtatgtga aggataggtt ctgtctgcca 1621 tgccacccgg agtgtcagcc
ccagaatggc tcagtgacct gcttgggctc ggaagctgac 1681 cagtgtgtgg
cctgtgccca ctacaaggac cctcctttct gtgtggctcg ctgccccagt 1741
ggggtgaaac ctgacctctc cttcatgccc atctggaagt tcgcagatga ggagggcacg
1801 tgccagccat gccccatcaa ctgcacccac tcctgtgcgg acctggacga
gaagggctgc 1861 cccgccgagc agagagccag ccctgtgacg tccatcattg
ctgctgtggt gggcattctg 1921 ctggtcgtgg ttgtggggct ggtccttggc
atcctaatca agcgaaggcg gcagaagatc 1981 cggaagtaca cgatg prehumHER2,
SEQ ID NO: 45 1 melaalcrwg lllallppga astqvctgtd mklrlpaspe
thldmlrhly qgcqvvqgnl 61 eltylptnas lsflqdiqev qgyvliahnq
vrqvplqrlr ivrgtqlfed nyalavldng 121 dpinnttpvt gaspgglrel
qlrslteilk ggvliqrnpq lcyqdtilwk difhknnqla 181 ltlidtnrsr
achpcspmck gsrcwgesse dcqsltrtvc aggcarckgp 1ptdccheqc 241
aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp
301 ynylstdvgs ctivcplhnq evtaedgtqr cekcskpcar vcyglgmehl
revravtsan 361 igefagckki fgslaflpes fdgdpasnta plqpeqlqvf
etleeitgyl yisawpdslp 421 dlsvfqnlqv irgrilhnga ysltlqglgi
swlglrslre lgsglalihh nthlcfvhtv 481 pwdqlfrnph qallhtanrp
edecvgegla chqlcarghc wgpgptqcvn csqflrgqec 541 veecrvlqgl
preyvnarhc lpchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc 601
psgvkpdlsy mpiwkfpdee gacqpcpinc thscvdlddk gcpaeqrasp ltsiisavvg
661 illvvvlgvv fgilikrrqq kirkytm mhumHER2, SEQ ID NO: 46 1
tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tylptnasls flqdiqevqg
61 yvliahnqvr qvplqrlriv rgtqlfedny alavldngdp lnnttpvtga
spgglrelql 121 rslteilkgg vliqrnpqlc yqdtilwkdi fhknnqlalt
lidtnrsrac hpcspmckgs 181 rcwgessedc qsltrtvcag gcarckgplp
tdccheqcaa gctgpkhsdc laclhfnhsg 241 icelhcpalv tyntdtfesm
pnpegrytfg ascvtacpyn ylstdvgsct lvcplhnqev 301 taedgtqrce
kcskpcarvc yglgmehlre vravtsaniq efagckkifg slaflpesfd 361
gdpasntapl qpeqlqvfet leeitgylyi sawpdslpdl svfqnlqvir grilhngays
421 ltlqglgisw lglrslrelg sglalihhnt hlcfvhtvpw dqlfrnphqa
llhtanrped 481 ecvgeglach qlcarghcwg pgptqcvncs qflrgqecve
ecrvlqglpr eyvnarhclp 541 chpecqpqng svtcfgpead qcvacahykd
ppfcvarcps gvkpdlsymp iwkfpdeega 601 cqpcpincth scvdlddkgc
paeqrasplt siisavvgil lvvvlgvvfg ilikrrqqki 661 rkytm premouseHER2,
SEQ ID NO: 47 1 melaawcrwg fllallspga agtqvctgtd mklrlpaspe
thldmlrhly qgcqvvqgnl 61 eltylpanas lsflqdiqev qgymliahnr
vkhvplqrlr ivrgtqlfed kyalavldnr 121 dpldnvttaa pgrtpeglre
lqlrslteil kggvlirgnp qlcyqdmvlw kdvlrknnql 181 apvdmdtnrs
racppcaptc kdnhcwgesp edcqiltgti ctsgcarckg rlptdccheq 241
caagctgpkh sdclaclhfn hsgicelhcp alityntdtf esmlnpegry tfgascvttc
301 pynylstevg sctivcppnn qevtaedgtq rcekcskpca gvcyglgmeh
lrgaraitsd 361 niqefagckk ifgslaflpe sfdgnpssgv aplkpehlqv
fetleeitgy lyisawpesf 421 qdlsvfqnlr virgrilhdg aysltlqglg
ihslglrslr elgsglalih rnthlcfvnt 481 vpwdqlfrnp hqallhsgnr
peeacglegl vcnslcargh cwgpgptqcv ncsqflrgqe 541 cveecrvwkg
1preyvrgkh clpchpecqp qnssetcygs eadqceacah ykdssscvar 601
cpsgvkpdls ympiwkypde egicqpcpin cthscvdlde rgcpaeqras pvtfiiatvv
661 gvllfliivv vigilikrrr qkirkytm mmouseHER2, SEQ ID NO: 48 1
tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tylpanasls flqdiqevqg
61 ymliahnrvk hvplqrlriv rgtqlfedky alavldnrdp ldnvttaapg
rtpeglrelq 121 lrslteilkg gvlirgnpql cyqdmvlwkd vlrknnqlap
vdmdtnrsra cppcaptckd 181 nhcwgesped cqiltgtict sgcarckgrl
ptdccheqca agctgpkhsd claclhfnhs 241 gicelhcpal ityntdtfes
mlnpegrytf gascvttcpy nylstevgsc tivcppnnqe 301 vtaedgtqrc
ekcskpcagv cyglgmehlr garaitsdni qefagokkif gslaflpesf 361
dgnpssgvap lkpehlqvfe tleeitgyly isawpesfqd lsvfqnlrvi rgrilhdgay
421 sltlqglgih slglrslrel gsglalihrn thlcfvntvp wdqlfrnphq
allhsgnrpe 481 eacgleglvc nslcarghcw gpgptqcvnc sqflrgqecv
eecrvwkglp reyvrgkhcl 541 pchpecqpqn ssetcygsea dqceacahyk
dssscvarcp sgvkpdlsym piwkypdeeg 601 icqpcpinct hscvdlderg
cpaeqraspv tfiiatvvgv llfliivvvi gilikrrrqk 661 irkytm preratHER2,
SEQ ID NO: 49 1 miimelaawc rwgfllallp pgiagtqvct gtdmklrlpa
spethldmlr hlyqgcqvvq 61 gnleltyvpa naslsflqdi qevqgymlia
hnqvkrvplq rlrivrgtql fedkyalavl 121 dnrdpqdnva astpgrtpeg
lrelqlrslt eilkggvlir gnpqlcyqdm vlwkdvfrkn 181 nqlapvdidt
nrsracppca packdnhcwg espedcqilt gtictsgcar ckgrlptdcc 241
heqcaagctg pkhsdclacl hfnhsgicel hcpalvtynt dtfesmhnpe grytfgascv
301 ttcpynylst evgsctivcp pnnqevtaed gtqrcekcsk pcarvcyglg
mehlrgarai 361 tsdnvqefdg ckkifgslaf lpesfdgdps sgiaplrpeq
lqvfetleei tgylyisawp 421 dslrdlsvfq nlriirgril hdgaysltlq
glgihslglr slrelgsgla lihrnahlcf 481 vhtvpwdqlf rnphqallhs
gnrpeedcgl eglvcnslca hghcwgpgpt qcvncshflr 541 gqecveecry
wkglpreyvs dkrclpchpe cqpqnssetc fgseadqcaa cahykdsssc 601
varcpsgvkp dlsympiwky pdeegicqpc pincthscvd ldergcpaeq raspvtfiia
661 tvvgvllfli lvvvvgilik rrrqkirkyt m mratHER2, SEQ ID NO: 50 1
tqvctgtdmk lrlpaspeth ldmlrhlyqg cqvvqgnlel tyvpanasls flqdiqevqg
61 ymliahnqvk rvplqrlriv rgtqlfedky alavldnrdp qdnvaastpg
rtpeglrelq 121 lrslteilkg gvlirgnpql cyqdmvlwkd vfrknnqlap
vdidtnrsra cppcapackd 181 nhcwgesped cqiltgtict sgcarckgrl
ptdccheqca agctgpkhsd claclhfnhs 241 gicelhcpal vtyntdtfes
mhnpegrytf gascvttcpy nylstevgsc tivcppnnqe 301 vtaedgtqrc
ekcskpcary cyglgmehlr garaitsdnv qefdgckkif gslaflpesf 361
dgdpssgiap lrpeqlqvfe tleeitgyly isawpdslrd lsvfqnlrii rgrilhdgay
421 sltlqglgih slglrslrel gsglalihrn ahlcfvhtvp wdqlfrnphq
allhsgnrpe 481 edcgleglvc nslcahghcw gpgptqcvnc shflrgqecv
eecrvwkglp reyvsdkrcl 541 pchpecqpqn ssetcfgsea dqcaacahyk
dssscvarcp sgvkpdlsym piwkypdeeg 601 icqpcpinct hscvdlderg
cpaeqraspv tfiiatvvgv llflilvvvv gilikrrrqk 661 irkytm Precursor:
preE2Neu SEQ ID NO: 51 1 melaalcrwg lllallppga astqvctgtd
mklrlpaspe thldmlrhly qgcqvvqgnl 61 eltylptnas lsflqdiqev
qgyvliahnq vrqvplqrlr ivrgtqlfed nyalavldng 121 dpinnttpvt
gaspgglrel qlrslteilk ggvliqrnpq lcyqdtilwk difhknnqla 181
ltlidtnrsr achpcspmck gsrcwgesse dcqsltrtvc aggcarckgp 1ptdccheqc
241 aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt
fgascvtacp 301 ynylstdvgs ctivcplhnq evtaedgtqr cekcskpcar
vcyglgmehl revravtsan
361 igefagckki fgslaflpes fdgdpasnta efaplrpeql qvfetleeit
gylyisawpd 421 slrdlsvfqn lriirgrilh dgaysltlqg lgihslglrs
lrelgsglal ihrnahlcfv 481 htvpwdqlfr nphqallhsg nrpeedcgle
glvcnslcah ghcwgpgptq cvncshflrg 541 qecveecrvw kglpreyvsd
krclpchpec qpqnssetcf gseadqcaac ahykdssscv 601 arcpsgvkpd
lsympiwkyp deegicqpcp incthscvdl dergcpaeqr aspvtfiiat 661
vvgvllflil vvvvgilikr rrqkirkytm mE2Neu, SEQ ID NO: 52 1 tqvctgtdmk
lrlpaspeth ldmlrhlyqg cqvvqgnlel tylptnasls flqdiqevqg 61
yvliahnqvr qvplqrlriv rgtqlfedny alavldngdp lnnttpvtga spgglrelql
121 rslteilkgg vliqrnpqlc yqdtilwkdi fhknnqlalt lidtnrsrac
hpcspmckgs 181 rcwgessedc qsltrtvcag gcarckgplp tdccheqcaa
gctgpkhsdc laclhfnhsg 241 icelhcpalv tyntdtfesm pnpegrytfg
ascvtacpyn ylstdvgsct lvcplhnqev 301 taedgtqrce kcskpcarvc
yglgmehlre vravtsaniq efagckkifg slaflpesfd 361 gdpasntaef
aplrpeqlqv fetleeitgy lyisawpdsl rdlsvfqnlr iirgrilhdg 421
aysltlqglg ihslglrslr elgsglalih rnahlcfvht vpwdqlfrnp hqallhsgnr
481 peedcglegl vcnslcahgh cwgpgptqcv ncshflrgqe cveecrvwkg
lpreyvsdkr 541 clpchpecqp qnssetcfgs eadqcaacah ykdssscvar
cpsgvkpdls ympiwkypde 601 egicqpcpin cthscvdlde rgcpaeqras
pvtfiiatvv gvllflilvv vvgilikrrr 661 qkirkytm prehumHER2, SEQ ID
NO: 53 1 atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc
ccccggagcc 61 gcgagcaccc aagtgtgcac cggcacagac atgaagctgc
ggctccctgc cagtcccgag 121 acccacctgg acatgctccg ccacctctac
cagggctgcc aggtggtgca gggaaacctg 181 gaactcacct acctgcccac
caatgccagc ctgtccttcc tgcaggatat ccaggaggtg 241 cagggctacg
tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca gaggctgcgg 301
attgtgcgag gcacccagct ctttgaggac aactatgccc tggccgtgct agacaatgga
361 gacccgctga acaataccac ccctgtcaca ggggcctccc caggaggcct
gcgggagctg 421 cagcttcgaa gcctcacaga gatcttgaaa ggaggggtct
tgatccagcg gaacccccag 481 ctctgctacc aggacacgat tttgtggaag
gacatcttcc acaagaacaa ccagctggct 541 ctcacactga tagacaccaa
ccgctctcgg gcctgccacc cctgttctcc gatgtgtaag 601 ggctcccgct
gctggggaga gagttctgag gattgtcaga gcctgacgcg cactgtctgt 661
gccggtggct gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt
721 gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca
cttcaaccac 781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacaga cacgtttgag 841 tccatgccca atcccgaggg ccggtataca
ttcggcgcca gctgtgtgac tgcctgtccc 901 tacaactacc tttctacgga
cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa 961 gaggtgacag
cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga 1021
gtgtgctatg gtctgggcat ggagcacttg cgagaggtga gggcagttac cagtgccaat
1081 atccaggagt ttgctggctg caagaagatc tttgggagcc tggcatttct
gccggagagc 1141 tttgatgggg acccagcctc caacactgcc ccgctccagc
cagagcagct ccaagtgttt 1201 gagactctgg aagagatcac aggttaccta
tacatctcag catggccgga cagcctgcct 1261 gacctcagcg tcttccagaa
cctgcaagta atccggggac gaattctgca caatggcgcc 1321 tactcgctga
ccctgcaagg gctgggcatc agctggctgg ggctgcgctc actgagggaa 1381
ctgggcagtg gactggccct catccaccat aacacccacc tctgcttcgt gcacacggtg
1441 ccctgggacc agctctttcg gaacccgcac caagctctgc tccacactgc
caaccggcca 1501 gaggacgagt gtgtgggcga gggcctggcc tgccaccagc
tgtgcgcccg agggcactgc 1561 tggggtccag ggcccaccca gtgtgtcaac
tgcagccagt tccttcgggg ccaggagtgc 1621 gtggaggaat gccgagtact
gcaggggctc cccagggagt atgtgaatgc caggcactgt 1681 ttgccgtgcc
accctgagtg tcagccccag aatggctcag tgacctgttt tggaccggag 1741
gctgaccagt gtgtggcctg tgcccactat aaggaccctc ccttctgcgt ggcccgctgc
1801 cccagcggtg tgaaacctga cctctcctac atgcccatct ggaagtttcc
agatgaggag 1861 ggcgcatgcc agccttgccc catcaactgc acccactcct
gtgtggacct ggatgacaag 1921 ggctgccccg ccgagcagag agccagccct
ctgacgtcca tcatctctgc ggtggttggc 1981 attctgctgg tcgtggtctt
gggggtggtc tttgggatcc tcatcaagcg acggcagcag 2041 aagatccgga
agtacacgat gtga mhumHER2, SEQ ID NO: 54 1 acccaagtgt gcaccggcac
agacatgaag ctgcggctcc ctgccagtcc cgagacccac 61 ctggacatgc
tccgccacct ctaccagggc tgccaggtgg tgcagggaaa cctggaactc 121
acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg atatccagga ggtgcagggc
181 tacgtgctca tcgctcacaa ccaagtgagg caggtcccac tgcagaggct
gcggattgtg 241 cgaggcaccc agctctttga ggacaactat gccctggccg
tgctagacaa tggagacccg 301 ctgaacaata ccacccctgt cacaggggcc
tccccaggag gcctgcggga gctgcagctt 361 cgaagcctca cagagatctt
gaaaggaggg gtcttgatcc agcggaaccc ccagctctgc 421 taccaggaca
cgattttgtg gaaggacatc ttccacaaga acaaccagct ggctctcaca 481
ctgatagaca ccaaccgctc tcgggcctgc cacccctgtt ctccgatgtg taagggctcc
541 cgctgctggg gagagagttc tgaggattgt cagagcctga cgcgcactgt
ctgtgccggt 601 ggctgtgccc gctgcaaggg gccactgccc actgactgct
gccatgagca gtgtgctgcc 661 ggctgcacgg gccccaagca ctctgactgc
ctggcctgcc tccacttcaa ccacagtggc 721 atctgtgagc tgcactgccc
agccctggtc acctacaaca cagacacgtt tgagtccatg 781 cccaatcccg
agggccggta tacattcggc gccagctgtg tgactgcctg tccctacaac 841
tacctttcta cggacgtggg atcctgcacc ctcgtctgcc ccctgcacaa ccaagaggtg
901 acagcagagg atggaacaca gcggtgtgag aagtgcagca agccctgtgc
ccgagtgtgc 961 tatggtctgg gcatggagca cttgcgagag gtgagggcag
ttaccagtgc caatatccag 1021 gagtttgctg gctgcaagaa gatctttggg
agcctggcat ttctgccgga gagctttgat 1081 ggggacccag cctccaacac
tgccccgctc cagccagagc agctccaagt gtttgagact 1141 ctggaagaga
tcacaggtta cctatacatc tcagcatggc cggacagcct gcctgacctc 1201
agcgtcttcc agaacctgca agtaatccgg ggacgaattc tgcacaatgg cgcctactcg
1261 ctgaccctgc aagggctggg catcagctgg ctggggctgc gctcactgag
ggaactgggc 1321 agtggactgg ccctcatcca ccataacacc cacctctgct
tcgtgcacac ggtgccctgg 1381 gaccagctct ttcggaaccc gcaccaagct
ctgctccaca ctgccaaccg gccagaggac 1441 gagtgtgtgg gcgagggcct
ggcctgccac cagctgtgcg cccgagggca ctgctggggt 1501 ccagggccca
cccagtgtgt caactgcagc cagttccttc ggggccagga gtgcgtggag 1561
gaatgccgag tactgcaggg gctccccagg gagtatgtga atgccaggca ctgtttgccg
1621 tgccaccctg agtgtcagcc ccagaatggc tcagtgacct gttttggacc
ggaggctgac 1681 cagtgtgtgg cctgtgccca ctataaggac cctcccttct
gcgtggcccg ctgccccagc 1741 ggtgtgaaac ctgacctctc ctacatgccc
atctggaagt ttccagatga ggagggcgca 1801 tgccagcctt gccccatcaa
ctgcacccac tcctgtgtgg acctggatga caagggctgc 1861 cccgccgagc
agagagccag ccctctgacg tccatcatct ctgcggtggt tggcattctg 1921
ctggtcgtgg tcttgggggt ggtctttggg atcctcatca agcgacggca gcagaagatc
1981 cggaagtaca cgatgtga premouseHER2, SEQ ID NO: 55 1 atggagctgg
cggcctggtg ccgttggggg ttcctcctcg ccctcctgtc ccccggagcc 61
gcgggtaccc aagtgtgtac cggtaccgac atgaagttgc gactccctgc cagtcctgag
121 acccacctgg acatgcttcg ccacctctac cagggctgtc aggtggtgca
gggcaatttg 181 gagcttacct acctgcccgc caatgccagc ctctcattcc
tgcaggacat ccaggaagtc 241 cagggataca tgctcatcgc tcacaaccga
gtgaaacacg tcccactgca gaggttgcgc 301 atcgtgagag ggactcagct
ctttgaggac aagtatgccc tggctgtgct agacaaccga 361 gaccctttgg
acaacgtcac caccgccgcc ccaggcagaa ccccagaagg gctgcgggag 421
ctgcagcttc gaagtctcac agagatcttg aagggaggag ttttgatccg tgggaaccct
481 cagctctgct accaggacat ggttttgtgg aaggatgtcc tccgtaagaa
taaccagctg 541 gctcctgtcg acatggacac caatcgttcc cgggcctgtc
caccttgtgc cccaacctgc 601 aaagacaatc actgttgggg tgagagtcct
gaagactgtc agatcttgac tggcaccatc 661 tgtactagtg gctgtgcccg
gtgcaagggc cggctgccca ctgactgttg ccatgagcag 721 tgtgctgcag
gctgcacggg tcccaagcat tctgactgcc tggcctgcct ccacttcaat 781
catagtggta tctgtgagct gcactgcccg gccctcatca cctacaacac agacaccttc
841 gagtccatgc tcaaccctga gggtcgctac acctttggtg ccagctgtgt
gaccacctgc 901 ccctacaact acctctccac ggaagtggga tcctgcactc
tggtctgtcc cccgaacaac 961 caagaggtca cagctgagga cggaacacag
cggtgtgaga aatgcagcaa gccctgtgct 1021 ggagtatgct atggtctggg
catggagcac ctccgagggg cgagggccat caccagtgac 1081 aatatccagg
agtttgctgg ctgcaagaag atctttggga gcctggcatt tttgccggag 1141
agctttgatg ggaacccctc ctccggcgtt gccccactga agccagagca tctccaagtg
1201 ttcgaaaccc tggaggagat cacaggttac ctatacattt cagcatggcc
agagagcttc 1261 caagacctca gtgtcttcca gaaccttcgg gtcattcggg
gacggattct ccatgatggt 1321 gcttactcat tgacgttgca aggcctgggg
attcactcac tggggctacg ctcactgcgg 1381 gagctgggca gtggattggc
tctcattcac cgcaacaccc atctctgctt tgtaaacact 1441 gtaccttggg
accagctctt ccggaacccg caccaggccc tactccacag tgggaaccgg 1501
ccagaagagg catgtggtct tgagggcttg gtctgtaact cactgtgtgc ccgtgggcac
1561 tgctgggggc cagggcccac ccagtgtgtc aactgcagtc agttcctccg
gggccaggag 1621 tgtgtggagg agtgccgagt atggaagggg ctccccaggg
agtatgtgag gggcaagcac 1681 tgtctgccat gccaccccga gtgtcagcct
caaaacagct cggagacctg ctatggatcg 1741 gaggctgacc agtgtgaggc
ttgtgcccac tacaaggact catcttcctg tgtggctcgc 1801 tgccccagtg
gtgtgaagcc agacctctcc tacatgccta tctggaagta cccggatgag 1861
gagggcatat gtcagccatg ccccatcaac tgcacccact catgtgtgga cctggacgaa
1921 cgaggctgcc cagcagagca gagagccagc ccagtgacat tcatcattgc
aactgtggtg 1981 ggcgtcctgt tgttcctgat catagtggtg gtcattggaa
tcctaatcaa acgaaggcga 2041 cagaagatcc ggaagtatac catg mmouseHER2,
SEQ ID NO: 56 1 acccaagtgt gtaccggtac cgacatgaag ttgcgactcc
ctgccagtcc tgagacccac
61 ctggacatgc ttcgccacct ctaccagggc tgtcaggtgg tgcagggcaa
tttggagctt 121 acctacctgc ccgccaatgc cagcctctca ttcctgcagg
acatccagga agtccaggga 181 tacatgctca tcgctcacaa ccgagtgaaa
cacgtcccac tgcagaggtt gcgcatcgtg 241 agagggactc agctctttga
ggacaagtat gccctggctg tgctagacaa ccgagaccct 301 ttggacaacg
tcaccaccgc cgccccaggc agaaccccag aagggctgcg ggagctgcag 361
cttcgaagtc tcacagagat cttgaaggga ggagttttga tccgtgggaa ccctcagctc
421 tgctaccagg acatggtttt gtggaaggat gtcctccgta agaataacca
gctggctcct 481 gtcgacatgg acaccaatcg ttcccgggcc tgtccacctt
gtgccccaac ctgcaaagac 541 aatcactgtt ggggtgagag tcctgaagac
tgtcagatct tgactggcac catctgtact 601 agtggctgtg cccggtgcaa
gggccggctg cccactgact gttgccatga gcagtgtgct 661 gcaggctgca
cgggtcccaa gcattctgac tgcctggcct gcctccactt caatcatagt 721
ggtatctgtg agctgcactg cccggccctc atcacctaca acacagacac cttcgagtcc
781 atgctcaacc ctgagggtcg ctacaccttt ggtgccagct gtgtgaccac
ctgcccctac 841 aactacctct ccacggaagt gggatcctgc actctggtct
gtcccccgaa caaccaagag 901 gtcacagctg aggacggaac acagcggtgt
gagaaatgca gcaagccctg tgctggagta 961 tgctatggtc tgggcatgga
gcacctccga ggggcgaggg ccatcaccag tgacaatatc 1021 caggagtttg
ctggctgcaa gaagatcttt gggagcctgg catttttgcc ggagagcttt 1081
gatgggaacc cctcctccgg cgttgcccca ctgaagccag agcatctcca agtgttcgaa
1141 accctggagg agatcacagg ttacctatac atttcagcat ggccagagag
cttccaagac 1201 ctcagtgtct tccagaacct tcgggtcatt cggggacgga
ttctccatga tggtgcttac 1261 tcattgacgt tgcaaggcct ggggattcac
tcactggggc tacgctcact gcgggagctg 1321 ggcagtggat tggctctcat
tcaccgcaac acccatctct gctttgtaaa cactgtacct 1381 tgggaccagc
tcttccggaa cccgcaccag gccctactcc acagtgggaa ccggccagaa 1441
gaggcatgtg gtcttgaggg cttggtctgt aactcactgt gtgcccgtgg gcactgctgg
1501 gggccagggc ccacccagtg tgtcaactgc agtcagttcc tccggggcca
ggagtgtgtg 1561 gaggagtgcc gagtatggaa ggggctcccc agggagtatg
tgaggggcaa gcactgtctg 1621 ccatgccacc ccgagtgtca gcctcaaaac
agctcggaga cctgctatgg atcggaggct 1681 gaccagtgtg aggcttgtgc
ccactacaag gactcatctt cctgtgtggc tcgctgcccc 1741 agtggtgtga
agccagacct ctcctacatg cctatctgga agtacccgga tgaggagggc 1801
atatgtcagc catgccccat caactgcacc cactcatgtg tggacctgga cgaacgaggc
1861 tgcccagcag agcagagagc cagcccagtg acattcatca ttgcaactgt
ggtgggcgtc 1921 ctgttgttcc tgatcatagt ggtggtcatt ggaatcctaa
tcaaacgaag gcgacagaag 1981 atccggaagt ataccatg preratHER2, SEQ ID
NO: 57 1 atgatcatca tggagctggc ggcctggtgc cgctgggggt tcctcctcgc
cctcctgccc 61 cccggaatcg cgggcaccca agtgtgtacc ggcacagaca
tgaagttgcg gctccctgcc 121 agtcctgaga cccacctgga catgctccgc
cacctgtacc agggctgtca ggtagtgcag 181 ggcaacttgg agcttaccta
cgtgcctgcc aatgccagcc tctcattcct gcaggacatc 241 caggaagttc
agggttacat gctcatcgct cacaaccagg tgaagcgcgt cccactgcaa 301
aggctgcgca tcgtgagagg gacccagctc tttgaggaca agtatgccct ggctgtgcta
361 gacaaccgag atcctcagga caatgtcgcc gcctccaccc caggcagaac
cccagagggg 421 ctgcgggagc tgcagcttcg aagtctcaca gagatcctga
agggaggagt tttgatccgt 481 gggaaccctc agctctgcta ccaggacatg
gttttgtgga aggacgtctt ccgcaagaat 541 aaccaactgg ctcctgtcga
tatagacacc aatcgttccc gggcctgtcc accttgtgcc 601 cccgcctgca
aagacaatca ctgttggggt gagagtccgg aagactgtca gatcttgact 661
ggcaccatct gtaccagtgg ttgtgcccgg tgcaagggcc ggctgcccac tgactgctgc
721 catgagcagt gtgccgcagg ctgcacgggc cccaagcatt ctgactgcct
ggcctgcctc 781 cacttcaatc atagtggtat ctgtgagctg cactgcccag
ccctcgtcac ctacaacaca 841 gacacctttg agtccatgca caaccctgag
ggtcgctaca cctttggtgc cagctgcgtg 901 accacctgcc cctacaacta
cctgtctacg gaagtgggat cctgcactct ggtgtgtccc 961 ccgaataacc
aagaggtcac agctgaggac ggaacacagc gttgtgagaa atgcagcaag 1021
ccctgtgctc gagtgtgcta tggtctgggc atggagcacc ttcgaggggc gagggccatc
1081 accagtgaca atgtccagga gtttgatggc tgcaagaaga tctttgggag
cctggcattt 1141 ttgccggaga gctttgatgg ggacccctcc tccggcattg
ctccgctgag gcctgagcag 1201 ctccaagtgt tcgaaaccct ggaggagatc
acaggttacc tgtacatctc agcatggcca 1261 gacagtctcc gtgacctcag
tgtcttccag aaccttcgaa tcattcgggg acggattctc 1321 cacgatggcg
cgtactcatt gacactgcaa ggcctgggga tccactcgct ggggctgcgc 1381
tcactgcggg agctgggcag tggattggct ctgattcacc gcaacgccca tctctgcttt
1441 gtacacactg taccttggga ccagctcttc cggaacccac atcaggccct
gctccacagt 1501 gggaaccggc cggaagagga ttgtggtctc gagggcttgg
tctgtaactc actgtgtgcc 1561 cacgggcact gctgggggcc agggcccacc
cagtgtgtca actgcagtca tttccttcgg 1621 ggccaggagt gtgtggagga
gtgccgagta tggaaggggc tcccccggga gtatgtgagt 1681 gacaagcgct
gtctgccgtg tcaccccgag tgtcagcctc aaaacagctc agagacctgc 1741
tttggatcgg aggctgatca gtgtgcagcc tgcgcccact acaaggactc gtcctcctgt
1801 gtggctcgct gccccagtgg tgtgaaaccg gacctctcct acatgcccat
ctggaagtac 1861 ccggatgagg agggcatatg ccagccgtgc cccatcaact
gcacccactc ctgtgtggat 1921 ctggatgaac gaggctgccc agcagagcag
agagccagcc cggtgacatt catcattgca 1981 actgtagtgg gcgtcctgct
gttcctgatc ttagtggtgg tcgttggaat cctaatcaaa 2041 cgaaggagac
agaagatccg gaagtatacg atg mratHER2, SEQ ID NO: 58 1 acccaagtgt
gtaccggcac agacatgaag ttgcggctcc ctgccagtcc tgagacccac 61
ctggacatgc tccgccacct gtaccagggc tgtcaggtag tgcagggcaa cttggagctt
121 acctacgtgc ctgccaatgc cagcctctca ttcctgcagg acatccagga
agttcagggt 181 tacatgctca tcgctcacaa ccaggtgaag cgcgtcccac
tgcaaaggct gcgcatcgtg 241 agagggaccc agctctttga ggacaagtat
gccctggctg tgctagacaa ccgagatcct 301 caggacaatg tcgccgcctc
caccccaggc agaaccccag aggggctgcg ggagctgcag 361 cttcgaagtc
tcacagagat cctgaaggga ggagttttga tccgtgggaa ccctcagctc 421
tgctaccagg acatggtttt gtggaaggac gtcttccgca agaataacca actggctcct
481 gtcgatatag acaccaatcg ttcccgggcc tgtccacctt gtgcccccgc
ctgcaaagac 541 aatcactgtt ggggtgagag tccggaagac tgtcagatct
tgactggcac catctgtacc 601 agtggttgtg cccggtgcaa gggccggctg
cccactgact gctgccatga gcagtgtgcc 661 gcaggctgca cgggccccaa
gcattctgac tgcctggcct gcctccactt caatcatagt 721 ggtatctgtg
agctgcactg cccagccctc gtcacctaca acacagacac ctttgagtcc 781
atgcacaacc ctgagggtcg ctacaccttt ggtgccagct gcgtgaccac ctgcccctac
841 aactacctgt ctacggaagt gggatcctgc actctggtgt gtcccccgaa
taaccaagag 901 gtcacagctg aggacggaac acagcgttgt gagaaatgca
gcaagccctg tgctcgagtg 961 tgctatggtc tgggcatgga gcaccttcga
ggggcgaggg ccatcaccag tgacaatgtc 1021 caggagtttg atggctgcaa
gaagatcttt gggagcctgg catttttgcc ggagagcttt 1081 gatggggacc
cctcctccgg cattgctccg ctgaggcctg agcagctcca agtgttcgaa 1141
accctggagg agatcacagg ttacctgtac atctcagcat ggccagacag tctccgtgac
1201 ctcagtgtct tccagaacct tcgaatcatt cggggacgga ttctccacga
tggcgcgtac 1261 tcattgacac tgcaaggcct ggggatccac tcgctggggc
tgcgctcact gcgggagctg 1321 ggcagtggat tggctctgat tcaccgcaac
gcccatctct gctttgtaca cactgtacct 1381 tgggaccagc tcttccggaa
cccacatcag gccctgctcc acagtgggaa ccggccggaa 1441 gaggattgtg
gtctcgaggg cttggtctgt aactcactgt gtgcccacgg gcactgctgg 1501
gggccagggc ccacccagtg tgtcaactgc agtcatttcc ttcggggcca ggagtgtgtg
1561 gaggagtgcc gagtatggaa ggggctcccc cgggagtatg tgagtgacaa
gcgctgtctg 1621 ccgtgtcacc ccgagtgtca gcctcaaaac agctcagaga
cctgctttgg atcggaggct 1681 gatcagtgtg cagcctgcgc ccactacaag
gactcgtcct cctgtgtggc tcgctgcccc 1741 agtggtgtga aaccggacct
ctcctacatg cccatctgga agtacccgga tgaggagggc 1801 atatgccagc
cgtgccccat caactgcacc cactcctgtg tggatctgga tgaacgaggc 1861
tgcccagcag agcagagagc cagcccggtg acattcatca ttgcaactgt agtgggcgtc
1921 ctgctgttcc tgatcttagt ggtggtcgtt ggaatcctaa tcaaacgaag
gagacagaag 1981 atccggaagt atacgatg preE2Neu, SEQ ID NO: 59 1
atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc ccccggagcc
61 gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc
cagtcccgag 121 acccacctgg acatgctccg ccacctctac cagggctgcc
aggtggtgca gggaaacctg 181 gaactcacct acctgcccac caatgccagc
ctgtccttcc tgcaggatat ccaggaggtg 241 cagggctacg tgctcatcgc
tcacaaccaa gtgaggcagg tcccactgca gaggctgcgg 301 attgtgcgag
gcacccagct ctttgaggac aactatgccc tggccgtgct agacaatgga 361
gacccgctga acaataccac ccctgtcaca ggggcctccc caggaggcct gcgggagctg
421 cagcttcgaa gcctcacaga gatcttgaaa ggaggggtct tgatccagcg
gaacccccag 481 ctctgctacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggct 541 ctcacactga tagacaccaa ccgctctcgg
gcctgccacc cctgttctcc gatgtgtaag 601 ggctcccgct gctggggaga
gagttctgag gattgtcaga gcctgacgcg cactgtctgt 661 gccggtggct
gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt 721
gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca cttcaaccac
781 agtggcatct gtgagctgca ctgcccagcc ctggtcacct acaacacaga
cacgtttgag 841 tccatgccca atcccgaggg ccggtataca ttcggcgcca
gctgtgtgac tgcctgtccc 901 tacaactacc tttctacgga cgtgggatcc
tgcaccctcg tctgccccct gcacaaccaa 961 gaggtgacag cagaggatgg
aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga 1021 gtgtgctatg
gtctgggcat ggagcacttg cgagaggtga gggcagttac cagtgccaat 1081
atccaggagt ttgctggctg caagaagatc tttgggagcc tggcatttct gccggagagc
1141 tttgatgggg acccagcctc caacactgcc gaattcgctc cgctgaggcc
tgagcagctc 1201 caagtgttcg aaaccctgga ggagatcaca ggttacctgt
acatctcagc atggccagac 1261 agtctccgtg acctcagtgt cttccagaac
cttcgaatca ttcggggacg gattctccac
1321 gatggcgcgt actcattgac actgcaaggc ctggggatcc actcgctggg
gctgcgctca 1381 ctgcgggagc tgggcagtgg attggctctg attcaccgca
acgcccatct ctgctttgta 1441 cacactgtac cttgggacca gctcttccgg
aacccacatc aggccctgct ccacagtggg 1501 aaccggccgg aagaggattg
tggtctcgag ggcttggtct gtaactcact gtgtgcccac 1561 gggcactgct
gggggccagg gcccacccag tgtgtcaact gcagtcattt ccttcggggc 1621
caggagtgtg tggaggagtg ccgagtatgg aaggggctcc cccgggagta tgtgagtgac
1681 aagcgctgtc tgccgtgtca ccccgagtgt cagcctcaaa acagctcaga
gacctgcttt 1741 ggatcggagg ctgatcagtg tgcagcctgc gcccactaca
aggactcgtc ctcctgtgtg 1801 gctcgctgcc ccagtggtgt gaaaccggac
ctctcctaca tgcccatctg gaagtacccg 1861 gatgaggagg gcatatgcca
gccgtgcccc atcaactgca cccactcctg tgtggatctg 1921 gatgaacgag
gctgcccagc agagcagaga gccagcccgg tgacattcat cattgcaact 1981
gtagtgggcg tcctgctgtt cctgatctta gtggtggtcg ttggaatcct aatcaaacga
2041 aggagacaga agatccggaa gtatacgatg mE2Neu, SEQ ID NO: 60 1
acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac
61 ctggacatgc tccgccacct ctaccagggc tgccaggtgg tgcagggaaa
cctggaactc 121 acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg
atatccagga ggtgcagggc 181 tacgtgctca tcgctcacaa ccaagtgagg
caggtcccac tgcagaggct gcggattgtg 241 cgaggcaccc agctctttga
ggacaactat gccctggccg tgctagacaa tggagacccg 301 ctgaacaata
ccacccctgt cacaggggcc tccccaggag gcctgcggga gctgcagctt 361
cgaagcctca cagagatctt gaaaggaggg gtcttgatcc agcggaaccc ccagctctgc
421 taccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggctctcaca 481 ctgatagaca ccaaccgctc tcgggcctgc cacccctgtt
ctccgatgtg taagggctcc 541 cgctgctggg gagagagttc tgaggattgt
cagagcctga cgcgcactgt ctgtgccggt 601 ggctgtgccc gctgcaaggg
gccactgccc actgactgct gccatgagca gtgtgctgcc 661 ggctgcacgg
gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc 721
atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt tgagtccatg
781 cccaatcccg agggccggta tacattcggc gccagctgtg tgactgcctg
tccctacaac 841 tacctttcta cggacgtggg atcctgcacc ctcgtctgcc
ccctgcacaa ccaagaggtg 901 acagcagagg atggaacaca gcggtgtgag
aagtgcagca agccctgtgc ccgagtgtgc 961 tatggtctgg gcatggagca
cttgcgagag gtgagggcag ttaccagtgc caatatccag 1021 gagtttgctg
gctgcaagaa gatctttggg agcctggcat ttctgccgga gagctttgat 1081
ggggacccag cctccaacac tgccgaattc gctccgctga ggcctgagca gctccaagtg
1141 ttcgaaaccc tggaggagat cacaggttac ctgtacatct cagcatggcc
agacagtctc 1201 cgtgacctca gtgtcttcca gaaccttcga atcattcggg
gacggattct ccacgatggc 1261 gcgtactcat tgacactgca aggcctgggg
atccactcgc tggggctgcg ctcactgcgg 1321 gagctgggca gtggattggc
tctgattcac cgcaacgccc atctctgctt tgtacacact 1381 gtaccttggg
accagctctt ccggaaccca catcaggccc tgctccacag tgggaaccgg 1441
ccggaagagg attgtggtct cgagggcttg gtctgtaact cactgtgtgc ccacgggcac
1501 tgctgggggc cagggcccac ccagtgtgtc aactgcagtc atttccttcg
gggccaggag 1561 tgtgtggagg agtgccgagt atggaagggg ctcccccggg
agtatgtgag tgacaagcgc 1621 tgtctgccgt gtcaccccga gtgtcagcct
caaaacagct cagagacctg ctttggatcg 1681 gaggctgatc agtgtgcagc
ctgcgcccac tacaaggact cgtcctcctg tgtggctcgc 1741 tgccccagtg
gtgtgaaacc ggacctctcc tacatgccca tctggaagta cccggatgag 1801
gagggcatat gccagccgtg ccccatcaac tgcacccact cctgtgtgga tctggatgaa
1861 cgaggctgcc cagcagagca gagagccagc ccggtgacat tcatcattgc
aactgtagtg 1921 ggcgtcctgc tgttcctgat cttagtggtg gtcgttggaa
tcctaatcaa acgaaggaga 1981 cagaagatcc ggaagtatac gatg
pprehumHER2-Q213K, SEQ ID NO: 84 1 melaalcrwg lllallppga astqvctgtd
mklrlpaspe thldmlrhly qgcqvvqgnl 61 eltylptnas lsflqdiqev
qgyvliahnq vrqvplqrlr ivrgtqlfed nyalavldng 121 dpinnttpvt
gaspgglrel qlrslteilk ggvliqrnpq lcyqdtilwk difhknnqla 181
ltlidtnrsr achpcspmck gsrcwgesse dcKsltrtvc aggcarckgp 1ptdccheqc
241 aagctgpkhs dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt
fgascvtacp 301 ynylstdvgs ctlvcplhnq evtaedgtqr cekcskpcar
vcyglgmehl revravtsan 361 igefagckki fgslaflpes fdgdpasnta
plqpeqlqvf etleeitgyl yisawpdslp 421 dlsvfqnlqv irgrilhnga
ysltlqglgi swlglrslre lgsglalihh nthlcfvhtv 481 pwdqlfrnph
qallhtanrp edecvgegla chqlcarghc wgpgptqcvn csqflrgqec 541
veecrvlqgl preyvnarhc 1pchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc
601 psgvkpdlsy mpiwkfpdee gacqpcpinc thscvdlddk gcpaeqrasp
