# Patent

## Title
Zone 3 necrosis associated markers and method of use thereof

## Abstract

Disclosed are methods of identifying toxic agents, e.g., hepatotoxic agents, using differential gene expression.

## Background
RELATED APPLICATION
This application claims priority to U.S. Ser. No. 60/410,763, filed Sep. 13, 2002. The contents of this application is incorporated herein by reference in its entirety.


FIELD OF THE INVENTION
The invention relates generally to the identification of genetic markers associated with toxicity.
BACKGROUND OF THE INVENTION
Necrotic cell death is a common response of the liver to hepatotoxic agents and represents an irreversible form of damage to individual liver cells. While the mechanisms by which hepatotoxic agents lead to necrosis remain to be fully understood, progress has been made in understanding the biochemical pathways involved. Necrotic cell death occurs when a chemical or it's metabolite react with critical cellular systems resulting in ion dysregulation, mitochondrial dysfunction and oxidative stress [1–3]. The acute disruption of these normal cellular events in effect leads to ATP depletion. This loss of energy within the cell distinguishes necrosis from the other classification of cell death known as apoptosis. When cells undergo apoptosis or programmed cell death, the cell requires an energy level capable of triggering special metabolic, signal transduction and gene regulation pathways that systematically shut down the cell. Necrosis occurs when the ATP levels falls below the threshold required for these processes and the cell is driven into a passive state of cellular chaos that culminates in cell death [4]. Thus, although these two forms of cell death are distinct, they can share initiating pathways depending on the how sharply the ATP levels decline. Such can be seen with the induction of the mitochondrial permeability transition (MPT), a mechanism that causes mitochondrial failure. The MPT will lead to necrosis if ATP is depleted or apoptosis if there are sufficient amounts available to initiate a caspase cascade [5].
Chemical insult that produces necrosis of the liver can be either nonzonal or zonal. Zonal necrosis is separated into zones 1, 2 and 3 based on the region of the lobule affected. Different hepatotoxic agents preferentially target specific zones [6]. This research project report specifically deals with those agents that produced zone 3 or centrilobular necrosis. This is the most commonly affected area of the liver for hepatotoxic agents producing zonal necrosis. Zones 1–3 are distinguishable in terms of blood flow, oxygen content, bile flow and ratio of intoxication versus detoxification pathways. Factors such as these explain the specificity of hepatotoxic agents for particular zones. Acetaminophen and carbon tetrachloride (CCL4) are examples of agents that produce mainly zone 3 necrosis once they are converted to reactive metabolites. This can be attributed to the high degree of regional organization of agent specific cytochrome P450's within the liver [7]. The supply of oxygen available to the cell has also been shown to be a factor for zone 3 necrosis producing agents. Zone 3 is the region of the liver that is furthest from the arterial blood supply receiving the least supply of oxygen. When CCl4 is metabolized to its reactive metabolite CCl3, the reduction reaction is inhibited by oxygen, favoring a necrotic response in the centrilobular area [7].
Liver cell necrosis can evoke a range of responses within the liver that depend on the severity of insult. These responses range from regeneration of necrotic tissue with restoration of full liver function to concomitant loss of liver function, liver failure and death [8]. In the process, necrosis may trigger the development of other liver diseases. Recurring bouts of necrosis and repair may result in disruption of the structure of the liver and result in subacute hepatitis, chronic hepatitis or even cirrhosis [9]. In this process, inflammatory cells stimulate the deposition of collagen around hepatocytes causing alteration in hepatic function and blood flow [10]. There is also evidence that necrosis may play a role in the induction of early hepatocellular carcinoma through compensatory liver regeneration. Diethylnitrosamine and Fumonisin B(1) are two examples of compounds that show evidence of producing hepatocellular carcinoma in rats through a sequence of events that begin with necrosis [11–13]. Thus the benefit of obtaining marker genes predictive of hepatic zone 3 necrosis stem from its participation in the pathogenesis of other liver diseases as well as it being an early indicator of hepatic toxicity.
SUMMARY OF THE INVENTION
The invention is based in part on the discovery that certain nucleic acids are differentially expressed in liver cells or liver tissue of animals treated with toxic compounds. These differentially expressed nucleic acids include novel sequences and nucleic acids sequences that, while previously described, have not heretofore been identified as associated with toxicity and are collectively referred to herein as “TOXMARKER nucleic acids” or “TOXMARKER polynucleotides” and the corresponding encoded polypeptides are referred to as “TOXMARKER polypeptides” or “TOXMARKER proteins”. The TOXMARKER genes are useful in high throughput screening of potential therapeutic compounds for toxicity.
In on aspect the invention provides methods of predicting the hepatotoxicity of a test agent. Hepatotoxicity is predicted by determining the level of expression of a toxicity-associated gene in a cell exposed to a test agent. The level of expression of the toxicity-associated gene is compared to the level of expression of the toxicity-associated gene in a control population exposed to a control agent. A test agent is predicted to be toxic if an alteration (e.g., increase or decrease) in the level of expression in the cell exposed to the test agent compared to the control population is identified.
Also provided by the invention are methods of screening a test agent for inducing changes in gene expression associated with a toxic agent. An agent is screened for inducing changes in gene expression associated with a toxic agent by determining the level of expression of a toxicity-associated gene in a cell exposed to a test agent. The level of expression of the toxicity-associated gene is compared to the level of expression of the toxicity-associated gene in a control population exposed to a control agent.
The alteration is statistically significant. By statistically significant is meant that the alteration is greater than what might be expected to happen by change alone. Statistical significance is determined by method known in the art. An alteration is statistically significant if the p-value is at least 0.05. Preferably, the p-value is 0.04, 0.03, 0.02, 0.01, 0.005, 0.001 or less.
By toxicity-associated gene is meant a gene that is characterized by a level of expression which differs in a cell exposed to a toxic compound compared to a control population. A toxicity-associated gene includes for example TOXMARKER 1–131. Preferably, the toxicity-associated gene is the genes listed on Table 5. More preferably, the toxicity-associated gene is TOXMARKER 42, 59, 65, 66, 71, 76, and 97.
A control population is a for example a cell not exposed to a toxic agent. Optionally, the control population is exposed to a control agent. A control agent is an agent that does not elicit a histology associated with a condition associated with liver toxicity such as Cholestasis; Steatosis; Reactive Inflamation; Necrosis, e.g., zone 3, general or multifocal; Genotoxic Carcinogenesis; Hepatocellular Hypertrophy; Non-Genotoxic Carcinogenesis; Appoptosis and Kupffer Cell Aggregation. Exemplary control agents are those listed in Table 1 and Table 2 below. A control level is a single expression pattern derived from a single control population or from a plurality of expression patterns. For example, the control level can be a database of expression patterns from previously tested cells.
The test cell is provided in vitro. Alternatively, the test cell is provided ex vivo or in vivo from a mammalian subject. The test cell is derived from liver tissue, such as for example a hepatocyte. Alternatively, the test cell is a subject derived cell sample. The subject derived tissue sample is any tissue from a test subject.
Expression is determined by for example detecting hybridization, e.g., on a chip, of a toxicity-associated gene probe to a gene transcript of the test cell.
The invention also provides a zone 3 necrosis reference expression profile of a gene expression level two or more of TOXMARKER 1–132. For example, the reference profile contains the expression levels of TOXMARKER 1–132. Alternatively, the reference profile contains the expression levels of TOXMARKER genes listed on Table 5. Preferably, the reference profile contains the expression levels of TOXMARKER 42, 59, 65, 66, 71, 76, and 97
The invention also provides a kit with a detection reagent which binds to two or more TOXMARKER nucleic acid sequences or which binds to a gene product encoded by the nucleic acid sequences. Also provided is an array of nucleic acids, e.g. oligonucleotides that binds to two or more TOXMARKER nucleic acids. For example, the array contains oligonucleotides that bind TOXMARKER 1–132. Alternatively, the array contains oligonucleotides that bind the TOXMARKER genes listed on Table 5. Preferably, the array contains oligonucleotides that bind TOXMARKER 42, 59, 65, 66, 71, 76, and 97 Most preferably, the array contains oligonucleotides that binds at least five TOXMARKER genes listed one Table 5, where the collection of TOXMARKER genes predict toxicity to a confidence level of a p-value of at least 0.05 or less.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.
Other features and advantages of the invention will be apparent from the following detailed description and claims.