ltsiisavvg 661 illvvvlgvv fgilikrrqq kirkytm pprehumHER2-Q239K, SEQ
ID NO: 85 1 melaalcrwg lllallppga astqvctgtd mklrlpaspe thldmlrhly
qgcqvvqgnl 61 eltylptnas lsflqdiqev qgyvliahnq vrqvplqrlr
ivrgtqlfed nyalavldng 121 dpinnttpvt gaspgglrel qlrslteilk
ggvliqrnpq lcyqdtilwk difhknnqla 181 ltlidtnrsr achpcspmck
gsrcwgesse dcqsltrtvc aggcarckgp 1ptdccheKc 241 aagctgpkhs
dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp 301
ynylstdvgs ctlvcplhnq evtaedgtqr cekcskpcar vcyglgmehl revravtsan
361 igefagckki fgslaflpes fdgdpasnta plqpeqlqvf etleeitgyl
yisawpdslp 421 dlsvfqnlqv irgrilhnga ysltlqglgi swlglrslre
lgsglalihh nthlcfvhtv 481 pwdqlfrnph qallhtanrp edecvgegla
chqlcarghc wgpgptqcvn csqflrgqec 541 veecrvlqgl preyvnarhc
lpchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc 601 psgvkpdlsy
mpiwkfpdee gacqpcpinc thscvdlddk gcpaeqrasp ltsiisavvg 661
illvvvlgvv fgilikrrqq kirkytm pprehumHER2-NNT 124-126 DSG, SEQ ID
NO: 86 1 melaalcrwg lllallppga astqvctgtd mklrlpaspe thldmlrhly
qgcqvvqgnl 61 eltylptnas lsflqdiqev qgyvliahnq vrqvplqrlr
ivrgtqlfed nyalavldng 121 dplDSGtpvt gaspgglrel qlrslteilk
ggvliqrnpq lcyqdtilwk difhknnqla 181 ltlidtnrsr achpcspmck
gsrcwgesse dcqsltrtvc aggcarckgp lptdccheqc 241 aagctgpkhs
dclaclhfnh sgicelhcpa lvtyntdtfe smpnpegryt fgascvtacp 301
ynylstdvgs ctlvcplhnq evtaedgtqr cekcskpcar vcyglgmehl revravtsan
361 igefagckki fgslaflpes fdgdpasnta plqpeqlqvf etleeitgyl
yisawpdslp 421 dlsvfqnlqv irgrilhnga ysltlqglgi swlglrslre
lgsglalihh nthlcfvhtv 481 pwdqlfrnph qallhtanrp edecvgegla
chqlcarghc wgpgptqcvn csqflrgqec 541 veecrvlqgl preyvnarhc
lpchpecqpq ngsvtcfgpe adqcvacahy kdppfcvarc 601 psgvkpdlsy
mpiwkfpdee gacqpcpinc thscvdlddk gcpaeqrasp ltsiisavvg 661
illvvvlgvv fgilikrrqq kirkytm
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Sequence CWU 1
1
1181665PRTFelis catus 1Thr Gln Val Cys Thr Gly Thr Asp Met Lys Leu
Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu Arg His
Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu Thr
Tyr Leu His Ala Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile Gln
Glu Val Gln Gly Tyr Val Leu Ile 50 55 60Ala His Asn Gln Val Lys Gln
Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu
Phe Glu Asp Asn Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro
Leu Asp Ser Gly Thr Pro Ala Thr Gly Ala Ala Leu 100 105 110Gly Gly
Leu Arg Glu Leu Lys Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120
125Gly Gly Val Leu Ile Gln Arg Asn Pro Gln Leu Cys His Gln Asp Thr
130 135 140Ile Leu Trp Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala
Leu Met145 150 155 160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys Gln
Pro Cys Ser Pro Ala 165 170 175Cys Lys Asp Ser His Cys Trp Gly Ala
Ser Ser Gly Asp Cys Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala
Gly Gly Cys Ala Arg Cys Lys Gly Pro 195 200 205Gln Pro Thr Asp Cys
Cys His Glu Gln Cys Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His
Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser Gly225 230 235
240Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr
245 250 255Phe Glu Ser Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly
Ala Ser 260 265 270Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr
Asp Val Gly Ser 275 280 285Cys Thr Leu Val Cys Pro Leu Asn Asn Gln
Glu Val Thr Ala Glu Asp 290 295 300Gly Thr Gln Arg Cys Glu Lys Cys
Ser Lys Pro Cys Ala Arg Val Cys305 310 315 320Tyr Gly Leu Gly Met
Glu His Leu Arg Glu Ala Arg Ala Val Thr Ser 325 330 335Ala Asn Ile
Gln Glu Phe Val Gly Cys Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala
Phe Leu Pro Glu Ser Phe Glu Gly Asp Pro Ala Ser Asn Thr Ala 355 360
365Pro Leu Gln Pro Glu Gln Leu Arg Val Phe Glu Ala Leu Glu Glu Ile
370 375 380Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu Pro
Asn Leu385 390 395 400Ser Val Phe Gln Asn Leu Arg Val Ile Arg Gly
Arg Val Leu His Asp 405 410 415Gly Ala Tyr Ser Leu Thr Leu Gln Gly
Leu Gly Ile Ser Trp Leu Gly 420 425 430Leu Arg Ser Leu Arg Glu Leu
Gly Ser Gly Leu Ala Leu Ile His Arg 435 440 445Asn Ser Arg Leu Cys
Phe Val His Thr Val Pro Trp Asp Gln Leu Phe 450 455 460Arg Asn Pro
His Gln Ala Leu Leu His Ser Ala Asn Arg Pro Glu Asp465 470 475
480Glu Cys Ala Gly Glu Gly Leu Ala Cys Tyr Pro Leu Cys Ala His Gly
485 490 495His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser
Gln Phe 500 505 510Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val
Leu Gln Gly Leu 515 520 525Pro Arg Glu Tyr Val Lys Asp Arg Phe Cys
Leu Pro Cys His Pro Glu 530 535 540Cys Gln Pro Gln Asn Gly Ser Val
Thr Cys Leu Gly Ser Glu Ala Asp545 550 555 560Gln Cys Val Ala Cys
Ala His Tyr Lys Asp Pro Pro Phe Cys Val Ala 565 570 575Arg Cys Pro
Ser Gly Val Lys Pro Asp Leu Ser Phe Met Pro Ile Trp 580 585 590Lys
Phe Ala Asp Glu Glu Gly Thr Cys Gln Pro Cys Pro Ile Asn Cys 595 600
605Thr His Ser Cys Ala Asp Leu Asp Glu Lys Gly Cys Pro Ala Glu Gln
610 615 620Arg Ala Ser Pro Val Thr Ser Ile Ile Ala Ala Val Val Gly
Ile Leu625 630 635 640Leu Val Val Val Val Gly Leu Val Leu Gly Ile
Leu Ile Lys Arg Arg 645 650 655Arg Gln Lys Ile Arg Lys Tyr Thr Met
660 6652665PRTUrsus americanus 2Thr Gln Val Cys Thr Gly Thr Asp Met
Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu
Arg His Leu Tyr Gln Ala Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu
Leu Thr Tyr Leu Pro Ala Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp
Ile Gln Glu Val Gln Gly Tyr Val Leu Ile 50 55 60Ala His Ser Gln Val
Arg Gln Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr
Gln Leu Phe Glu Asp Asn Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Gly
Glu Pro Pro Lys Gly Asp Thr Ser Val Ala Gly Ala Thr Pro 100 105
110Gly Gly Leu Arg Glu Leu Lys Leu Arg Ser Leu Thr Glu Ile Leu Lys
115 120 125Gly Gly Val Leu Ile Gln Arg Asn Pro Gln Leu Cys His Gln
Asp Thr 130 135 140Ile Leu Trp Lys Asp Ile Phe His Lys Asn Asn Gln
Leu Ala Leu Thr145 150 155 160Leu Ile Asp Thr Asn Arg Ser Arg Ala
Cys Gln Pro Cys Ser Pro Ala 165 170 175Cys Lys Asp Pro His Cys Trp
Gly Ala Ser Ser Gly Asp Cys Gln Ser 180 185 190Leu Thr Arg Thr Val
Cys Ala Gly Gly Cys Ala Arg Cys Lys Gly Pro 195 200 205Lys Pro Thr
Asp Cys Cys His Glu Gln Cys Ala Ala Gly Cys Thr Gly 210 215 220Pro
Lys His Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser Gly225 230
235 240Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp
Thr 245 250 255Phe Glu Ser Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe
Gly Ala Ser 260 265 270Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser
Thr Asp Val Gly Ser 275 280 285Cys Thr Leu Val Cys Pro Leu Asn Asn
Gln Glu Val Thr Ala Glu Asp 290 295 300Gly Thr Gln Arg Cys Glu Lys
Cys Ser Arg Pro Cys Ala Arg Val Cys305 310 315 320Tyr Gly Leu Gly
Met Glu His Leu Arg Glu Ala Arg Ala Val Thr Ser 325 330 335Ala Asn
Ile Gln Glu Phe Ala Gly Cys Lys Lys Ile Phe Gly Ser Leu 340 345
350Ala Phe Leu Pro Glu Ser Phe Glu Gly Asp Pro Ala Ser Asn Thr Ala
355 360 365Pro Leu Gln Pro Glu Gln Leu Arg Val Phe Glu Ala Leu Glu
Glu Ile 370 375 380Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser
Leu Pro Asn Leu385 390 395 400Ser Val Phe Gln Asn Leu Arg Val Ile
Arg Gly Arg Val Leu His Asp 405 410 415Gly Ala Tyr Ser Leu Thr Leu
Gln Gly Leu Gly Ile Ser Trp Leu Gly 420 425 430Leu Arg Ser Leu Arg
Glu Leu Gly Ser Gly Leu Ala Leu Ile His Arg 435 440 445Asn Ala Arg
Leu Cys Phe Ile His Thr Val Pro Trp Glu Gln Leu Phe 450 455 460Arg
Asn Pro His Gln Ala Leu Leu His Ser Ala Asn Arg Pro Glu Ala465 470
475 480Glu Cys Val Gly Glu Gly Leu Ala Cys Tyr Pro Leu Cys Ala His
Gly 485 490 495His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys
Ser Gln Phe 500 505 510Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg
Glu Leu His Gly Leu 515 520 525Pro Arg Glu Tyr Val Lys Asp Arg Tyr
Cys Leu Pro Cys His Pro Glu 530 535 540Cys Arg Pro Gln Asn Gly Ser
Val Thr Cys Phe Gly Ser Glu Ala Asp545 550 555 560Gln Cys Val Ala
Cys Ala His Tyr Lys Asp Pro Pro Ser Cys Val Ala 565 570 575Arg Cys
Pro Ser Gly Val Lys Pro Asp Leu Ser Phe Met Pro Ile Trp 580 585
590Lys Phe Ala Asp Glu Glu Gly Thr Cys Gln Pro Cys Pro Ile Asn Cys
595 600 605Thr His Ser Cys Gly Asp Leu Asp Glu Arg Gly Cys Pro Ala
Glu Gln 610 615 620Arg Ala Ser Pro Val Thr Ser Ile Ile Ala Ala Val
Val Gly Ile Leu625 630 635 640Leu Ala Val Val Met Gly Leu Val Leu
Gly Ile Leu Ile Lys Arg Arg 645 650 655Arg Gln Lys Ile Arg Lys Tyr
Thr Met 660 6653665PRTHomo sapiens 3Thr Gln Val Cys Thr Gly Thr Asp
Met Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met
Leu Arg His Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu
Glu Leu Thr Tyr Leu Pro Thr Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln
Asp Ile Gln Glu Val Gln Gly Tyr Val Leu Ile 50 55 60Ala His Asn Gln
Val Arg Gln Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly
Thr Gln Leu Phe Glu Asp Asn Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn
Gly Asp Pro Leu Asn Asn Thr Thr Pro Val Thr Gly Ala Ser Pro 100 105
110Gly Gly Leu Arg Glu Leu Lys Leu Arg Ser Leu Thr Glu Ile Leu Lys
115 120 125Gly Gly Val Leu Ile Gln Arg Asn Pro Gln Leu Cys Tyr Gln
Asp Thr 130 135 140Ile Leu Trp Lys Asp Ile Phe His Lys Asn Asn Gln
Leu Ala Leu Thr145 150 155 160Leu Ile Asp Thr Asn Arg Ser Arg Ala
Cys His Pro Cys Ser Pro Met 165 170 175Cys Lys Gly Ser Arg Cys Trp
Gly Glu Ser Ser Glu Asp Cys Gln Ser 180 185 190Leu Thr Arg Thr Val
Cys Ala Gly Gly Cys Ala Arg Cys Lys Gly Pro 195 200 205Leu Pro Thr
Asp Cys Cys His Glu Gln Cys Ala Ala Gly Cys Thr Gly 210 215 220Pro
Lys His Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser Gly225 230
235 240Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp
Thr 245 250 255Phe Glu Ser Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe
Gly Ala Ser 260 265 270Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser
Thr Asp Val Gly Ser 275 280 285Cys Thr Leu Val Cys Pro Leu His Asn
Gln Glu Val Thr Ala Glu Asp 290 295 300Gly Thr Gln Arg Cys Glu Lys
Cys Ser Lys Pro Cys Ala Arg Val Cys305 310 315 320Tyr Gly Leu Gly
Met Glu His Leu Arg Glu Val Arg Ala Val Thr Ser 325 330 335Ala Asn
Ile Gln Glu Phe Ala Gly Cys Lys Lys Ile Phe Gly Ser Leu 340 345
350Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp Pro Ala Ser Asn Thr Ala
355 360 365Pro Leu Gln Pro Glu Gln Leu Gln Val Phe Glu Thr Leu Glu
Glu Ile 370 375 380Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser
Leu Pro Asp Leu385 390 395 400Ser Val Phe Gln Asn Leu Gln Val Ile
Arg Gly Arg Ile Leu His Asn 405 410 415Gly Ala Tyr Ser Leu Thr Leu
Gln Gly Leu Gly Ile Ser Trp Leu Gly 420 425 430Leu Arg Ser Leu Arg
Glu Leu Gly Ser Gly Leu Ala Leu Ile His His 435 440 445Asn Thr His
Leu Cys Phe Val His Thr Val Pro Trp Asp Gln Leu Phe 450 455 460Arg
Asn Pro His Gln Ala Leu Leu His Thr Ala Asn Arg Pro Glu Asp465 470
475 480Glu Cys Val Gly Glu Gly Leu Ala Cys His Gln Leu Cys Ala Arg
Gly 485 490 495His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys
Ser Gln Phe 500 505 510Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg
Val Leu Gln Gly Leu 515 520 525Pro Arg Glu Tyr Val Asn Ala Arg His
Cys Leu Pro Cys His Pro Glu 530 535 540Cys Gln Pro Gln Asn Gly Ser
Val Thr Cys Phe Gly Pro Glu Ala Asp545 550 555 560Gln Cys Val Ala
Cys Ala His Tyr Lys Asp Pro Pro Phe Cys Val Ala 565 570 575Arg Cys
Pro Ser Gly Val Lys Pro Asp Leu Ser Tyr Met Pro Ile Trp 580 585
590Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln Pro Cys Pro Ile Asn Cys
595 600 605Thr His Ser Cys Val Asp Leu Asp Asp Lys Gly Cys Pro Ala
Glu Gln 610 615 620Arg Ala Ser Pro Leu Thr Ser Ile Ile Ser Ala Val
Val Gly Ile Leu625 630 635 640Leu Val Val Val Leu Gly Val Val Phe
Gly Ile Leu Ile Lys Arg Arg 645 650 655Gln Gln Lys Ile Arg Lys Tyr
Thr Met 660 6654666PRTMus musculus 4Thr Gln Val Cys Thr Gly Thr Asp
Met Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met
Leu Arg His Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu
Glu Leu Thr Tyr Leu Pro Ala Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln
Asp Ile Gln Glu Val Gln Gly Tyr Met Leu Ile 50 55 60Ala His Asn Arg
Val Lys His Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly
Thr Gln Leu Phe Glu Asp Lys Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn
Arg Asp Pro Leu Asp Asn Val Thr Thr Ala Ala Pro Gly Arg Thr 100 105
110Pro Glu Gly Leu Arg Glu Leu Lys Leu Arg Ser Leu Thr Glu Ile Leu
115 120 125Lys Gly Gly Val Leu Ile Arg Gly Asn Pro Gln Leu Cys Tyr
Gln Asp 130 135 140Met Val Leu Trp Lys Asp Val Leu Arg Lys Asn Asn
Gln Leu Ala Pro145 150 155 160Val Asp Met Asp Thr Asn Arg Ser Arg
Ala Cys Pro Pro Cys Ala Pro 165 170 175Thr Cys Lys Asp Asn His Cys
Trp Gly Glu Ser Pro Glu Asp Cys Gln 180 185 190Ile Leu Thr Gly Thr
Ile Cys Thr Ser Gly Cys Ala Arg Cys Lys Gly 195 200 205Arg Leu Pro
Thr Asp Cys Cys His Glu Gln Cys Ala Ala Gly Cys Thr 210 215 220Gly
Pro Lys His Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser225 230
235 240Gly Ile Cys Glu Leu His Cys Pro Ala Leu Ile Thr Tyr Asn Thr
Asp 245 250 255Thr Phe Glu Ser Met Leu Asn Pro Glu Gly Arg Tyr Thr
Phe Gly Ala 260 265 270Ser Cys Val Thr Thr Cys Pro Tyr Asn Tyr Leu
Ser Thr Glu Val Gly 275 280 285Ser Cys Thr Leu Val Cys Pro Pro Asn
Asn Gln Glu Val Thr Ala Glu 290 295 300Asp Gly Thr Gln Arg Cys Glu
Lys Cys Ser Lys Pro Cys Ala Gly Val305 310 315 320Cys Tyr Gly Leu
Gly Met Glu His Leu Arg Gly Ala Arg Ala Ile Thr 325 330 335Ser Asp
Asn Ile Gln Glu Phe Ala Gly Cys Lys Lys Ile Phe Gly Ser 340 345
350Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asn Pro Ser Ser Gly Val
355 360 365Ala Pro Leu Lys Pro Glu His Leu Gln Val Phe Glu Thr Leu
Glu Glu 370 375 380Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Glu
Ser Phe Gln Asp385 390 395 400Leu Ser Val Phe Gln Asn Leu Arg Val
Ile Arg Gly Arg Ile Leu His 405 410 415Asp Gly Ala Tyr Ser Leu Thr
Leu Gln Gly Leu Gly Ile His Ser Leu 420 425 430Gly Leu Arg Ser Leu
Arg Glu Leu Gly Ser Gly Leu Ala Leu Ile His 435 440 445Arg Asn Thr
His Leu Cys Phe Val Asn Thr Val Pro Trp Asp Gln Leu 450 455 460Phe
Arg Asn Pro His
Gln Ala Leu Leu His Ser Gly Asn Arg Pro Glu465 470 475 480Glu Ala
Cys Gly Leu Glu Gly Leu Val Cys Asn Ser Leu Cys Ala Arg 485 490
495Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser Gln
500 505 510Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Trp
Lys Gly 515 520 525Leu Pro Arg Glu Tyr Val Arg Gly Lys His Cys Leu
Pro Cys His Pro 530 535 540Glu Cys Gln Pro Gln Asn Ser Ser Glu Thr
Cys Tyr Gly Ser Glu Ala545 550 555 560Asp Gln Cys Glu Ala Cys Ala
His Tyr Lys Asp Ser Ser Ser Cys Val 565 570 575Ala Arg Cys Pro Ser
Gly Val Lys Pro Asp Leu Ser Tyr Met Pro Ile 580 585 590Trp Lys Tyr
Pro Asp Glu Glu Gly Ile Cys Gln Pro Cys Pro Ile Asn 595 600 605Cys
Thr His Ser Cys Val Asp Leu Asp Glu Arg Gly Cys Pro Ala Glu 610 615
620Gln Arg Ala Ser Pro Val Thr Phe Ile Ile Ala Thr Val Val Gly
Val625 630 635 640Leu Leu Phe Leu Ile Ile Val Val Val Ile Gly Ile
Leu Ile Lys Arg 645 650 655Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met
660 6655666PRTRattus norvegicus 5Thr Gln Val Cys Thr Gly Thr Asp
Met Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met
Leu Arg His Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu
Glu Leu Thr Tyr Val Pro Ala Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln
Asp Ile Gln Glu Val Gln Gly Tyr Met Leu Ile 50 55 60Ala His Asn Gln
Val Lys Arg Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly
Thr Gln Leu Phe Glu Asp Lys Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn
Arg Asp Pro Gln Asp Asn Val Ala Ala Ser Thr Pro Gly Arg Thr 100 105
110Pro Glu Gly Leu Arg Glu Leu Lys Leu Arg Ser Leu Thr Glu Ile Leu
115 120 125Lys Gly Gly Val Leu Ile Arg Gly Asn Pro Gln Leu Cys Tyr
Gln Asp 130 135 140Met Val Leu Trp Lys Asp Val Phe Arg Lys Asn Asn
Gln Leu Ala Pro145 150 155 160Val Asp Ile Asp Thr Asn Arg Ser Arg
Ala Cys Pro Pro Cys Ala Pro 165 170 175Ala Cys Lys Asp Asn His Cys
Trp Gly Glu Ser Pro Glu Asp Cys Gln 180 185 190Ile Leu Thr Gly Thr
Ile Cys Thr Ser Gly Cys Ala Arg Cys Lys Gly 195 200 205Arg Leu Pro
Thr Asp Cys Cys His Glu Gln Cys Ala Ala Gly Cys Thr 210 215 220Gly
Pro Lys His Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser225 230
235 240Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr
Asp 245 250 255Thr Phe Glu Ser Met His Asn Pro Glu Gly Arg Tyr Thr
Phe Gly Ala 260 265 270Ser Cys Val Thr Thr Cys Pro Tyr Asn Tyr Leu
Ser Thr Glu Val Gly 275 280 285Ser Cys Thr Leu Val Cys Pro Pro Asn
Asn Gln Glu Val Thr Ala Glu 290 295 300Asp Gly Thr Gln Arg Cys Glu
Lys Cys Ser Lys Pro Cys Ala Arg Val305 310 315 320Cys Tyr Gly Leu
Gly Met Glu His Leu Arg Gly Ala Arg Ala Ile Thr 325 330 335Ser Asp
Asn Val Gln Glu Phe Asp Gly Cys Lys Lys Ile Phe Gly Ser 340 345
350Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp Pro Ser Ser Gly Ile
355 360 365Ala Pro Leu Arg Pro Glu Gln Leu Gln Val Phe Glu Thr Leu
Glu Glu 370 375 380Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp
Ser Leu Arg Asp385 390 395 400Leu Ser Val Phe Gln Asn Leu Arg Ile
Ile Arg Gly Arg Ile Leu His 405 410 415Asp Gly Ala Tyr Ser Leu Thr
Leu Gln Gly Leu Gly Ile His Ser Leu 420 425 430Gly Leu Arg Ser Leu
Arg Glu Leu Gly Ser Gly Leu Ala Leu Ile His 435 440 445Arg Asn Ala
His Leu Cys Phe Val His Thr Val Pro Trp Asp Gln Leu 450 455 460Phe
Arg Asn Pro His Gln Ala Leu Leu His Ser Gly Asn Arg Pro Glu465 470
475 480Glu Asp Cys Gly Leu Glu Gly Leu Val Cys Asn Ser Leu Cys Ala
His 485 490 495Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn
Cys Ser His 500 505 510Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys
Arg Val Trp Lys Gly 515 520 525Leu Pro Arg Glu Tyr Val Ser Asp Lys
Arg Cys Leu Pro Cys His Pro 530 535 540Glu Cys Gln Pro Gln Asn Ser
Ser Glu Thr Cys Phe Gly Ser Glu Ala545 550 555 560Asp Gln Cys Ala
Ala Cys Ala His Tyr Lys Asp Ser Ser Ser Cys Val 565 570 575Ala Arg
Cys Pro Ser Gly Val Lys Pro Asp Leu Ser Tyr Met Pro Ile 580 585
590Trp Lys Tyr Pro Asp Glu Glu Gly Ile Cys Gln Pro Cys Pro Ile Asn
595 600 605Cys Thr His Ser Cys Val Asp Leu Asp Glu Arg Gly Cys Pro
Ala Glu 610 615 620Gln Arg Ala Ser Pro Val Thr Phe Ile Ile Ala Thr
Val Val Gly Val625 630 635 640Leu Leu Phe Leu Ile Leu Val Val Val
Val Gly Ile Leu Ile Lys Arg 645 650 655Arg Arg Gln Lys Ile Arg Lys
Tyr Thr Met 660 6656668PRTArtificial SequenceChimeric human-rat
E2Neu (HER2) 6Thr Gln Val Cys Thr Gly Thr Asp Met Lys Leu Arg Leu
Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu Arg His Leu Tyr
Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu Thr Tyr Leu
Pro Thr Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile Gln Glu Val
Gln Gly Tyr Val Leu Ile 50 55 60Ala His Asn Gln Val Arg Gln Val Pro
Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu
Asp Asn Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro Leu Asn
Asn Thr Thr Pro Val Thr Gly Ala Ser Pro 100 105 110Gly Gly Leu Arg
Glu Leu Lys Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120 125Gly Gly
Val Leu Ile Gln Arg Asn Pro Gln Leu Cys Tyr Gln Asp Thr 130 135
140Ile Leu Trp Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala Leu
Thr145 150 155 160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys His Pro
Cys Ser Pro Met 165 170 175Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser
Ser Glu Asp Cys Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala Gly
Gly Cys Ala Arg Cys Lys Gly Pro 195 200 205Leu Pro Thr Asp Cys Cys
His Glu Gln Cys Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His Ser
Asp Cys Leu Ala Cys Leu His Phe Asn His Ser Gly225 230 235 240Ile
Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr 245 250
255Phe Glu Ser Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala Ser
260 265 270Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr Asp Val
Gly Ser 275 280 285Cys Thr Leu Val Cys Pro Leu His Asn Gln Glu Val
Thr Ala Glu Asp 290 295 300Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys
Pro Cys Ala Arg Val Cys305 310 315 320Tyr Gly Leu Gly Met Glu His
Leu Arg Glu Val Arg Ala Val Thr Ser 325 330 335Ala Asn Ile Gln Glu
Phe Ala Gly Cys Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala Phe Leu
Pro Glu Ser Phe Asp Gly Asp Pro Ala Ser Asn Thr Ala 355 360 365Glu
Phe Ala Pro Leu Arg Pro Glu Gln Leu Gln Val Phe Glu Thr Leu 370 375
380Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser
Leu385 390 395 400Arg Asp Leu Ser Val Phe Gln Asn Leu Arg Ile Ile
Arg Gly Arg Ile 405 410 415Leu His Asp Gly Ala Tyr Ser Leu Thr Leu
Gln Gly Leu Gly Ile His 420 425 430Ser Leu Gly Leu Arg Ser Leu Arg
Glu Leu Gly Ser Gly Leu Ala Leu 435 440 445Ile His Arg Asn Ala His
Leu Cys Phe Val His Thr Val Pro Trp Asp 450 455 460Gln Leu Phe Arg
Asn Pro His Gln Ala Leu Leu His Ser Gly Asn Arg465 470 475 480Pro
Glu Glu Asp Cys Gly Leu Glu Gly Leu Val Cys Asn Ser Leu Cys 485 490
495Ala His Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys
500 505 510Ser His Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg
Val Trp 515 520 525Lys Gly Leu Pro Arg Glu Tyr Val Ser Asp Lys Arg
Cys Leu Pro Cys 530 535 540His Pro Glu Cys Gln Pro Gln Asn Ser Ser
Glu Thr Cys Phe Gly Ser545 550 555 560Glu Ala Asp Gln Cys Ala Ala
Cys Ala His Tyr Lys Asp Ser Ser Ser 565 570 575Cys Val Ala Arg Cys
Pro Ser Gly Val Lys Pro Asp Leu Ser Tyr Met 580 585 590Pro Ile Trp
Lys Tyr Pro Asp Glu Glu Gly Ile Cys Gln Pro Cys Pro 595 600 605Ile
Asn Cys Thr His Ser Cys Val Asp Leu Asp Glu Arg Gly Cys Pro 610 615
620Ala Glu Gln Arg Ala Ser Pro Val Thr Phe Ile Ile Ala Thr Val
Val625 630 635 640Gly Val Leu Leu Phe Leu Ile Leu Val Val Val Val
Gly Ile Leu Ile 645 650 655Lys Arg Arg Arg Gln Lys Ile Arg Lys Tyr
Thr Met 660 6657687PRTFelis catus 7Met Glu Leu Ala Ala Trp Cys Arg
Trp Gly Leu Leu Leu Ala Leu Leu1 5 10 15Pro Ser Gly Ala Thr Gly Thr
Gln Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser
Pro Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys
Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu His Ala Asn
Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly
Tyr Val Leu Ile Ala His Asn Gln Val Lys Gln Val Pro Leu 85 90 95Gln
Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105
110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asp Ser Gly Thr Pro
115 120 125Ala Thr Gly Ala Ala Leu Gly Gly Leu Arg Glu Leu Lys Leu
Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln
Arg Asn Pro Gln145 150 155 160Leu Cys His Gln Asp Thr Ile Leu Trp
Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu Met Leu
Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190Gln Pro Cys Ser Pro
Ala Cys Lys Asp Ser His Cys Trp Gly Ala Ser 195 200 205Ser Gly Asp
Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220Ala
Arg Cys Lys Gly Pro Gln Pro Thr Asp Cys Cys His Glu Gln Cys225 230
235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys
Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro
Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro
Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr
Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys
Thr Leu Val Cys Pro Leu Asn Asn Gln305 310 315 320Glu Val Thr Ala
Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys
Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345
350Ala Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Val Gly Cys Lys
355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Glu
Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln
Leu Arg Val Phe385 390 395 400Glu Ala Leu Glu Glu Ile Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asn Leu Ser
Val Phe Gln Asn Leu Arg Val Ile Arg 420 425 430Gly Arg Val Leu His
Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser
Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu
Ala Leu Ile His Arg Asn Ser Arg Leu Cys Phe Val His Thr Val465 470
475 480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His
Ser 485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Ala Gly Glu Gly Leu
Ala Cys Tyr 500 505 510Pro Leu Cys Ala His Gly His Cys Trp Gly Pro
Gly Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly
Gln Glu Cys Val Glu Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro
Arg Glu Tyr Val Lys Asp Arg Phe Cys545 550 555 560Leu Pro Cys His
Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570 575Leu Gly
Ser Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585
590Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu
595 600 605Ser Phe Met Pro Ile Trp Lys Phe Ala Asp Glu Glu Gly Thr
Cys Gln 610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys Ala Asp
Leu Asp Glu Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser
Pro Val Thr Ser Ile Ile Ala 645 650 655Ala Val Val Gly Ile Leu Leu
Val Val Val Val Gly Leu Val Leu Gly 660 665 670Ile Leu Ile Lys Arg
Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met 675 680 6858687PRTUrsus
americanus 8Met Glu Leu Ala Ala Trp Cys Arg Trp Gly Leu Leu Leu Ala
Leu Leu1 5 10 15Pro Ser Gly Ala Ala Gly Thr Gln Val Cys Thr Gly Thr
Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp
Met Leu Arg His 35 40 45Leu Tyr Gln Ala Cys Gln Val Val Gln Gly Asn
Leu Glu Leu Thr Tyr 50 55 60Leu Pro Ala Asn Ala Ser Leu Ser Phe Leu
Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val Leu Ile Ala His
Ser Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg
Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110Ala Leu Ala Val Leu
Asp Asn Gly Glu Pro Pro Lys Gly Asp Thr Ser 115 120 125Val Ala Gly
Ala Thr Pro Gly Gly Leu Arg Glu Leu Lys Leu Arg Ser 130 135 140Leu
Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln145 150
155 160Leu Cys His Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys
Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser
Arg Ala Cys 180 185 190Gln Pro Cys Ser Pro Ala Cys Lys Asp Pro His
Cys Trp Gly Ala Ser 195 200 205Ser Gly Asp Cys Gln Ser Leu Thr Arg
Thr Val Cys Ala Gly Gly Cys 210 215 220Ala Arg Cys Lys Gly Pro Lys
Pro Thr Asp Cys Cys His Glu Gln Cys225
230 235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala
Cys Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys
Pro Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met
Pro Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val
Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser
Cys Thr Leu Val Cys Pro Leu Asn Asn Gln305 310 315 320Glu Val Thr
Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Arg 325 330 335Pro
Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345
350Ala Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys
355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Glu
Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln
Leu Arg Val Phe385 390 395 400Glu Ala Leu Glu Glu Ile Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asn Leu Ser
Val Phe Gln Asn Leu Arg Val Ile Arg 420 425 430Gly Arg Val Leu His
Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser
Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu
Ala Leu Ile His Arg Asn Ala Arg Leu Cys Phe Ile His Thr Val465 470
475 480Pro Trp Glu Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His
Ser 485 490 495Ala Asn Arg Pro Glu Ala Glu Cys Val Gly Glu Gly Leu
Ala Cys Tyr 500 505 510Pro Leu Cys Ala His Gly His Cys Trp Gly Pro
Gly Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly
Gln Glu Cys Val Glu Glu Cys 530 535 540Arg Glu Leu His Gly Leu Pro
Arg Glu Tyr Val Lys Asp Arg Tyr Cys545 550 555 560Leu Pro Cys His
Pro Glu Cys Arg Pro Gln Asn Gly Ser Val Thr Cys 565 570 575Phe Gly
Ser Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585
590Pro Pro Ser Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu
595 600 605Ser Phe Met Pro Ile Trp Lys Phe Ala Asp Glu Glu Gly Thr