BRIEF DESCRIPTION OF THE DRAWING
FIG. 1 is a schematic of the hepatotoxicity prediction screening method of the invention.
FIG. 2 is a illustration of a chart showing the linear discriminant model.



DETAILED DESCRIPTION OF THE INVENTION
The present invention is based in part on the discovery of changes in expression patterns of multiple nucleic acid sequences in response to compounds known to elicit a histopathologic condition (i.e., a pathologic change to the liver visible on examination that causes the liver to function less effectively) associated with liver toxicity. The compounds are for example, compounds listed in Table 1 or 2 below. Examples of histopathologic conditions include, Cholestasis; Steatosis; Reactive Inflamation; Necrosis, e.g., zone 3, general or multifocal; Genotoxic Carcinogenesis; Hepatocellular Hypertrophy; Non-Genotoxic Carcinogenesis; Appoptosis and Kupffer Cell Aggregation. The histopathological conditions are identified by methods know in the art. For example, zone 3 necrosis is identified by choleastasis and hypertrophy. The identification of genes that are differentially expressed in response to toxic compounds are useful in screening potential therapeutic compositions for toxicity.
The genes whose expression levels are modulated (i.e., increased or decreased) in response to exposure to a toxic compound are summarized in Tables 3–5 (see EXAMPLES 4 and 5) and are collectively referred to herein as “toxicity-associated gene”, “TOXMARKER nucleic acids” or “TOXMARKER polynucleotides” and the corresponding encoded polypeptides are referred to as “TOXMARKER polypeptides” or “TOXMARKER proteins.” Unless indicated otherwise, “TOXMARKER” or “toxicity-associated gene” is meant to refer to any of the sequences disclosed herein.
For a given TOXMARKER sequence, its expression can be measured in the methods described herein. For previously described sequences, database accession numbers are provided. This information allows for one of ordinary skill in the art to deduce information necessary for detecting and measuring expression of the TOXMARKER nucleic acid sequences.
General Methods
The TOXMARKER nucleic acids and encoded polypeptides can be identified using the information provided in the EXAMPLES below. In some embodiments, the TOXMARKER nucleic acids and polypeptides correspond to the nucleic acids or polypeptides which include the various sequences (referenced by SEQ ID NOs) disclosed for each TOXMARKER.
The invention includes providing a test cell population which includes at least one cell that is capable of expressing one or more of the sequences TOXMARKER 1–132. By “capable of expressing” is meant that the gene is present in an intact form in the cell and can be expressed. Expression of one, some, or all of the TOXMARKER sequences is then detected, if present, and, preferably, measured to yield an expression profile, e.g., subject expression profile or a test cell expression profile. By “expression profile” is meant a pattern of the level of expression of at least two toxicity-associated genes.
“Similarity of expression profile” is a similarity of expression profile between two samples exists when the linear combination of the genes in the profile has a linear discriminant score that is more similar to one of the training classes than the other. Linear discriminant analysis (LDA) identifies a linear combination of markers that best separates the defined classes. In the training data (i.e., control population) of this invention linear discriminant score could be determined by the following equation:

Linear disc. score=aGene1+bGene2+ . . . +nGeneN

where a, b . . . n are the coefficients identified by least squares that best separate the phenotypes under investigation. Thus, similarity in expression profile is a similarity in gene combinations. Interpretation of raw data is difficult since the samples are plotted in more than 3 dimensions, one dimension for each gene, which makes it difficult to visualize the data. LDA compresses this information into a single dimension.
By “toxicity-associated gene” is meant a gene, which the level of expression differs in a cell or subject exposed to a known toxic compound as compared to a cell or subject not exposed to a toxic compound (i.e., control). Preferably, the TOXMARKER genes 42 (IFNAR-2), 59 (Transaldolase), 65 (Clp-1), 66 (Hex), 71 (cszr—204152648—191521095), 76 (scr_gb-aa899865—3), and 97 (scr_gb-bm986259—1).
Using sequence information provided by the database entries for the known sequences, or the sequence information provided herein for the newly described sequences, expression of the TOXMARKER sequences are detected (if present) and measured using techniques well known to one of ordinary skill in the art. For example, sequences within the sequence database entries corresponding to TOXMARKER sequences, or within the sequences disclosed herein, can be used to construct probes for detecting TOXMARKER RNA sequences in, e.g., northern blot hybridization analyses or methods which specifically, and, preferably, quantitatively amplify specific nucleic acid sequences. As another example, the sequences can be used to construct primers for specifically amplifying the TOXMARKER sequences in, e.g., amplification-based detection methods such as reverse-transcription based polymerase chain reaction. When alterations in gene expression are associated with gene amplification or deletion, sequence comparisons in test and reference populations can be made by comparing relative amounts of the examined DNA sequences in the test and reference cell populations.
Expression of the genes disclosed herein can be measured at the RNA level using any method known in the art. For example, northern hybridization analysis using probes which specifically recognize one or more of these sequences can be used to determine gene expression. Alternatively, expression can be measured using reverse-transcription-based PCR assays, e.g., using primers specific for the differentially expressed sequences.
Expression is also measured at the protein level, i.e., by measuring the levels of polypeptides encoded by the gene products described herein. Such methods are well known in the art and include, e.g., immunoassays based on antibodies to proteins encoded by the genes.
Expression levels of one or more of the TOXMARKER sequences in the test cell population are then compared to expression levels of the sequences in one or more cells from a reference (i.e., control) cell population. If desired, a reference expression profile is generated. A reference profile is a single expression pattern derived from a single reference population or from a plurality of expression patterns. For example, the reference cell population can be a database of expression patterns from previously tested cells for which one of the herein-described parameters or conditions (e.g., toxicity) is known.
The reference profile is obtained from the training data. Training data is a collection of data from the in vitro or in vivo samples that were exposed to compounds that produce a known pathology. (i.e., pathology present or pathology absent) Profile is defined here to indicate the absolute estimate of the expression level of any one TOXMARKER gene fragment (e.g. Intensity).
Expression of sequences in test and reference populations of cells are compared using any art-recognized method for comparing expression of nucleic acid sequences. For example, expression can be compared using GENECALLING® methods as described in U.S. Pat. No. 5,871,697 and in Shimkets et al., Nat. Biotechnol. 17:798–803.
In various embodiments, the expression of one or more sequences encoding genes of related function, as listed in Tables 3–5, is compared. In various embodiments, the expression of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 35, 40, 50, 100 or all of the sequences represented by TOXMARKER 1–132 are measured. If desired, expression of these sequences can be measured along with other sequences whose expression is known to be altered according to one of the herein described parameters or conditions.
The reference cell population includes one or more cells for which the compared parameter is known. The compared parameter can be, e.g. toxic agent expression status. By “toxic agent expression status” is meant that it is known whether the reference cell has had contact with a toxic agent. Whether or not comparison of the gene expression profile in the test cell population to the reference cell population reveals the presence, or degree, of the measured parameter depends on the composition of the reference cell population. For example, if the reference cell population is composed of cells that have not been treated with a known toxic agent, a similar gene expression level in the test cell population and a reference cell population indicates the test agent is not a toxic agent. Conversely, if the reference cell population is made up of cells that have been treated with a toxic agent, a similar gene expression profile between the test cell population and the reference cell population indicates the test agent is a toxic agent.
In various embodiments, a TOXMARKER sequence in a test cell population is considered comparable in expression level to the expression level of the TOXMARKER sequence if its expression level varies within a factor of 2.0, 1.5, or 1.0 fold to the level of the TOXMARKER transcript in the reference cell population. In various embodiments, a TOXMARKER sequence in a test cell population can be considered altered in levels of expression if its expression level varies from the reference cell population by more than 3.0, 4.0, 5.0 or more fold from the expression level of the corresponding TOXMARKER sequence in the reference cell population.
If desired, comparison of differentially expressed sequences between a test cell population and a reference cell population can be done with respect to a control nucleic acid whose expression is independent of the parameter or condition being measured. Expression levels of the control nucleic acid in the test and reference nucleic acid can be used to normalize signal levels in the compared populations.
In some embodiments, the test cell population is compared to multiple reference cell populations. Each of the multiple reference populations may differ in the known parameter. Thus, a test cell population may be compared to a first reference cell population known to have been exposed to a toxic agent, as well as a second reference population known to have not been exposed to a toxic agent.
The test cell population that is exposed to, i.e., contacted with, the test toxic agent can be any number of cells, i.e., one or more cells, and can be provided in vitro, in vivo, or ex vivo. The test cell is obtained from a bodily fluid, e.g., biological fluid (such as blood, serum, urine, saliva, milk, ductal fluid, or tears). For example, the test cell is purified from blood or another tissue, i.e., liver tissue.
In other embodiments, the test cell population can be divided into two or more subpopulations. The subpopulations can be created by dividing the first population of cells to create as identical a subpopulation as possible. This will be suitable, in, for example, in vitro or ex vivo screening methods. In some embodiments, various subpopulations can be exposed to a control agent, and/or a test agent, multiple test agents, or, e.g., varying dosages of one or multiple test agents administered together, or in various combinations.
Preferably, cells in the reference cell population are derived from a tissue type as similar as possible to the test cell, e.g., liver tissue. Alternatively the cells are derived from an established cell line. Preferably, the cell is a hepatocyte. In some embodiments, the control cell is derived from the same subject as the test cell, e.g., from a region proximal to the region of origin of the test cell. In other embodiments, the reference cell population is derived from a plurality of cells. For example, the reference cell population can be a database of expression patterns from previously tested cells for which one of the herein-described parameters or conditions (toxic agent expression status) is known.
The test agent can be a compound not previously described or can be a previously known compound but which is not known to be a toxic agent.
The subject is preferably a mammal. The mammal can be, e.g., a human, non-human primate, mouse, rat, dog, cat, horse, or cow.
Prediction of Toxicity
In one aspect, the invention provides a method of predicting the toxicity e.g., hepatotoxicity of a test agent or identifying a toxic agents, e.g., a hepatotoxic agent. The method is an in vivo method. Alternatively, the method is an in vitro method.
Differences in the genetic makeup of individuals can result in differences in their relative abilities to metabolize various drugs. Accordingly, the differentially expressed TOXMARKER sequences disclosed herein allow for a putative therapeutic or prophylactic agent to be tested in a test cell population from a selected subject in order to predict if the agent causes toxicity in the subject.
By predicting the toxicity is meant that the test compound is more likely to be hepatotoxic that not be hepatotoxic. Hepatotoxicity is predicted by determining the level of expression of a toxicity-associated gene in a cell exposed to a test agent. The level of expression of the toxicity-associated gene is compared to the level of expression of the toxicity-associated gene in a control population exposed to a control agent. A test agent is predicted to be toxic if an alteration (e.g., increase or decrease) in the level of expression in the cell exposed to the test agent compared to the control population is identified.
The toxicity-associated gene is for example TOXMARKER 1–132. Alternatively, the toxicity-associated gene is the TOXMARKER genes listed on Table 5. Optionally, the toxicity-associated gene is TOXMARKER 42, 59, 65, 66, 71, 76, and 97 The toxicity-associated gene is a nucleic acid sequences homologous to those listed in Tables 3–5 as TOXMARKER 1–132. The sequences need not be identical to sequences including TOXMARKER 1–132, as long as the sequence is sufficiently similar that specific hybridization can be detected. Preferably, the cell includes sequences that are identical, or nearly identical to those identifying the TOXMARKER nucleic acids shown in Tables 3–5.
By hepatotoxicity is meant that that the compound causes a hispathological change in the live tissue associate with toxicity. By “toxicity” is meant that the agent is damaging or destructive to liver when administered to a subject. Damage to the liver is measured for example, histologically. Hepatotoxicity is determined, for example as described in the examples below.
The cell population is contacted in vitro, or in vivo. Optionally, the cell population is contacted ex vivo with the agent or activated form of the agent.
Expression of the nucleic acid sequences in the test cell population is then compared to the expression of the nucleic acid sequences in a control population, which is a cell population that has not been exposed to the test agent, or, in some embodiments, a cell population exposed to the test agent. Comparison can be performed on test and reference samples measured concurrently or at temporally distinct times. An example of the latter is the use of compiled expression information, e.g., a sequence database, which assembles information about expression levels of known sequences following administration of various agents. For example, alteration of expression levels following administration of test agent can be compared to the expression changes observed in the nucleic acid sequences following administration of a control agent. A control agent is a compound that elicits the histopathology. Alternatively, the control agent is a compound that does not elicit the histopathology. Exemplary control compounds are listed in Tables 1 and 2.