Cys Gln 610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys Gly Asp
Leu Asp Glu Arg625 630 635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser
Pro Val Thr Ser Ile Ile Ala 645 650 655Ala Val Val Gly Ile Leu Leu
Ala Val Val Met Gly Leu Val Leu Gly 660 665 670Ile Leu Ile Lys Arg
Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met 675 680 6859687PRTHomo
sapiens 9Met Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu Ala
Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly Thr
Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp
Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn
Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu
Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val Leu Ile Ala His
Asn Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg
Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110Ala Leu Ala Val Leu
Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro 115 120 125Val Thr Gly
Ala Ser Pro Gly Gly Leu Arg Glu Leu Lys Leu Arg Ser 130 135 140Leu
Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln145 150
155 160Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys
Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser
Arg Ala Cys 180 185 190His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg
Cys Trp Gly Glu Ser 195 200 205Ser Glu Asp Cys Gln Ser Leu Thr Arg
Thr Val Cys Ala Gly Gly Cys 210 215 220Ala Arg Cys Lys Gly Pro Leu
Pro Thr Asp Cys Cys His Glu Gln Cys225 230 235 240Ala Ala Gly Cys
Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu 245 250 255His Phe
Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val 260 265
270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg
275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn
Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro
Leu His Asn Gln305 310 315 320Glu Val Thr Ala Glu Asp Gly Thr Gln
Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys Ala Arg Val Cys Tyr
Gly Leu Gly Met Glu His Leu Arg Glu 340 345 350Val Arg Ala Val Thr
Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys 355 360 365Lys Ile Phe
Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp 370 375 380Pro
Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln Val Phe385 390
395 400Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp
Pro 405 410 415Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln
Val Ile Arg 420 425 430Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu
Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser Trp Leu Gly Leu Arg Ser
Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu Ala Leu Ile His His Asn
Thr His Leu Cys Phe Val His Thr Val465 470 475 480Pro Trp Asp Gln
Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Thr 485 490 495Ala Asn
Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala Cys His 500 505
510Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys
515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu
Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn
Ala Arg His Cys545 550 555 560Leu Pro Cys His Pro Glu Cys Gln Pro
Gln Asn Gly Ser Val Thr Cys 565 570 575Phe Gly Pro Glu Ala Asp Gln
Cys Val Ala Cys Ala His Tyr Lys Asp 580 585 590Pro Pro Phe Cys Val
Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu 595 600 605Ser Tyr Met
Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln 610 615 620Pro
Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp Lys625 630
635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu Thr Ser Ile Ile
Ser 645 650 655Ala Val Val Gly Ile Leu Leu Val Val Val Leu Gly Val
Val Phe Gly 660 665 670Ile Leu Ile Lys Arg Arg Gln Gln Lys Ile Arg
Lys Tyr Thr Met 675 680 68510688PRTMus Musculus 10Met Glu Leu Ala
Ala Trp Cys Arg Trp Gly Phe Leu Leu Ala Leu Leu1 5 10 15Ser Pro Gly
Ala Ala Gly Thr Gln Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg
Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu
Tyr Gln Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55
60Leu Pro Ala Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65
70 75 80Gln Gly Tyr Met Leu Ile Ala His Asn Arg Val Lys His Val Pro
Leu 85 90 95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp
Lys Tyr 100 105 110Ala Leu Ala Val Leu Asp Asn Arg Asp Pro Leu Asp
Asn Val Thr Thr 115 120 125Ala Ala Pro Gly Arg Thr Pro Glu Gly Leu
Arg Glu Leu Lys Leu Arg 130 135 140Ser Leu Thr Glu Ile Leu Lys Gly
Gly Val Leu Ile Arg Gly Asn Pro145 150 155 160Gln Leu Cys Tyr Gln
Asp Met Val Leu Trp Lys Asp Val Leu Arg Lys 165 170 175Asn Asn Gln
Leu Ala Pro Val Asp Met Asp Thr Asn Arg Ser Arg Ala 180 185 190Cys
Pro Pro Cys Ala Pro Thr Cys Lys Asp Asn His Cys Trp Gly Glu 195 200
205Ser Pro Glu Asp Cys Gln Ile Leu Thr Gly Thr Ile Cys Thr Ser Gly
210 215 220Cys Ala Arg Cys Lys Gly Arg Leu Pro Thr Asp Cys Cys His
Glu Gln225 230 235 240Cys Ala Ala Gly Cys Thr Gly Pro Lys His Ser
Asp Cys Leu Ala Cys 245 250 255Leu His Phe Asn His Ser Gly Ile Cys
Glu Leu His Cys Pro Ala Leu 260 265 270Ile Thr Tyr Asn Thr Asp Thr
Phe Glu Ser Met Leu Asn Pro Glu Gly 275 280 285Arg Tyr Thr Phe Gly
Ala Ser Cys Val Thr Thr Cys Pro Tyr Asn Tyr 290 295 300Leu Ser Thr
Glu Val Gly Ser Cys Thr Leu Val Cys Pro Pro Asn Asn305 310 315
320Gln Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser
325 330 335Lys Pro Cys Ala Gly Val Cys Tyr Gly Leu Gly Met Glu His
Leu Arg 340 345 350Gly Ala Arg Ala Ile Thr Ser Asp Asn Ile Gln Glu
Phe Ala Gly Cys 355 360 365Lys Lys Ile Phe Gly Ser Leu Ala Phe Leu
Pro Glu Ser Phe Asp Gly 370 375 380Asn Pro Ser Ser Gly Val Ala Pro
Leu Lys Pro Glu His Leu Gln Val385 390 395 400Phe Glu Thr Leu Glu
Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp 405 410 415Pro Glu Ser
Phe Gln Asp Leu Ser Val Phe Gln Asn Leu Arg Val Ile 420 425 430Arg
Gly Arg Ile Leu His Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly 435 440
445Leu Gly Ile His Ser Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser
450 455 460Gly Leu Ala Leu Ile His Arg Asn Thr His Leu Cys Phe Val
Asn Thr465 470 475 480Val Pro Trp Asp Gln Leu Phe Arg Asn Pro His
Gln Ala Leu Leu His 485 490 495Ser Gly Asn Arg Pro Glu Glu Ala Cys
Gly Leu Glu Gly Leu Val Cys 500 505 510Asn Ser Leu Cys Ala Arg Gly
His Cys Trp Gly Pro Gly Pro Thr Gln 515 520 525Cys Val Asn Cys Ser
Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu 530 535 540Cys Arg Val
Trp Lys Gly Leu Pro Arg Glu Tyr Val Arg Gly Lys His545 550 555
560Cys Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Ser Ser Glu Thr
565 570 575Cys Tyr Gly Ser Glu Ala Asp Gln Cys Glu Ala Cys Ala His
Tyr Lys 580 585 590Asp Ser Ser Ser Cys Val Ala Arg Cys Pro Ser Gly
Val Lys Pro Asp 595 600 605Leu Ser Tyr Met Pro Ile Trp Lys Tyr Pro
Asp Glu Glu Gly Ile Cys 610 615 620Gln Pro Cys Pro Ile Asn Cys Thr
His Ser Cys Val Asp Leu Asp Glu625 630 635 640Arg Gly Cys Pro Ala
Glu Gln Arg Ala Ser Pro Val Thr Phe Ile Ile 645 650 655Ala Thr Val
Val Gly Val Leu Leu Phe Leu Ile Ile Val Val Val Ile 660 665 670Gly
Ile Leu Ile Lys Arg Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met 675 680
68511691PRTRattus norvegicus 11Met Ile Ile Met Glu Leu Ala Ala Trp
Cys Arg Trp Gly Phe Leu Leu1 5 10 15Ala Leu Leu Pro Pro Gly Ile Ala
Gly Thr Gln Val Cys Thr Gly Thr 20 25 30Asp Met Lys Leu Arg Leu Pro
Ala Ser Pro Glu Thr His Leu Asp Met 35 40 45Leu Arg His Leu Tyr Gln
Gly Cys Gln Val Val Gln Gly Asn Leu Glu 50 55 60Leu Thr Tyr Val Pro
Ala Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile65 70 75 80Gln Glu Val
Gln Gly Tyr Met Leu Ile Ala His Asn Gln Val Lys Arg 85 90 95Val Pro
Leu Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu 100 105
110Asp Lys Tyr Ala Leu Ala Val Leu Asp Asn Arg Asp Pro Gln Asp Asn
115 120 125Val Ala Ala Ser Thr Pro Gly Arg Thr Pro Glu Gly Leu Arg
Glu Leu 130 135 140Lys Leu Arg Ser Leu Thr Glu Ile Leu Lys Gly Gly
Val Leu Ile Arg145 150 155 160Gly Asn Pro Gln Leu Cys Tyr Gln Asp
Met Val Leu Trp Lys Asp Val 165 170 175Phe Arg Lys Asn Asn Gln Leu
Ala Pro Val Asp Ile Asp Thr Asn Arg 180 185 190Ser Arg Ala Cys Pro
Pro Cys Ala Pro Ala Cys Lys Asp Asn His Cys 195 200 205Trp Gly Glu
Ser Pro Glu Asp Cys Gln Ile Leu Thr Gly Thr Ile Cys 210 215 220Thr
Ser Gly Cys Ala Arg Cys Lys Gly Arg Leu Pro Thr Asp Cys Cys225 230
235 240His Glu Gln Cys Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp
Cys 245 250 255Leu Ala Cys Leu His Phe Asn His Ser Gly Ile Cys Glu
Leu His Cys 260 265 270Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr Phe
Glu Ser Met His Asn 275 280 285Pro Glu Gly Arg Tyr Thr Phe Gly Ala
Ser Cys Val Thr Thr Cys Pro 290 295 300Tyr Asn Tyr Leu Ser Thr Glu
Val Gly Ser Cys Thr Leu Val Cys Pro305 310 315 320Pro Asn Asn Gln
Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu 325 330 335Lys Cys
Ser Lys Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu 340 345
350His Leu Arg Gly Ala Arg Ala Ile Thr Ser Asp Asn Val Gln Glu Phe
355 360 365Asp Gly Cys Lys Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro
Glu Ser 370 375 380Phe Asp Gly Asp Pro Ser Ser Gly Ile Ala Pro Leu
Arg Pro Glu Gln385 390 395 400Leu Gln Val Phe Glu Thr Leu Glu Glu
Ile Thr Gly Tyr Leu Tyr Ile 405 410 415Ser Ala Trp Pro Asp Ser Leu
Arg Asp Leu Ser Val Phe Gln Asn Leu 420 425 430Arg Ile Ile Arg Gly
Arg Ile Leu His Asp Gly Ala Tyr Ser Leu Thr 435 440 445Leu Gln Gly
Leu Gly Ile His Ser Leu Gly Leu Arg Ser Leu Arg Glu 450 455 460Leu
Gly Ser Gly Leu Ala Leu Ile His Arg Asn Ala His Leu Cys Phe465 470
475 480Val His Thr Val Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln
Ala 485 490 495Leu Leu His Ser Gly Asn Arg Pro Glu Glu Asp Cys Gly
Leu Glu Gly 500 505 510Leu Val Cys Asn Ser Leu Cys Ala His Gly His
Cys Trp Gly Pro Gly 515 520 525Pro Thr Gln Cys Val Asn Cys Ser His
Phe Leu Arg Gly Gln Glu Cys 530 535 540Val Glu Glu Cys Arg Val Trp
Lys Gly Leu Pro Arg Glu Tyr Val Ser545 550 555 560Asp Lys Arg Cys
Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Ser 565 570 575Ser Glu
Thr Cys Phe Gly Ser Glu Ala Asp Gln Cys Ala Ala Cys Ala 580 585
590His Tyr Lys Asp Ser Ser Ser Cys Val Ala Arg Cys Pro Ser Gly Val
595 600 605Lys Pro Asp Leu Ser Tyr Met Pro Ile Trp Lys Tyr Pro Asp
Glu Glu 610 615 620Gly Ile Cys Gln Pro Cys Pro Ile Asn Cys Thr His
Ser Cys Val Asp625 630
635 640Leu Asp Glu Arg Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Val
Thr 645 650 655Phe Ile Ile Ala Thr Val Val Gly Val Leu Leu Phe Leu
Ile Leu Val 660 665 670Val Val Val Gly Ile Leu Ile Lys Arg Arg Arg
Gln Lys Ile Arg Lys 675 680 685Tyr Thr Met 69012690PRTArtificial
SequenceChimeric human-rat E2Neu (HER2) 12Met Glu Leu Ala Ala Leu
Cys Arg Trp Gly Leu Leu Leu Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala
Ser Thr Gln Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro
Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln
Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro
Thr Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75
80Gln Gly Tyr Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu
85 90 95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn
Tyr 100 105 110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn
Thr Thr Pro 115 120 125Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu
Leu Lys Leu Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val
Leu Ile Gln Arg Asn Pro Gln145 150 155 160Leu Cys Tyr Gln Asp Thr
Ile Leu Trp Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala
Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190His Pro
Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser 195 200
205Ser Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys
210 215 220Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu
Gln Cys225 230 235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp
Cys Leu Ala Cys Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu
Leu His Cys Pro Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe
Glu Ser Met Pro Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala
Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp
Val Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn Gln305 310 315
320Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys
325 330 335Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu
Arg Glu 340 345 350Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe
Ala Gly Cys Lys 355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro
Glu Ser Phe Asp Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Glu Phe
Ala Pro Leu Arg Pro Glu Gln Leu385 390 395 400Gln Val Phe Glu Thr
Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser 405 410 415Ala Trp Pro
Asp Ser Leu Arg Asp Leu Ser Val Phe Gln Asn Leu Arg 420 425 430Ile
Ile Arg Gly Arg Ile Leu His Asp Gly Ala Tyr Ser Leu Thr Leu 435 440
445Gln Gly Leu Gly Ile His Ser Leu Gly Leu Arg Ser Leu Arg Glu Leu
450 455 460Gly Ser Gly Leu Ala Leu Ile His Arg Asn Ala His Leu Cys
Phe Val465 470 475 480His Thr Val Pro Trp Asp Gln Leu Phe Arg Asn
Pro His Gln Ala Leu 485 490 495Leu His Ser Gly Asn Arg Pro Glu Glu
Asp Cys Gly Leu Glu Gly Leu 500 505 510Val Cys Asn Ser Leu Cys Ala
His Gly His Cys Trp Gly Pro Gly Pro 515 520 525Thr Gln Cys Val Asn
Cys Ser His Phe Leu Arg Gly Gln Glu Cys Val 530 535 540Glu Glu Cys
Arg Val Trp Lys Gly Leu Pro Arg Glu Tyr Val Ser Asp545 550 555
560Lys Arg Cys Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Ser Ser
565 570 575Glu Thr Cys Phe Gly Ser Glu Ala Asp Gln Cys Ala Ala Cys
Ala His 580 585 590Tyr Lys Asp Ser Ser Ser Cys Val Ala Arg Cys Pro
Ser Gly Val Lys 595 600 605Pro Asp Leu Ser Tyr Met Pro Ile Trp Lys
Tyr Pro Asp Glu Glu Gly 610 615 620Ile Cys Gln Pro Cys Pro Ile Asn
Cys Thr His Ser Cys Val Asp Leu625 630 635 640Asp Glu Arg Gly Cys
Pro Ala Glu Gln Arg Ala Ser Pro Val Thr Phe 645 650 655Ile Ile Ala
Thr Val Val Gly Val Leu Leu Phe Leu Ile Leu Val Val 660 665 670Val
Val Gly Ile Leu Ile Lys Arg Arg Arg Gln Lys Ile Arg Lys Tyr 675 680
685Thr Met 690131995DNAFelis catus 13acccaagtgt gcaccggcac
agacatgaag ctgcggctcc cagccagtcc cgagacccac 60ctggacatgc tccgccacct
ctaccagggc tgtcaagtgg tacagggcaa cctggagctc 120acctacctgc
atgccaatgc cagcctctcc ttcctgcagg atatccagga ggtgcaaggc
180tatgtgctca ttgcccacaa ccaagtgaaa caggtcccac tgcagaggct
acgaatcgtg 240cgaggcaccc agctctttga ggacaactac gccctggccg
tgctggacaa cggagaccca 300ctggacagtg gcacccctgc tacaggggct
gccctaggag ggctgcggga gctgaagctc 360cgaagcctca cagagatcct
gaagggaggg gtcctcattc agcggaaccc gcagctctgc 420caccaggaca
cgattctgtg gaaggacatc ttccacaaga acaaccagct ggccctcatg
480ctgatagaca ccaaccgctc tcgggcctgc caaccctgtt ctccagcttg
taaagactcc 540cactgctggg gagcaagttc cggggactgt cagagcttga
ctcgaactgt ctgtgctggc 600ggctgtgccc gctgcaaggg cccgcagccc
accgactgct gccacgagca atgtgctgct 660ggctgcacgg gccccaagca
ttctgactgc ctggcctgcc tccacttcaa ccacagtggc 720atctgtgagc
tgcactgccc agccctggtc acctacaaca cggacacctt cgaatccatg
780cccaaccctg agggccgtta taccttcggt gccagctgtg tgactgcctg
tccctacaac 840tacctgtcta cggacgtggg atcctgcacc ctggtctgtc
ccctgaacaa ccaagaggtg 900acagctgagg atggaacaca gcggtgtgag
aaatgcagca agccctgtgc ccgagtgtgc 960tacggcctag gcatggagca
cctgcgggag gcgagggcag tcaccagtgc caacatccaa 1020gaatttgtcg
gctgcaagaa gatctttggg agcctggcgt ttctgccaga gagctttgag
1080ggggacccag cctccaacac tgcccccctg cagcctgagc agctcagagt
gtttgaggct 1140ctggaggaga ttacaggtta cctgtacatc tcagcgtggc
cagacagctt gcctaacctc 1200agtgtcttcc agaacctcag agtgatccgg
ggccgagttc tgcatgacgg tgcttactcg 1260ctgacccttc aagggctggg
catcagctgg ctggggctgc gctcgctgcg ggagctgggc 1320agtgggctgg
ccctcatcca ccgcaactcc cgcctctgct tcgtacacac ggtgccctgg
1380gaccagctct tccggaaccc ccaccaggcc ctgctccaca gcgccaaccg
gccagaggac 1440gagtgcgcgg gtgagggcct ggcctgctat ccgctgtgtg
cccacgggca ctgctggggt 1500ccgggaccca cccagtgtgt caactgcagc
cagttccttc ggggccagga gtgcgtggag 1560gaatgccgag tattgcaggg
gcttccccgg gagtatgtga aggataggtt ctgtctgcca 1620tgccacccgg
agtgtcagcc ccagaatggc tcagtgacct gcttgggctc ggaagctgac
1680cagtgtgtgg cctgtgccca ctacaaggac cctcctttct gtgtggctcg
ctgccccagt 1740ggggtgaaac ctgacctctc cttcatgccc atctggaagt
tcgcagatga ggagggcacg 1800tgccagccat gccccatcaa ctgcacccac
tcctgtgcgg acctggacga gaagggctgc 1860cccgccgagc agagagccag
ccctgtgacg tccatcattg ctgctgtggt gggcattctg 1920ctggtcgtgg
ttgtggggct ggtccttggc atcctaatca agcgaaggcg gcagaagatc
1980cggaagtaca cgatg 1995141995DNAUrsus americanus 14acccaagtgt
gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac 60ctggatatgc
tccgccacct ctaccaggcc tgtcaagtgg tacagggtaa cctggagctc
120acctacctgc ccgccaatgc cagcctgtcc ttcctgcagg atatccagga
ggtacagggc 180tatgtgctca ttgctcacag ccaagtgaga caggtcccgc
tgcagaggct ccgaatcgtg 240cgaggcaccc agctctttga ggacaactac
gccctggccg tgctggacaa tggagagccg 300cccaaggggg acacctctgt
ggcaggggct accccaggag ggctgcggga gctgaagctt 360cgaagcctca
cagagatcct gaagggaggg gtcttgattc agcggaaccc acagctctgc
420caccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggccctcacg 480ctgatagaca ccaaccgctc tcgggcctgc caaccctgtt
ctccagcctg taaagacccc 540cactgctggg gagcaagttc cggggactgt
cagagcttga cacgaaccgt ctgtgccggc 600ggctgtgccc gctgcaaggg
cccaaaaccc actgactgct gccatgagca gtgcgcggct 660ggctgcacgg
gccccaagca ctcggactgc ctggcctgcc ttcacttcaa ccacagtggc
720atctgtgagc tgcactgccc agccctggtc acctacaaca cggacacgtt
cgaatccatg 780cccaaccctg agggccgata caccttcggt gccagctgtg
tgaccgcctg tccctacaac 840tacctgtcca cggacgtggg atcctgcacc
ctggtctgtc ccctgaacaa ccaagaggtg 900acggctgagg atggcaccca
gcggtgtgag aaatgcagca gaccctgtgc ccgagtgtgc 960tatggtctgg
gcatggagca cctgcgggag gcgagggcgg tcaccagcgc caacatccaa
1020gagttcgccg gctgcaagaa gatctttggg agcctggcgt ttctgccaga
gagcttcgag 1080ggagacccag cctccaacac tgcccccctg cagcctgaac
agctcagagt gttcgaggcc 1140ctggaggaga tcacaggtta cctgtatatc
tcagcgtggc cggacagctt gcctaacctc 1200agtgtcttcc agaacctgcg
agtaatccgg ggacgagttc tgcatgatgg cgcctactcg 1260ctgaccctgc
aagggctggg catcagctgg ctggggctgc gctcgctgcg ggaactgggc
1320agcgggctgg ccctcatcca ccgcaacgcc cgcctctgct tcatccacac
ggtgccctgg 1380gagcagctct tccggaaccc ccaccaagcc ctgctgcaca
gtgccaaccg gccggaggcc 1440gagtgcgtgg gcgagggcct ggcctgctac
ccgctgtgcg cccatgggca ctgctggggt 1500ccggggccca cccagtgcgt
caactgcagc caattccttc ggggccagga gtgcgtggag 1560gaatgccgag
aactgcacgg gctaccccgg gaatatgtga aggacagata ctgtctgcca
1620tgccaccccg agtgtcggcc ccagaatggc tcagtgacct gctttgggtc
ggaggctgac 1680cagtgtgtgg cctgcgccca ctacaaggac cctccctcct
gcgtggctcg ctgccccagt 1740ggtgtgaaac ccgacctctc tttcatgccc
atttggaagt ttgcagatga ggagggcaca 1800tgccagccgt gccccatcaa
ctgcacccac tcctgtgggg acctggacga gaggggctgc 1860cccgccgaac
agagagccag ccctgtgaca tccatcattg ccgctgtggt gggcattctg
1920ctggccgtgg tcatggggct ggtcctcggc atcctgatca agcgaaggcg
acagaagatc 1980cggaagtaca cgatg 1995151998DNAHomo sapiens
15acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac
60ctggacatgc tccgccacct ctaccagggc tgccaggtgg tgcagggaaa cctggaactc
120acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg atatccagga
ggtgcagggc 180tacgtgctca tcgctcacaa ccaagtgagg caggtcccac
tgcagaggct gcggattgtg 240cgaggcaccc agctctttga ggacaactat
gccctggccg tgctagacaa tggagacccg 300ctgaacaata ccacccctgt
cacaggggcc tccccaggag gcctgcggga gctgaagctt 360cgaagcctca
cagagatctt gaaaggaggg gtcttgatcc agcggaaccc ccagctctgc
420taccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggctctcaca 480ctgatagaca ccaaccgctc tcgggcctgc cacccctgtt
ctccgatgtg taagggctcc 540cgctgctggg gagagagttc tgaggattgt
cagagcctga cgcgcactgt ctgtgccggt 600ggctgtgccc gctgcaaggg
gccactgccc actgactgct gccatgagca gtgtgctgcc 660ggctgcacgg
gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc
720atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt
tgagtccatg 780cccaatcccg agggccggta tacattcggc gccagctgtg
tgactgcctg tccctacaac 840tacctttcta cggacgtggg atcctgcacc
ctcgtctgcc ccctgcacaa ccaagaggtg 900acagcagagg atggaacaca
gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc 960tatggtctgg
gcatggagca cttgcgagag gtgagggcag ttaccagtgc caatatccag
1020gagtttgctg gctgcaagaa gatctttggg agcctggcat ttctgccgga
gagctttgat 1080ggggacccag cctccaacac tgccccgctc cagccagagc
agctccaagt gtttgagact 1140ctggaagaga tcacaggtta cctatacatc
tcagcatggc cggacagcct gcctgacctc 1200agcgtcttcc agaacctgca
agtaatccgg ggacgaattc tgcacaatgg cgcctactcg 1260ctgaccctgc
aagggctggg catcagctgg ctggggctgc gctcactgag ggaactgggc
1320agtggactgg ccctcatcca ccataacacc cacctctgct tcgtgcacac
ggtgccctgg 1380gaccagctct ttcggaaccc gcaccaagct ctgctccaca
ctgccaaccg gccagaggac 1440gagtgtgtgg gcgagggcct ggcctgccac
cagctgtgcg cccgagggca ctgctggggt 1500ccagggccca cccagtgtgt
caactgcagc cagttccttc ggggccagga gtgcgtggag 1560gaatgccgag
tactgcaggg gctccccagg gagtatgtga atgccaggca ctgtttgccg
1620tgccaccctg agtgtcagcc ccagaatggc tcagtgacct gttttggacc
ggaggctgac 1680cagtgtgtgg cctgtgccca ctataaggac cctcccttct
gcgtggcccg ctgccccagc 1740ggtgtgaaac ctgacctctc ctacatgccc
atctggaagt ttccagatga ggagggcgca 1800tgccagcctt gccccatcaa
ctgcacccac tcctgtgtgg acctggatga caagggctgc 1860cccgccgagc
agagagccag ccctctgacg tccatcatct ctgcggtggt tggcattctg
1920ctggtcgtgg tcttgggggt ggtctttggg atcctcatca agcgacggca
gcagaagatc 1980cggaagtaca cgatgtag 1998161998DNAMus musculus
16acccaagtgt gtaccggtac cgacatgaag ttgcgactcc ctgccagtcc tgagacccac
60ctggacatgc ttcgccacct ctaccagggc tgtcaggtgg tgcagggcaa tttggagctt
120acctacctgc ccgccaatgc cagcctctca ttcctgcagg acatccagga
agtccaggga 180tacatgctca tcgctcacaa ccgagtgaaa cacgtcccac
tgcagaggtt gcgcatcgtg 240agagggactc agctctttga ggacaagtat
gccctggctg tgctagacaa ccgagaccct 300ttggacaacg tcaccaccgc
cgccccaggc agaaccccag aagggctgcg ggagctgaag 360cttcgaagtc
tcacagagat cttgaaggga ggagttttga tccgtgggaa ccctcagctc
420tgctaccagg acatggtttt gtggaaggat gtcctccgta agaataacca
gctggctcct 480gtcgacatgg acaccaatcg ttcccgggcc tgtccacctt
gtgccccaac ctgcaaagac 540aatcactgtt ggggtgagag tcctgaagac
tgtcagatct tgactggcac catctgtact 600agtggctgtg cccggtgcaa
gggccggctg cccactgact gttgccatga gcagtgtgct 660gcaggctgca
cgggtcccaa gcattctgac tgcctggcct gcctccactt caatcatagt
720ggtatctgtg agctgcactg cccggccctc atcacctaca acacagacac
cttcgagtcc 780atgctcaacc ctgagggtcg ctacaccttt ggtgccagct
gtgtgaccac ctgcccctac 840aactacctct ccacggaagt gggatcctgc
actctggtct gtcccccgaa caaccaagag 900gtcacagctg aggacggaac
acagcggtgt gagaaatgca gcaagccctg tgctggagta 960tgctatggtc
tgggcatgga gcacctccga ggggcgaggg ccatcaccag tgacaatatc
1020caggagtttg ctggctgcaa gaagatcttt gggagcctgg catttttgcc
ggagagcttt 1080gatgggaacc cctcctccgg cgttgcccca ctgaagccag
agcatctcca agtgttcgaa 1140accctggagg agatcacagg ttacctatac
atttcagcat ggccagagag cttccaagac 1200ctcagtgtct tccagaacct
tcgggtcatt cggggacgga ttctccatga tggtgcttac 1260tcattgacgt
tgcaaggcct ggggattcac tcactggggc tacgctcact gcgggagctg
1320ggcagtggat tggctctcat tcaccgcaac acccatctct gctttgtaaa
cactgtacct 1380tgggaccagc tcttccggaa cccgcaccag gccctactcc
acagtgggaa ccggccagaa 1440gaggcatgtg gtcttgaggg cttggtctgt
aactcactgt gtgcccgtgg gcactgctgg 1500gggccagggc ccacccagtg
tgtcaactgc agtcagttcc tccggggcca ggagtgtgtg 1560gaggagtgcc
gagtatggaa ggggctcccc agggagtatg tgaggggcaa gcactgtctg
1620ccatgccacc ccgagtgtca gcctcaaaac agctcggaga cctgctatgg
atcggaggct 1680gaccagtgtg aggcttgtgc ccactacaag gactcatctt
cctgtgtggc tcgctgcccc 1740agtggtgtga agccagacct ctcctacatg
cctatctgga agtacccgga tgaggagggc 1800atatgtcagc catgccccat
caactgcacc cactcatgtg tggacctgga cgaacgaggc 1860tgcccagcag
agcagagagc cagcccagtg acattcatca ttgcaactgt ggtgggcgtc
1920ctgttgttcc tgatcatagt ggtggtcatt ggaatcctaa tcaaacgaag
gcgacagaag 1980atccggaagt ataccatg 1998171998DNARattus norvegicus
17acccaagtgt gtaccggcac agacatgaag ttgcggctcc ctgccagtcc tgagacccac
60ctggacatgc tccgccacct gtaccagggc tgtcaggtag tgcagggcaa cttggagctt
120acctacgtgc ctgccaatgc cagcctctca ttcctgcagg acatccagga
agttcagggt 180tacatgctca tcgctcacaa ccaggtgaag cgcgtcccac
tgcaaaggct gcgcatcgtg 240agagggaccc agctctttga ggacaagtat
gccctggctg tgctagacaa ccgagatcct 300caggacaatg tcgccgcctc
caccccaggc agaaccccag aggggctgcg ggagctgaag 360cttcgaagtc
tcacagagat cctgaaggga ggagttttga tccgtgggaa ccctcagctc
420tgctaccagg acatggtttt gtggaaggac gtcttccgca agaataacca
actggctcct 480gtcgatatag acaccaatcg ttcccgggcc tgtccacctt
gtgcccccgc ctgcaaagac 540aatcactgtt ggggtgagag tccggaagac
tgtcagatct tgactggcac catctgtacc 600agtggttgtg cccggtgcaa
gggccggctg cccactgact gctgccatga gcagtgtgcc 660gcaggctgca
cgggccccaa gcattctgac tgcctggcct gcctccactt caatcatagt
720ggtatctgtg agctgcactg cccagccctc gtcacctaca acacagacac
ctttgagtcc 780atgcacaacc ctgagggtcg ctacaccttt ggtgccagct
gcgtgaccac ctgcccctac 840aactacctgt ctacggaagt gggatcctgc
actctggtgt gtcccccgaa taaccaagag 900gtcacagctg aggacggaac
acagcgttgt gagaaatgca gcaagccctg tgctcgagtg 960tgctatggtc
tgggcatgga gcaccttcga ggggcgaggg ccatcaccag tgacaatgtc
1020caggagtttg atggctgcaa gaagatcttt gggagcctgg catttttgcc
ggagagcttt 1080gatggggacc cctcctccgg cattgctccg ctgaggcctg
agcagctcca agtgttcgaa 1140accctggagg agatcacagg ttacctgtac
atctcagcat ggccagacag tctccgtgac 1200ctcagtgtct tccagaacct
tcgaatcatt cggggacgga ttctccacga tggcgcgtac 1260tcattgacac
tgcaaggcct ggggatccac tcgctggggc tgcgctcact gcgggagctg
1320ggcagtggat tggctctgat tcaccgcaac gcccatctct gctttgtaca
cactgtacct 1380tgggaccagc tcttccggaa cccacatcag gccctgctcc
acagtgggaa ccggccggaa 1440gaggattgtg gtctcgaggg cttggtctgt
aactcactgt gtgcccacgg gcactgctgg 1500gggccagggc ccacccagtg
tgtcaactgc agtcatttcc ttcggggcca ggagtgtgtg 1560gaggagtgcc
gagtatggaa ggggctcccc cgggagtatg tgagtgacaa gcgctgtctg
1620ccgtgtcacc ccgagtgtca gcctcaaaac agctcagaga cctgctttgg
atcggaggct 1680gatcagtgtg cagcctgcgc ccactacaag gactcgtcct
cctgtgtggc tcgctgcccc 1740agtggtgtga aaccggacct ctcctacatg
cccatctgga agtacccgga tgaggagggc 1800atatgccagc cgtgccccat
caactgcacc cactcctgtg tggatctgga tgaacgaggc 1860tgcccagcag
agcagagagc cagcccggtg acattcatca ttgcaactgt agtgggcgtc
1920ctgctgttcc tgatcttagt ggtggtcgtt ggaatcctaa tcaaacgaag
gagacagaag 1980atccggaagt atacgatg 1998182004DNAArtificial
SequenceChimeric human-rat E2Neu (HER2) 18acccaagtgt gcaccggcac
agacatgaag ctgcggctcc ctgccagtcc cgagacccac 60ctggacatgc tccgccacct
ctaccagggc tgccaggtgg tgcagggaaa
cctggaactc 120acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg
atatccagga ggtgcagggc 180tacgtgctca tcgctcacaa ccaagtgagg
caggtcccac tgcagaggct gcggattgtg 240cgaggcaccc agctctttga
ggacaactat gccctggccg tgctagacaa tggagacccg 300ctgaacaata
ccacccctgt cacaggggcc tccccaggag gcctgcggga gctgaagctt
360cgaagcctca cagagatctt gaaaggaggg gtcttgatcc agcggaaccc
ccagctctgc 420taccaggaca cgattttgtg gaaggacatc ttccacaaga
acaaccagct ggctctcaca 480ctgatagaca ccaaccgctc tcgggcctgc
cacccctgtt ctccgatgtg taagggctcc 540cgctgctggg gagagagttc
tgaggattgt cagagcctga cgcgcactgt ctgtgccggt 600ggctgtgccc
gctgcaaggg gccactgccc actgactgct gccatgagca gtgtgctgcc
660ggctgcacgg gccccaagca ctctgactgc ctggcctgcc tccacttcaa
ccacagtggc 720atctgtgagc tgcactgccc agccctggtc acctacaaca
cagacacgtt tgagtccatg 780cccaatcccg agggccggta tacattcggc
gccagctgtg tgactgcctg tccctacaac 840tacctttcta cggacgtggg
atcctgcacc ctcgtctgcc ccctgcacaa ccaagaggtg 900acagcagagg
atggaacaca gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc
960tatggtctgg gcatggagca cttgcgagag gtgagggcag ttaccagtgc
caatatccag 1020gagtttgctg gctgcaagaa gatctttggg agcctggcat
ttctgccgga gagctttgat 1080ggggacccag cctccaacac tgccgaattc
gctccgctga ggcctgagca gctccaagtg 1140ttcgaaaccc tggaggagat
cacaggttac ctgtacatct cagcatggcc agacagtctc 1200cgtgacctca
gtgtcttcca gaaccttcga atcattcggg gacggattct ccacgatggc
1260gcgtactcat tgacactgca aggcctgggg atccactcgc tggggctgcg
ctcactgcgg 1320gagctgggca gtggattggc tctgattcac cgcaacgccc
atctctgctt tgtacacact 1380gtaccttggg accagctctt ccggaaccca
catcaggccc tgctccacag tgggaaccgg 1440ccggaagagg attgtggtct
cgagggcttg gtctgtaact cactgtgtgc ccacgggcac 1500tgctgggggc
cagggcccac ccagtgtgtc aactgcagtc atttccttcg gggccaggag