An alteration in expression of the nucleic acid sequence in the test cell population compared to the expression of the nucleic acid sequence in the control cell population that has not been exposed to the test agent indicates the test agent is a toxic agent.
The alteration is statistically significant. By statistically significant is meant that the alteration is greater than what might be expected to happen by change alone. Statistical significance is determined by method known in the art. For example statistical significance is determined by p-value. The p-values is a measure of probability that a difference between groups during an experiment happened by chance. (P(z≧zobserved)). For example, a p-value of 0.01 means that there is a 1 in 100 chance the result occurred by chance. The lower the p-value, the more likely it is that the difference between groups was caused by treatment. An alteration is statistically significant if the p-value is at least 0.05. Preferably, the p-value is 0.04, 0.03, 0.02, 0.01, 0.005, 0.001 or less.
The invention also includes a toxic agent identified according to this screening method.
The differentially expressed TOXMARKER sequences identified herein also allow for the toxicity of a toxic agent to be determined or monitored. In this method, a test cell population from a subject is exposed to a test agent, i.e. a toxic agent. If desired, test cell populations can be taken from the subject at various time points before, during, or after exposure to the test agent. Expression of one or more of the TOXMARKER sequences, e.g., TOXMARKER: 1–132, in the cell population is then measured and compared to a control population which includes cells whose toxic agent expression status is known.
Kits
The invention also includes a TOXMARKER-detection reagent, e.g., nucleic acids that specifically identify one or more TOXMARKER nucleic acids by having homologous nucleic acid sequences, such as oligonucleotide sequences, complementary to a portion of the TOXMARKER nucleic acids or antibodies to proteins encoded by the TOXMARKER nucleic acids packaged together in the form of a kit. The oligonucleotides are fragments of the the TOXMARKER genes. For example the olignucleitides are 200, 150, 100, 50, 25, 10 or less nucleotides in length. The kit may contain in separate containers a nucleic acid or antibody (either already bound to a solid matrix or packaged separately with reagents for binding them to the matrix), control formulations (positive and/or negative), and/or a detectable label. Instructions (e.g., written, tape, VCR, CD-ROM, etc.) for carrying out the assay may be included in the kit. The assay may for example be in the form of a Northern hybridization or a sandwich ELISA as known in the art.
For example, TOXMARKER detection reagent, is immobilized on a solid matrix such as a porous strip to form at least one TOXMARKER detection site. The measurement or detection region of the porous strip may include a plurality of sites containing a nucleic acid. A test strip may also contain sites for negative and/or positive controls. Alternatively, control sites are located on a separate strip from the test strip. Optionally, the different detection sites may contain different amounts of immobilized nucleic acids, i.e., a higher amount in the first detection site and lesser amounts in subsequent sites. Upon the addition of test sample, the number of sites displaying a detectable signal provides a quantitative indication of the amount of TOXMARKER present in the sample. The detection sites may be configured in any suitably detectable shape and are typically in the shape of a bar or dot spanning the width of a test strip.
Alternatively, the kit contains a nucleic acid substrate array comprising one or more nucleic acid sequences. The nucleic acids on the array specifically identify one or more nucleic acid sequences represented by TOXMARKER 1–132. In various embodiments, the expression of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 40 or 50 or more of the sequences represented by TOXMARKER 1–132. are identified by virtue of binding to the array. The substrate array can be on, e.g., a solid substrate, e.g., a “chip” as described in U.S. Pat. No. 5,744,305.
Arrays and Pluralities
The invention also includes a nucleic acid substrate array comprising one or more nucleic acid sequences. The nucleic acids on the array specifically identify one or more nucleic acid sequences represented by TOXMARKER 1–132. In various embodiments, the expression of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 40 or 50 or more of the sequences represented by TOXMARKER 1–132 are identified.
The nucleic acids in the array can identify the enumerated nucleic acids by, e.g., having homologous nucleic acid sequences, such as oligonucleotide sequences, complementary to a portion of the recited nucleic acids. The substrate array can be on, e.g., a solid substrate, e.g., a “chip” as described in U.S. Pat. No. 5,744,305.
The invention also includes an isolated plurality (i.e., a mixture of two or more nucleic acids) of nucleic acid sequences. The nucleic acid sequence can be in a liquid phase or a solid phase, e.g., immobilized on a solid support such as a nitrocellulose membrane. The plurality typically includes one or more of the nucleic acid sequences represented by TOXMARKER 1–132. In various embodiments, the plurality includes 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 40 or 50 or more of the sequences represented by TOXMARKER 1–132.
Nucleic Acids
One aspect of the invention pertains to isolated nucleic acid molecules that encode TOXMARKER proteins or biologically active portions thereof, as well as nucleic acid fragments sufficient for use as hybridization probes to identify TOXMARKER-encoding nucleic acids (e.g., TOXMARKER mRNA) and fragments for use as PCR primers for the amplification or mutation of TOXMARKER nucleic acid molecules. As used herein, the term “nucleic acid molecule” is intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), analogs of the DNA or RNA generated using nucleotide analogs, and derivatives, fragments and homologs thereof. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA.
“Probes” refer to nucleic acid sequences of variable length, preferably between at least about 10 nucleotides (nt), 100 nt, or as many as about, e.g., 6,000 nt, depending on use. Probes are used in the detection of identical, similar, or complementary nucleic acid sequences. Longer length probes are usually obtained from a natural or recombinant source, are highly specific and much slower to hybridize than oligomers. Probes may be single- or double-stranded and designed to have specificity in PCR, membrane-based hybridization technologies, or ELISA-like technologies.
An “isolated” nucleic acid molecule is one that is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid. Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated TOXMARKER nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived (e.g., testis, lung, B-cells). Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or of chemical precursors or other chemicals when chemically synthesized.
A nucleic acid molecule of the present invention, e.g., a nucleic acid molecule having the nucleotide sequence of SEQ ID NO: 1–171 or a complement of any of these nucleotide sequences, can be isolated using standard molecular biology techniques and the sequence information provided herein. Using all or a portion of the nucleic acid sequences of SEQ ID NO: 1–171 as a hybridization probe, TOXMARKER molecules can be isolated using standard hybridization and cloning techniques (e.g., as described in Sambrook et al., (eds.), MOLECULAR CLONING: A LABORATORY MANUAL 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; and Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, N.Y., 1993.)
A nucleic acid of the invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to TOXMARKER nucleotide sequences can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.
As used herein, the term “oligonucleotide” refers to a series of linked nucleotide residues, which oligonucleotide has a sufficient number of nucleotide bases to be used in a PCR reaction. A short oligonucleotide sequence may be based on, or designed from, a genomic or cDNA sequence and is used to amplify, confirm, or reveal the presence of an identical, similar or complementary DNA or RNA in a particular cell or tissue. Oligonucleotides comprise portions of a nucleic acid sequence having about 10 nt, 50 nt, or 100 nt in length, preferably about 15 nt to 30 nt in length. In one embodiment, an oligonucleotide comprising a nucleic acid molecule less than 100 nt in length would further comprise at lease 6 contiguous nucleotides of SEQ ID NO: 1–171, or a complement thereof. Oligonucleotides may be chemically synthesized and may be used as probes.
In another embodiment, an isolated nucleic acid molecule of the invention comprises a nucleic acid molecule that is a complement of the nucleotide sequence shown in SEQ ID NO: 1–171. In another embodiment, an isolated nucleic acid molecule of the invention comprises a nucleic acid molecule that is a complement of the nucleotide sequence shown in SEQ ID NO: 1–171, or a portion of this nucleotide sequence. A nucleic acid molecule that is complementary to the nucleotide sequence shown in SEQ ID NO: 1–171 is one that is sufficiently complementary to the nucleotide sequence shown in SEQ ID NO: 1–171 that it can hydrogen bond with little or no mismatches to the nucleotide sequence shown in SEQ ID NO: 1–171, thereby forming a stable duplex.
As used herein, the term “complementary” refers to Watson-Crick or Hoogsteen base pairing between nucleotides units of a nucleic acid molecule, and the term “binding” means the physical or chemical interaction between two polypeptides or compounds or associated polypeptides or compounds or combinations thereof. Binding includes ionic, non-ionic, Von der Waals, hydrophobic interactions, etc. A physical interaction can be either direct or indirect. Indirect interactions may be through or due to the effects of another polypeptide or compound. Direct binding refers to interactions that do not take place through, or due to, the effect of another polypeptide or compound, but instead are without other substantial chemical intermediates.
Moreover, the nucleic acid molecule of the invention can comprise only a portion of the nucleic acid sequence of SEQ ID NO: 1–171, e.g., a fragment that can be used as a probe or primer or a fragment encoding a biologically active portion of TOXMARKER.
Fragments provided herein are defined as sequences of at least 6 (contiguous) nucleic acids or at least 4 (contiguous) amino acids, a length sufficient to allow for specific hybridization in the case of nucleic acids or for specific recognition of an epitope in the case of amino acids, respectively, and are at most some portion less than a full length sequence. Fragments may be derived from any contiguous portion of a nucleic acid or amino acid sequence of choice. Derivatives are nucleic acid sequences or amino acid sequences formed from the native compounds either directly or by modification or partial substitution. Analogs are nucleic acid sequences or amino acid sequences that have a structure similar to, but not identical to, the native compound but differs from it in respect to certain components or side chains. Analogs may be synthetic or from a different evolutionary origin and may have a similar or opposite metabolic activity compared to wild type. Homologs are nucleic acid sequences or amino acid sequences of a particular gene that are derived from different species.