1560tgtgtggagg agtgccgagt atggaagggg ctcccccggg agtatgtgag
tgacaagcgc 1620tgtctgccgt gtcaccccga gtgtcagcct caaaacagct
cagagacctg ctttggatcg 1680gaggctgatc agtgtgcagc ctgcgcccac
tacaaggact cgtcctcctg tgtggctcgc 1740tgccccagtg gtgtgaaacc
ggacctctcc tacatgccca tctggaagta cccggatgag 1800gagggcatat
gccagccgtg ccccatcaac tgcacccact cctgtgtgga tctggatgaa
1860cgaggctgcc cagcagagca gagagccagc ccggtgacat tcatcattgc
aactgtagtg 1920ggcgtcctgc tgttcctgat cttagtggtg gtcgttggaa
tcctaatcaa acgaaggaga 1980cagaagatcc ggaagtatac gatg
2004192061DNAFelis catus 19atggagctgg cggcctggtg ccgctggggg
ctcctcctcg ccctcctgcc ctccggagcc 60acgggcaccc aagtgtgcac cggcacagac
atgaagctgc ggctcccagc cagtcccgag 120acccacctgg acatgctccg
ccacctctac cagggctgtc aagtggtaca gggcaacctg 180gagctcacct
acctgcatgc caatgccagc ctctccttcc tgcaggatat ccaggaggtg
240caaggctatg tgctcattgc ccacaaccaa gtgaaacagg tcccactgca
gaggctacga 300atcgtgcgag gcacccagct ctttgaggac aactacgccc
tggccgtgct ggacaacgga 360gacccactgg acagtggcac ccctgctaca
ggggctgccc taggagggct gcgggagctg 420aagctccgaa gcctcacaga
gatcctgaag ggaggggtcc tcattcagcg gaacccgcag 480ctctgccacc
aggacacgat tctgtggaag gacatcttcc acaagaacaa ccagctggcc
540ctcatgctga tagacaccaa ccgctctcgg gcctgccaac cctgttctcc
agcttgtaaa 600gactcccact gctggggagc aagttccggg gactgtcaga
gcttgactcg aactgtctgt 660gctggcggct gtgcccgctg caagggcccg
cagcccaccg actgctgcca cgagcaatgt 720gctgctggct gcacgggccc
caagcattct gactgcctgg cctgcctcca cttcaaccac 780agtggcatct
gtgagctgca ctgcccagcc ctggtcacct acaacacgga caccttcgaa
840tccatgccca accctgaggg ccgttatacc ttcggtgcca gctgtgtgac
tgcctgtccc 900tacaactacc tgtctacgga cgtgggatcc tgcaccctgg
tctgtcccct gaacaaccaa 960gaggtgacag ctgaggatgg aacacagcgg
tgtgagaaat gcagcaagcc ctgtgcccga 1020gtgtgctacg gcctaggcat
ggagcacctg cgggaggcga gggcagtcac cagtgccaac 1080atccaagaat
ttgtcggctg caagaagatc tttgggagcc tggcgtttct gccagagagc
1140tttgaggggg acccagcctc caacactgcc cccctgcagc ctgagcagct
cagagtgttt 1200gaggctctgg aggagattac aggttacctg tacatctcag
cgtggccaga cagcttgcct 1260aacctcagtg tcttccagaa cctcagagtg
atccggggcc gagttctgca tgacggtgct 1320tactcgctga cccttcaagg
gctgggcatc agctggctgg ggctgcgctc gctgcgggag 1380ctgggcagtg
ggctggccct catccaccgc aactcccgcc tctgcttcgt acacacggtg
1440ccctgggacc agctcttccg gaacccccac caggccctgc tccacagcgc
caaccggcca 1500gaggacgagt gcgcgggtga gggcctggcc tgctatccgc
tgtgtgccca cgggcactgc 1560tggggtccgg gacccaccca gtgtgtcaac
tgcagccagt tccttcgggg ccaggagtgc 1620gtggaggaat gccgagtatt
gcaggggctt ccccgggagt atgtgaagga taggttctgt 1680ctgccatgcc
acccggagtg tcagccccag aatggctcag tgacctgctt gggctcggaa
1740gctgaccagt gtgtggcctg tgcccactac aaggaccctc ctttctgtgt
ggctcgctgc 1800cccagtgggg tgaaacctga cctctccttc atgcccatct
ggaagttcgc agatgaggag 1860ggcacgtgcc agccatgccc catcaactgc
acccactcct gtgcggacct ggacgagaag 1920ggctgccccg ccgagcagag
agccagccct gtgacgtcca tcattgctgc tgtggtgggc 1980attctgctgg
tcgtggttgt ggggctggtc cttggcatcc taatcaagcg aaggcggcag
2040aagatccgga agtacacgat g 2061202061DNAUrsus americanus
20atggagctgg cggcctggtg ccgctggggg ctcctcctcg ccctcctgcc ctccggagcc
60gcgggcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc cagtcccgag
120acccacctgg atatgctccg ccacctctac caggcctgtc aagtggtaca
gggtaacctg 180gagctcacct acctgcccgc caatgccagc ctgtccttcc
tgcaggatat ccaggaggta 240cagggctatg tgctcattgc tcacagccaa
gtgagacagg tcccgctgca gaggctccga 300atcgtgcgag gcacccagct
ctttgaggac aactacgccc tggccgtgct ggacaatgga 360gagccgccca
agggggacac ctctgtggca ggggctaccc caggagggct gcgggagctg
420aagcttcgaa gcctcacaga gatcctgaag ggaggggtct tgattcagcg
gaacccacag 480ctctgccacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggcc 540ctcacgctga tagacaccaa ccgctctcgg
gcctgccaac cctgttctcc agcctgtaaa 600gacccccact gctggggagc
aagttccggg gactgtcaga gcttgacacg aaccgtctgt 660gccggcggct
gtgcccgctg caagggccca aaacccactg actgctgcca tgagcagtgc
720gcggctggct gcacgggccc caagcactcg gactgcctgg cctgccttca
cttcaaccac 780agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacgga cacgttcgaa 840tccatgccca accctgaggg ccgatacacc
ttcggtgcca gctgtgtgac cgcctgtccc 900tacaactacc tgtccacgga
cgtgggatcc tgcaccctgg tctgtcccct gaacaaccaa 960gaggtgacgg
ctgaggatgg cacccagcgg tgtgagaaat gcagcagacc ctgtgcccga
1020gtgtgctatg gtctgggcat ggagcacctg cgggaggcga gggcggtcac
cagcgccaac 1080atccaagagt tcgccggctg caagaagatc tttgggagcc
tggcgtttct gccagagagc 1140ttcgagggag acccagcctc caacactgcc
cccctgcagc ctgaacagct cagagtgttc 1200gaggccctgg aggagatcac
aggttacctg tatatctcag cgtggccgga cagcttgcct 1260aacctcagtg
tcttccagaa cctgcgagta atccggggac gagttctgca tgatggcgcc
1320tactcgctga ccctgcaagg gctgggcatc agctggctgg ggctgcgctc
gctgcgggaa 1380ctgggcagcg ggctggccct catccaccgc aacgcccgcc
tctgcttcat ccacacggtg 1440ccctgggagc agctcttccg gaacccccac
caagccctgc tgcacagtgc caaccggccg 1500gaggccgagt gcgtgggcga
gggcctggcc tgctacccgc tgtgcgccca tgggcactgc 1560tggggtccgg
ggcccaccca gtgcgtcaac tgcagccaat tccttcgggg ccaggagtgc
1620gtggaggaat gccgagaact gcacgggcta ccccgggaat atgtgaagga
cagatactgt 1680ctgccatgcc accccgagtg tcggccccag aatggctcag
tgacctgctt tgggtcggag 1740gctgaccagt gtgtggcctg cgcccactac
aaggaccctc cctcctgcgt ggctcgctgc 1800cccagtggtg tgaaacccga
cctctctttc atgcccattt ggaagtttgc agatgaggag 1860ggcacatgcc
agccgtgccc catcaactgc acccactcct gtggggacct ggacgagagg
1920ggctgccccg ccgaacagag agccagccct gtgacatcca tcattgccgc
tgtggtgggc 1980attctgctgg ccgtggtcat ggggctggtc ctcggcatcc
tgatcaagcg aaggcgacag 2040aagatccgga agtacacgat g 2061212064DNAHomo
sapiens 21atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc
ccccggagcc 60gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc
cagtcccgag 120acccacctgg acatgctccg ccacctctac cagggctgcc
aggtggtgca gggaaacctg 180gaactcacct acctgcccac caatgccagc
ctgtccttcc tgcaggatat ccaggaggtg 240cagggctacg tgctcatcgc
tcacaaccaa gtgaggcagg tcccactgca gaggctgcgg 300attgtgcgag
gcacccagct ctttgaggac aactatgccc tggccgtgct agacaatgga
360gacccgctga acaataccac ccctgtcaca ggggcctccc caggaggcct
gcgggagctg 420aagcttcgaa gcctcacaga gatcttgaaa ggaggggtct
tgatccagcg gaacccccag 480ctctgctacc aggacacgat tttgtggaag
gacatcttcc acaagaacaa ccagctggct 540ctcacactga tagacaccaa
ccgctctcgg gcctgccacc cctgttctcc gatgtgtaag 600ggctcccgct
gctggggaga gagttctgag gattgtcaga gcctgacgcg cactgtctgt
660gccggtggct gtgcccgctg caaggggcca ctgcccactg actgctgcca
tgagcagtgt 720gctgccggct gcacgggccc caagcactct gactgcctgg
cctgcctcca cttcaaccac 780agtggcatct gtgagctgca ctgcccagcc
ctggtcacct acaacacaga cacgtttgag 840tccatgccca atcccgaggg
ccggtataca ttcggcgcca gctgtgtgac tgcctgtccc 900tacaactacc
tttctacgga cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa
960gaggtgacag cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc
ctgtgcccga 1020gtgtgctatg gtctgggcat ggagcacttg cgagaggtga
gggcagttac cagtgccaat 1080atccaggagt ttgctggctg caagaagatc
tttgggagcc tggcatttct gccggagagc 1140tttgatgggg acccagcctc
caacactgcc ccgctccagc cagagcagct ccaagtgttt 1200gagactctgg
aagagatcac aggttaccta tacatctcag catggccgga cagcctgcct
1260gacctcagcg tcttccagaa cctgcaagta atccggggac gaattctgca
caatggcgcc 1320tactcgctga ccctgcaagg gctgggcatc agctggctgg
ggctgcgctc actgagggaa 1380ctgggcagtg gactggccct catccaccat
aacacccacc tctgcttcgt gcacacggtg 1440ccctgggacc agctctttcg
gaacccgcac caagctctgc tccacactgc caaccggcca 1500gaggacgagt
gtgtgggcga gggcctggcc tgccaccagc tgtgcgcccg agggcactgc
1560tggggtccag ggcccaccca gtgtgtcaac tgcagccagt tccttcgggg
ccaggagtgc 1620gtggaggaat gccgagtact gcaggggctc cccagggagt
atgtgaatgc caggcactgt 1680ttgccgtgcc accctgagtg tcagccccag
aatggctcag tgacctgttt tggaccggag 1740gctgaccagt gtgtggcctg
tgcccactat aaggaccctc ccttctgcgt ggcccgctgc 1800cccagcggtg
tgaaacctga cctctcctac atgcccatct ggaagtttcc agatgaggag
1860ggcgcatgcc agccttgccc catcaactgc acccactcct gtgtggacct
ggatgacaag 1920ggctgccccg ccgagcagag agccagccct ctgacgtcca
tcatctctgc ggtggttggc 1980attctgctgg tcgtggtctt gggggtggtc
tttgggatcc tcatcaagcg acggcagcag 2040aagatccgga agtacacgat gtag
2064222064DNAMus musculus 22atggagctgg cggcctggtg ccgttggggg
ttcctcctcg ccctcctgtc ccccggagcc 60gcgggtaccc aagtgtgtac cggtaccgac
atgaagttgc gactccctgc cagtcctgag 120acccacctgg acatgcttcg
ccacctctac cagggctgtc aggtggtgca gggcaatttg 180gagcttacct
acctgcccgc caatgccagc ctctcattcc tgcaggacat ccaggaagtc
240cagggataca tgctcatcgc tcacaaccga gtgaaacacg tcccactgca
gaggttgcgc 300atcgtgagag ggactcagct ctttgaggac aagtatgccc
tggctgtgct agacaaccga 360gaccctttgg acaacgtcac caccgccgcc
ccaggcagaa ccccagaagg gctgcgggag 420ctgaagcttc gaagtctcac
agagatcttg aagggaggag ttttgatccg tgggaaccct 480cagctctgct
accaggacat ggttttgtgg aaggatgtcc tccgtaagaa taaccagctg
540gctcctgtcg acatggacac caatcgttcc cgggcctgtc caccttgtgc
cccaacctgc 600aaagacaatc actgttgggg tgagagtcct gaagactgtc
agatcttgac tggcaccatc 660tgtactagtg gctgtgcccg gtgcaagggc
cggctgccca ctgactgttg ccatgagcag 720tgtgctgcag gctgcacggg
tcccaagcat tctgactgcc tggcctgcct ccacttcaat 780catagtggta
tctgtgagct gcactgcccg gccctcatca cctacaacac agacaccttc
840gagtccatgc tcaaccctga gggtcgctac acctttggtg ccagctgtgt
gaccacctgc 900ccctacaact acctctccac ggaagtggga tcctgcactc
tggtctgtcc cccgaacaac 960caagaggtca cagctgagga cggaacacag
cggtgtgaga aatgcagcaa gccctgtgct 1020ggagtatgct atggtctggg
catggagcac ctccgagggg cgagggccat caccagtgac 1080aatatccagg
agtttgctgg ctgcaagaag atctttggga gcctggcatt tttgccggag
1140agctttgatg ggaacccctc ctccggcgtt gccccactga agccagagca
tctccaagtg 1200ttcgaaaccc tggaggagat cacaggttac ctatacattt
cagcatggcc agagagcttc 1260caagacctca gtgtcttcca gaaccttcgg
gtcattcggg gacggattct ccatgatggt 1320gcttactcat tgacgttgca
aggcctgggg attcactcac tggggctacg ctcactgcgg 1380gagctgggca
gtggattggc tctcattcac cgcaacaccc atctctgctt tgtaaacact
1440gtaccttggg accagctctt ccggaacccg caccaggccc tactccacag
tgggaaccgg 1500ccagaagagg catgtggtct tgagggcttg gtctgtaact
cactgtgtgc ccgtgggcac 1560tgctgggggc cagggcccac ccagtgtgtc
aactgcagtc agttcctccg gggccaggag 1620tgtgtggagg agtgccgagt
atggaagggg ctccccaggg agtatgtgag gggcaagcac 1680tgtctgccat
gccaccccga gtgtcagcct caaaacagct cggagacctg ctatggatcg
1740gaggctgacc agtgtgaggc ttgtgcccac tacaaggact catcttcctg
tgtggctcgc 1800tgccccagtg gtgtgaagcc agacctctcc tacatgccta
tctggaagta cccggatgag 1860gagggcatat gtcagccatg ccccatcaac
tgcacccact catgtgtgga cctggacgaa 1920cgaggctgcc cagcagagca
gagagccagc ccagtgacat tcatcattgc aactgtggtg 1980ggcgtcctgt
tgttcctgat catagtggtg gtcattggaa tcctaatcaa acgaaggcga
2040cagaagatcc ggaagtatac catg 2064232073DNARattus norvegicus
23atgatcatca tggagctggc ggcctggtgc cgctgggggt tcctcctcgc cctcctgccc
60cccggaatcg cgggcaccca agtgtgtacc ggcacagaca tgaagttgcg gctccctgcc
120agtcctgaga cccacctgga catgctccgc cacctgtacc agggctgtca
ggtagtgcag 180ggcaacttgg agcttaccta cgtgcctgcc aatgccagcc
tctcattcct gcaggacatc 240caggaagttc agggttacat gctcatcgct
cacaaccagg tgaagcgcgt cccactgcaa 300aggctgcgca tcgtgagagg
gacccagctc tttgaggaca agtatgccct ggctgtgcta 360gacaaccgag
atcctcagga caatgtcgcc gcctccaccc caggcagaac cccagagggg
420ctgcgggagc tgaagcttcg aagtctcaca gagatcctga agggaggagt
tttgatccgt 480gggaaccctc agctctgcta ccaggacatg gttttgtgga
aggacgtctt ccgcaagaat 540aaccaactgg ctcctgtcga tatagacacc
aatcgttccc gggcctgtcc accttgtgcc 600cccgcctgca aagacaatca
ctgttggggt gagagtccgg aagactgtca gatcttgact 660ggcaccatct
gtaccagtgg ttgtgcccgg tgcaagggcc ggctgcccac tgactgctgc
720catgagcagt gtgccgcagg ctgcacgggc cccaagcatt ctgactgcct
ggcctgcctc 780cacttcaatc atagtggtat ctgtgagctg cactgcccag
ccctcgtcac ctacaacaca 840gacacctttg agtccatgca caaccctgag
ggtcgctaca cctttggtgc cagctgcgtg 900accacctgcc cctacaacta
cctgtctacg gaagtgggat cctgcactct ggtgtgtccc 960ccgaataacc
aagaggtcac agctgaggac ggaacacagc gttgtgagaa atgcagcaag
1020ccctgtgctc gagtgtgcta tggtctgggc atggagcacc ttcgaggggc
gagggccatc 1080accagtgaca atgtccagga gtttgatggc tgcaagaaga
tctttgggag cctggcattt 1140ttgccggaga gctttgatgg ggacccctcc
tccggcattg ctccgctgag gcctgagcag 1200ctccaagtgt tcgaaaccct
ggaggagatc acaggttacc tgtacatctc agcatggcca 1260gacagtctcc
gtgacctcag tgtcttccag aaccttcgaa tcattcgggg acggattctc
1320cacgatggcg cgtactcatt gacactgcaa ggcctgggga tccactcgct
ggggctgcgc 1380tcactgcggg agctgggcag tggattggct ctgattcacc
gcaacgccca tctctgcttt 1440gtacacactg taccttggga ccagctcttc
cggaacccac atcaggccct gctccacagt 1500gggaaccggc cggaagagga
ttgtggtctc gagggcttgg tctgtaactc actgtgtgcc 1560cacgggcact
gctgggggcc agggcccacc cagtgtgtca actgcagtca tttccttcgg
1620ggccaggagt gtgtggagga gtgccgagta tggaaggggc tcccccggga
gtatgtgagt 1680gacaagcgct gtctgccgtg tcaccccgag tgtcagcctc
aaaacagctc agagacctgc 1740tttggatcgg aggctgatca gtgtgcagcc
tgcgcccact acaaggactc gtcctcctgt 1800gtggctcgct gccccagtgg
tgtgaaaccg gacctctcct acatgcccat ctggaagtac 1860ccggatgagg
agggcatatg ccagccgtgc cccatcaact gcacccactc ctgtgtggat
1920ctggatgaac gaggctgccc agcagagcag agagccagcc cggtgacatt
catcattgca 1980actgtagtgg gcgtcctgct gttcctgatc ttagtggtgg
tcgttggaat cctaatcaaa 2040cgaaggagac agaagatccg gaagtatacg atg
2073242070DNAArtificial SequenceChimeric human-rat E2Neu (HER2)
24atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc ccccggagcc
60gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc cagtcccgag
120acccacctgg acatgctccg ccacctctac cagggctgcc aggtggtgca
gggaaacctg 180gaactcacct acctgcccac caatgccagc ctgtccttcc
tgcaggatat ccaggaggtg 240cagggctacg tgctcatcgc tcacaaccaa
gtgaggcagg tcccactgca gaggctgcgg 300attgtgcgag gcacccagct
ctttgaggac aactatgccc tggccgtgct agacaatgga 360gacccgctga
acaataccac ccctgtcaca ggggcctccc caggaggcct gcgggagctg
420aagcttcgaa gcctcacaga gatcttgaaa ggaggggtct tgatccagcg
gaacccccag 480ctctgctacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggct 540ctcacactga tagacaccaa ccgctctcgg
gcctgccacc cctgttctcc gatgtgtaag 600ggctcccgct gctggggaga
gagttctgag gattgtcaga gcctgacgcg cactgtctgt 660gccggtggct
gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt
720gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca
cttcaaccac 780agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacaga cacgtttgag 840tccatgccca atcccgaggg ccggtataca
ttcggcgcca gctgtgtgac tgcctgtccc 900tacaactacc tttctacgga
cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa 960gaggtgacag
cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga
1020gtgtgctatg gtctgggcat ggagcacttg cgagaggtga gggcagttac
cagtgccaat 1080atccaggagt ttgctggctg caagaagatc tttgggagcc
tggcatttct gccggagagc 1140tttgatgggg acccagcctc caacactgcc
gaattcgctc cgctgaggcc tgagcagctc 1200caagtgttcg aaaccctgga
ggagatcaca ggttacctgt acatctcagc atggccagac 1260agtctccgtg
acctcagtgt cttccagaac cttcgaatca ttcggggacg gattctccac
1320gatggcgcgt actcattgac actgcaaggc ctggggatcc actcgctggg
gctgcgctca 1380ctgcgggagc tgggcagtgg attggctctg attcaccgca
acgcccatct ctgctttgta 1440cacactgtac cttgggacca gctcttccgg
aacccacatc aggccctgct ccacagtggg 1500aaccggccgg aagaggattg
tggtctcgag ggcttggtct gtaactcact gtgtgcccac 1560gggcactgct
gggggccagg gcccacccag tgtgtcaact gcagtcattt ccttcggggc
1620caggagtgtg tggaggagtg ccgagtatgg aaggggctcc cccgggagta
tgtgagtgac 1680aagcgctgtc tgccgtgtca ccccgagtgt cagcctcaaa
acagctcaga gacctgcttt 1740ggatcggagg ctgatcagtg tgcagcctgc
gcccactaca aggactcgtc ctcctgtgtg 1800gctcgctgcc ccagtggtgt
gaaaccggac ctctcctaca tgcccatctg gaagtacccg 1860gatgaggagg
gcatatgcca gccgtgcccc atcaactgca cccactcctg tgtggatctg
1920gatgaacgag gctgcccagc agagcagaga gccagcccgg tgacattcat
cattgcaact 1980gtagtgggcg tcctgctgtt cctgatctta gtggtggtcg
ttggaatcct aatcaaacga 2040aggagacaga agatccggaa gtatacgatg
207025665PRTHomo sapiens 25Thr Gln Val Cys Thr Gly Thr Asp Met Lys
Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu Arg
His Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu
Thr Tyr Leu Pro Thr Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile
Gln Glu Val Gln Gly
Tyr Val Leu Ile 50 55 60Ala His Asn Gln Val Arg Gln Val Pro Leu Gln
Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu Asp Asn
Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro Leu Asn Asn Thr
Thr Pro Val Thr Gly Ala Ser Pro 100 105 110Gly Gly Leu Arg Glu Leu
Gln Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120 125Gly Gly Val Leu
Ile Gln Arg Asn Pro Gln Leu Cys Tyr Gln Asp Thr 130 135 140Ile Leu
Trp Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala Leu Thr145 150 155
160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys His Pro Cys Ser Pro Met
165 170 175Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser Ser Glu Asp Cys
Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala Gly Gly Cys Ala Arg
Cys Lys Gly Pro 195 200 205Leu Pro Thr Asp Cys Cys His Glu Gln Cys
Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His Ser Asp Cys Leu Ala
Cys Leu His Phe Asn His Ser Gly225 230 235 240Ile Cys Glu Leu His
Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr 245 250 255Phe Glu Ser
Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala Ser 260 265 270Cys
Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr Asp Val Gly Ser 275 280
285Cys Thr Leu Val Cys Pro Leu His Asn Gln Glu Val Thr Ala Glu Asp
290 295 300Gly Thr Lys Arg Cys Glu Lys Cys Ser Lys Pro Cys Ala Arg
Val Cys305 310 315 320Tyr Gly Leu Gly Met Glu His Leu Arg Glu Val
Arg Ala Val Thr Ser 325 330 335Ala Asn Ile Gln Glu Phe Ala Gly Cys
Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala Phe Leu Pro Glu Ser Phe
Asp Gly Asp Pro Ala Ser Asn Thr Ala 355 360 365Pro Leu Gln Pro Glu
Gln Leu Gln Val Phe Glu Thr Leu Glu Glu Ile 370 375 380Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu Pro Asp Leu385 390 395
400Ser Val Phe Gln Asn Leu Gln Val Ile Arg Gly Arg Ile Leu His Asn
405 410 415Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu Gly Ile Ser Trp
Leu Gly 420 425 430Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly Leu Ala
Leu Ile His His 435 440 445Asn Thr His Leu Cys Phe Val His Thr Val
Pro Trp Asp Gln Leu Phe 450 455 460Arg Asn Pro His Gln Ala Leu Leu
His Thr Ala Asn Arg Pro Glu Asp465 470 475 480Glu Cys Val Gly Glu
Gly Leu Ala Cys His Gln Leu Cys Ala Arg Gly 485 490 495His Cys Trp
Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser Gln Phe 500 505 510Leu
Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Leu Gln Gly Leu 515 520
525Pro Arg Glu Tyr Val Asn Ala Arg His Cys Leu Pro Cys His Pro Glu
530 535 540Cys Gln Pro Gln Asn Gly Ser Val Thr Cys Phe Gly Pro Glu
Ala Asp545 550 555 560Gln Cys Val Ala Cys Ala His Tyr Lys Asp Pro
Pro Phe Cys Val Ala 565 570 575Arg Cys Pro Ser Gly Val Lys Pro Asp
Leu Ser Tyr Met Pro Ile Trp 580 585 590Lys Phe Pro Asp Glu Glu Gly
Ala Cys Gln Pro Cys Pro Ile Asn Cys 595 600 605Thr His Ser Cys Val
Asp Leu Asp Asp Lys Gly Cys Pro Ala Glu Gln 610 615 620Arg Ala Ser
Pro Leu Thr Ser Ile Ile Ser Ala Val Val Gly Ile Leu625 630 635
640Leu Val Val Val Leu Gly Val Val Phe Gly Ile Leu Ile Lys Arg Arg
645 650 655Gln Gln Lys Ile Arg Lys Tyr Thr Met 660 66526665PRTHomo
sapiens 26Thr Gln Val Cys Thr Gly Thr Asp Met Lys Leu Arg Leu Pro
Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu Arg His Leu Tyr Gln
Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu Thr Tyr Leu Pro
Thr Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile Gln Glu Val Gln
Gly Tyr Val Leu Ile 50 55 60Ala His Asn Gln Val Arg Gln Val Pro Leu
Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu Asp
Asn Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro Leu Asn Asn
Thr Thr Pro Val Thr Gly Ala Ser Pro 100 105 110Gly Gly Leu Arg Glu
Leu Gln Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120 125Gly Gly Val
Leu Ile Gln Arg Asn Pro Gln Leu Cys Tyr Gln Asp Thr 130 135 140Ile
Leu Trp Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala Leu Thr145 150
155 160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys His Pro Cys Ser Pro
Met 165 170 175Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser Ser Glu Asp
Cys Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala Gly Gly Cys Ala
Arg Cys Lys Gly Pro 195 200 205Leu Pro Thr Asp Cys Cys His Glu Gln
Cys Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His Ser Asp Cys Leu
Ala Cys Leu His Phe Asn His Ser Gly225 230 235 240Ile Cys Glu Leu
His Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr 245 250 255Phe Glu
Ser Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala Ser 260 265
270Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr Asp Val Gly Ser
275 280 285Cys Thr Leu Val Cys Pro Leu His Asn Gln Glu Val Thr Ala
Glu Asp 290 295 300Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys Pro Cys
Ala Arg Val Cys305 310 315 320Tyr Gly Leu Gly Met Glu His Leu Arg
Glu Val Arg Ala Val Thr Ser 325 330 335Ala Asn Ile Gln Glu Phe Ala
Gly Cys Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala Phe Leu Pro Glu
Ser Phe Asp Gly Asp Pro Ala Ser Asn Thr Ala 355 360 365Pro Leu Gln
Pro Glu Gln Leu Gln Val Phe Glu Thr Leu Glu Glu Ile 370 375 380Thr
Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu Pro Asp Leu385 390
395 400Ser Val Phe Gln Asn Leu Arg Val Ile Arg Gly Arg Ile Leu His
Asn 405 410 415Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu Gly Ile Ser
Trp Leu Gly 420 425 430Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly Leu
Ala Leu Ile His His 435 440 445Asn Thr His Leu Cys Phe Val His Thr
Val Pro Trp Asp Gln Leu Phe 450 455 460Arg Asn Pro His Gln Ala Leu
Leu His Thr Ala Asn Arg Pro Glu Asp465 470 475 480Glu Cys Val Gly
Glu Gly Leu Ala Cys His Gln Leu Cys Ala Arg Gly 485 490 495His Cys
Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser Gln Phe 500 505
510Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Leu Gln Gly Leu
515 520 525Pro Arg Glu Tyr Val Asn Ala Arg His Cys Leu Pro Cys His
Pro Glu 530 535 540Cys Gln Pro Gln Asn Gly Ser Val Thr Cys Phe Gly
Pro Glu Ala Asp545 550 555 560Gln Cys Val Ala Cys Ala His Tyr Lys
Asp Pro Pro Phe Cys Val Ala 565 570 575Arg Cys Pro Ser Gly Val Lys
Pro Asp Leu Ser Tyr Met Pro Ile Trp 580 585 590Lys Phe Pro Asp Glu
Glu Gly Ala Cys Gln Pro Cys Pro Ile Asn Cys 595 600 605Thr His Ser
Cys Val Asp Leu Asp Asp Lys Gly Cys Pro Ala Glu Gln 610 615 620Arg
Ala Ser Pro Leu Thr Ser Ile Ile Ser Ala Val Val Gly Ile Leu625 630
635 640Leu Val Val Val Leu Gly Val Val Phe Gly Ile Leu Ile Lys Arg
Arg 645 650 655Gln Gln Lys Ile Arg Lys Tyr Thr Met 660
66527665PRTHomo sapiens 27Thr Gln Val Cys Thr Gly Thr Asp Met Lys
Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu Arg
His Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu
Thr Tyr Leu Pro Thr Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile
Gln Glu Val Gln Gly Tyr Val Leu Ile 50 55 60Ala His Asn Gln Val Arg
Gln Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln
Leu Phe Glu Asp Asn Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp
Pro Leu Asn Asn Thr Thr Pro Val Thr Gly Ala Ser Pro 100 105 110Gly
Gly Leu Arg Glu Leu Gln Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120
125Gly Gly Val Leu Ile Gln Arg Asn Pro Gln Leu Cys Tyr Gln Asp Thr
130 135 140Ile Leu Trp Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala
Leu Thr145 150 155 160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys His
Pro Cys Ser Pro Met 165 170 175Cys Lys Gly Ser Arg Cys Trp Gly Glu
Ser Ser Glu Asp Cys Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala
Gly Gly Cys Ala Arg Cys Lys Gly Pro 195 200 205Leu Pro Thr Asp Cys
Cys His Glu Gln Cys Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His
Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser Gly225 230 235
240Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr
245 250 255Phe Glu Ser Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly
Ala Ser 260 265 270Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr
Asp Val Gly Ser 275 280 285Cys Thr Leu Val Cys Pro Leu His Asn Gln
Glu Val Thr Ala Glu Asp 290 295 300Gly Thr Gln Arg Cys Glu Lys Cys
Ser Lys Pro Cys Ala Arg Val Cys305 310 315 320Tyr Gly Leu Gly Met
Glu His Leu Arg Glu Val Arg Ala Val Thr Ser 325 330 335Ala Asn Ile
Gln Glu Phe Ala Gly Cys Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala
Phe Leu Pro Glu Ser Phe Asp Gly Asp Pro Ala Ser Asn Thr Ala 355 360
365Pro Leu Gln Pro Glu Gln Leu Gln Val Phe Glu Thr Leu Glu Glu Ile
370 375 380Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu Pro
Asp Leu385 390 395 400Ser Val Phe Gln Asn Leu Gln Val Ile Arg Gly
Arg Ile Leu His Asp 405 410 415Gly Ala Tyr Ser Leu Thr Leu Gln Gly
Leu Gly Ile Ser Trp Leu Gly 420 425 430Leu Arg Ser Leu Arg Glu Leu
Gly Ser Gly Leu Ala Leu Ile His His 435 440 445Asn Thr His Leu Cys
Phe Val His Thr Val Pro Trp Asp Gln Leu Phe 450 455 460Arg Asn Pro
His Gln Ala Leu Leu His Thr Ala Asn Arg Pro Glu Asp465 470 475
480Glu Cys Val Gly Glu Gly Leu Ala Cys His Gln Leu Cys Ala Arg Gly
485 490 495His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser
Gln Phe 500 505 510Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val
Leu Gln Gly Leu 515 520 525Pro Arg Glu Tyr Val Asn Ala Arg His Cys
Leu Pro Cys His Pro Glu 530 535 540Cys Gln Pro Gln Asn Gly Ser Val
Thr Cys Phe Gly Pro Glu Ala Asp545 550 555 560Gln Cys Val Ala Cys
Ala His Tyr Lys Asp Pro Pro Phe Cys Val Ala 565 570 575Arg Cys Pro
Ser Gly Val Lys Pro Asp Leu Ser Tyr Met Pro Ile Trp 580 585 590Lys
Phe Pro Asp Glu Glu Gly Ala Cys Gln Pro Cys Pro Ile Asn Cys 595 600
605Thr His Ser Cys Val Asp Leu Asp Asp Lys Gly Cys Pro Ala Glu Gln
610 615 620Arg Ala Ser Pro Leu Thr Ser Ile Ile Ser Ala Val Val Gly
Ile Leu625 630 635 640Leu Val Val Val Leu Gly Val Val Phe Gly Ile
Leu Ile Lys Arg Arg 645 650 655Gln Gln Lys Ile Arg Lys Tyr Thr Met
660 66528687PRTHomo sapiens 28Met Glu Leu Ala Ala Leu Cys Arg Trp
Gly Leu Leu Leu Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln
Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro
Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln
Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala
Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr
Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg
Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105
110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro
115 120 125Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu
Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln
Arg Asn Pro Gln145 150 155 160Leu Cys Tyr Gln Asp Thr Ile Leu Trp
Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu
Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190His Pro Cys Ser Pro
Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser 195 200 205Ser Glu Asp
Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220Ala
Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln Cys225 230
235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys
Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro
Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro
Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr
Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys
Thr Leu Val Cys Pro Leu His Asn Gln305 310 315 320Glu Val Thr Ala
Glu Asp Gly Thr Lys Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys
Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345
350Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys
355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp
Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln
Leu Gln Val Phe385 390 395 400Glu Thr Leu Glu Glu Ile Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asp Leu Ser
Val Phe Gln Asn Leu Gln Val Ile Arg 420 425 430Gly Arg Ile Leu His
Asn Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser
Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu
Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His Thr Val465 470
475 480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His