Derivatives and analogs may be full length or other than full length, if the derivative or analog contains a modified nucleic acid or amino acid, as described below. Derivatives or analogs of the nucleic acids or proteins of the invention include, but are not limited to, molecules comprising regions that are substantially homologous to the nucleic acids or proteins of the invention, in various embodiments, by at least about 30%, 50%, 70%, 80%, or 95% identity (with a preferred identity of 80–95%) over a nucleic acid or amino acid sequence of identical size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art, or whose encoding nucleic acid is capable of hybridizing to the complement of a sequence encoding the aforementioned proteins under stringent, moderately stringent, or low stringent conditions. See e.g. Ausubel, et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, N.Y., 1993, and below.
A “homologous nucleic acid sequence” or “homologous amino acid sequence,” or variations thereof, refer to sequences characterized by a homology at the nucleotide level or amino acid level as discussed above. Homologous nucleotide sequences encode those sequences coding for isoforms of TOXMARKER polypeptide. Isoforms can be expressed in different tissues of the same organism as a result of, for example, alternative splicing of RNA. Alternatively, isoforms can be encoded by different genes. In the present invention, homologous nucleotide sequences include nucleotide sequences encoding for a TOXMARKER polypeptide of species other than humans, including, but not limited to, mammals, and thus can include, e.g., mouse, rat, rabbit, dog, cat cow, horse, and other organisms. Homologous nucleotide sequences also include, but are not limited to, naturally occurring allelic variations and mutations of the nucleotide sequences set forth herein. A homologous nucleotide sequence does not, however, include the nucleotide sequence encoding human TOXMARKER protein. Homologous nucleic acid sequences include those nucleic acid sequences that encode conservative amino acid substitutions (see below) in SEQ ID NO: 1–171, as well as a polypeptide having TOXMARKER activity. Biological activities of the TOXMARKER proteins are described below. A homologous amino acid sequence does not encode the amino acid sequence of a human TOXMARKER polypeptide.
An TOXMARKER polypeptide is encoded by the open reading frame (“ORF”) of a TOXMARKER nucleic acid. An “open reading frame” (“ORF”) corresponds to a nucleotide sequence that could potentially be translated into a polypeptide. A stretch of nucleic acids comprising an ORF is uninterrupted by a stop codon. An ORF that represents the coding sequence for a full protein begins with an ATG “start” codon and terminates with one of the three “stop” codons, namely, TAA, TAG, or TGA. For the purposes of this invention, an ORF may be any part of a coding sequence, with or without a start codon, a stop codon, or both. For an ORF to be considered as a good candidate for coding for a bona fide cellular protein, a minimum size requirement is often set, for example, a stretch of DNA that would encode a protein of 50 amino acids or more.
The nucleotide sequence determined from the cloning of the human TOXMARKER gene allows for the generation of probes and primers designed for use in identifying and/or cloning TOXMARKER homologues in other cell types, e.g. from other tissues, as well as TOXMARKER homologues from other mammals. The probe/primer typically comprises substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 25, 50, 100, 150, 200, 250, 300, 350 or 400 consecutive sense strand nucleotide sequence of SEQ ID NO: 1–171, or an anti-sense strand nucleotide sequence of SEQ ID NO: 1–171 or of a naturally occurring mutant of SEQ ID NO: 1–171.
Probes based on the human TOXMARKER nucleotide sequence can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In various embodiments, the probe further comprises a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a part of a diagnostic test kit for identifying cells or tissue which misexpress a TOXMARKER protein, such as by measuring a level of a TOXMARKER-encoding nucleic acid in a sample of cells from a subject e.g., detecting TOXMARKER mRNA levels or determining whether a genomic TOXMARKER gene has been mutated or deleted.
“A polypeptide having a biologically active portion of TOXMARKER” refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of a polypeptide of the present invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. A nucleic acid fragment encoding a “biologically active portion of TOXMARKER” can be prepared by isolating a portion of SEQ ID NO: 1–171 that encodes a polypeptide having a TOXMARKER biological activity (the biological activities of the TOXMARKER proteins are described below), expressing the encoded portion of TOXMARKER protein (e.g., by recombinant expression in vitro) and assessing the activity of the encoded portion of TOXMARKER.
TOXMARKER Variants
The invention further encompasses nucleic acid molecules that differ from the nucleotide sequence shown in SEQ ID NO: 1–171 due to degeneracy of the genetic code and thus encode the same TOXMARKER protein as that encoded by the nucleotide sequence shown in SEQ ID NO: 1–171.
In addition to the human TOXMARKER nucleotide sequence shown in SEQ ID NO: 1–171 it will be appreciated by those skilled in the art that DNA sequence polymorphisms that lead to changes in the amino acid sequences of TOXMARKER may exist within a population (e.g., the human population). Such genetic polymorphism in the TOXMARKER gene may exist among individuals within a population due to natural allelic variation. As used herein, the terms “gene” and “recombinant gene” refer to nucleic acid molecules comprising an open reading frame encoding a TOXMARKER protein, preferably a mammalian TOXMARKER protein. Such natural allelic variations can typically result in 1–5% variance in the nucleotide sequence of the TOXMARKER gene. Any and all such nucleotide variations and resulting amino acid polymorphisms in TOXMARKER that are the result of natural allelic variation and that do not alter the functional activity of TOXMARKER are intended to be within the scope of the invention.
Moreover, nucleic acid molecules encoding TOXMARKER proteins from other species, and thus that have a nucleotide sequence that differs from the human sequence of SEQ ID NO: 1–171 are intended to be within the scope of the invention. Nucleic acid molecules corresponding to natural allelic variants and homologues of the TOXMARKER cDNAs of the invention can be isolated based on their homology to the human TOXMARKER nucleic acids disclosed herein using the human cDNAs, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions. For example, a soluble human TOXMARKER cDNA can be isolated based on its homology to human membrane-bound TOXMARKER. Likewise, a membrane-bound human TOXMARKER cDNA can be isolated based on its homology to soluble human TOXMARKER.
Accordingly, in another embodiment, an isolated nucleic acid molecule of the invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1–171. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 750, 1000 or 1250 nucleotides in length. In another embodiment, an isolated nucleic acid molecule of the invention hybridizes to the coding region. As used herein, the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% homologous to each other typically remain hybridized to each other.
Homologs (i.e., nucleic acids encoding TOXMARKER proteins derived from species other than human) or other related sequences (e.g., paralogs) can be obtained by low, moderate or high stringency hybridization with all or a portion of the particular human sequence as a probe using methods well known in the art for nucleic acid hybridization and cloning.
As used herein, the phrase “stringent hybridization conditions” refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target sequence hybridize to the target sequence at equilibrium. Since the target sequences are generally present at excess, at Tm, 50% of the probes are occupied at equilibrium. Typically, stringent conditions will be those in which the salt concentration is less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 60° C. for longer probes, primers and oligonucleotides. Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide.
Stringent conditions are known to those skilled in the art and can be found in Ausubel et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y. (1989), 6.3.1–6.3.6. Preferably, the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other. A non-limiting example of stringent hybridization conditions are hybridization in a high salt buffer comprising 6× SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 65° C., followed by one or more washes in 0.2× SSC, 0.01% BSA at 50° C. An isolated nucleic acid molecule of the invention that hybridizes under stringent conditions to the sequence of SEQ ID NO: 1–171 corresponds to a naturally-occurring nucleic acid molecule. As used herein, a “naturally-occurring” nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).
In a second embodiment, a nucleic acid sequence that is hybridizable to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1–171, or fragments, analogs or derivatives thereof, under conditions of moderate stringency is provided. A non-limiting example of moderate stringency hybridization conditions are hybridization in 6× SSC, 5× Denhardt's solution, 0.5% SDS and 100 mg/ml denatured salmon sperm DNA at 55° C., followed by one or more washes in 1× SSC, 0.1% SDS at 37° C. Other conditions of moderate stringency that may be used are well-known in the art. See, e.g., Ausubel et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY.
In a third embodiment, a nucleic acid that is hybridizable to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1–171, or fragments, analogs or derivatives thereof, under conditions of low stringency, is provided. A non-limiting example of low stringency hybridization conditions are hybridization in 35% formamide, 5× SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 mg/ml denatured salmon sperm DNA, 10% (wt/vol) dextran sulfate at 40° C., followed by one or more washes in 2× SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS at 50° C. Other conditions of low stringency that may be used are well known in the art (e.g., as employed for cross-species hybridizations). See, e.g., Ausubel et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY; Shilo and Weinberg, 1981, Proc Natl Acad Sci USA 78: 6789–6792.
EXAMPLES
Example 1
Induction of Zone 3 Necrosis In Vivo
Over 100 compounds were chosen based on evidence that they elicit one of eleven selected histopathology subtypes. The criteria for inclusion of a compound into the nongenotoxic carcinogens mode included evidence of parenchymal changes and an increase in mitosis in vivo. Compounds assigned to this group must also have strong historical documentation. Compounds from other pathology modes were not added to this histopathology subtype. The compounds included in nongenotoxic carcinogenesis can been seen in Table 1. Each compound was delivered orally on a daily basis at a high dose (tox dose) and a 1/10 low dose (mode dose) for up to 14 days. Five male rats/dose/time were randomly assigned to sacrifice on days 1, 3, 7, and 14. In order to best identify genes characteristic of the histopathology subtype, total RNA for all rat livers from a given dose time point were pooled and converted to mRNA and cDNA for GeneCalling®. In GeneCalling, the cDNA is cut with a battery of restriction enzyme pairs in different combinations followed by amplification by PCR using specific primers linked to specific adaptors. After gel electrophoresis, the resulting fragments are identified based on the inherent information in the cDNA fragment: The flanking restriction site sequences on the ends, the size of the fragment and the species (and sometimes the tissue) origin of the DNA. This information is used to query public and proprietary databases. The fragments that do not match any sequences in the database are isolated, sequenced and identified as novel.







TABLE 1







Zone 3 Necrosis in vivo


















Pathology
Pathology


Compound
Vehicle
Dose
Concentration
Time Points
Present
Absent





1,3-Dibromobenzene
Corn Oil
High
600 mg/kg/d
1, 3, 7, 14 d
1 d



1,3-Dibromobenzene
Corn Oil
Low
60 mg/kg/d
1, 3, 7, 14 d


1,4-dichlorobenzene
Corn Oil
High
300 mg/kg/d
1, 3, 7, 14 d


1,4-dichlorobenzene
Corn Oil
Low
30 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


17α-Ethynyl-19-
Corn Oil
High
30 mg/kg/d
1, 3, 7, 14 d


nortestosterone


17α-Ethynyl-19-
Corn Oil
Low
3 mg/kg/d
1, 3, 7, 14 d


nortestosterone


2,4-diaminotoluene
Methylcellulose
High
15 mg/kg/d
1, 3, 7, 14 d


2,4-diaminotoluene
Methylcellulose
Low
1.5 mg/kg/d
1, 3, 7, 14 d


2-acetylaminofluorene
Methylcellulose
High
12 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


2-acetylaminofluorene
Methylcellulose
Low
1.2 mg/kg/d
1, 3, 7, 14 d


2-acetylaminofluorene
Methylcellulose
High
120 mg/kg (ip)
6, 12, 24, 48 h


2-acetylaminofluorene
Methylcellulose
Low
12 mg/kg (ip)
6, 12, 24, 48 h


2-nitrofluorene
Corn oil
High
44 mg/kg/d
1, 3, 7, 14 d


2-nitrofluorene
Corn oil
Low
4.4 mg/kg/d
1, 3, 7, 14 d


3-methyl-4-
Methylcellulose
High
36 mg/kg/d
1, 3, 7, 14 d


(dimethylamino)


azobenzene


3-methyl-4-
Methylcellulose
Low
3.6 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


(dimethylamino)


azobenzene


3-methylcholanthrene
Corn Oil
High
25 mg/kg/d (ip)
1, 3, 7, 14 d


3-methylcholanthrene
Corn Oil
Low
2.5 mg/kg/d (ip)
1, 3, 7, 14 d

1, 3, 7, 14 d


Acetamide
Methylcellulose
High
3000 mg/kg/d
1, 3, 7, 14 d


Acetamide
Methylcellulose
Low
300 mg/kg/d
1, 3, 7, 14 d


Acetaminophen
Saline
High
4.25 g/kg (po)
6, 12, 24, 48 h
24, 48 h


Acetaminophen
Saline
Low
425 mg/kg (po)
6, 12, 24, 48 h


Aflatoxin B1
Methylcellulose
High
0.24 mg/kg/d
1, 3, 7, 14 d


Aflatoxin B1
Methylcellulose
Low
0.024 mg/kg/d
1, 3, 7, 14 d


Allyl Alcohol
Methylcellulose
High
36 mg/kg/d
1, 3, 7, 14 d


Allyl Alcohol
Methylcellulose
Low
3.6 mg/kg/d
1, 3, 7, 14 d


Allyl Formate
Corn oil
High
94.8 mg/kg (ip)
3, 6, 12, 24 h


Allyl Formate
Corn oil
Low
9.48 mg/kg (ip)
3, 6, 12, 24 h


Amiodarone
Methylcellulose
High
500 mg/kg/d
1, 3, 7, 14 d


Amiodarone
Methylcellulose
Low
50 mg/kg/d
1, 3, 7, 14 d


ANIT
Corn Oil
High
60 mg/kg/d
1, 3, 7, 14 d


ANIT
Corn Oil
Low
6 mg/kg/d
1, 3, 7, 14 d


Azaserine
Saline
High
100 mg/kg (ip)
1, 3, 7, 14 d
1,3 d


Azaserine
Saline
Low
10 mg/kg (ip)
1, 3, 7, 14 d


BCNU
Corn Oil
High
20 mg/kg/d
1, 3, 7, 14 d


BCNU
Corn Oil
Low
2 mg/kg/d
1, 3, 7, 14 d


BHT
Corn Oil
High
500 mg/kg/d
1, 3, 7, 14 d


BHT
Corn Oil
Low
50 mg/kg/d
1, 3, 7, 14 d


Bromobenzene
Saline
High
1200 mg/kg (ip)
6, 12, 24, 48 h
12, 48 h


Bromobenzene
Saline
Low
120 mg/kg (ip)
6, 12, 24, 48 h


C.I. Direct Black
Corn oil
High
146 mg/kg/d
1, 3, 7, 14 d


C.I. Direct Black
Corn oil
Low
14.6 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Cadmium chloride
Saline
High
3.9 mg/kg (iv)
6, 12, 24, 48 h


Cadmium chloride
Saline
Low
0.39 mg/kg (iv)
6, 12, 24, 48 h


Carbamazepine
Methylcellulose
High
200 mg/kg/d
1, 3, 7, 14 d


Carbamazepine
Methylcellulose
Low
20 mg/kg/d
1, 3, 7, 14 d


CCl4
Corn Oil
High
50 mg/kg/d
1, 3, 7, 14 d
7 d


CCl4
Methylcellulose
High
956 mg/kg (ip)
6, 12, 24, 48 h
6, 12, 48 h


CCl4
Corn Oil
Low
5 mg/kg/d
1, 3, 7, 14 d


CCl4
Methylcellulose
Low
95.6 mg/kg (ip)
6, 12, 24, 48 h


CCNU
Corn Oil
High
20 mg/kg/d
1, 3, 7, 14 d


CCNU
Corn Oil
Low
2 mg/kg/d
1, 3, 7, 14 d


Cefuroxime
Methylcellulose
Safe
125 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Cerium (III) Chloride
Saline
High
10 mg/kg (iv)
1, 2, 3, 4 d
1, 2, 3, 4 d


Cerium (III) Chloride
Saline
Low
1 mg/kg (iv)
1, 2, 3, 4 d


Chlordane
Corn Oil
High
25 mg/kg/d
1, 3, 7, 14 d


Chlordane
Corn Oil
Low
2.5 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Ciprofibrate
Corn Oil
High
10 mg/kg/d
1, 3, 7, 14 d


Ciprofibrate
Corn Oil
Low
1 mg/kg/d
1, 3, 7, 14 d


Ciprofloxacin
Methylcellulose
Safe
40 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Clofibrate
Methylcellulose
High
300 mg/kg/d
1, 3, 7, 14 d