Thr 485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu
Ala Cys His 500 505 510Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro
Gly Pro Thr
Gln Cys 515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys
Val Glu Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr
Val Asn Ala Arg His Cys545 550 555 560Leu Pro Cys His Pro Glu Cys
Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570 575Phe Gly Pro Glu Ala
Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585 590Pro Pro Phe
Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu 595 600 605Ser
Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln 610 615
620Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp
Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu Thr
Ser Ile Ile Ser 645 650 655Ala Val Val Gly Ile Leu Leu Val Val Val
Leu Gly Val Val Phe Gly 660 665 670Ile Leu Ile Lys Arg Arg Gln Gln
Lys Ile Arg Lys Tyr Thr Met 675 680 68529687PRTHomo sapiens 29Met
Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu Ala Leu Leu1 5 10
15Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly Thr Asp Met Lys
20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg
His 35 40 45Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu
Thr Tyr 50 55 60Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile
Gln Glu Val65 70 75 80Gln Gly Tyr Val Leu Ile Ala His Asn Gln Val
Arg Gln Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln
Leu Phe Glu Asp Asn Tyr 100 105 110Ala Leu Ala Val Leu Asp Asn Gly
Asp Pro Leu Asn Asn Thr Thr Pro 115 120 125Val Thr Gly Ala Ser Pro
Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser 130 135 140Leu Thr Glu Ile
Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln145 150 155 160Leu
Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys Asn 165 170
175Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys
180 185 190His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly
Glu Ser 195 200 205Ser Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys
Ala Gly Gly Cys 210 215 220Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp
Cys Cys His Glu Gln Cys225 230 235 240Ala Ala Gly Cys Thr Gly Pro
Lys His Ser Asp Cys Leu Ala Cys Leu 245 250 255His Phe Asn His Ser
Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val 260 265 270Thr Tyr Asn
Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg 275 280 285Tyr
Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295
300Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn
Gln305 310 315 320Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu
Lys Cys Ser Lys 325 330 335Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly
Met Glu His Leu Arg Glu 340 345 350Val Arg Ala Val Thr Ser Ala Asn
Ile Gln Glu Phe Ala Gly Cys Lys 355 360 365Lys Ile Phe Gly Ser Leu
Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp 370 375 380Pro Ala Ser Asn
Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln Val Phe385 390 395 400Glu
Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro 405 410
415Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Arg Val Ile Arg
420 425 430Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu Thr Leu Gln
Gly Leu 435 440 445Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu
Leu Gly Ser Gly 450 455 460Leu Ala Leu Ile His His Asn Thr His Leu
Cys Phe Val His Thr Val465 470 475 480Pro Trp Asp Gln Leu Phe Arg
Asn Pro His Gln Ala Leu Leu His Thr 485 490 495Ala Asn Arg Pro Glu
Asp Glu Cys Val Gly Glu Gly Leu Ala Cys His 500 505 510Gln Leu Cys
Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys 515 520 525Val
Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys 530 535
540Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg His
Cys545 550 555 560Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly
Ser Val Thr Cys 565 570 575Phe Gly Pro Glu Ala Asp Gln Cys Val Ala
Cys Ala His Tyr Lys Asp 580 585 590Pro Pro Phe Cys Val Ala Arg Cys
Pro Ser Gly Val Lys Pro Asp Leu 595 600 605Ser Tyr Met Pro Ile Trp
Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln 610 615 620Pro Cys Pro Ile
Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp Lys625 630 635 640Gly
Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu Thr Ser Ile Ile Ser 645 650
655Ala Val Val Gly Ile Leu Leu Val Val Val Leu Gly Val Val Phe Gly
660 665 670Ile Leu Ile Lys Arg Arg Gln Gln Lys Ile Arg Lys Tyr Thr
Met 675 680 68530687PRTHomo sapiens 30Met Glu Leu Ala Ala Leu Cys
Arg Trp Gly Leu Leu Leu Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser
Thr Gln Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala
Ser Pro Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly
Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr
Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln
Gly Tyr Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu 85 90
95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr
100 105 110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr
Thr Pro 115 120 125Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu
Gln Leu Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu
Ile Gln Arg Asn Pro Gln145 150 155 160Leu Cys Tyr Gln Asp Thr Ile
Leu Trp Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu
Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190His Pro Cys
Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser 195 200 205Ser
Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215
220Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln
Cys225 230 235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys
Leu Ala Cys Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu
His Cys Pro Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu
Ser Met Pro Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser
Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val
Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn Gln305 310 315 320Glu
Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330
335Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu
340 345 350Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly
Cys Lys 355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser
Phe Asp Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro
Glu Gln Leu Gln Val Phe385 390 395 400Glu Thr Leu Glu Glu Ile Thr
Gly Tyr Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asp
Leu Ser Val Phe Gln Asn Leu Gln Val Ile Arg 420 425 430Gly Arg Ile
Leu His Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly
Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455
460Leu Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His Thr
Val465 470 475 480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala
Leu Leu His Thr 485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Val Gly
Glu Gly Leu Ala Cys His 500 505 510Gln Leu Cys Ala Arg Gly His Cys
Trp Gly Pro Gly Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe
Leu Arg Gly Gln Glu Cys Val Glu Glu Cys 530 535 540Arg Val Leu Gln
Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg His Cys545 550 555 560Leu
Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570
575Phe Gly Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp
580 585 590Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro
Asp Leu 595 600 605Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu
Gly Ala Cys Gln 610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys
Val Asp Leu Asp Asp Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg
Ala Ser Pro Leu Thr Ser Ile Ile Ser 645 650 655Ala Val Val Gly Ile
Leu Leu Val Val Val Leu Gly Val Val Phe Gly 660 665 670Ile Leu Ile
Lys Arg Arg Gln Gln Lys Ile Arg Lys Tyr Thr Met 675 680
685311998DNAHomo sapiens 31acccaagtgt gcaccggcac agacatgaag
ctgcggctcc ctgccagtcc cgagacccac 60ctggacatgc tccgccacct ctaccagggc
tgccaggtgg tgcagggaaa cctggaactc 120acctacctgc ccaccaatgc
cagcctgtcc ttcctgcagg atatccagga ggtgcagggc 180tacgtgctca
tcgctcacaa ccaagtgagg caggtcccac tgcagaggct gcggattgtg
240cgaggcaccc agctctttga ggacaactat gccctggccg tgctagacaa
tggagacccg 300ctgaacaata ccacccctgt cacaggggcc tccccaggag
gcctgcggga gctgcagctt 360cgaagcctca cagagatctt gaaaggaggg
gtcttgatcc agcggaaccc ccagctctgc 420taccaggaca cgattttgtg
gaaggacatc ttccacaaga acaaccagct ggctctcaca 480ctgatagaca
ccaaccgctc tcgggcctgc cacccctgtt ctccgatgtg taagggctcc
540cgctgctggg gagagagttc tgaggattgt cagagcctga cgcgcactgt
ctgtgccggt 600ggctgtgccc gctgcaaggg gccactgccc actgactgct
gccatgagca gtgtgctgcc 660ggctgcacgg gccccaagca ctctgactgc
ctggcctgcc tccacttcaa ccacagtggc 720atctgtgagc tgcactgccc
agccctggtc acctacaaca cagacacgtt tgagtccatg 780cccaatcccg
agggccggta tacattcggc gccagctgtg tgactgcctg tccctacaac
840tacctttcta cggacgtggg atcctgcacc ctcgtctgcc ccctgcacaa
ccaagaggtg 900acagcagagg atggaacaaa gcggtgtgag aagtgcagca
agccctgtgc ccgagtgtgc 960tatggtctgg gcatggagca cttgcgagag
gtgagggcag ttaccagtgc caatatccag 1020gagtttgctg gctgcaagaa
gatctttggg agcctggcat ttctgccgga gagctttgat 1080ggggacccag
cctccaacac tgccccgctc cagccagagc agctccaagt gtttgagact
1140ctggaagaga tcacaggtta cctatacatc tcagcatggc cggacagcct
gcctgacctc 1200agcgtcttcc agaacctgca agtaatccgg ggacgaattc
tgcacaatgg cgcctactcg 1260ctgaccctgc aagggctggg catcagctgg
ctggggctgc gctcactgag ggaactgggc 1320agtggactgg ccctcatcca
ccataacacc cacctctgct tcgtgcacac ggtgccctgg 1380gaccagctct
ttcggaaccc gcaccaagct ctgctccaca ctgccaaccg gccagaggac
1440gagtgtgtgg gcgagggcct ggcctgccac cagctgtgcg cccgagggca
ctgctggggt 1500ccagggccca cccagtgtgt caactgcagc cagttccttc
ggggccagga gtgcgtggag 1560gaatgccgag tactgcaggg gctccccagg
gagtatgtga atgccaggca ctgtttgccg 1620tgccaccctg agtgtcagcc
ccagaatggc tcagtgacct gttttggacc ggaggctgac 1680cagtgtgtgg
cctgtgccca ctataaggac cctcccttct gcgtggcccg ctgccccagc
1740ggtgtgaaac ctgacctctc ctacatgccc atctggaagt ttccagatga
ggagggcgca 1800tgccagcctt gccccatcaa ctgcacccac tcctgtgtgg
acctggatga caagggctgc 1860cccgccgagc agagagccag ccctctgacg
tccatcatct ctgcggtggt tggcattctg 1920ctggtcgtgg tcttgggggt
ggtctttggg atcctcatca agcgacggca gcagaagatc 1980cggaagtaca cgatgtga
1998321998DNAHomo sapiens 32acccaagtgt gcaccggcac agacatgaag
ctgcggctcc ctgccagtcc cgagacccac 60ctggacatgc tccgccacct ctaccagggc
tgccaggtgg tgcagggaaa cctggaactc 120acctacctgc ccaccaatgc
cagcctgtcc ttcctgcagg atatccagga ggtgcagggc 180tacgtgctca
tcgctcacaa ccaagtgagg caggtcccac tgcagaggct gcggattgtg
240cgaggcaccc agctctttga ggacaactat gccctggccg tgctagacaa
tggagacccg 300ctgaacaata ccacccctgt cacaggggcc tccccaggag
gcctgcggga gctgcagctt 360cgaagcctca cagagatctt gaaaggaggg
gtcttgatcc agcggaaccc ccagctctgc 420taccaggaca cgattttgtg
gaaggacatc ttccacaaga acaaccagct ggctctcaca 480ctgatagaca
ccaaccgctc tcgggcctgc cacccctgtt ctccgatgtg taagggctcc
540cgctgctggg gagagagttc tgaggattgt cagagcctga cgcgcactgt
ctgtgccggt 600ggctgtgccc gctgcaaggg gccactgccc actgactgct
gccatgagca gtgtgctgcc 660ggctgcacgg gccccaagca ctctgactgc
ctggcctgcc tccacttcaa ccacagtggc 720atctgtgagc tgcactgccc
agccctggtc acctacaaca cagacacgtt tgagtccatg 780cccaatcccg
agggccggta tacattcggc gccagctgtg tgactgcctg tccctacaac
840tacctttcta cggacgtggg atcctgcacc ctcgtctgcc ccctgcacaa
ccaagaggtg 900acagcagagg atggaacaca gcggtgtgag aagtgcagca
agccctgtgc ccgagtgtgc 960tatggtctgg gcatggagca cttgcgagag
gtgagggcag ttaccagtgc caatatccag 1020gagtttgctg gctgcaagaa
gatctttggg agcctggcat ttctgccgga gagctttgat 1080ggggacccag
cctccaacac tgccccgctc cagccagagc agctccaagt gtttgagact
1140ctggaagaga tcacaggtta cctatacatc tcagcatggc cggacagcct
gcctgacctc 1200agcgtcttcc agaacctgag agtaatccgg ggacgaattc
tgcacaatgg cgcctactcg 1260ctgaccctgc aagggctggg catcagctgg
ctggggctgc gctcactgag ggaactgggc 1320agtggactgg ccctcatcca
ccataacacc cacctctgct tcgtgcacac ggtgccctgg 1380gaccagctct
ttcggaaccc gcaccaagct ctgctccaca ctgccaaccg gccagaggac
1440gagtgtgtgg gcgagggcct ggcctgccac cagctgtgcg cccgagggca
ctgctggggt 1500ccagggccca cccagtgtgt caactgcagc cagttccttc
ggggccagga gtgcgtggag 1560gaatgccgag tactgcaggg gctccccagg
gagtatgtga atgccaggca ctgtttgccg 1620tgccaccctg agtgtcagcc
ccagaatggc tcagtgacct gttttggacc ggaggctgac 1680cagtgtgtgg
cctgtgccca ctataaggac cctcccttct gcgtggcccg ctgccccagc
1740ggtgtgaaac ctgacctctc ctacatgccc atctggaagt ttccagatga
ggagggcgca 1800tgccagcctt gccccatcaa ctgcacccac tcctgtgtgg
acctggatga caagggctgc 1860cccgccgagc agagagccag ccctctgacg
tccatcatct ctgcggtggt tggcattctg 1920ctggtcgtgg tcttgggggt
ggtctttggg atcctcatca agcgacggca gcagaagatc 1980cggaagtaca cgatgtga
1998331998DNAHomo sapiens 33acccaagtgt gcaccggcac agacatgaag
ctgcggctcc ctgccagtcc cgagacccac 60ctggacatgc tccgccacct ctaccagggc
tgccaggtgg tgcagggaaa cctggaactc 120acctacctgc ccaccaatgc
cagcctgtcc ttcctgcagg atatccagga ggtgcagggc 180tacgtgctca
tcgctcacaa ccaagtgagg caggtcccac tgcagaggct gcggattgtg
240cgaggcaccc agctctttga ggacaactat gccctggccg tgctagacaa
tggagacccg 300ctgaacaata ccacccctgt cacaggggcc tccccaggag
gcctgcggga gctgcagctt 360cgaagcctca cagagatctt gaaaggaggg
gtcttgatcc agcggaaccc ccagctctgc 420taccaggaca cgattttgtg
gaaggacatc ttccacaaga acaaccagct ggctctcaca 480ctgatagaca
ccaaccgctc tcgggcctgc cacccctgtt ctccgatgtg taagggctcc
540cgctgctggg gagagagttc tgaggattgt cagagcctga cgcgcactgt
ctgtgccggt 600ggctgtgccc gctgcaaggg gccactgccc actgactgct
gccatgagca gtgtgctgcc 660ggctgcacgg gccccaagca ctctgactgc
ctggcctgcc tccacttcaa ccacagtggc 720atctgtgagc tgcactgccc
agccctggtc acctacaaca cagacacgtt tgagtccatg 780cccaatcccg
agggccggta tacattcggc gccagctgtg tgactgcctg tccctacaac
840tacctttcta cggacgtggg atcctgcacc ctcgtctgcc ccctgcacaa
ccaagaggtg 900acagcagagg atggaacaca gcggtgtgag aagtgcagca
agccctgtgc ccgagtgtgc 960tatggtctgg gcatggagca cttgcgagag
gtgagggcag ttaccagtgc caatatccag 1020gagtttgctg gctgcaagaa
gatctttggg agcctggcat ttctgccgga gagctttgat 1080ggggacccag
cctccaacac tgccccgctc cagccagagc agctccaagt gtttgagact
1140ctggaagaga tcacaggtta cctatacatc tcagcatggc cggacagcct
gcctgacctc 1200agcgtcttcc agaacctgca agtaatccgg ggacgaattc
tgcacgatgg cgcctactcg 1260ctgaccctgc aagggctggg catcagctgg
ctggggctgc gctcactgag ggaactgggc 1320agtggactgg ccctcatcca
ccataacacc cacctctgct tcgtgcacac ggtgccctgg 1380gaccagctct
ttcggaaccc gcaccaagct ctgctccaca ctgccaaccg gccagaggac
1440gagtgtgtgg gcgagggcct ggcctgccac cagctgtgcg cccgagggca
ctgctggggt
1500ccagggccca cccagtgtgt caactgcagc cagttccttc ggggccagga
gtgcgtggag 1560gaatgccgag tactgcaggg gctccccagg gagtatgtga
atgccaggca ctgtttgccg 1620tgccaccctg agtgtcagcc ccagaatggc
tcagtgacct gttttggacc ggaggctgac 1680cagtgtgtgg cctgtgccca
ctataaggac cctcccttct gcgtggcccg ctgccccagc 1740ggtgtgaaac
ctgacctctc ctacatgccc atctggaagt ttccagatga ggagggcgca
1800tgccagcctt gccccatcaa ctgcacccac tcctgtgtgg acctggatga
caagggctgc 1860cccgccgagc agagagccag ccctctgacg tccatcatct
ctgcggtggt tggcattctg 1920ctggtcgtgg tcttgggggt ggtctttggg
atcctcatca agcgacggca gcagaagatc 1980cggaagtaca cgatgtga
1998342064DNAHomo sapiens 34atggagctgg cggccttgtg ccgctggggg
ctcctcctcg ccctcttgcc ccccggagcc 60gcgagcaccc aagtgtgcac cggcacagac
atgaagctgc ggctccctgc cagtcccgag 120acccacctgg acatgctccg
ccacctctac cagggctgcc aggtggtgca gggaaacctg 180gaactcacct
acctgcccac caatgccagc ctgtccttcc tgcaggatat ccaggaggtg
240cagggctacg tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca
gaggctgcgg 300attgtgcgag gcacccagct ctttgaggac aactatgccc
tggccgtgct agacaatgga 360gacccgctga acaataccac ccctgtcaca
ggggcctccc caggaggcct gcgggagctg 420cagcttcgaa gcctcacaga
gatcttgaaa ggaggggtct tgatccagcg gaacccccag 480ctctgctacc
aggacacgat tttgtggaag gacatcttcc acaagaacaa ccagctggct
540ctcacactga tagacaccaa ccgctctcgg gcctgccacc cctgttctcc
gatgtgtaag 600ggctcccgct gctggggaga gagttctgag gattgtcaga
gcctgacgcg cactgtctgt 660gccggtggct gtgcccgctg caaggggcca
ctgcccactg actgctgcca tgagcagtgt 720gctgccggct gcacgggccc
caagcactct gactgcctgg cctgcctcca cttcaaccac 780agtggcatct
gtgagctgca ctgcccagcc ctggtcacct acaacacaga cacgtttgag
840tccatgccca atcccgaggg ccggtataca ttcggcgcca gctgtgtgac
tgcctgtccc 900tacaactacc tttctacgga cgtgggatcc tgcaccctcg
tctgccccct gcacaaccaa 960gaggtgacag cagaggatgg aacaaagcgg
tgtgagaagt gcagcaagcc ctgtgcccga 1020gtgtgctatg gtctgggcat
ggagcacttg cgagaggtga gggcagttac cagtgccaat 1080atccaggagt
ttgctggctg caagaagatc tttgggagcc tggcatttct gccggagagc
1140tttgatgggg acccagcctc caacactgcc ccgctccagc cagagcagct
ccaagtgttt 1200gagactctgg aagagatcac aggttaccta tacatctcag
catggccgga cagcctgcct 1260gacctcagcg tcttccagaa cctgcaagta
atccggggac gaattctgca caatggcgcc 1320tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc actgagggaa 1380ctgggcagtg
gactggccct catccaccat aacacccacc tctgcttcgt gcacacggtg
1440ccctgggacc agctctttcg gaacccgcac caagctctgc tccacactgc
caaccggcca 1500gaggacgagt gtgtgggcga gggcctggcc tgccaccagc
tgtgcgcccg agggcactgc 1560tggggtccag ggcccaccca gtgtgtcaac
tgcagccagt tccttcgggg ccaggagtgc 1620gtggaggaat gccgagtact
gcaggggctc cccagggagt atgtgaatgc caggcactgt 1680ttgccgtgcc
accctgagtg tcagccccag aatggctcag tgacctgttt tggaccggag
1740gctgaccagt gtgtggcctg tgcccactat aaggaccctc ccttctgcgt
ggcccgctgc 1800cccagcggtg tgaaacctga cctctcctac atgcccatct
ggaagtttcc agatgaggag 1860ggcgcatgcc agccttgccc catcaactgc
acccactcct gtgtggacct ggatgacaag 1920ggctgccccg ccgagcagag
agccagccct ctgacgtcca tcatctctgc ggtggttggc 1980attctgctgg
tcgtggtctt gggggtggtc tttgggatcc tcatcaagcg acggcagcag
2040aagatccgga agtacacgat gtga 2064352064DNAHomo sapiens
35atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc ccccggagcc
60gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc cagtcccgag
120acccacctgg acatgctccg ccacctctac cagggctgcc aggtggtgca
gggaaacctg 180gaactcacct acctgcccac caatgccagc ctgtccttcc
tgcaggatat ccaggaggtg 240cagggctacg tgctcatcgc tcacaaccaa
gtgaggcagg tcccactgca gaggctgcgg 300attgtgcgag gcacccagct
ctttgaggac aactatgccc tggccgtgct agacaatgga 360gacccgctga
acaataccac ccctgtcaca ggggcctccc caggaggcct gcgggagctg
420cagcttcgaa gcctcacaga gatcttgaaa ggaggggtct tgatccagcg
gaacccccag 480ctctgctacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggct 540ctcacactga tagacaccaa ccgctctcgg
gcctgccacc cctgttctcc gatgtgtaag 600ggctcccgct gctggggaga
gagttctgag gattgtcaga gcctgacgcg cactgtctgt 660gccggtggct
gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt
720gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca
cttcaaccac 780agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacaga cacgtttgag 840tccatgccca atcccgaggg ccggtataca
ttcggcgcca gctgtgtgac tgcctgtccc 900tacaactacc tttctacgga
cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa 960gaggtgacag
cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga
1020gtgtgctatg gtctgggcat ggagcacttg cgagaggtga gggcagttac
cagtgccaat 1080atccaggagt ttgctggctg caagaagatc tttgggagcc
tggcatttct gccggagagc 1140tttgatgggg acccagcctc caacactgcc
ccgctccagc cagagcagct ccaagtgttt 1200gagactctgg aagagatcac
aggttaccta tacatctcag catggccgga cagcctgcct 1260gacctcagcg
tcttccagaa cctgagagta atccggggac gaattctgca caatggcgcc
1320tactcgctga ccctgcaagg gctgggcatc agctggctgg ggctgcgctc
actgagggaa 1380ctgggcagtg gactggccct catccaccat aacacccacc
tctgcttcgt gcacacggtg 1440ccctgggacc agctctttcg gaacccgcac
caagctctgc tccacactgc caaccggcca 1500gaggacgagt gtgtgggcga
gggcctggcc tgccaccagc tgtgcgcccg agggcactgc 1560tggggtccag
ggcccaccca gtgtgtcaac tgcagccagt tccttcgggg ccaggagtgc
1620gtggaggaat gccgagtact gcaggggctc cccagggagt atgtgaatgc
caggcactgt 1680ttgccgtgcc accctgagtg tcagccccag aatggctcag
tgacctgttt tggaccggag 1740gctgaccagt gtgtggcctg tgcccactat
aaggaccctc ccttctgcgt ggcccgctgc 1800cccagcggtg tgaaacctga
cctctcctac atgcccatct ggaagtttcc agatgaggag 1860ggcgcatgcc
agccttgccc catcaactgc acccactcct gtgtggacct ggatgacaag
1920ggctgccccg ccgagcagag agccagccct ctgacgtcca tcatctctgc
ggtggttggc 1980attctgctgg tcgtggtctt gggggtggtc tttgggatcc
tcatcaagcg acggcagcag 2040aagatccgga agtacacgat gtga
2064362064DNAHomo sapiens 36atggagctgg cggccttgtg ccgctggggg
ctcctcctcg ccctcttgcc ccccggagcc 60gcgagcaccc aagtgtgcac cggcacagac
atgaagctgc ggctccctgc cagtcccgag 120acccacctgg acatgctccg
ccacctctac cagggctgcc aggtggtgca gggaaacctg 180gaactcacct
acctgcccac caatgccagc ctgtccttcc tgcaggatat ccaggaggtg
240cagggctacg tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca
gaggctgcgg 300attgtgcgag gcacccagct ctttgaggac aactatgccc
tggccgtgct agacaatgga 360gacccgctga acaataccac ccctgtcaca
ggggcctccc caggaggcct gcgggagctg 420cagcttcgaa gcctcacaga
gatcttgaaa ggaggggtct tgatccagcg gaacccccag 480ctctgctacc
aggacacgat tttgtggaag gacatcttcc acaagaacaa ccagctggct
540ctcacactga tagacaccaa ccgctctcgg gcctgccacc cctgttctcc
gatgtgtaag 600ggctcccgct gctggggaga gagttctgag gattgtcaga
gcctgacgcg cactgtctgt 660gccggtggct gtgcccgctg caaggggcca
ctgcccactg actgctgcca tgagcagtgt 720gctgccggct gcacgggccc
caagcactct gactgcctgg cctgcctcca cttcaaccac 780agtggcatct
gtgagctgca ctgcccagcc ctggtcacct acaacacaga cacgtttgag
840tccatgccca atcccgaggg ccggtataca ttcggcgcca gctgtgtgac
tgcctgtccc 900tacaactacc tttctacgga cgtgggatcc tgcaccctcg
tctgccccct gcacaaccaa 960gaggtgacag cagaggatgg aacacagcgg
tgtgagaagt gcagcaagcc ctgtgcccga 1020gtgtgctatg gtctgggcat
ggagcacttg cgagaggtga gggcagttac cagtgccaat 1080atccaggagt
ttgctggctg caagaagatc tttgggagcc tggcatttct gccggagagc
1140tttgatgggg acccagcctc caacactgcc ccgctccagc cagagcagct
ccaagtgttt 1200gagactctgg aagagatcac aggttaccta tacatctcag
catggccgga cagcctgcct 1260gacctcagcg tcttccagaa cctgcaagta
atccggggac gaattctgca cgatggcgcc 1320tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc actgagggaa 1380ctgggcagtg
gactggccct catccaccat aacacccacc tctgcttcgt gcacacggtg
1440ccctgggacc agctctttcg gaacccgcac caagctctgc tccacactgc
caaccggcca 1500gaggacgagt gtgtgggcga gggcctggcc tgccaccagc
tgtgcgcccg agggcactgc 1560tggggtccag ggcccaccca gtgtgtcaac
tgcagccagt tccttcgggg ccaggagtgc 1620gtggaggaat gccgagtact
gcaggggctc cccagggagt atgtgaatgc caggcactgt 1680ttgccgtgcc
accctgagtg tcagccccag aatggctcag tgacctgttt tggaccggag
1740gctgaccagt gtgtggcctg tgcccactat aaggaccctc ccttctgcgt
ggcccgctgc 1800cccagcggtg tgaaacctga cctctcctac atgcccatct
ggaagtttcc agatgaggag 1860ggcgcatgcc agccttgccc catcaactgc
acccactcct gtgtggacct ggatgacaag 1920ggctgccccg ccgagcagag
agccagccct ctgacgtcca tcatctctgc ggtggttggc 1980attctgctgg
tcgtggtctt gggggtggtc tttgggatcc tcatcaagcg acggcagcag
2040aagatccgga agtacacgat gtga 206437687PRTUrsus americanus 37Met
Glu Leu Ala Ala Trp Cys Arg Trp Gly Leu Leu Leu Ala Leu Leu1 5 10
15Pro Ser Gly Ala Ala Gly Thr Gln Val Cys Thr Gly Thr Asp Met Lys
20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg
His 35 40 45Leu Tyr Gln Ala Cys Gln Val Val Gln Gly Asn Leu Glu Leu
Thr Tyr 50 55 60Leu Pro Ala Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile
Gln Glu Val65 70 75 80Gln Gly Tyr Val Leu Ile Ala His Ser Gln Val
Arg Gln Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln
Leu Phe Glu Asp Asn Tyr 100 105 110Ala Leu Ala Val Leu Asp Asn Gly
Glu Pro Pro Lys Gly Asp Thr Ser 115 120 125Val Ala Gly Ala Thr Pro
Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser 130 135 140Leu Thr Glu Ile
Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln145 150 155 160Leu
Cys His Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys Asn 165 170
175Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys
180 185 190Gln Pro Cys Ser Pro Ala Cys Lys Asp Pro His Cys Trp Gly
Ala Ser 195 200 205Ser Gly Asp Cys Gln Ser Leu Thr Arg Thr Val Cys
Ala Gly Gly Cys 210 215 220Ala Arg Cys Lys Gly Pro Lys Pro Thr Asp
Cys Cys His Glu Gln Cys225 230 235 240Ala Ala Gly Cys Thr Gly Pro
Lys His Ser Asp Cys Leu Ala Cys Leu 245 250 255His Phe Asn His Ser
Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val 260 265 270Thr Tyr Asn
Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg 275 280 285Tyr
Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295
300Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu Asn Asn
Gln305 310 315 320Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu
Lys Cys Ser Arg 325 330 335Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly
Met Glu His Leu Arg Glu 340 345 350Ala Arg Ala Val Thr Ser Ala Asn
Ile Gln Glu Phe Ala Gly Cys Lys 355 360 365Lys Ile Phe Gly Ser Leu
Ala Phe Leu Pro Glu Ser Phe Glu Gly Asp 370 375 380Pro Ala Ser Asn
Thr Ala Pro Leu Gln Pro Glu Gln Leu Arg Val Phe385 390 395 400Glu
Ala Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro 405 410
415Asp Ser Leu Pro Asn Leu Ser Val Phe Gln Asn Leu Arg Val Ile Arg
420 425 430Gly Arg Val Leu His Asp Gly Ala Tyr Ser Leu Thr Leu Gln
Gly Leu 435 440 445Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu
Leu Gly Ser Gly 450 455 460Leu Ala Leu Ile His Arg Asn Ala Arg Leu
Cys Phe Ile His Thr Val465 470 475 480Pro Trp Glu Gln Leu Phe Arg
Asn Pro His Gln Ala Leu Leu His Ser 485 490 495Ala Asn Arg Pro Glu
Ala Glu Cys Val Gly Glu Gly Leu Ala Cys Tyr 500 505 510Pro Leu Cys
Ala His Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys 515 520 525Val
Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys 530 535
540Arg Glu Leu His Gly Leu Pro Arg Glu Tyr Val Lys Asp Arg Tyr
Cys545 550 555 560Leu Pro Cys His Pro Glu Cys Arg Pro Gln Asn Gly
Ser Val Thr Cys 565 570 575Phe Gly Ser Glu Ala Asp Gln Cys Val Ala
Cys Ala His Tyr Lys Asp 580 585 590Pro Pro Ser Cys Val Ala Arg Cys
Pro Ser Gly Val Lys Pro Asp Leu 595 600 605Ser Phe Met Pro Ile Trp
Lys Phe Ala Asp Glu Glu Gly Thr Cys Gln 610 615 620Pro Cys Pro Ile
Asn Cys Thr His Ser Cys Gly Asp Leu Asp Glu Arg625 630 635 640Gly
Cys Pro Ala Glu Gln Arg Ala Ser Pro Val Thr Ser Ile Ile Ala 645 650
655Ala Val Val Gly Ile Leu Leu Ala Val Val Met Gly Leu Val Leu Gly
660 665 670Ile Leu Ile Lys Arg Arg Arg Gln Lys Ile Arg Lys Tyr Thr
Met 675 680 68538665PRTUrsus americanus 38Thr Gln Val Cys Thr Gly
Thr Asp Met Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu
Asp Met Leu Arg His Leu Tyr Gln Ala Cys Gln 20 25 30Val Val Gln Gly
Asn Leu Glu Leu Thr Tyr Leu Pro Ala Asn Ala Ser 35 40 45Leu Ser Phe
Leu Gln Asp Ile Gln Glu Val Gln Gly Tyr Val Leu Ile 50 55 60Ala His
Ser Gln Val Arg Gln Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75
80Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr Ala Leu Ala Val Leu Asp
85 90 95Asn Gly Glu Pro Pro Lys Gly Asp Thr Ser Val Ala Gly Ala Thr
Pro 100 105 110Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser Leu Thr Glu
Ile Leu Lys 115 120 125Gly Gly Val Leu Ile Gln Arg Asn Pro Gln Leu
Cys His Gln Asp Thr 130 135 140Ile Leu Trp Lys Asp Ile Phe His Lys
Asn Asn Gln Leu Ala Leu Thr145 150 155 160Leu Ile Asp Thr Asn Arg
Ser Arg Ala Cys Gln Pro Cys Ser Pro Ala 165 170 175Cys Lys Asp Pro
His Cys Trp Gly Ala Ser Ser Gly Asp Cys Gln Ser 180 185 190Leu Thr
Arg Thr Val Cys Ala Gly Gly Cys Ala Arg Cys Lys Gly Pro 195 200
205Lys Pro Thr Asp Cys Cys His Glu Gln Cys Ala Ala Gly Cys Thr Gly
210 215 220Pro Lys His Ser Asp Cys Leu Ala Cys Leu His Phe Asn His
Ser Gly225 230 235 240Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr
Tyr Asn Thr Asp Thr 245 250 255Phe Glu Ser Met Pro Asn Pro Glu Gly
Arg Tyr Thr Phe Gly Ala Ser 260 265 270Cys Val Thr Ala Cys Pro Tyr
Asn Tyr Leu Ser Thr Asp Val Gly Ser 275 280 285Cys Thr Leu Val Cys
Pro Leu Asn Asn Gln Glu Val Thr Ala Glu Asp 290 295 300Gly Thr Gln
Arg Cys Glu Lys Cys Ser Arg Pro Cys Ala Arg Val Cys305 310 315
320Tyr Gly Leu Gly Met Glu His Leu Arg Glu Ala Arg Ala Val Thr Ser
325 330 335Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys Lys Ile Phe Gly
Ser Leu 340 345 350Ala Phe Leu Pro Glu Ser Phe Glu Gly Asp Pro Ala
Ser Asn Thr Ala 355 360 365Pro Leu Gln Pro Glu Gln Leu Arg Val Phe
Glu Ala Leu Glu Glu Ile 370 375 380Thr Gly Tyr Leu Tyr Ile Ser Ala
Trp Pro Asp Ser Leu Pro Asn Leu385 390 395 400Ser Val Phe Gln Asn
Leu Arg Val Ile Arg Gly Arg Val Leu His Asp 405 410 415Gly Ala Tyr