Clofibrate
Methylcellulose
Low
30 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Clonidine
Methylcellulose
Safe
0.1 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Colchicine
Saline
High
5 mg/kg × 2 (ip)
6, 12, 24, 48 h


Colchicine
Saline
Low
0.5 mg/kg × 2 (ip)
6, 12, 24, 48 h


Concanavalin A
Saline
High
20 mg/kg (iv)
6, 12, 24, 48 h


Concanavalin A
Saline
Low
2 mg/kg (iv)
6, 12, 24, 48 h


Corn Oil

Control

1, 3, 7, 14 d

1, 3, 7, 14 d


Corn Oil (ip)

Control

1, 3, 7, 14 d

1, 3, 7, 14 d


Corn Oil (ip)

Control

3, 6, 12, 24 h

3, 6, 12, 24 h


Coumarin
Corn Oil
High
150 mg/kg/d
1, 3, 7, 14 d
3 d


Coumarin
Corn Oil
Low
15 mg/kg/d
1, 3, 7, 14 d


CTFT
Corn Oil
High
1 g/kg/d
1, 3, 7, 14 d


CTFT
Corn Oil
Low
100 mg/kg/d
1, 3, 7, 14 d


Cyclosporine A
Corn Oil
High
50 mg/kg/day
1, 3, 7, 14 d


Cyclosporine A
Corn Oil
Low
5 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Cyproterone acetate
Corn Oil
High
100 mg/kg/d
1, 3, 7, 14 d


Cyproterone acetate
Corn Oil
Low
10 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Dehydroepiandrosterone
Methylcellulose
High
600 mg/kg/d
1, 3, 7, 14 d


Dehydroepiandrosterone
Methylcellulose
Low
60 mg/kg/d
1, 3, 7, 14 d


Deoxycholic Acid
Methylcellulose
High
300 mg/kg/d
1, 3, 7, 14 d


Deoxycholic Acid
Methylcellulose
Low
30 mg/kg/d
1, 3, 7, 14 d


Dexamethasone
Corn oil
High
50 mg/kg/d (ip)
1, 3, 7, 14 d


Dexamethasone
Corn oil
Low
5 mg/kg/d (ip)
1, 3, 7, 14 d

1, 3, 7, 14 d


D-galactosamine
Saline
High
500 mg/kg (ip)
6, 12, 24, 48 h


D-galactosamine
Saline
Low
50 mg/kg (ip)
6, 12, 24, 48 h


Di(2-Ethylhexyl)
Methylcellulose
High
1200 mg/kg/d
1, 3, 7, 14 d


phthalate


Di(2-Ethylhexyl)
Methylcellulose
Low
120 mg/kg/d
1, 3, 7, 14 d


phthalate


Dibutyltin chloride
Corn oil
High
20 mg/kg/d
1, 3, 7, 14 d


Dibutyltin chloride
Corn oil
Low
2 mg/kg/d
1, 3, 7, 14 d


Dichloropropane
Corn oil
High
1000 mg/kg/d
1, 3, 7, 14 d
1, 3 d


Dichloropropane
Corn oil
Low
100 mg/kg/d
1, 3, 7, 14 d


Diethylnitrosamine
Saline
High
150 mg/kg (ip)
1, 3, 7, 14 d
1, 3, 7 d


Diethylnitrosamine
Saline
Low
15 mg/kg (ip)
1, 3, 7, 14 d


Diethylstilbestrol
Methylcellulose
High
10 mg/kg/d
1, 3, 7, 14 d


Diethylstilbestrol
Methylcellulose
Low
1 mg/kg/d
1, 3, 7, 14 d


Dimethylformamide
Saline
High
850 mg/kg (ip)
6, 12, 24, 48 h
48 h


Dimethylformamide
Saline
Low
85 mg/kg (ip)
6, 12, 24, 48 h


Dimethylnitrosamine
Corn Oil
High
4 mg/kg/d
1, 3, 7, 14 d
7, 14 d


Dimethylnitrosamine
Corn Oil
Low
0.4 mg/kg/d
1, 3, 7, 14 d


Diquat
Saline
High
36 mg/kg (ip)
6, 12, 24, 48 h


Diquat
Saline
Low
3.6 mg/kg (ip)
6, 12, 24, 48 h

6, 12, 24, 48 h


Disopyramide
Methylcellulose
Safe
20 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Endotoxin
Saline
High
8 mg/kg (ip)
6, 12, 24, 48 h


Endotoxin
Saline
Low
0.8 mg/kg (ip)
6, 12, 24, 48 h


Erythromycin Estolate
Methylcellulose
High
800 mg/kg/d
1, 3, 7, 14 d


(EE)


Erythromycin Estolate
Methylcellulose
Low
80 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


(EE)


Ethanol
Corn Oil
High
15 g/kg/d
1, 3, 7, 14 d


Ethanol
Corn Oil
Low
1.5 g/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Ethinylestradiol
Corn Oil
High
15 mg/kg/d
1, 3, 7, 14 d


Ethinylestradiol
Corn Oil
Low
1.5 mg/kg/d
1, 3, 7, 14 d


Ethionine
Methylcellulose
High
200 mg/kg/d
1, 3, 7, 14 d
1, 3 d


Ethionine
Methylcellulose
Low
20 mg/kg/d
1, 3, 7, 14 d


Ethosuximide
Methylcellulose
Safe
100 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Ethylenethiourea
Saline
High
920 mg/kg (po)
6, 12, 24, 48 h


Ethylenethiourea
Saline
Low
92 mg/kg (po)
6, 12, 24, 48 h


Fenarimol
Corn Oil
High
62.5 mg/kg/d
1, 3, 7, 14 d


Fenarimol
Corn Oil
Low
6.25 mg/kg/d
1, 3, 7, 14 d


Fenbendazole
Methylcellulose
High
3000 mg/kg/d
1, 3, 7, 14 d


Fenbendazole
Methylcellulose
Low
300 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Furan
Corn Oil
High
15 mg/kg/d
1, 3, 7, 14 d


Furan
Corn Oil
Low
1.5 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Hydrazine
Methylcellulose
High
100 mg/kg/d
1, 3, 7, 14 d


Hydrazine
Methylcellulose
Low
10 mg/kg/d
1, 3, 7, 14 d


Ibuprofen
Methylcellulose
Safe
94 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Ketoconazole
Methylcellulose
High
100 mg/kg/d
1, 3, 7, 14 d


Ketoconazole
Methylcellulose
Low
10 mg/kg/d
1, 3, 7, 14 d

1, 3 d


Lead nitrate
Saline
High
33 g/kg (iv)
1, 3, 7, 14 d


Lead nitrate
Saline
Low
3.3 g/kg (iv)
1, 3, 7, 14 d


Methapyrilene
Methylcellulose
High
60 mg/kg/d
1, 3, 7, 14 d


Methapyrilene
Methylcellulose
Low
6 mg/kg/d
1, 3, 7, 14 d


Methionine-choline
In feed
High
60 g/kg/d
1, 3, 7, 14 d


deficient diet


Methyl Carbamate
Methylcellulose
High
400 mg/kg/d
1, 3, 7, 14 d


Methyl Carbamate
Methylcellulose
Low
40 mg/kg/d
1, 3, 7, 14 d


Methylcellulose

Control

1, 3, 7, 14 d

1, 3, 7, 14 d


Methylcellulose (ip)