Ser Leu Thr Leu Gln Gly Leu Gly Ile Ser Trp Leu Gly 420 425 430Leu
Arg Ser Leu Arg Glu Leu Gly Ser Gly Leu Ala Leu Ile His Arg 435 440
445Asn Ala Arg Leu Cys Phe Ile His Thr Val Pro Trp Glu Gln Leu Phe
450 455 460Arg Asn Pro His Gln Ala Leu Leu His Ser Ala Asn Arg Pro
Glu Ala465 470 475 480Glu Cys Val Gly Glu Gly Leu Ala Cys Tyr Pro
Leu Cys Ala His Gly 485 490 495His Cys Trp Gly Pro Gly Pro Thr Gln
Cys Val Asn Cys Ser Gln Phe 500 505 510Leu Arg Gly Gln Glu Cys Val
Glu Glu Cys Arg Glu Leu His Gly Leu 515 520 525Pro Arg Glu Tyr Val
Lys Asp Arg Tyr Cys Leu Pro Cys His Pro Glu 530 535 540Cys Arg Pro
Gln Asn Gly Ser Val Thr Cys Phe Gly Ser Glu Ala Asp545 550 555
560Gln Cys Val Ala Cys Ala His Tyr Lys Asp Pro Pro Ser Cys Val Ala
565 570 575Arg Cys Pro Ser Gly Val Lys Pro Asp Leu Ser Phe Met Pro
Ile Trp 580 585 590Lys Phe Ala Asp Glu Glu Gly Thr Cys Gln Pro Cys
Pro Ile Asn Cys 595 600 605Thr His Ser Cys Gly Asp Leu Asp Glu Arg
Gly Cys Pro Ala Glu Gln 610 615 620Arg Ala Ser Pro Val Thr Ser Ile
Ile Ala Ala Val Val Gly Ile Leu625 630 635 640Leu Ala Val Val Met
Gly Leu Val Leu Gly Ile Leu
Ile Lys Arg Arg 645 650 655Arg Gln Lys Ile Arg Lys Tyr Thr Met 660
66539687PRTFelis catus 39Met Glu Leu Ala Ala Trp Cys Arg Trp Gly
Leu Leu Leu Ala Leu Leu1 5 10 15Pro Ser Gly Ala Thr Gly Thr Gln Val
Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu
Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val
Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu His Ala Asn Ala Ser
Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val
Leu Ile Ala His Asn Gln Val Lys Gln Val Pro Leu 85 90 95Gln Arg Leu
Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110Ala
Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asp Ser Gly Thr Pro 115 120
125Ala Thr Gly Ala Ala Leu Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser
130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn
Pro Gln145 150 155 160Leu Cys His Gln Asp Thr Ile Leu Trp Lys Asp
Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu Met Leu Ile Asp
Thr Asn Arg Ser Arg Ala Cys 180 185 190Gln Pro Cys Ser Pro Ala Cys
Lys Asp Ser His Cys Trp Gly Ala Ser 195 200 205Ser Gly Asp Cys Gln
Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220Ala Arg Cys
Lys Gly Pro Gln Pro Thr Asp Cys Cys His Glu Gln Cys225 230 235
240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu
245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala
Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn
Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala
Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys Thr
Leu Val Cys Pro Leu Asn Asn Gln305 310 315 320Glu Val Thr Ala Glu
Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys Ala
Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345 350Ala
Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Val Gly Cys Lys 355 360
365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Glu Gly Asp
370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Arg
Val Phe385 390 395 400Glu Ala Leu Glu Glu Ile Thr Gly Tyr Leu Tyr
Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asn Leu Ser Val Phe
Gln Asn Leu Arg Val Ile Arg 420 425 430Gly Arg Val Leu His Asp Gly
Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser Trp Leu
Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu Ala Leu
Ile His Arg Asn Ser Arg Leu Cys Phe Val His Thr Val465 470 475
480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Ser
485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Ala Gly Glu Gly Leu Ala
Cys Tyr 500 505 510Pro Leu Cys Ala His Gly His Cys Trp Gly Pro Gly
Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly Gln
Glu Cys Val Glu Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro Arg
Glu Tyr Val Lys Asp Arg Phe Cys545 550 555 560Leu Pro Cys His Pro
Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570 575Leu Gly Ser
Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585 590Pro
Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu 595 600
605Ser Phe Met Pro Ile Trp Lys Phe Ala Asp Glu Glu Gly Thr Cys Gln
610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys Ala Asp Leu Asp
Glu Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Val
Thr Ser Ile Ile Ala 645 650 655Ala Val Val Gly Ile Leu Leu Val Val
Val Val Gly Leu Val Leu Gly 660 665 670Ile Leu Ile Lys Arg Arg Arg
Gln Lys Ile Arg Lys Tyr Thr Met 675 680 68540665PRTFelis catus
40Thr Gln Val Cys Thr Gly Thr Asp Met Lys Leu Arg Leu Pro Ala Ser1
5 10 15Pro Glu Thr His Leu Asp Met Leu Arg His Leu Tyr Gln Gly Cys
Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu Thr Tyr Leu His Ala Asn
Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile Gln Glu Val Gln Gly Tyr
Val Leu Ile 50 55 60Ala His Asn Gln Val Lys Gln Val Pro Leu Gln Arg
Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr
Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro Leu Asp Ser Gly Thr
Pro Ala Thr Gly Ala Ala Leu 100 105 110Gly Gly Leu Arg Glu Leu Gln
Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120 125Gly Gly Val Leu Ile
Gln Arg Asn Pro Gln Leu Cys His Gln Asp Thr 130 135 140Ile Leu Trp
Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala Leu Met145 150 155
160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys Gln Pro Cys Ser Pro Ala
165 170 175Cys Lys Asp Ser His Cys Trp Gly Ala Ser Ser Gly Asp Cys
Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala Gly Gly Cys Ala Arg
Cys Lys Gly Pro 195 200 205Gln Pro Thr Asp Cys Cys His Glu Gln Cys
Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His Ser Asp Cys Leu Ala
Cys Leu His Phe Asn His Ser Gly225 230 235 240Ile Cys Glu Leu His
Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr 245 250 255Phe Glu Ser
Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala Ser 260 265 270Cys
Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr Asp Val Gly Ser 275 280
285Cys Thr Leu Val Cys Pro Leu Asn Asn Gln Glu Val Thr Ala Glu Asp
290 295 300Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys Pro Cys Ala Arg
Val Cys305 310 315 320Tyr Gly Leu Gly Met Glu His Leu Arg Glu Ala
Arg Ala Val Thr Ser 325 330 335Ala Asn Ile Gln Glu Phe Val Gly Cys
Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala Phe Leu Pro Glu Ser Phe
Glu Gly Asp Pro Ala Ser Asn Thr Ala 355 360 365Pro Leu Gln Pro Glu
Gln Leu Arg Val Phe Glu Ala Leu Glu Glu Ile 370 375 380Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu Pro Asn Leu385 390 395
400Ser Val Phe Gln Asn Leu Arg Val Ile Arg Gly Arg Val Leu His Asp
405 410 415Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu Gly Ile Ser Trp
Leu Gly 420 425 430Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly Leu Ala
Leu Ile His Arg 435 440 445Asn Ser Arg Leu Cys Phe Val His Thr Val
Pro Trp Asp Gln Leu Phe 450 455 460Arg Asn Pro His Gln Ala Leu Leu
His Ser Ala Asn Arg Pro Glu Asp465 470 475 480Glu Cys Ala Gly Glu
Gly Leu Ala Cys Tyr Pro Leu Cys Ala His Gly 485 490 495His Cys Trp
Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser Gln Phe 500 505 510Leu
Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Leu Gln Gly Leu 515 520
525Pro Arg Glu Tyr Val Lys Asp Arg Phe Cys Leu Pro Cys His Pro Glu
530 535 540Cys Gln Pro Gln Asn Gly Ser Val Thr Cys Leu Gly Ser Glu
Ala Asp545 550 555 560Gln Cys Val Ala Cys Ala His Tyr Lys Asp Pro
Pro Phe Cys Val Ala 565 570 575Arg Cys Pro Ser Gly Val Lys Pro Asp
Leu Ser Phe Met Pro Ile Trp 580 585 590Lys Phe Ala Asp Glu Glu Gly
Thr Cys Gln Pro Cys Pro Ile Asn Cys 595 600 605Thr His Ser Cys Ala
Asp Leu Asp Glu Lys Gly Cys Pro Ala Glu Gln 610 615 620Arg Ala Ser
Pro Val Thr Ser Ile Ile Ala Ala Val Val Gly Ile Leu625 630 635
640Leu Val Val Val Val Gly Leu Val Leu Gly Ile Leu Ile Lys Arg Arg
645 650 655Arg Gln Lys Ile Arg Lys Tyr Thr Met 660
665412061DNAUrsus americanus 41atggagctgg cggcctggtg ccgctggggg
ctcctcctcg ccctcctgcc ctccggagcc 60gcgggcaccc aagtgtgcac cggcacagac
atgaagctgc ggctccctgc cagtcccgag 120acccacctgg atatgctccg
ccacctctac caggcctgtc aagtggtaca gggtaacctg 180gagctcacct
acctgcccgc caatgccagc ctgtccttcc tgcaggatat ccaggaggta
240cagggctatg tgctcattgc tcacagccaa gtgagacagg tcccgctgca
gaggctccga 300atcgtgcgag gcacccagct ctttgaggac aactacgccc
tggccgtgct ggacaatgga 360gagccgccca agggggacac ctctgtggca
ggggctaccc caggagggct gcgggagctg 420cagcttcgaa gcctcacaga
gatcctgaag ggaggggtct tgattcagcg gaacccacag 480ctctgccacc
aggacacgat tttgtggaag gacatcttcc acaagaacaa ccagctggcc
540ctcacgctga tagacaccaa ccgctctcgg gcctgccaac cctgttctcc
agcctgtaaa 600gacccccact gctggggagc aagttccggg gactgtcaga
gcttgacacg aaccgtctgt 660gccggcggct gtgcccgctg caagggccca
aaacccactg actgctgcca tgagcagtgc 720gcggctggct gcacgggccc
caagcactcg gactgcctgg cctgccttca cttcaaccac 780agtggcatct
gtgagctgca ctgcccagcc ctggtcacct acaacacgga cacgttcgaa
840tccatgccca accctgaggg ccgatacacc ttcggtgcca gctgtgtgac
cgcctgtccc 900tacaactacc tgtccacgga cgtgggatcc tgcaccctgg
tctgtcccct gaacaaccaa 960gaggtgacgg ctgaggatgg cacccagcgg
tgtgagaaat gcagcagacc ctgtgcccga 1020gtgtgctatg gtctgggcat
ggagcacctg cgggaggcga gggcggtcac cagcgccaac 1080atccaagagt
tcgccggctg caagaagatc tttgggagcc tggcgtttct gccagagagc
1140ttcgagggag acccagcctc caacactgcc cccctgcagc ctgaacagct
cagagtgttc 1200gaggccctgg aggagatcac aggttacctg tatatctcag
cgtggccgga cagcttgcct 1260aacctcagtg tcttccagaa cctgcgagta
atccggggac gagttctgca tgatggcgcc 1320tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc gctgcgggaa 1380ctgggcagcg
ggctggccct catccaccgc aacgcccgcc tctgcttcat ccacacggtg
1440ccctgggagc agctcttccg gaacccccac caagccctgc tgcacagtgc
caaccggccg 1500gaggccgagt gcgtgggcga gggcctggcc tgctacccgc
tgtgcgccca tgggcactgc 1560tggggtccgg ggcccaccca gtgcgtcaac
tgcagccaat tccttcgggg ccaggagtgc 1620gtggaggaat gccgagaact
gcacgggcta ccccgggaat atgtgaagga cagatactgt 1680ctgccatgcc
accccgagtg tcggccccag aatggctcag tgacctgctt tgggtcggag
1740gctgaccagt gtgtggcctg cgcccactac aaggaccctc cctcctgcgt
ggctcgctgc 1800cccagtggtg tgaaacccga cctctctttc atgcccattt
ggaagtttgc agatgaggag 1860ggcacatgcc agccgtgccc catcaactgc
acccactcct gtggggacct ggacgagagg 1920ggctgccccg ccgaacagag
agccagccct gtgacatcca tcattgccgc tgtggtgggc 1980attctgctgg
ccgtggtcat ggggctggtc ctcggcatcc tgatcaagcg aaggcgacag
2040aagatccgga agtacacgat g 2061421995DNAUrsus americanus
42acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac
60ctggatatgc tccgccacct ctaccaggcc tgtcaagtgg tacagggtaa cctggagctc
120acctacctgc ccgccaatgc cagcctgtcc ttcctgcagg atatccagga
ggtacagggc 180tatgtgctca ttgctcacag ccaagtgaga caggtcccgc
tgcagaggct ccgaatcgtg 240cgaggcaccc agctctttga ggacaactac
gccctggccg tgctggacaa tggagagccg 300cccaaggggg acacctctgt
ggcaggggct accccaggag ggctgcggga gctgcagctt 360cgaagcctca
cagagatcct gaagggaggg gtcttgattc agcggaaccc acagctctgc
420caccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggccctcacg 480ctgatagaca ccaaccgctc tcgggcctgc caaccctgtt
ctccagcctg taaagacccc 540cactgctggg gagcaagttc cggggactgt
cagagcttga cacgaaccgt ctgtgccggc 600ggctgtgccc gctgcaaggg
cccaaaaccc actgactgct gccatgagca gtgcgcggct 660ggctgcacgg
gccccaagca ctcggactgc ctggcctgcc ttcacttcaa ccacagtggc
720atctgtgagc tgcactgccc agccctggtc acctacaaca cggacacgtt
cgaatccatg 780cccaaccctg agggccgata caccttcggt gccagctgtg
tgaccgcctg tccctacaac 840tacctgtcca cggacgtggg atcctgcacc
ctggtctgtc ccctgaacaa ccaagaggtg 900acggctgagg atggcaccca
gcggtgtgag aaatgcagca gaccctgtgc ccgagtgtgc 960tatggtctgg
gcatggagca cctgcgggag gcgagggcgg tcaccagcgc caacatccaa
1020gagttcgccg gctgcaagaa gatctttggg agcctggcgt ttctgccaga
gagcttcgag 1080ggagacccag cctccaacac tgcccccctg cagcctgaac
agctcagagt gttcgaggcc 1140ctggaggaga tcacaggtta cctgtatatc
tcagcgtggc cggacagctt gcctaacctc 1200agtgtcttcc agaacctgcg
agtaatccgg ggacgagttc tgcatgatgg cgcctactcg 1260ctgaccctgc
aagggctggg catcagctgg ctggggctgc gctcgctgcg ggaactgggc
1320agcgggctgg ccctcatcca ccgcaacgcc cgcctctgct tcatccacac
ggtgccctgg 1380gagcagctct tccggaaccc ccaccaagcc ctgctgcaca
gtgccaaccg gccggaggcc 1440gagtgcgtgg gcgagggcct ggcctgctac
ccgctgtgcg cccatgggca ctgctggggt 1500ccggggccca cccagtgcgt
caactgcagc caattccttc ggggccagga gtgcgtggag 1560gaatgccgag
aactgcacgg gctaccccgg gaatatgtga aggacagata ctgtctgcca
1620tgccaccccg agtgtcggcc ccagaatggc tcagtgacct gctttgggtc
ggaggctgac 1680cagtgtgtgg cctgcgccca ctacaaggac cctccctcct
gcgtggctcg ctgccccagt 1740ggtgtgaaac ccgacctctc tttcatgccc
atttggaagt ttgcagatga ggagggcaca 1800tgccagccgt gccccatcaa
ctgcacccac tcctgtgggg acctggacga gaggggctgc 1860cccgccgaac
agagagccag ccctgtgaca tccatcattg ccgctgtggt gggcattctg
1920ctggccgtgg tcatggggct ggtcctcggc atcctgatca agcgaaggcg
acagaagatc 1980cggaagtaca cgatg 1995432061DNAFelis catus
43atggagctgg cggcctggtg ccgctggggg ctcctcctcg ccctcctgcc ctccggagcc
60acgggcaccc aagtgtgcac cggcacagac atgaagctgc ggctcccagc cagtcccgag
120acccacctgg acatgctccg ccacctctac cagggctgtc aagtggtaca
gggcaacctg 180gagctcacct acctgcatgc caatgccagc ctctccttcc
tgcaggatat ccaggaggtg 240caaggctatg tgctcattgc ccacaaccaa
gtgaaacagg tcccactgca gaggctacga 300atcgtgcgag gcacccagct
ctttgaggac aactacgccc tggccgtgct ggacaacgga 360gacccactgg
acagtggcac ccctgctaca ggggctgccc taggagggct gcgggagctg
420cagctccgaa gcctcacaga gatcctgaag ggaggggtcc tcattcagcg
gaacccgcag 480ctctgccacc aggacacgat tctgtggaag gacatcttcc
acaagaacaa ccagctggcc 540ctcatgctga tagacaccaa ccgctctcgg
gcctgccaac cctgttctcc agcttgtaaa 600gactcccact gctggggagc
aagttccggg gactgtcaga gcttgactcg aactgtctgt 660gctggcggct
gtgcccgctg caagggcccg cagcccaccg actgctgcca cgagcaatgt
720gctgctggct gcacgggccc caagcattct gactgcctgg cctgcctcca
cttcaaccac 780agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacgga caccttcgaa 840tccatgccca accctgaggg ccgttatacc
ttcggtgcca gctgtgtgac tgcctgtccc 900tacaactacc tgtctacgga
cgtgggatcc tgcaccctgg tctgtcccct gaacaaccaa 960gaggtgacag
ctgaggatgg aacacagcgg tgtgagaaat gcagcaagcc ctgtgcccga
1020gtgtgctacg gcctaggcat ggagcacctg cgggaggcga gggcagtcac
cagtgccaac 1080atccaagaat ttgtcggctg caagaagatc tttgggagcc
tggcgtttct gccagagagc 1140tttgaggggg acccagcctc caacactgcc
cccctgcagc ctgagcagct cagagtgttt 1200gaggctctgg aggagattac
aggttacctg tacatctcag cgtggccaga cagcttgcct 1260aacctcagtg
tcttccagaa cctcagagtg atccggggcc gagttctgca tgacggtgct
1320tactcgctga cccttcaagg gctgggcatc agctggctgg ggctgcgctc
gctgcgggag 1380ctgggcagtg ggctggccct catccaccgc aactcccgcc
tctgcttcgt acacacggtg 1440ccctgggacc agctcttccg gaacccccac
caggccctgc tccacagcgc caaccggcca 1500gaggacgagt gcgcgggtga
gggcctggcc tgctatccgc tgtgtgccca cgggcactgc 1560tggggtccgg
gacccaccca gtgtgtcaac tgcagccagt tccttcgggg ccaggagtgc
1620gtggaggaat gccgagtatt gcaggggctt ccccgggagt atgtgaagga
taggttctgt 1680ctgccatgcc acccggagtg tcagccccag aatggctcag
tgacctgctt gggctcggaa 1740gctgaccagt gtgtggcctg tgcccactac
aaggaccctc ctttctgtgt ggctcgctgc 1800cccagtgggg tgaaacctga
cctctccttc atgcccatct ggaagttcgc agatgaggag 1860ggcacgtgcc
agccatgccc catcaactgc acccactcct gtgcggacct ggacgagaag
1920ggctgccccg ccgagcagag agccagccct gtgacgtcca tcattgctgc
tgtggtgggc 1980attctgctgg tcgtggttgt ggggctggtc cttggcatcc
taatcaagcg aaggcggcag 2040aagatccgga agtacacgat g
2061441995DNAFelis catus 44acccaagtgt gcaccggcac agacatgaag
ctgcggctcc cagccagtcc cgagacccac 60ctggacatgc tccgccacct ctaccagggc
tgtcaagtgg tacagggcaa cctggagctc 120acctacctgc atgccaatgc
cagcctctcc ttcctgcagg atatccagga ggtgcaaggc 180tatgtgctca
ttgcccacaa ccaagtgaaa caggtcccac tgcagaggct acgaatcgtg
240cgaggcaccc agctctttga ggacaactac gccctggccg tgctggacaa
cggagaccca 300ctggacagtg gcacccctgc tacaggggct gccctaggag
ggctgcggga gctgcagctc 360cgaagcctca cagagatcct gaagggaggg
gtcctcattc agcggaaccc gcagctctgc 420caccaggaca cgattctgtg
gaaggacatc ttccacaaga acaaccagct ggccctcatg 480ctgatagaca
ccaaccgctc tcgggcctgc caaccctgtt ctccagcttg taaagactcc
540cactgctggg gagcaagttc cggggactgt cagagcttga ctcgaactgt
ctgtgctggc 600ggctgtgccc gctgcaaggg cccgcagccc accgactgct
gccacgagca atgtgctgct 660ggctgcacgg gccccaagca ttctgactgc
ctggcctgcc tccacttcaa ccacagtggc 720atctgtgagc tgcactgccc
agccctggtc acctacaaca cggacacctt cgaatccatg 780cccaaccctg
agggccgtta taccttcggt gccagctgtg tgactgcctg tccctacaac
840tacctgtcta cggacgtggg atcctgcacc ctggtctgtc ccctgaacaa
ccaagaggtg 900acagctgagg atggaacaca gcggtgtgag aaatgcagca
agccctgtgc ccgagtgtgc 960tacggcctag gcatggagca cctgcgggag
gcgagggcag tcaccagtgc caacatccaa 1020gaatttgtcg gctgcaagaa
gatctttggg agcctggcgt ttctgccaga gagctttgag 1080ggggacccag
cctccaacac tgcccccctg cagcctgagc agctcagagt gtttgaggct
1140ctggaggaga ttacaggtta cctgtacatc tcagcgtggc cagacagctt
gcctaacctc 1200agtgtcttcc agaacctcag agtgatccgg ggccgagttc
tgcatgacgg tgcttactcg 1260ctgacccttc aagggctggg catcagctgg
ctggggctgc gctcgctgcg ggagctgggc 1320agtgggctgg ccctcatcca
ccgcaactcc cgcctctgct tcgtacacac ggtgccctgg 1380gaccagctct
tccggaaccc ccaccaggcc ctgctccaca gcgccaaccg gccagaggac
1440gagtgcgcgg gtgagggcct ggcctgctat ccgctgtgtg cccacgggca
ctgctggggt 1500ccgggaccca cccagtgtgt caactgcagc cagttccttc
ggggccagga gtgcgtggag 1560gaatgccgag tattgcaggg gcttccccgg
gagtatgtga aggataggtt ctgtctgcca 1620tgccacccgg agtgtcagcc
ccagaatggc tcagtgacct gcttgggctc ggaagctgac 1680cagtgtgtgg
cctgtgccca ctacaaggac cctcctttct gtgtggctcg ctgccccagt
1740ggggtgaaac ctgacctctc cttcatgccc atctggaagt tcgcagatga
ggagggcacg 1800tgccagccat gccccatcaa ctgcacccac tcctgtgcgg
acctggacga gaagggctgc 1860cccgccgagc agagagccag ccctgtgacg
tccatcattg ctgctgtggt gggcattctg 1920ctggtcgtgg ttgtggggct
ggtccttggc atcctaatca agcgaaggcg gcagaagatc 1980cggaagtaca cgatg
199545687PRTHomo sapiens 45Met Glu Leu Ala Ala Leu Cys Arg Trp Gly
Leu Leu Leu Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln Val
Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu
Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val
Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala Ser
Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val
Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg Leu
Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110Ala
Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro 115 120
125Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser
130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn
Pro Gln145 150 155 160Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp
Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu Ile Asp
Thr Asn Arg Ser Arg Ala Cys 180 185 190His Pro Cys Ser Pro Met Cys
Lys Gly Ser Arg Cys Trp Gly Glu Ser 195 200 205Ser Glu Asp Cys Gln
Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220Ala Arg Cys
Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln Cys225 230 235
240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu
245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala
Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn
Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala
Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys Thr
Leu Val Cys Pro Leu His Asn Gln305 310 315 320Glu Val Thr Ala Glu
Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys Ala
Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345 350Val
Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys 355 360
365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp
370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln
Val Phe385 390 395 400Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr
Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asp Leu Ser Val Phe
Gln Asn Leu Gln Val Ile Arg 420 425 430Gly Arg Ile Leu His Asn Gly
Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser Trp Leu
Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu Ala Leu
Ile His His Asn Thr His Leu Cys Phe Val His Thr Val465 470 475
480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Thr
485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala
Cys His 500 505 510Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly
Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly Gln
Glu Cys Val Glu Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro Arg
Glu Tyr Val Asn Ala Arg His Cys545 550 555 560Leu Pro Cys His Pro
Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570 575Phe Gly Pro
Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585 590Pro
Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu 595 600
605Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln
610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp
Asp Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu
Thr Ser Ile Ile Ser 645 650 655Ala Val Val Gly Ile Leu Leu Val Val
Val Leu Gly Val Val Phe Gly 660 665 670Ile Leu Ile Lys Arg Arg Gln
Gln Lys Ile Arg Lys Tyr Thr Met 675 680 68546665PRTHomo sapiens
46Thr Gln Val Cys Thr Gly Thr Asp Met Lys Leu Arg Leu Pro Ala Ser1
5 10 15Pro Glu Thr His Leu Asp Met Leu Arg His Leu Tyr Gln Gly Cys
Gln 20 25 30Val Val Gln Gly Asn Leu Glu Leu Thr Tyr Leu Pro Thr Asn
Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile Gln Glu Val Gln Gly Tyr
Val Leu Ile 50 55 60Ala His Asn Gln Val Arg Gln Val Pro Leu Gln Arg
Leu Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr
Ala Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro Leu Asn Asn Thr Thr
Pro Val Thr Gly Ala Ser Pro 100 105 110Gly Gly Leu Arg Glu Leu Gln
Leu Arg Ser Leu Thr Glu Ile Leu Lys 115 120 125Gly Gly Val Leu Ile
Gln Arg Asn Pro Gln Leu Cys Tyr Gln Asp Thr 130 135 140Ile Leu Trp
Lys Asp Ile Phe His Lys Asn Asn Gln Leu Ala Leu Thr145 150 155
160Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys His Pro Cys Ser Pro Met
165 170 175Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser Ser Glu Asp Cys
Gln Ser 180 185 190Leu Thr Arg Thr Val Cys Ala Gly Gly Cys Ala Arg
Cys Lys Gly Pro 195 200 205Leu Pro Thr Asp Cys Cys His Glu Gln Cys
Ala Ala Gly Cys Thr Gly 210 215 220Pro Lys His Ser Asp Cys Leu Ala
Cys Leu His Phe Asn His Ser Gly225 230 235 240Ile Cys Glu Leu His
Cys Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr 245 250 255Phe Glu Ser
Met Pro Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala Ser 260 265 270Cys
Val Thr Ala Cys Pro Tyr Asn Tyr Leu Ser Thr Asp Val Gly Ser 275 280
285Cys Thr Leu Val Cys Pro Leu His Asn Gln Glu Val Thr Ala Glu Asp
290 295 300Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys Pro Cys Ala Arg
Val Cys305 310 315 320Tyr Gly Leu Gly Met Glu His Leu Arg Glu Val
Arg Ala Val Thr Ser 325 330 335Ala Asn Ile Gln Glu Phe Ala Gly Cys
Lys Lys Ile Phe Gly Ser Leu 340 345 350Ala Phe Leu Pro Glu Ser Phe
Asp Gly Asp Pro Ala Ser Asn Thr Ala 355 360 365Pro Leu Gln Pro Glu
Gln Leu Gln Val Phe Glu Thr Leu Glu Glu Ile 370 375 380Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu Pro Asp Leu385 390 395
400Ser Val Phe Gln Asn Leu Gln Val Ile Arg Gly Arg Ile Leu His Asn
405 410 415Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu Gly Ile Ser Trp
Leu Gly 420 425 430Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly Leu Ala
Leu Ile His His 435 440 445Asn Thr His Leu Cys Phe Val His Thr Val
Pro Trp Asp Gln Leu Phe 450 455 460Arg Asn Pro His Gln Ala Leu Leu
His Thr Ala Asn Arg Pro Glu Asp465 470 475 480Glu Cys Val Gly Glu
Gly Leu Ala Cys His Gln Leu Cys Ala Arg Gly 485 490 495His Cys Trp
Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser Gln Phe 500 505 510Leu
Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Leu Gln Gly Leu 515 520
525Pro Arg Glu Tyr Val Asn Ala Arg His Cys Leu Pro Cys His Pro Glu
530 535 540Cys Gln Pro Gln Asn Gly Ser Val Thr Cys Phe Gly Pro Glu
Ala Asp545 550 555 560Gln Cys Val Ala Cys Ala His Tyr Lys Asp Pro
Pro Phe Cys Val Ala 565 570 575Arg Cys Pro Ser Gly Val Lys Pro Asp
Leu Ser Tyr Met Pro Ile Trp 580 585 590Lys Phe Pro Asp Glu Glu Gly
Ala Cys Gln Pro Cys Pro Ile Asn Cys 595 600 605Thr His Ser Cys Val
Asp Leu Asp Asp Lys Gly Cys Pro Ala Glu Gln 610 615 620Arg Ala Ser
Pro Leu Thr Ser Ile Ile Ser Ala Val Val Gly Ile Leu625 630 635
640Leu Val Val Val Leu Gly Val Val Phe Gly Ile Leu Ile Lys Arg Arg
645 650 655Gln Gln Lys Ile Arg Lys Tyr Thr Met 660 66547688PRTMus
musculus 47Met Glu Leu Ala Ala Trp Cys Arg Trp Gly Phe Leu Leu Ala
Leu Leu1 5 10 15Ser Pro Gly Ala Ala Gly Thr Gln Val Cys Thr Gly Thr
Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp
Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn
Leu Glu Leu Thr Tyr 50 55 60Leu Pro Ala Asn Ala Ser Leu Ser Phe Leu
Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Met Leu Ile Ala His
Asn Arg Val Lys His Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg
Gly Thr Gln Leu Phe Glu Asp Lys Tyr 100 105 110Ala Leu Ala Val Leu
Asp Asn Arg Asp Pro Leu Asp Asn Val Thr Thr 115 120 125Ala Ala Pro
Gly Arg Thr Pro Glu Gly Leu Arg Glu Leu Gln Leu Arg 130 135 140Ser
Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Arg Gly Asn Pro145 150
155 160Gln Leu Cys Tyr Gln Asp Met Val Leu Trp Lys Asp Val Leu Arg
Lys 165 170 175Asn Asn Gln Leu Ala Pro Val Asp Met Asp Thr Asn Arg
Ser Arg Ala 180 185 190Cys Pro Pro Cys Ala Pro Thr Cys Lys Asp Asn
His Cys Trp Gly Glu 195 200 205Ser Pro Glu Asp Cys Gln Ile Leu Thr
Gly Thr Ile Cys Thr Ser Gly 210 215 220Cys Ala Arg Cys Lys Gly Arg
Leu Pro Thr Asp Cys Cys His Glu Gln225 230 235 240Cys Ala Ala Gly
Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys 245 250 255Leu His
Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu 260 265
270Ile Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Leu Asn Pro Glu Gly
275 280 285Arg Tyr Thr Phe Gly Ala Ser Cys Val Thr Thr Cys Pro Tyr
Asn Tyr 290 295 300Leu Ser Thr Glu Val Gly Ser Cys Thr Leu Val Cys
Pro Pro Asn Asn305 310 315 320Gln Glu Val Thr Ala Glu Asp Gly Thr
Gln Arg Cys Glu Lys Cys Ser 325 330 335Lys Pro Cys Ala Gly Val Cys
Tyr Gly Leu Gly Met Glu His Leu Arg 340 345 350Gly Ala Arg Ala Ile
Thr Ser Asp Asn Ile Gln Glu Phe Ala Gly Cys 355 360 365Lys Lys Ile
Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly 370 375 380Asn
Pro Ser Ser Gly Val Ala Pro Leu Lys Pro Glu His Leu Gln Val385 390
395 400Phe Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala
Trp 405 410 415Pro Glu Ser Phe Gln Asp Leu Ser Val Phe Gln Asn Leu
Arg Val Ile 420 425 430Arg Gly Arg Ile Leu His Asp Gly Ala Tyr Ser
Leu Thr Leu Gln Gly 435 440 445Leu Gly Ile His Ser Leu Gly Leu Arg
Ser Leu Arg Glu Leu Gly Ser 450 455 460Gly Leu Ala Leu Ile His Arg
Asn Thr His Leu Cys Phe Val Asn Thr465 470 475 480Val Pro Trp Asp
Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His 485 490 495Ser Gly
Asn Arg Pro Glu Glu Ala Cys Gly Leu Glu Gly Leu Val Cys 500 505
510Asn Ser Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln
515 520 525Cys Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val
Glu Glu 530 535 540Cys Arg Val Trp Lys Gly Leu Pro Arg Glu Tyr Val
Arg Gly Lys His545 550 555 560Cys Leu Pro Cys His Pro Glu Cys Gln
Pro Gln Asn Ser Ser Glu Thr 565 570 575Cys Tyr Gly Ser Glu Ala Asp
Gln Cys Glu Ala Cys Ala His Tyr Lys 580 585 590Asp Ser Ser Ser Cys
Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp 595 600 605Leu Ser Tyr
Met Pro Ile Trp Lys Tyr Pro Asp Glu Glu Gly Ile Cys 610 615 620Gln
Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Glu625 630
635 640Arg Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Val Thr Phe Ile
Ile 645 650 655Ala Thr Val Val Gly Val Leu Leu Phe Leu Ile Ile Val
Val Val Ile 660 665 670Gly Ile Leu Ile Lys Arg Arg Arg Gln Lys Ile
Arg Lys Tyr Thr Met 675 680 68548666PRTMus musculus 48Thr Gln Val
Cys Thr Gly Thr Asp Met Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu
Thr His Leu Asp Met Leu Arg His Leu Tyr Gln Gly Cys Gln 20 25 30Val
Val Gln Gly Asn Leu Glu Leu Thr Tyr Leu Pro Ala Asn Ala Ser 35 40
45Leu Ser Phe Leu Gln Asp Ile Gln Glu Val Gln Gly Tyr Met Leu Ile
50 55 60Ala His Asn Arg Val Lys His Val Pro Leu Gln Arg Leu Arg Ile
Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu Asp Lys Tyr Ala Leu Ala
Val Leu Asp 85 90 95Asn Arg Asp Pro Leu Asp Asn Val Thr Thr Ala Ala
Pro Gly Arg Thr 100 105 110Pro Glu Gly Leu Arg Glu Leu Gln Leu Arg
Ser Leu Thr Glu Ile Leu 115 120 125Lys Gly Gly Val Leu Ile Arg Gly
Asn Pro Gln Leu Cys Tyr Gln Asp 130 135 140Met
Val Leu Trp Lys Asp Val Leu Arg Lys Asn Asn Gln Leu Ala Pro145 150
155 160Val Asp Met Asp Thr Asn Arg Ser Arg Ala Cys Pro Pro Cys Ala
Pro 165 170 175Thr Cys Lys Asp Asn His Cys Trp Gly Glu Ser Pro Glu
Asp Cys Gln 180 185 190Ile Leu Thr Gly Thr Ile Cys Thr Ser Gly Cys
Ala Arg Cys Lys Gly 195 200 205Arg Leu Pro Thr Asp Cys Cys His Glu
Gln Cys Ala Ala Gly Cys Thr 210 215 220Gly Pro Lys His Ser Asp Cys
Leu Ala Cys Leu His Phe Asn His Ser225 230 235 240Gly Ile Cys Glu
Leu His Cys Pro Ala Leu Ile Thr Tyr Asn Thr Asp 245 250 255Thr Phe
Glu Ser Met Leu Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala 260 265
270Ser Cys Val Thr Thr Cys Pro Tyr Asn Tyr Leu Ser Thr Glu Val Gly
275 280 285Ser Cys Thr Leu Val Cys Pro Pro Asn Asn Gln Glu Val Thr
Ala Glu 290 295 300Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys Pro
Cys Ala Gly Val305 310 315 320Cys Tyr Gly Leu Gly Met Glu His Leu
Arg Gly Ala Arg Ala Ile Thr 325 330 335Ser Asp Asn Ile Gln Glu Phe
Ala Gly Cys Lys Lys Ile Phe Gly Ser 340 345 350Leu Ala Phe Leu Pro
Glu Ser Phe Asp Gly Asn Pro Ser Ser Gly Val 355 360 365Ala Pro Leu
Lys Pro Glu His Leu Gln Val Phe Glu Thr Leu Glu Glu 370 375 380Ile
Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Glu Ser Phe Gln Asp385 390
395 400Leu Ser Val Phe Gln Asn Leu Arg Val Ile Arg Gly Arg Ile Leu
His 405 410 415Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu Gly Ile
His Ser Leu 420 425 430Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly
Leu Ala Leu Ile His 435 440 445Arg Asn Thr His Leu Cys Phe Val Asn
Thr Val Pro Trp Asp Gln Leu 450 455 460Phe Arg Asn Pro His Gln Ala
Leu Leu His Ser Gly Asn Arg Pro Glu465 470 475 480Glu Ala Cys Gly
Leu Glu Gly Leu Val Cys Asn Ser Leu Cys Ala Arg 485 490 495Gly His
Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys Ser Gln 500 505
510Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Trp Lys Gly
515 520 525Leu Pro Arg Glu Tyr Val Arg Gly Lys His Cys Leu Pro Cys
His Pro 530 535 540Glu Cys Gln Pro Gln Asn Ser Ser Glu Thr Cys Tyr
Gly Ser Glu Ala545 550 555 560Asp Gln Cys Glu Ala Cys Ala His Tyr
Lys Asp Ser Ser Ser Cys Val 565 570 575Ala Arg Cys Pro Ser Gly Val
Lys Pro Asp Leu Ser Tyr Met Pro Ile 580 585 590Trp Lys Tyr Pro Asp
Glu Glu Gly Ile Cys Gln Pro Cys Pro Ile Asn 595 600 605Cys Thr His
Ser Cys Val Asp Leu Asp Glu Arg Gly Cys Pro Ala Glu 610 615 620Gln
Arg Ala Ser Pro Val Thr Phe Ile Ile Ala Thr Val Val Gly Val625 630
635 640Leu Leu Phe Leu Ile Ile Val Val Val Ile Gly Ile Leu Ile Lys
Arg 645 650 655Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met 660
66549691PRTRattus norvegicus 49Met Ile Ile Met Glu Leu Ala Ala Trp
Cys Arg Trp Gly Phe Leu Leu1 5 10 15Ala Leu Leu Pro Pro Gly Ile Ala
Gly Thr Gln Val Cys Thr Gly Thr 20 25 30Asp Met Lys Leu Arg Leu Pro
Ala Ser Pro Glu Thr His Leu Asp Met 35 40 45Leu Arg His Leu Tyr Gln
Gly Cys Gln Val Val Gln Gly Asn Leu Glu 50 55 60Leu Thr Tyr Val Pro
Ala Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile65 70 75 80Gln Glu Val
Gln Gly Tyr Met Leu Ile Ala His Asn Gln Val Lys Arg 85 90 95Val Pro
Leu Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu 100 105
110Asp Lys Tyr Ala Leu Ala Val Leu Asp Asn Arg Asp Pro Gln Asp Asn
115 120 125Val Ala Ala Ser Thr Pro Gly Arg Thr Pro Glu Gly Leu Arg
Glu Leu 130 135 140Gln Leu Arg Ser Leu Thr Glu Ile Leu Lys Gly Gly
Val Leu Ile Arg145 150 155 160Gly Asn Pro Gln Leu Cys Tyr Gln Asp
Met Val Leu Trp Lys Asp Val 165 170 175Phe Arg Lys Asn Asn Gln Leu
Ala Pro Val Asp Ile Asp Thr Asn Arg 180 185 190Ser Arg Ala Cys Pro
Pro Cys Ala Pro Ala Cys Lys Asp Asn His Cys 195 200 205Trp Gly Glu
Ser Pro Glu Asp Cys Gln Ile Leu Thr Gly Thr Ile Cys 210 215 220Thr
Ser Gly Cys Ala Arg Cys Lys Gly Arg Leu Pro Thr Asp Cys Cys225 230
235 240His Glu Gln Cys Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp
Cys 245 250 255Leu Ala Cys Leu His Phe Asn His Ser Gly Ile Cys Glu
Leu His Cys 260 265 270Pro Ala Leu Val Thr Tyr Asn Thr Asp Thr Phe
Glu Ser Met His Asn 275 280 285Pro Glu Gly Arg Tyr Thr Phe Gly Ala
Ser Cys Val Thr Thr Cys Pro 290 295 300Tyr Asn Tyr Leu Ser Thr Glu
Val Gly Ser Cys Thr Leu Val Cys Pro305 310 315 320Pro Asn Asn Gln
Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu 325 330 335Lys Cys
Ser Lys Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu 340 345
350His Leu Arg Gly Ala Arg Ala Ile Thr Ser Asp Asn Val Gln Glu Phe
355 360 365Asp Gly Cys Lys Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro
Glu Ser 370 375 380Phe Asp Gly Asp Pro Ser Ser Gly Ile Ala Pro Leu
Arg Pro Glu Gln385 390 395 400Leu Gln Val Phe Glu Thr Leu Glu Glu
Ile Thr Gly Tyr Leu Tyr Ile 405 410 415Ser Ala Trp Pro Asp Ser Leu
Arg Asp Leu Ser Val Phe Gln Asn Leu 420 425 430Arg Ile Ile Arg Gly
Arg Ile Leu His Asp Gly Ala Tyr Ser Leu Thr 435 440 445Leu Gln Gly
Leu Gly Ile His Ser Leu Gly Leu Arg Ser Leu Arg Glu 450 455 460Leu
Gly Ser Gly Leu Ala Leu Ile His Arg Asn Ala His Leu Cys Phe465 470
475 480Val His Thr Val Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln
Ala 485 490 495Leu Leu His Ser Gly Asn Arg Pro Glu Glu Asp Cys Gly
Leu Glu Gly 500 505 510Leu Val Cys Asn Ser Leu Cys Ala His Gly His
Cys Trp Gly Pro Gly 515 520 525Pro Thr Gln Cys Val Asn Cys Ser His
Phe Leu Arg Gly Gln Glu Cys 530 535 540Val Glu Glu Cys Arg Val Trp
Lys Gly Leu Pro Arg Glu Tyr Val Ser545 550 555 560Asp Lys Arg Cys
Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Ser 565 570 575Ser Glu
Thr Cys Phe Gly Ser Glu Ala Asp Gln Cys Ala Ala Cys Ala 580 585
590His Tyr Lys Asp Ser Ser Ser Cys Val Ala Arg Cys Pro Ser Gly Val
595 600 605Lys Pro Asp Leu Ser Tyr Met Pro Ile Trp Lys Tyr Pro Asp
Glu Glu 610 615 620Gly Ile Cys Gln Pro Cys Pro Ile Asn Cys Thr His
Ser Cys Val Asp625 630 635 640Leu Asp Glu Arg Gly Cys Pro Ala Glu
Gln Arg Ala Ser Pro Val Thr 645 650 655Phe Ile Ile Ala Thr Val Val
Gly Val Leu Leu Phe Leu Ile Leu Val 660 665 670Val Val Val Gly Ile
Leu Ile Lys Arg Arg Arg Gln Lys Ile Arg Lys 675 680 685Tyr Thr Met
69050666PRTRattus norvegicus 50Thr Gln Val Cys Thr Gly Thr Asp Met
Lys Leu Arg Leu Pro Ala Ser1 5 10 15Pro Glu Thr His Leu Asp Met Leu
Arg His Leu Tyr Gln Gly Cys Gln 20 25 30Val Val Gln Gly Asn Leu Glu
Leu Thr Tyr Val Pro Ala Asn Ala Ser 35 40 45Leu Ser Phe Leu Gln Asp
Ile Gln Glu Val Gln Gly Tyr Met Leu Ile 50 55 60Ala His Asn Gln Val
Lys Arg Val Pro Leu Gln Arg Leu Arg Ile Val65 70 75 80Arg Gly Thr
Gln Leu Phe Glu Asp Lys Tyr Ala Leu Ala Val Leu Asp 85 90 95Asn Arg
Asp Pro Gln Asp Asn Val Ala Ala Ser Thr Pro Gly Arg Thr 100 105
110Pro Glu Gly Leu Arg Glu Leu Gln Leu Arg Ser Leu Thr Glu Ile Leu
115 120 125Lys Gly Gly Val Leu Ile Arg Gly Asn Pro Gln Leu Cys Tyr
Gln Asp 130 135 140Met Val Leu Trp Lys Asp Val Phe Arg Lys Asn Asn
Gln Leu Ala Pro145 150 155 160Val Asp Ile Asp Thr Asn Arg Ser Arg
Ala Cys Pro Pro Cys Ala Pro 165 170 175Ala Cys Lys Asp Asn His Cys
Trp Gly Glu Ser Pro Glu Asp Cys Gln 180 185 190Ile Leu Thr Gly Thr
Ile Cys Thr Ser Gly Cys Ala Arg Cys Lys Gly 195 200 205Arg Leu Pro
Thr Asp Cys Cys His Glu Gln Cys Ala Ala Gly Cys Thr 210 215 220Gly
Pro Lys His Ser Asp Cys Leu Ala Cys Leu His Phe Asn His Ser225 230
235 240Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val Thr Tyr Asn Thr
Asp 245 250 255Thr Phe Glu Ser Met His Asn Pro Glu Gly Arg Tyr Thr
Phe Gly Ala 260 265 270Ser Cys Val Thr Thr Cys Pro Tyr Asn Tyr Leu
Ser Thr Glu Val Gly 275 280 285Ser Cys Thr Leu Val Cys Pro Pro Asn
Asn Gln Glu Val Thr Ala Glu 290 295 300Asp Gly Thr Gln Arg Cys Glu
Lys Cys Ser Lys Pro Cys Ala Arg Val305 310 315 320Cys Tyr Gly Leu
Gly Met Glu His Leu Arg Gly Ala Arg Ala Ile Thr 325 330 335Ser Asp
Asn Val Gln Glu Phe Asp Gly Cys Lys Lys Ile Phe Gly Ser 340 345
350Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp Pro Ser Ser Gly Ile
355 360 365Ala Pro Leu Arg Pro Glu Gln Leu Gln Val Phe Glu Thr Leu
Glu Glu 370 375 380Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp
Ser Leu Arg Asp385 390 395 400Leu Ser Val Phe Gln Asn Leu Arg Ile
Ile Arg Gly Arg Ile Leu His 405 410 415Asp Gly Ala Tyr Ser Leu Thr
Leu Gln Gly Leu Gly Ile His Ser Leu 420 425 430Gly Leu Arg Ser Leu
Arg Glu Leu Gly Ser Gly Leu Ala Leu Ile His 435 440 445Arg Asn Ala
His Leu Cys Phe Val His Thr Val Pro Trp Asp Gln Leu 450 455 460Phe
Arg Asn Pro His Gln Ala Leu Leu His Ser Gly Asn Arg Pro Glu465 470
475 480Glu Asp Cys Gly Leu Glu Gly Leu Val Cys Asn Ser Leu Cys Ala
His 485 490 495Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys Val Asn
Cys Ser His 500 505 510Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys
Arg Val Trp Lys Gly 515 520 525Leu Pro Arg Glu Tyr Val Ser Asp Lys
Arg Cys Leu Pro Cys His Pro 530 535 540Glu Cys Gln Pro Gln Asn Ser
Ser Glu Thr Cys Phe Gly Ser Glu Ala545 550 555 560Asp Gln Cys Ala
Ala Cys Ala His Tyr Lys Asp Ser Ser Ser Cys Val 565 570 575Ala Arg
Cys Pro Ser Gly Val Lys Pro Asp Leu Ser Tyr Met Pro Ile 580 585
590Trp Lys Tyr Pro Asp Glu Glu Gly Ile Cys Gln Pro Cys Pro Ile Asn
595 600 605Cys Thr His Ser Cys Val Asp Leu Asp Glu Arg Gly Cys Pro
Ala Glu 610 615 620Gln Arg Ala Ser Pro Val Thr Phe Ile Ile Ala Thr
Val Val Gly Val625 630 635 640Leu Leu Phe Leu Ile Leu Val Val Val
Val Gly Ile Leu Ile Lys Arg 645 650 655Arg Arg Gln Lys Ile Arg Lys
Tyr Thr Met 660 66551690PRTArtificial SequenceChimeric human-rat
E2Neu (HER2) 51Met Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu
Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly
Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu
Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val Val Gln Gly
Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala Ser Leu Ser Phe
Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val Leu Ile Ala
His Asn Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val
Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110Ala Leu Ala Val
Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro 115 120 125Val Thr
Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser 130 135
140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro
Gln145 150 155 160Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile
Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr
Asn Arg Ser Arg Ala Cys 180 185 190His Pro Cys Ser Pro Met Cys Lys
Gly Ser Arg Cys Trp Gly Glu Ser 195 200 205Ser Glu Asp Cys Gln Ser
Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220Ala Arg Cys Lys
Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln Cys225 230 235 240Ala
Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu 245 250
255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val
260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu
Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro
Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys Thr Leu Val
Cys Pro Leu His Asn Gln305 310 315 320Glu Val Thr Ala Glu Asp Gly
Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys Ala Arg Val
Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345 350Val Arg Ala
Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys 355 360 365Lys
Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp 370 375
380Pro Ala Ser Asn Thr Ala Glu Phe Ala Pro Leu Arg Pro Glu Gln
Leu385 390 395 400Gln Val Phe Glu Thr Leu Glu Glu Ile Thr Gly Tyr
Leu Tyr Ile Ser 405 410 415Ala Trp Pro Asp Ser Leu Arg Asp Leu Ser
Val Phe Gln Asn Leu Arg 420 425 430Ile Ile Arg Gly Arg Ile Leu His
Asp Gly Ala Tyr Ser Leu Thr Leu 435 440 445Gln Gly Leu Gly Ile His
Ser Leu Gly Leu Arg Ser Leu Arg Glu Leu 450 455 460Gly Ser Gly Leu
Ala Leu Ile His Arg Asn Ala His Leu Cys Phe Val465 470 475 480His
Thr Val Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu 485 490
495Leu His Ser Gly Asn Arg Pro Glu Glu Asp Cys Gly Leu Glu Gly Leu
500 505 510Val Cys Asn Ser Leu Cys Ala His Gly His Cys Trp Gly Pro
Gly Pro 515 520 525Thr Gln Cys Val Asn Cys Ser His Phe Leu Arg Gly
Gln Glu Cys Val 530 535 540Glu Glu Cys Arg Val Trp Lys Gly Leu Pro
Arg Glu Tyr Val Ser Asp545 550 555 560Lys Arg Cys Leu Pro Cys His
Pro Glu Cys Gln Pro Gln Asn Ser Ser 565 570 575Glu Thr Cys Phe
Gly
Ser Glu Ala Asp Gln Cys Ala Ala Cys Ala His 580 585 590Tyr Lys Asp
Ser Ser Ser Cys Val Ala Arg Cys Pro Ser Gly Val Lys 595 600 605Pro
Asp Leu Ser Tyr Met Pro Ile Trp Lys Tyr Pro Asp Glu Glu Gly 610 615
620Ile Cys Gln Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp
Leu625 630 635 640Asp Glu Arg Gly Cys Pro Ala Glu Gln Arg Ala Ser
Pro Val Thr Phe 645 650 655Ile Ile Ala Thr Val Val Gly Val Leu Leu
Phe Leu Ile Leu Val Val 660 665 670Val Val Gly Ile Leu Ile Lys Arg
Arg Arg Gln Lys Ile Arg Lys Tyr 675 680 685Thr Met
69052668PRTArtificial SequenceChimeric human-rat E2Neu (HER2) 52Thr
Gln Val Cys Thr Gly Thr Asp Met Lys Leu Arg Leu Pro Ala Ser1 5 10
15Pro Glu Thr His Leu Asp Met Leu Arg His Leu Tyr Gln Gly Cys Gln
20 25 30Val Val Gln Gly Asn Leu Glu Leu Thr Tyr Leu Pro Thr Asn Ala
Ser 35 40 45Leu Ser Phe Leu Gln Asp Ile Gln Glu Val Gln Gly Tyr Val
Leu Ile 50 55 60Ala His Asn Gln Val Arg Gln Val Pro Leu Gln Arg Leu
Arg Ile Val65 70 75 80Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr Ala
Leu Ala Val Leu Asp 85 90 95Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro
Val Thr Gly Ala Ser Pro 100 105 110Gly Gly Leu Arg Glu Leu Gln Leu
Arg Ser Leu Thr Glu Ile Leu Lys 115 120 125Gly Gly Val Leu Ile Gln
Arg Asn Pro Gln Leu Cys Tyr Gln Asp Thr 130 135 140Ile Leu Trp Lys
Asp Ile Phe His Lys Asn Asn Gln Leu Ala Leu Thr145 150 155 160Leu
Ile Asp Thr Asn Arg Ser Arg Ala Cys His Pro Cys Ser Pro Met 165 170
175Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser Ser Glu Asp Cys Gln Ser
180 185 190Leu Thr Arg Thr Val Cys Ala Gly Gly Cys Ala Arg Cys Lys
Gly Pro 195 200 205Leu Pro Thr Asp Cys Cys His Glu Gln Cys Ala Ala
Gly Cys Thr Gly 210 215 220Pro Lys His Ser Asp Cys Leu Ala Cys Leu
His Phe Asn His Ser Gly225 230 235 240Ile Cys Glu Leu His Cys Pro
Ala Leu Val Thr Tyr Asn Thr Asp Thr 245 250 255Phe Glu Ser Met Pro
Asn Pro Glu Gly Arg Tyr Thr Phe Gly Ala Ser 260 265 270Cys Val Thr
Ala Cys Pro Tyr Asn Tyr Leu Ser Thr Asp Val Gly Ser 275 280 285Cys
Thr Leu Val Cys Pro Leu His Asn Gln Glu Val Thr Ala Glu Asp 290 295
300Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys Pro Cys Ala Arg Val
Cys305 310 315 320Tyr Gly Leu Gly Met Glu His Leu Arg Glu Val Arg
Ala Val Thr Ser 325 330 335Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys
Lys Ile Phe Gly Ser Leu 340 345 350Ala Phe Leu Pro Glu Ser Phe Asp
Gly Asp Pro Ala Ser Asn Thr Ala 355 360 365Glu Phe Ala Pro Leu Arg
Pro Glu Gln Leu Gln Val Phe Glu Thr Leu 370 375 380Glu Glu Ile Thr
Gly Tyr Leu Tyr Ile Ser Ala Trp Pro Asp Ser Leu385 390 395 400Arg
Asp Leu Ser Val Phe Gln Asn Leu Arg Ile Ile Arg Gly Arg Ile 405 410
415Leu His Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu Gly Ile His
420 425 430Ser Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly Leu
Ala Leu 435 440 445Ile His Arg Asn Ala His Leu Cys Phe Val His Thr
Val Pro Trp Asp 450 455 460Gln Leu Phe Arg Asn Pro His Gln Ala Leu
Leu His Ser Gly Asn Arg465 470 475 480Pro Glu Glu Asp Cys Gly Leu
Glu Gly Leu Val Cys Asn Ser Leu Cys 485 490 495Ala His Gly His Cys
Trp Gly Pro Gly Pro Thr Gln Cys Val Asn Cys 500 505 510Ser His Phe
Leu Arg Gly Gln Glu Cys Val Glu Glu Cys Arg Val Trp 515 520 525Lys
Gly Leu Pro Arg Glu Tyr Val Ser Asp Lys Arg Cys Leu Pro Cys 530 535
540His Pro Glu Cys Gln Pro Gln Asn Ser Ser Glu Thr Cys Phe Gly
Ser545 550 555 560Glu Ala Asp Gln Cys Ala Ala Cys Ala His Tyr Lys
Asp Ser Ser Ser 565 570 575Cys Val Ala Arg Cys Pro Ser Gly Val Lys
Pro Asp Leu Ser Tyr Met 580 585 590Pro Ile Trp Lys Tyr Pro Asp Glu
Glu Gly Ile Cys Gln Pro Cys Pro 595 600 605Ile Asn Cys Thr His Ser
Cys Val Asp Leu Asp Glu Arg Gly Cys Pro 610 615 620Ala Glu Gln Arg
Ala Ser Pro Val Thr Phe Ile Ile Ala Thr Val Val625 630 635 640Gly
Val Leu Leu Phe Leu Ile Leu Val Val Val Val Gly Ile Leu Ile 645 650
655Lys Arg Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met 660
665532064DNAHomo sapiens 53atggagctgg cggccttgtg ccgctggggg
ctcctcctcg ccctcttgcc ccccggagcc 60gcgagcaccc aagtgtgcac cggcacagac
atgaagctgc ggctccctgc cagtcccgag 120acccacctgg acatgctccg
ccacctctac cagggctgcc aggtggtgca gggaaacctg 180gaactcacct
acctgcccac caatgccagc ctgtccttcc tgcaggatat ccaggaggtg
240cagggctacg tgctcatcgc tcacaaccaa gtgaggcagg tcccactgca
gaggctgcgg 300attgtgcgag gcacccagct ctttgaggac aactatgccc
tggccgtgct agacaatgga 360gacccgctga acaataccac ccctgtcaca
ggggcctccc caggaggcct gcgggagctg 420cagcttcgaa gcctcacaga
gatcttgaaa ggaggggtct tgatccagcg gaacccccag 480ctctgctacc
aggacacgat tttgtggaag gacatcttcc acaagaacaa ccagctggct
540ctcacactga tagacaccaa ccgctctcgg gcctgccacc cctgttctcc
gatgtgtaag 600ggctcccgct gctggggaga gagttctgag gattgtcaga
gcctgacgcg cactgtctgt 660gccggtggct gtgcccgctg caaggggcca
ctgcccactg actgctgcca tgagcagtgt 720gctgccggct gcacgggccc
caagcactct gactgcctgg cctgcctcca cttcaaccac 780agtggcatct
gtgagctgca ctgcccagcc ctggtcacct acaacacaga cacgtttgag
840tccatgccca atcccgaggg ccggtataca ttcggcgcca gctgtgtgac
tgcctgtccc 900tacaactacc tttctacgga cgtgggatcc tgcaccctcg
tctgccccct gcacaaccaa 960gaggtgacag cagaggatgg aacacagcgg
tgtgagaagt gcagcaagcc ctgtgcccga 1020gtgtgctatg gtctgggcat
ggagcacttg cgagaggtga gggcagttac cagtgccaat 1080atccaggagt
ttgctggctg caagaagatc tttgggagcc tggcatttct gccggagagc
1140tttgatgggg acccagcctc caacactgcc ccgctccagc cagagcagct
ccaagtgttt 1200gagactctgg aagagatcac aggttaccta tacatctcag
catggccgga cagcctgcct 1260gacctcagcg tcttccagaa cctgcaagta
atccggggac gaattctgca caatggcgcc 1320tactcgctga ccctgcaagg
gctgggcatc agctggctgg ggctgcgctc actgagggaa 1380ctgggcagtg
gactggccct catccaccat aacacccacc tctgcttcgt gcacacggtg
1440ccctgggacc agctctttcg gaacccgcac caagctctgc tccacactgc
caaccggcca 1500gaggacgagt gtgtgggcga gggcctggcc tgccaccagc
tgtgcgcccg agggcactgc 1560tggggtccag ggcccaccca gtgtgtcaac
tgcagccagt tccttcgggg ccaggagtgc 1620gtggaggaat gccgagtact
gcaggggctc cccagggagt atgtgaatgc caggcactgt 1680ttgccgtgcc
accctgagtg tcagccccag aatggctcag tgacctgttt tggaccggag
1740gctgaccagt gtgtggcctg tgcccactat aaggaccctc ccttctgcgt
ggcccgctgc 1800cccagcggtg tgaaacctga cctctcctac atgcccatct
ggaagtttcc agatgaggag 1860ggcgcatgcc agccttgccc catcaactgc
acccactcct gtgtggacct ggatgacaag 1920ggctgccccg ccgagcagag
agccagccct ctgacgtcca tcatctctgc ggtggttggc 1980attctgctgg
tcgtggtctt gggggtggtc tttgggatcc tcatcaagcg acggcagcag
2040aagatccgga agtacacgat gtga 2064541998DNAHomo sapiens
54acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac
60ctggacatgc tccgccacct ctaccagggc tgccaggtgg tgcagggaaa cctggaactc
120acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg atatccagga
ggtgcagggc 180tacgtgctca tcgctcacaa ccaagtgagg caggtcccac
tgcagaggct gcggattgtg 240cgaggcaccc agctctttga ggacaactat
gccctggccg tgctagacaa tggagacccg 300ctgaacaata ccacccctgt
cacaggggcc tccccaggag gcctgcggga gctgcagctt 360cgaagcctca
cagagatctt gaaaggaggg gtcttgatcc agcggaaccc ccagctctgc
420taccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggctctcaca 480ctgatagaca ccaaccgctc tcgggcctgc cacccctgtt
ctccgatgtg taagggctcc 540cgctgctggg gagagagttc tgaggattgt
cagagcctga cgcgcactgt ctgtgccggt 600ggctgtgccc gctgcaaggg
gccactgccc actgactgct gccatgagca gtgtgctgcc 660ggctgcacgg
gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc
720atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt
tgagtccatg 780cccaatcccg agggccggta tacattcggc gccagctgtg
tgactgcctg tccctacaac 840tacctttcta cggacgtggg atcctgcacc
ctcgtctgcc ccctgcacaa ccaagaggtg 900acagcagagg atggaacaca
gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc 960tatggtctgg
gcatggagca cttgcgagag gtgagggcag ttaccagtgc caatatccag
1020gagtttgctg gctgcaagaa gatctttggg agcctggcat ttctgccgga
gagctttgat 1080ggggacccag cctccaacac tgccccgctc cagccagagc
agctccaagt gtttgagact 1140ctggaagaga tcacaggtta cctatacatc
tcagcatggc cggacagcct gcctgacctc 1200agcgtcttcc agaacctgca
agtaatccgg ggacgaattc tgcacaatgg cgcctactcg 1260ctgaccctgc
aagggctggg catcagctgg ctggggctgc gctcactgag ggaactgggc
1320agtggactgg ccctcatcca ccataacacc cacctctgct tcgtgcacac
ggtgccctgg 1380gaccagctct ttcggaaccc gcaccaagct ctgctccaca
ctgccaaccg gccagaggac 1440gagtgtgtgg gcgagggcct ggcctgccac
cagctgtgcg cccgagggca ctgctggggt 1500ccagggccca cccagtgtgt
caactgcagc cagttccttc ggggccagga gtgcgtggag 1560gaatgccgag
tactgcaggg gctccccagg gagtatgtga atgccaggca ctgtttgccg
1620tgccaccctg agtgtcagcc ccagaatggc tcagtgacct gttttggacc
ggaggctgac 1680cagtgtgtgg cctgtgccca ctataaggac cctcccttct
gcgtggcccg ctgccccagc 1740ggtgtgaaac ctgacctctc ctacatgccc
atctggaagt ttccagatga ggagggcgca 1800tgccagcctt gccccatcaa
ctgcacccac tcctgtgtgg acctggatga caagggctgc 1860cccgccgagc
agagagccag ccctctgacg tccatcatct ctgcggtggt tggcattctg
1920ctggtcgtgg tcttgggggt ggtctttggg atcctcatca agcgacggca
gcagaagatc 1980cggaagtaca cgatgtga 1998552064DNAMus musculus
55atggagctgg cggcctggtg ccgttggggg ttcctcctcg ccctcctgtc ccccggagcc
60gcgggtaccc aagtgtgtac cggtaccgac atgaagttgc gactccctgc cagtcctgag
120acccacctgg acatgcttcg ccacctctac cagggctgtc aggtggtgca
gggcaatttg 180gagcttacct acctgcccgc caatgccagc ctctcattcc
tgcaggacat ccaggaagtc 240cagggataca tgctcatcgc tcacaaccga
gtgaaacacg tcccactgca gaggttgcgc 300atcgtgagag ggactcagct
ctttgaggac aagtatgccc tggctgtgct agacaaccga 360gaccctttgg
acaacgtcac caccgccgcc ccaggcagaa ccccagaagg gctgcgggag
420ctgcagcttc gaagtctcac agagatcttg aagggaggag ttttgatccg
tgggaaccct 480cagctctgct accaggacat ggttttgtgg aaggatgtcc
tccgtaagaa taaccagctg 540gctcctgtcg acatggacac caatcgttcc
cgggcctgtc caccttgtgc cccaacctgc 600aaagacaatc actgttgggg
tgagagtcct gaagactgtc agatcttgac tggcaccatc 660tgtactagtg
gctgtgcccg gtgcaagggc cggctgccca ctgactgttg ccatgagcag
720tgtgctgcag gctgcacggg tcccaagcat tctgactgcc tggcctgcct
ccacttcaat 780catagtggta tctgtgagct gcactgcccg gccctcatca
cctacaacac agacaccttc 840gagtccatgc tcaaccctga gggtcgctac
acctttggtg ccagctgtgt gaccacctgc 900ccctacaact acctctccac
ggaagtggga tcctgcactc tggtctgtcc cccgaacaac 960caagaggtca
cagctgagga cggaacacag cggtgtgaga aatgcagcaa gccctgtgct
1020ggagtatgct atggtctggg catggagcac ctccgagggg cgagggccat
caccagtgac 1080aatatccagg agtttgctgg ctgcaagaag atctttggga
gcctggcatt tttgccggag 1140agctttgatg ggaacccctc ctccggcgtt
gccccactga agccagagca tctccaagtg 1200ttcgaaaccc tggaggagat
cacaggttac ctatacattt cagcatggcc agagagcttc 1260caagacctca
gtgtcttcca gaaccttcgg gtcattcggg gacggattct ccatgatggt
1320gcttactcat tgacgttgca aggcctgggg attcactcac tggggctacg
ctcactgcgg 1380gagctgggca gtggattggc tctcattcac cgcaacaccc
atctctgctt tgtaaacact 1440gtaccttggg accagctctt ccggaacccg
caccaggccc tactccacag tgggaaccgg 1500ccagaagagg catgtggtct
tgagggcttg gtctgtaact cactgtgtgc ccgtgggcac 1560tgctgggggc
cagggcccac ccagtgtgtc aactgcagtc agttcctccg gggccaggag
1620tgtgtggagg agtgccgagt atggaagggg ctccccaggg agtatgtgag
gggcaagcac 1680tgtctgccat gccaccccga gtgtcagcct caaaacagct
cggagacctg ctatggatcg 1740gaggctgacc agtgtgaggc ttgtgcccac
tacaaggact catcttcctg tgtggctcgc 1800tgccccagtg gtgtgaagcc
agacctctcc tacatgccta tctggaagta cccggatgag 1860gagggcatat
gtcagccatg ccccatcaac tgcacccact catgtgtgga cctggacgaa
1920cgaggctgcc cagcagagca gagagccagc ccagtgacat tcatcattgc
aactgtggtg 1980ggcgtcctgt tgttcctgat catagtggtg gtcattggaa
tcctaatcaa acgaaggcga 2040cagaagatcc ggaagtatac catg
2064561998DNAMus musculus 56acccaagtgt gtaccggtac cgacatgaag
ttgcgactcc ctgccagtcc tgagacccac 60ctggacatgc ttcgccacct ctaccagggc
tgtcaggtgg tgcagggcaa tttggagctt 120acctacctgc ccgccaatgc
cagcctctca ttcctgcagg acatccagga agtccaggga 180tacatgctca
tcgctcacaa ccgagtgaaa cacgtcccac tgcagaggtt gcgcatcgtg
240agagggactc agctctttga ggacaagtat gccctggctg tgctagacaa
ccgagaccct 300ttggacaacg tcaccaccgc cgccccaggc agaaccccag
aagggctgcg ggagctgcag 360cttcgaagtc tcacagagat cttgaaggga
ggagttttga tccgtgggaa ccctcagctc 420tgctaccagg acatggtttt
gtggaaggat gtcctccgta agaataacca gctggctcct 480gtcgacatgg
acaccaatcg ttcccgggcc tgtccacctt gtgccccaac ctgcaaagac
540aatcactgtt ggggtgagag tcctgaagac tgtcagatct tgactggcac
catctgtact 600agtggctgtg cccggtgcaa gggccggctg cccactgact
gttgccatga gcagtgtgct 660gcaggctgca cgggtcccaa gcattctgac
tgcctggcct gcctccactt caatcatagt 720ggtatctgtg agctgcactg
cccggccctc atcacctaca acacagacac cttcgagtcc 780atgctcaacc
ctgagggtcg ctacaccttt ggtgccagct gtgtgaccac ctgcccctac
840aactacctct ccacggaagt gggatcctgc actctggtct gtcccccgaa
caaccaagag 900gtcacagctg aggacggaac acagcggtgt gagaaatgca
gcaagccctg tgctggagta 960tgctatggtc tgggcatgga gcacctccga
ggggcgaggg ccatcaccag tgacaatatc 1020caggagtttg ctggctgcaa
gaagatcttt gggagcctgg catttttgcc ggagagcttt 1080gatgggaacc
cctcctccgg cgttgcccca ctgaagccag agcatctcca agtgttcgaa
1140accctggagg agatcacagg ttacctatac atttcagcat ggccagagag
cttccaagac 1200ctcagtgtct tccagaacct tcgggtcatt cggggacgga
ttctccatga tggtgcttac 1260tcattgacgt tgcaaggcct ggggattcac
tcactggggc tacgctcact gcgggagctg 1320ggcagtggat tggctctcat
tcaccgcaac acccatctct gctttgtaaa cactgtacct 1380tgggaccagc
tcttccggaa cccgcaccag gccctactcc acagtgggaa ccggccagaa
1440gaggcatgtg gtcttgaggg cttggtctgt aactcactgt gtgcccgtgg
gcactgctgg 1500gggccagggc ccacccagtg tgtcaactgc agtcagttcc
tccggggcca ggagtgtgtg 1560gaggagtgcc gagtatggaa ggggctcccc
agggagtatg tgaggggcaa gcactgtctg 1620ccatgccacc ccgagtgtca
gcctcaaaac agctcggaga cctgctatgg atcggaggct 1680gaccagtgtg
aggcttgtgc ccactacaag gactcatctt cctgtgtggc tcgctgcccc
1740agtggtgtga agccagacct ctcctacatg cctatctgga agtacccgga
tgaggagggc 1800atatgtcagc catgccccat caactgcacc cactcatgtg
tggacctgga cgaacgaggc 1860tgcccagcag agcagagagc cagcccagtg
acattcatca ttgcaactgt ggtgggcgtc 1920ctgttgttcc tgatcatagt
ggtggtcatt ggaatcctaa tcaaacgaag gcgacagaag 1980atccggaagt ataccatg
1998572073DNARattus norvegicus 57atgatcatca tggagctggc ggcctggtgc
cgctgggggt tcctcctcgc cctcctgccc 60cccggaatcg cgggcaccca agtgtgtacc
ggcacagaca tgaagttgcg gctccctgcc 120agtcctgaga cccacctgga
catgctccgc cacctgtacc agggctgtca ggtagtgcag 180ggcaacttgg
agcttaccta cgtgcctgcc aatgccagcc tctcattcct gcaggacatc
240caggaagttc agggttacat gctcatcgct cacaaccagg tgaagcgcgt
cccactgcaa 300aggctgcgca tcgtgagagg gacccagctc tttgaggaca
agtatgccct ggctgtgcta 360gacaaccgag atcctcagga caatgtcgcc
gcctccaccc caggcagaac cccagagggg 420ctgcgggagc tgcagcttcg
aagtctcaca gagatcctga agggaggagt tttgatccgt 480gggaaccctc
agctctgcta ccaggacatg gttttgtgga aggacgtctt ccgcaagaat
540aaccaactgg ctcctgtcga tatagacacc aatcgttccc gggcctgtcc
accttgtgcc 600cccgcctgca aagacaatca ctgttggggt gagagtccgg
aagactgtca gatcttgact 660ggcaccatct gtaccagtgg ttgtgcccgg
tgcaagggcc ggctgcccac tgactgctgc 720catgagcagt gtgccgcagg
ctgcacgggc cccaagcatt ctgactgcct ggcctgcctc 780cacttcaatc
atagtggtat ctgtgagctg cactgcccag ccctcgtcac ctacaacaca
840gacacctttg agtccatgca caaccctgag ggtcgctaca cctttggtgc
cagctgcgtg 900accacctgcc cctacaacta cctgtctacg gaagtgggat
cctgcactct ggtgtgtccc 960ccgaataacc aagaggtcac agctgaggac
ggaacacagc gttgtgagaa atgcagcaag 1020ccctgtgctc gagtgtgcta
tggtctgggc atggagcacc ttcgaggggc gagggccatc 1080accagtgaca
atgtccagga gtttgatggc tgcaagaaga tctttgggag cctggcattt
1140ttgccggaga gctttgatgg ggacccctcc tccggcattg ctccgctgag
gcctgagcag 1200ctccaagtgt tcgaaaccct ggaggagatc acaggttacc
tgtacatctc agcatggcca 1260gacagtctcc gtgacctcag tgtcttccag
aaccttcgaa tcattcgggg acggattctc 1320cacgatggcg cgtactcatt
gacactgcaa ggcctgggga tccactcgct ggggctgcgc 1380tcactgcggg
agctgggcag tggattggct ctgattcacc gcaacgccca tctctgcttt
1440gtacacactg taccttggga ccagctcttc cggaacccac atcaggccct
gctccacagt 1500gggaaccggc cggaagagga ttgtggtctc gagggcttgg
tctgtaactc actgtgtgcc 1560cacgggcact gctgggggcc agggcccacc
cagtgtgtca actgcagtca tttccttcgg 1620ggccaggagt gtgtggagga
gtgccgagta tggaaggggc tcccccggga gtatgtgagt 1680gacaagcgct
gtctgccgtg tcaccccgag tgtcagcctc aaaacagctc agagacctgc
1740tttggatcgg aggctgatca gtgtgcagcc tgcgcccact acaaggactc
gtcctcctgt 1800gtggctcgct gccccagtgg tgtgaaaccg gacctctcct
acatgcccat ctggaagtac 1860ccggatgagg agggcatatg
ccagccgtgc cccatcaact gcacccactc ctgtgtggat 1920ctggatgaac
gaggctgccc agcagagcag agagccagcc cggtgacatt catcattgca
1980actgtagtgg gcgtcctgct gttcctgatc ttagtggtgg tcgttggaat
cctaatcaaa 2040cgaaggagac agaagatccg gaagtatacg atg
2073581998DNARattus norvegicus 58acccaagtgt gtaccggcac agacatgaag
ttgcggctcc ctgccagtcc tgagacccac 60ctggacatgc tccgccacct gtaccagggc
tgtcaggtag tgcagggcaa cttggagctt 120acctacgtgc ctgccaatgc
cagcctctca ttcctgcagg acatccagga agttcagggt 180tacatgctca
tcgctcacaa ccaggtgaag cgcgtcccac tgcaaaggct gcgcatcgtg
240agagggaccc agctctttga ggacaagtat gccctggctg tgctagacaa
ccgagatcct 300caggacaatg tcgccgcctc caccccaggc agaaccccag
aggggctgcg ggagctgcag 360cttcgaagtc tcacagagat cctgaaggga
ggagttttga tccgtgggaa ccctcagctc 420tgctaccagg acatggtttt
gtggaaggac gtcttccgca agaataacca actggctcct 480gtcgatatag
acaccaatcg ttcccgggcc tgtccacctt gtgcccccgc ctgcaaagac
540aatcactgtt ggggtgagag tccggaagac tgtcagatct tgactggcac
catctgtacc 600agtggttgtg cccggtgcaa gggccggctg cccactgact
gctgccatga gcagtgtgcc 660gcaggctgca cgggccccaa gcattctgac
tgcctggcct gcctccactt caatcatagt 720ggtatctgtg agctgcactg
cccagccctc gtcacctaca acacagacac ctttgagtcc 780atgcacaacc
ctgagggtcg ctacaccttt ggtgccagct gcgtgaccac ctgcccctac
840aactacctgt ctacggaagt gggatcctgc actctggtgt gtcccccgaa
taaccaagag 900gtcacagctg aggacggaac acagcgttgt gagaaatgca
gcaagccctg tgctcgagtg 960tgctatggtc tgggcatgga gcaccttcga
ggggcgaggg ccatcaccag tgacaatgtc 1020caggagtttg atggctgcaa
gaagatcttt gggagcctgg catttttgcc ggagagcttt 1080gatggggacc
cctcctccgg cattgctccg ctgaggcctg agcagctcca agtgttcgaa
1140accctggagg agatcacagg ttacctgtac atctcagcat ggccagacag
tctccgtgac 1200ctcagtgtct tccagaacct tcgaatcatt cggggacgga
ttctccacga tggcgcgtac 1260tcattgacac tgcaaggcct ggggatccac
tcgctggggc tgcgctcact gcgggagctg 1320ggcagtggat tggctctgat
tcaccgcaac gcccatctct gctttgtaca cactgtacct 1380tgggaccagc
tcttccggaa cccacatcag gccctgctcc acagtgggaa ccggccggaa
1440gaggattgtg gtctcgaggg cttggtctgt aactcactgt gtgcccacgg
gcactgctgg 1500gggccagggc ccacccagtg tgtcaactgc agtcatttcc
ttcggggcca ggagtgtgtg 1560gaggagtgcc gagtatggaa ggggctcccc
cgggagtatg tgagtgacaa gcgctgtctg 1620ccgtgtcacc ccgagtgtca
gcctcaaaac agctcagaga cctgctttgg atcggaggct 1680gatcagtgtg
cagcctgcgc ccactacaag gactcgtcct cctgtgtggc tcgctgcccc
1740agtggtgtga aaccggacct ctcctacatg cccatctgga agtacccgga
tgaggagggc 1800atatgccagc cgtgccccat caactgcacc cactcctgtg
tggatctgga tgaacgaggc 1860tgcccagcag agcagagagc cagcccggtg
acattcatca ttgcaactgt agtgggcgtc 1920ctgctgttcc tgatcttagt
ggtggtcgtt ggaatcctaa tcaaacgaag gagacagaag 1980atccggaagt atacgatg
1998592070DNAArtificial SequenceChimeric human-rat E2Neu (HER2)
59atggagctgg cggccttgtg ccgctggggg ctcctcctcg ccctcttgcc ccccggagcc
60gcgagcaccc aagtgtgcac cggcacagac atgaagctgc ggctccctgc cagtcccgag
120acccacctgg acatgctccg ccacctctac cagggctgcc aggtggtgca
gggaaacctg 180gaactcacct acctgcccac caatgccagc ctgtccttcc
tgcaggatat ccaggaggtg 240cagggctacg tgctcatcgc tcacaaccaa
gtgaggcagg tcccactgca gaggctgcgg 300attgtgcgag gcacccagct
ctttgaggac aactatgccc tggccgtgct agacaatgga 360gacccgctga
acaataccac ccctgtcaca ggggcctccc caggaggcct gcgggagctg
420cagcttcgaa gcctcacaga gatcttgaaa ggaggggtct tgatccagcg
gaacccccag 480ctctgctacc aggacacgat tttgtggaag gacatcttcc
acaagaacaa ccagctggct 540ctcacactga tagacaccaa ccgctctcgg
gcctgccacc cctgttctcc gatgtgtaag 600ggctcccgct gctggggaga
gagttctgag gattgtcaga gcctgacgcg cactgtctgt 660gccggtggct
gtgcccgctg caaggggcca ctgcccactg actgctgcca tgagcagtgt
720gctgccggct gcacgggccc caagcactct gactgcctgg cctgcctcca
cttcaaccac 780agtggcatct gtgagctgca ctgcccagcc ctggtcacct
acaacacaga cacgtttgag 840tccatgccca atcccgaggg ccggtataca
ttcggcgcca gctgtgtgac tgcctgtccc 900tacaactacc tttctacgga
cgtgggatcc tgcaccctcg tctgccccct gcacaaccaa 960gaggtgacag
cagaggatgg aacacagcgg tgtgagaagt gcagcaagcc ctgtgcccga
1020gtgtgctatg gtctgggcat ggagcacttg cgagaggtga gggcagttac
cagtgccaat 1080atccaggagt ttgctggctg caagaagatc tttgggagcc
tggcatttct gccggagagc 1140tttgatgggg acccagcctc caacactgcc
gaattcgctc cgctgaggcc tgagcagctc 1200caagtgttcg aaaccctgga
ggagatcaca ggttacctgt acatctcagc atggccagac 1260agtctccgtg
acctcagtgt cttccagaac cttcgaatca ttcggggacg gattctccac
1320gatggcgcgt actcattgac actgcaaggc ctggggatcc actcgctggg
gctgcgctca 1380ctgcgggagc tgggcagtgg attggctctg attcaccgca
acgcccatct ctgctttgta 1440cacactgtac cttgggacca gctcttccgg
aacccacatc aggccctgct ccacagtggg 1500aaccggccgg aagaggattg
tggtctcgag ggcttggtct gtaactcact gtgtgcccac 1560gggcactgct
gggggccagg gcccacccag tgtgtcaact gcagtcattt ccttcggggc
1620caggagtgtg tggaggagtg ccgagtatgg aaggggctcc cccgggagta
tgtgagtgac 1680aagcgctgtc tgccgtgtca ccccgagtgt cagcctcaaa
acagctcaga gacctgcttt 1740ggatcggagg ctgatcagtg tgcagcctgc
gcccactaca aggactcgtc ctcctgtgtg 1800gctcgctgcc ccagtggtgt
gaaaccggac ctctcctaca tgcccatctg gaagtacccg 1860gatgaggagg
gcatatgcca gccgtgcccc atcaactgca cccactcctg tgtggatctg
1920gatgaacgag gctgcccagc agagcagaga gccagcccgg tgacattcat
cattgcaact 1980gtagtgggcg tcctgctgtt cctgatctta gtggtggtcg
ttggaatcct aatcaaacga 2040aggagacaga agatccggaa gtatacgatg
2070602004DNAArtificial SequenceChimeric human-rat E2Neu (HER2)
60acccaagtgt gcaccggcac agacatgaag ctgcggctcc ctgccagtcc cgagacccac
60ctggacatgc tccgccacct ctaccagggc tgccaggtgg tgcagggaaa cctggaactc
120acctacctgc ccaccaatgc cagcctgtcc ttcctgcagg atatccagga
ggtgcagggc 180tacgtgctca tcgctcacaa ccaagtgagg caggtcccac
tgcagaggct gcggattgtg 240cgaggcaccc agctctttga ggacaactat
gccctggccg tgctagacaa tggagacccg 300ctgaacaata ccacccctgt
cacaggggcc tccccaggag gcctgcggga gctgcagctt 360cgaagcctca
cagagatctt gaaaggaggg gtcttgatcc agcggaaccc ccagctctgc
420taccaggaca cgattttgtg gaaggacatc ttccacaaga acaaccagct
ggctctcaca 480ctgatagaca ccaaccgctc tcgggcctgc cacccctgtt
ctccgatgtg taagggctcc 540cgctgctggg gagagagttc tgaggattgt
cagagcctga cgcgcactgt ctgtgccggt 600ggctgtgccc gctgcaaggg
gccactgccc actgactgct gccatgagca gtgtgctgcc 660ggctgcacgg
gccccaagca ctctgactgc ctggcctgcc tccacttcaa ccacagtggc
720atctgtgagc tgcactgccc agccctggtc acctacaaca cagacacgtt
tgagtccatg 780cccaatcccg agggccggta tacattcggc gccagctgtg
tgactgcctg tccctacaac 840tacctttcta cggacgtggg atcctgcacc
ctcgtctgcc ccctgcacaa ccaagaggtg 900acagcagagg atggaacaca
gcggtgtgag aagtgcagca agccctgtgc ccgagtgtgc 960tatggtctgg
gcatggagca cttgcgagag gtgagggcag ttaccagtgc caatatccag
1020gagtttgctg gctgcaagaa gatctttggg agcctggcat ttctgccgga
gagctttgat 1080ggggacccag cctccaacac tgccgaattc gctccgctga
ggcctgagca gctccaagtg 1140ttcgaaaccc tggaggagat cacaggttac
ctgtacatct cagcatggcc agacagtctc 1200cgtgacctca gtgtcttcca
gaaccttcga atcattcggg gacggattct ccacgatggc 1260gcgtactcat
tgacactgca aggcctgggg atccactcgc tggggctgcg ctcactgcgg
1320gagctgggca gtggattggc tctgattcac cgcaacgccc atctctgctt
tgtacacact 1380gtaccttggg accagctctt ccggaaccca catcaggccc
tgctccacag tgggaaccgg 1440ccggaagagg attgtggtct cgagggcttg
gtctgtaact cactgtgtgc ccacgggcac 1500tgctgggggc cagggcccac
ccagtgtgtc aactgcagtc atttccttcg gggccaggag 1560tgtgtggagg
agtgccgagt atggaagggg ctcccccggg agtatgtgag tgacaagcgc
1620tgtctgccgt gtcaccccga gtgtcagcct caaaacagct cagagacctg
ctttggatcg 1680gaggctgatc agtgtgcagc ctgcgcccac tacaaggact
cgtcctcctg tgtggctcgc 1740tgccccagtg gtgtgaaacc ggacctctcc
tacatgccca tctggaagta cccggatgag 1800gagggcatat gccagccgtg
ccccatcaac tgcacccact cctgtgtgga tctggatgaa 1860cgaggctgcc
cagcagagca gagagccagc ccggtgacat tcatcattgc aactgtagtg
1920ggcgtcctgc tgttcctgat cttagtggtg gtcgttggaa tcctaatcaa
acgaaggaga 1980cagaagatcc ggaagtatac gatg 20046121DNAArtificial
SequenceFCA733 forward primer 61aggctcctct aggtttactg d
216224DNAArtificial SequenceFCA733 reverse primer 62ccaccgataa
gtgaaatggt atgg 246321DNAArtificial SequenceFCA749 forward primer
63gggtgatgtg acctcttgta g 216421DNAArtificial SequenceFCA749
reverse primer 64ggacagaggc tccttaaaca g 216520DNAArtificial
SequenceMACROD2 foward primer 65tcttccctcc tccgtgtatg
206624DNAArtificial SequenceMACROD2 reverse primer 66agacgctcaa
tcaactgagc catc 246723DNAArtificial SequencemtCR forward primer
67tcttctcgct ccgggcccat ttc 236820DNAArtificial SequencemtCR
reverse primer 68agtctggcga ctcatctagg 2069885PRTArtificial
Sequencefusion protein feHER2ecd-hFc 69Met Glu Leu Ala Ala Trp Cys
Arg Trp Gly Leu Leu Leu Ala Leu Leu1 5 10 15Pro Ser Gly Ala Thr Gly
Thr Gln Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala
Ser Pro Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly
Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu His Ala
Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln
Gly Tyr Val Leu Ile Ala His Asn Gln Val Lys Gln Val Pro Leu 85 90
95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr
100 105 110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asp Ser Gly
Thr Pro 115 120 125Ala Thr Gly Ala Ala Leu Gly Gly Leu Arg Glu Leu
Gln Leu Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu
Ile Gln Arg Asn Pro Gln145 150 155 160Leu Cys His Gln Asp Thr Ile
Leu Trp Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu
Met Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190Gln Pro Cys
Ser Pro Ala Cys Lys Asp Ser His Cys Trp Gly Ala Ser 195 200 205Ser
Gly Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215
220Ala Arg Cys Lys Gly Pro Gln Pro Thr Asp Cys Cys His Glu Gln
Cys225 230 235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys
Leu Ala Cys Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu
His Cys Pro Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu
Ser Met Pro Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser
Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val
Gly Ser Cys Thr Leu Val Cys Pro Leu Asn Asn Gln305 310 315 320Glu
Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330
335Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu
340 345 350Ala Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Val Gly
Cys Lys 355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser
Phe Glu Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro
Glu Gln Leu Arg Val Phe385 390 395 400Glu Ala Leu Glu Glu Ile Thr
Gly Tyr Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asn
Leu Ser Val Phe Gln Asn Leu Arg Val Ile Arg 420 425 430Gly Arg Val
Leu His Asp Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly
Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455
460Leu Ala Leu Ile His Arg Asn Ser Arg Leu Cys Phe Val His Thr
Val465 470 475 480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala
Leu Leu His Ser 485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Ala Gly
Glu Gly Leu Ala Cys Tyr 500 505 510Pro Leu Cys Ala His Gly His Cys
Trp Gly Pro Gly Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe
Leu Arg Gly Gln Glu Cys Val Glu Glu Cys 530 535 540Arg Val Leu Gln
Gly Leu Pro Arg Glu Tyr Val Lys Asp Arg Phe Cys545 550 555 560Leu
Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570
575Leu Gly Ser Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp
580 585 590Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro
Asp Leu 595 600 605Ser Phe Met Pro Ile Trp Lys Phe Ala Asp Glu Glu
Gly Thr Cys Gln 610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys
Ala Asp Leu Asp Glu Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg
Ala Ser Pro Val Thr Ser Glu Pro Lys 645 650 655Ser Cys Asp Lys Thr
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 660 665 670Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 675 680 685Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 690 695
700Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val705 710 715 720Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser 725 730 735Thr Tyr Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu 740 745 750Asn Gly Lys Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ala 755 760 765Pro Ile Glu Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 770 775 780Gln Val Tyr Thr
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln785 790 795 800Val
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 805 810
815Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
820 825 830Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu 835 840 845Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser 850 855 860Val Met His Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser865 870 875 880Leu Ser Pro Gly Lys
8857039DNAArtificial SequenceFeline HER2 ECTM (prefeHER2) FORWARD
PRIMER 70attactacaa gcttgagacc atggagctgg cggcctggt
397139DNAArtificial SequenceFeline HER2 ECTM (prefeHER2) Reverse
PRIMER 71tactaatcta gatcacatcg tgtacttccg gatcttctg
397226DNAArtificial SequenceFeline HER2 ECD-Fc ECD FORWARD PRIMER
72caccaagctt gagaccatgg agctgg 267328DNAArtificial SequenceFeline
HER2 ECD-Fc ECD Reverse PRIMER 73gatttgggct cggacgtcac agggctgg
287429DNAArtificial SequenceFeline HER2 ECD-Fc tag FORWARD PRIMER
74ctgtgacgtc cgagcccaaa tcttgtgac 297534DNAArtificial
SequenceFeline HER2 ECD-Fc tag Reverse PRIMER 75tctagattat
ttacccggag acagggagag gctc 347633DNAArtificial SequenceBlack bear
HER2 ECTM (prebearHER2) FORWARD PRIMER 76taagcttgag accatggagc
tggcggcctg gtg 337733DNAArtificial SequenceBlack bear HER2 ECTM
(prebearHER2) Reverse PRIMER 77ctctagattc acatcgtgta cttccggatc ttc
337831DNAArtificial SequenceFeline GM-CSF FORWARD PRIMER
78caccatgtgg ctgcagaacc tgcttttcct g 317927DNAArtificial
SequenceFeline GM-CSF Reverse PRIMER 79ttacttctgg tctggtcccc
agcagtc 278027DNAArtificial SequencefeCSF2 1-67 FORWARD PRIMER
80atgtggctgc agaacctgct tttcctg 278124DNAArtificial SequencefeCSF2
1-67 Reverse PRIMER 81ctcagggtca aacatttcag agac
248225DNAArtificial SequencefeCSF2 60-145 FORWARD PRIMER
82gtctctgaaa tgtttgaccc tgagg 258327DNAArtificial SequencefeCSF2
60-145 Reverse PRIMER 83ttacttctgg tctggtcccc agcagtc
2784687PRTHomo sapiens 84Met Glu Leu Ala Ala Leu Cys Arg Trp Gly
Leu Leu Leu Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln Val
Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu
Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val
Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala Ser
Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val
Leu Ile Ala His Asn Gln Val Arg Gln Val
Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu
Asp Asn Tyr 100 105 110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu
Asn Asn Thr Thr Pro 115 120 125Val Thr Gly Ala Ser Pro Gly Gly Leu
Arg Glu Leu Gln Leu Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly
Gly Val Leu Ile Gln Arg Asn Pro Gln145 150 155 160Leu Cys Tyr Gln
Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln
Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185
190His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser
195 200 205Ser Glu Asp Cys Lys Ser Leu Thr Arg Thr Val Cys Ala Gly
Gly Cys 210 215 220Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys
His Glu Gln Cys225 230 235 240Ala Ala Gly Cys Thr Gly Pro Lys His
Ser Asp Cys Leu Ala Cys Leu 245 250 255His Phe Asn His Ser Gly Ile
Cys Glu Leu His Cys Pro Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp
Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe
Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser
Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn Gln305 310
315 320Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser
Lys 325 330 335Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His
Leu Arg Glu 340 345 350Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu
Phe Ala Gly Cys Lys 355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu
Pro Glu Ser Phe Asp Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro
Leu Gln Pro Glu Gln Leu Gln Val Phe385 390 395 400Glu Thr Leu Glu
Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser
Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln Val Ile Arg 420 425
430Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu
435 440 445Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly
Ser Gly 450 455 460Leu Ala Leu Ile His His Asn Thr His Leu Cys Phe
Val His Thr Val465 470 475 480Pro Trp Asp Gln Leu Phe Arg Asn Pro
His Gln Ala Leu Leu His Thr 485 490 495Ala Asn Arg Pro Glu Asp Glu
Cys Val Gly Glu Gly Leu Ala Cys His 500 505 510Gln Leu Cys Ala Arg
Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys 515 520 525Val Asn Cys
Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys 530 535 540Arg
Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg His Cys545 550
555 560Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr
Cys 565 570 575Phe Gly Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His
Tyr Lys Asp 580 585 590Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly
Val Lys Pro Asp Leu 595 600 605Ser Tyr Met Pro Ile Trp Lys Phe Pro
Asp Glu Glu Gly Ala Cys Gln 610 615 620Pro Cys Pro Ile Asn Cys Thr
His Ser Cys Val Asp Leu Asp Asp Lys625 630 635 640Gly Cys Pro Ala
Glu Gln Arg Ala Ser Pro Leu Thr Ser Ile Ile Ser 645 650 655Ala Val
Val Gly Ile Leu Leu Val Val Val Leu Gly Val Val Phe Gly 660 665
670Ile Leu Ile Lys Arg Arg Gln Gln Lys Ile Arg Lys Tyr Thr Met 675
680 68585687PRTHomo sapiens 85Met Glu Leu Ala Ala Leu Cys Arg Trp
Gly Leu Leu Leu Ala Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln
Val Cys Thr Gly Thr Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro
Glu Thr His Leu Asp Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln
Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala
Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr
Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg
Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105
110Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro
115 120 125Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu
Arg Ser 130 135 140Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln
Arg Asn Pro Gln145 150 155 160Leu Cys Tyr Gln Asp Thr Ile Leu Trp
Lys Asp Ile Phe His Lys Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu
Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190His Pro Cys Ser Pro
Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser 195 200 205Ser Glu Asp
Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220Ala
Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Lys Cys225 230
235 240Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys
Leu 245 250 255His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro
Ala Leu Val 260 265 270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro
Asn Pro Glu Gly Arg 275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr
Ala Cys Pro Tyr Asn Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys
Thr Leu Val Cys Pro Leu His Asn Gln305 310 315 320Glu Val Thr Ala
Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys
Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345
350Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys
355 360 365Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp
Gly Asp 370 375 380Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln
Leu Gln Val Phe385 390 395 400Glu Thr Leu Glu Glu Ile Thr Gly Tyr
Leu Tyr Ile Ser Ala Trp Pro 405 410 415Asp Ser Leu Pro Asp Leu Ser
Val Phe Gln Asn Leu Gln Val Ile Arg 420 425 430Gly Arg Ile Leu His
Asn Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser
Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu
Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His Thr Val465 470
475 480Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His
Thr 485 490 495Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu
Ala Cys His 500 505 510Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro
Gly Pro Thr Gln Cys 515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly
Gln Glu Cys Val Glu Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro
Arg Glu Tyr Val Asn Ala Arg His Cys545 550 555 560Leu Pro Cys His
Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570 575Phe Gly
Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585
590Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu
595 600 605Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala
Cys Gln 610 615 620Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp
Leu Asp Asp Lys625 630 635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser
Pro Leu Thr Ser Ile Ile Ser 645 650 655Ala Val Val Gly Ile Leu Leu
Val Val Val Leu Gly Val Val Phe Gly 660 665 670Ile Leu Ile Lys Arg
Arg Gln Gln Lys Ile Arg Lys Tyr Thr Met 675 680 68586687PRTHomo
sapiens 86Met Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu Ala
Leu Leu1 5 10 15Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly Thr
Asp Met Lys 20 25 30Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp
Met Leu Arg His 35 40 45Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn
Leu Glu Leu Thr Tyr 50 55 60Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu
Gln Asp Ile Gln Glu Val65 70 75 80Gln Gly Tyr Val Leu Ile Ala His
Asn Gln Val Arg Gln Val Pro Leu 85 90 95Gln Arg Leu Arg Ile Val Arg
Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110Ala Leu Ala Val Leu
Asp Asn Gly Asp Pro Leu Asp Ser Gly Thr Pro 115 120 125Val Thr Gly
Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser 130 135 140Leu
Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln145 150
155 160Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys
Asn 165 170 175Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser
Arg Ala Cys 180 185 190His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg
Cys Trp Gly Glu Ser 195 200 205Ser Glu Asp Cys Gln Ser Leu Thr Arg
Thr Val Cys Ala Gly Gly Cys 210 215 220Ala Arg Cys Lys Gly Pro Leu
Pro Thr Asp Cys Cys His Glu Gln Cys225 230 235 240Ala Ala Gly Cys
Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu 245 250 255His Phe
Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val 260 265
270Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg
275 280 285Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn
Tyr Leu 290 295 300Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro
Leu His Asn Gln305 310 315 320Glu Val Thr Ala Glu Asp Gly Thr Gln
Arg Cys Glu Lys Cys Ser Lys 325 330 335Pro Cys Ala Arg Val Cys Tyr
Gly Leu Gly Met Glu His Leu Arg Glu 340 345 350Val Arg Ala Val Thr
Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys 355 360 365Lys Ile Phe
Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp 370 375 380Pro
Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln Val Phe385 390
395 400Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp
Pro 405 410 415Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln
Val Ile Arg 420 425 430Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu
Thr Leu Gln Gly Leu 435 440 445Gly Ile Ser Trp Leu Gly Leu Arg Ser
Leu Arg Glu Leu Gly Ser Gly 450 455 460Leu Ala Leu Ile His His Asn
Thr His Leu Cys Phe Val His Thr Val465 470 475 480Pro Trp Asp Gln
Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Thr 485 490 495Ala Asn
Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala Cys His 500 505
510Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys
515 520 525Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu
Glu Cys 530 535 540Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn
Ala Arg His Cys545 550 555 560Leu Pro Cys His Pro Glu Cys Gln Pro
Gln Asn Gly Ser Val Thr Cys 565 570 575Phe Gly Pro Glu Ala Asp Gln
Cys Val Ala Cys Ala His Tyr Lys Asp 580 585 590Pro Pro Phe Cys Val
Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu 595 600 605Ser Tyr Met
Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln 610 615 620Pro
Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp Lys625 630
635 640Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu Thr Ser Ile Ile
Ser 645 650 655Ala Val Val Gly Ile Leu Leu Val Val Val Leu Gly Val
Val Phe Gly 660 665 670Ile Leu Ile Lys Arg Arg Gln Gln Lys Ile Arg
Lys Tyr Thr Met 675 680 6858725DNAArtificial
SequenceprehumHER2-Q141K FORWARD PRIMER 87gctgaagctt cgaagcctca
cagag 258823DNAArtificial SequenceprehumHER2-Q141K reverse PRIMER
88tcccgcaggc ctcctgggga ggc 238921DNAArtificial
SequenceprehumHER2-Q213K FORWARD 89tgaggattgt aagagcctga c
219018DNAArtificial SequenceprehumHER2-Q213K reverse 90gaactctctc
cccagcag 189121DNAArtificial SequenceprehumHER2-Q239K FORWARD
91ctgccatgag aagtgtgctg c 219218DNAArtificial
SequenceprehumHER2-Q239K reverse PRIMER 92cagtcagtgg gcagtggc
189325DNAArtificial SequenceprehumHER2-Q320K FORWARD PRIMER
93aagaggtgac agcagaggat ggaac 259422DNAArtificial
SequenceprehumHER2-Q320K reverse PRIMER 94tgttgtgcag ggggcagacg ag
229521DNAArtificial SequenceprehumHER2-Q329K FORWARD PRIMER
95ggatggaaca aagcggtgtg a 219618DNAArtificial
SequenceprehumHER2-Q329K reverse PRIMER 96tctgctgtca cctcttgg
189723DNAArtificial SequenceprehumHER2-Q429R FORWARD PRIMER
97ccagaacctg agagtaatcc ggg 239818DNAArtificial
SequenceprehumHER2-Q429R reverse PRIMER 98aagacgctga ggtcaggc
189921DNAArtificial SequenceprehumHER2-N438D FORWARD 99aattctgcac
gatggcgcct a 2110018DNAArtificial SequenceprehumHER2-N438D reverse
100cgtccccgga ttacttgc 1810128DNAArtificial
SequenceprehumHER2-NNT124DSG FORWARD PRIMER 101ggcacacctg
tcacaggggc ctccccag 2810230DNAArtificial
SequenceprehumHER2-NNT124DSG reverse PRIMER 102actgtccagc
gggtctccat tgtctagcac 3010321DNAArtificial SequenceprefeHER2-Q141K
FORWARD PRIMER 103gcgggagctg aagctccgaa g 2110425DNAArtificial
SequenceprefeHER2-Q141K reverse PRIMER 104agccctccta gggcagcccc
tgtag 2510531DNAArtificial SequenceprefeHER2-R398Q FORWARD PRIMER
105tgagcagctc caagtgtttg aggctctgga g 3110618DNAArtificial
SequenceprefeHER2-R398Q reverse PRIMER 106ggctgcaggg gggcagtg
1810722DNAArtificial SequenceprefeHER2-N421D FORWARD PRIMER
107cagcttgcct gacctcagtg tc 2210818DNAArtificial
SequenceprefeHER2-N421D reverse PRIMER 108tctggccacg ctgagatg
1810915PRTArtificial SequenceConserved sequence in HER2,
unsubstituted 109Gln Leu Arg Ser Leu Thr Glu Ile Leu Lys Gly Gly
Val Leu Ile1 5 10 1511015PRTArtificial SequenceConserved sequence
in HER2, with Q-K substitution 110Lys Leu Arg Ser Leu Thr Glu Ile
Leu Lys Gly Gly Val Leu Ile1 5 10 1511122DNAArtificial
SequenceprebearHER2-Q141K and mbearHER2-Q119K FORWARD primer
111aagcctcaca gagatcctga ag 2211225DNAArtificial
SequenceprebearHER2-Q141K and mbearHER2-Q119K REVERSE primer
112cgaagcttca gctcccgcag ccctc 2511322DNAArtificial
SequencepremouseHER2-Q142K and mmouseHER2-Q120K FORWARD primer
113agtctcacag agatcttgaa gg 2211424DNAArtificial
SequencepremouseHER2-Q142K and mmouseHER2-Q120K REVERSE primer
114tcgaagcttc agctcccgca gccc 2411522DNAArtificial
SequencepreratHER2-Q145K and mratHER2-Q120K FORWARD primer
115gcttcgaagt ctcacagaga tc 2211622DNAArtificial
SequencepreratHER2-Q145K and mratHER2-Q120K REVERSE primer
116ttcagctccc gcagcccctc tg 2211723DNAArtificial
SequenceprehumHER2-Q141K and mhumHER2-Q119K FORWARD primer
117gctgaagctt cgaagcctca cag 2311822DNAArtificial
SequenceprehumHER2-Q141K and mhumHER2-Q119K REVERSE primer
118tcccgcaggc ctcctgggga gg 22
* * * * *
References