Control

6, 12, 24, 48 h


Methylenedianiline
Corn Oil
High
50 mg/kg/d
1, 3, 7, 14 d


Methylenedianiline
Corn Oil
Low
5 mg/kg/d
1, 3, 7, 14 d


Methyleugenol
Methylcellulose
High
1000 mg/kg/d
1, 3, 7, 14 d


Methyleugenol
Methylcellulose
Low
100 mg/kg/d
1, 3, 7, 14 d


Methyl-tert-butyl ether
Corn Oil
High
1500 mg/kg/d
1, 3, 7, 14 d


Methyl-tert-butyl ether
Corn Oil
Low
150 mg/kg/d
1, 3, 7, 14 d


Microcystin-LR
Saline
High
20 μg/kg (iv)
6, 12, 24, 48 h


Microcystin-LR
Saline
Low
2 μg/kg (iv)
6, 12, 24, 48 h


Mirex
Corn Oil
High
10 mg/kg/d
1, 3, 7, 14 d


Mirex
Corn Oil
Low
1 mg/kg/d
1, 3, 7, 14 d


Molybdenum
Methylcellulose
High
500 mg/kg/d
1, 3, 7, 14 d
3 d


Molybdenum
Methylcellulose
Low
50 mg/kg/d
1, 3, 7, 14 d


Monocrotaline
H2O
High
160 mg/kg/d
1, 3, 7, 14 d
1, 3 d


Monocrotaline
H2O
Low
16 mg/kg/d
1, 3, 7, 14 d


N-diethylnitrosamine
Methylcellulose
High
12 mg/kg/d
1, 3, 7, 14 d


N-diethylnitrosamine
Methylcellulose
Low
1.2 mg/kg/d
1, 3, 7, 14 d


Nifedipine
Methylcellulose
Safe
3 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Nitrofurantoin
Saline
High
150 mg/kg (ip)
1, 3, 7, 14 d


Nitrofurantoin
Saline
Low
15 mg/kg (ip)
1, 3, 7, 14 d

1, 3, 7, 14 d


Nitrosodiethanolamine
Methylcellulose
High
200 mg/kg/d
1, 3, 7, 14 d


Nitrosodiethanolamine
Methylcellulose
Low
20 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Nitrosomethylethylamine
Methylcellulose
High
75 mg/kg/d
1, 3, 7, 14 d
1, 3 d


Nitrosomethylethylamine
Methylcellulose
Low
7.5 mg/kg/d
1, 3, 7, 14 d
3, 7, 14 d


N-nitrosodibutylamine
Methylcellulose
High
25 mg/kg/d
1, 3, 7, 14 d


N-nitrosodibutylamine
Methylcellulose
Low
2.5 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


N-nitrosomorpholine
Methylcellulose
High
35 mg/kg/d
1, 3, 7 d
1, 3, 7, 14 d


N-nitrosomorpholine
Methylcellulose
Low
3.5 mg/kg/d
1, 3, 7, 14 d


N-Nitrosopiperidine
Methylcellulose
High
200 mg/kg/d
1, 3, 7, 14 d
1, 3 d


N-Nitrosopiperidine
Methylcellulose
Low
20 mg/kg/d
1, 3, 7, 14 d


NNK
Methylcellulose
High
20 mg/kg/d
1, 3, 7, 14 d


NNK
Methylcellulose
Low
2 mg/kg/d
1, 3, 7, 14 d


Pentachlorophenol
Methylcellulose
High
50 mg/kg/d
1, 3, 7, 14 d


Pentachlorophenol
Methylcellulose
Low
5 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Phenobarbital
Methylcellulose
High
80 mg/kg/d
1, 3, 7, 14 d


Phenobarbital
Methylcellulose
Low
8 mg/kg/d
1, 3, 7, 14 d


Piperonyl Butoxide
Methylcellulose
High
1200 mg/kg/d
1, 3, 7, 14 d


Piperonyl Butoxide
Methylcellulose
Low
120 mg/kg/d
1, 3, 7, 14 d


Potassium bichromate
Methylcellulose
High
10 mg/kg (ip)
6, 12, 24, 48 h


Potassium bichromate
Methylcellulose
Low
1 mg/kg (ip)
6, 12, 24, 48 h


Prazosin
Methylcellulose
Safe
1 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Pregnenolone-16α-
Methylcellulose
High
100 mg/kg/d
1, 3, 7, 14 d


Carbonitrile


Pregnenolone-16α-
Methylcellulose
Low
10 mg/kg/d
1, 3, 7, 14 d


Carbonitrile


Propranolol
Methylcellulose
Safe
40 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Pyridine
Methylcellulose
High
300 mg/kg/d
1, 3, 7, 14 d
14 d


Pyridine
Methylcellulose
Low
30 mg/kg/d
1, 3, 7, 14 d


Ranitidine
Methylcellulose
Safe
5 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Rifampicin
Methylcellulose
High
250 mg/kg/d
1, 3, 7, 14 d


Rifampicin
Methylcellulose
Low
25 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Saline (ip)

Control

6, 12, 24, 48 h

6, 12, 24, 48 h


Saline (ip)

Control

1, 3, 7, 14 d

1, 3, 7, 14 d


Saline (iv)

Control

6, 12, 24, 48 h

6, 12, 24, 48 h


Saline (iv)

Control

1, 2, 3, 4 d

1, 2, 3, 4 d


Saline (iv)

Control

1, 3, 7, 14 d

1, 3, 7, 14 d


Saline (po)

Control

6, 12, 24, 48 h

6, 12, 24, 48 h


Terfenadine
Methylcellulose
Safe
10 mg/kg/d
1, 3, 7, 14 d

1, 3, 7, 14 d


Thioacetamide
Saline
High
200 mg/kg (ip)
6, 12, 24, 48 h
12, 24, 48 h


Thioacetamide
Saline
Low
20 mg/kg (ip)
6, 12, 24, 48 h
6, 12, 24, 48 h


Thioacetamide
Methylcellulose
High
19.2 mg/kg/d
1, 3, 7, 14 d


Thioacetamide
Methylcellulose
Low
1.92 mg/kg/d
1, 3, 7, 14 d


Thiobenzamide
In Feed
High
ad libitum 1 g/kg
1, 3, 7, 14 d
1, 3, 7, 14 d


Thiobenzamide
In Feed
Low
ad libitum 0.1 g/kg
1, 3, 7, 14 d


Untreated

Control

1, 3, 7, 14 d

1, 3, 7, 14 d


Untreated

Control

6, 12, 24, 48 h

6, 12, 24, 48 h


Untreated

Control

1, 2, 3, 4 d

1, 2, 3, 4 d


WY-14643
Methylcellulose
High
60 mg/kg/d
1, 3, 7, 14 d


WY-14643
Methylcellulose
Low
6 mg/kg/d
1, 3, 7, 14 d


Xylidine
Corn Oil
High
600 mg/kg/d
1, 3, 7, 14 d


Xyidine
Corn Oil
Low
60 mg/kg/d
1, 3, 7, 14 d









Example 2
Induction of Zone 3 Necrosis In Vitro
Over 100 compounds were chosen based on evidence that they elicit one of eleven selected histopathology subtypes. The criteria for inclusion of a compound into the zone 3 necrosis mode was necrosis or individual cell necrosis in centrilobular zone with an increase in some or all serum transaminases. The compounds included in the zone 3 necrosis mode can been seen in Table 2. Rat primary hepatocytes were treated with the same compounds used in the in vivo experiments. In all cases, compounds were run in any given two-day period with an untreated and DMSO vehicle as negative controls. Each compound was delivered daily at a reference dose derived from the literature (when available), a 10× high dose, and 1/10 and 1/100 low doses for up to 72 hours. Three hepatocyte cultures for each dose were randomly assigned for harvest at 6, 24, 48 and 72 hours. In order to best identify genes characteristic of zone 3 necrosis, total RNA for all hepatocyte cultures from a given dose time point were pooled and converted to mRNA and cDNA for GeneCalling®. In GeneCalling, the cDNA is cut with a battery of restriction enzyme pairs in different combinations followed by amplification by PCR using specific primers linked to specific adaptors. After gel electrophoresis, the resulting fragments are identified based on the inherent information in the cDNA fragment: The flanking restriction site sequences on the ends, the size of the fragment and the species (and sometimes the tissue) origin of the DNA. This information is used to query public and proprietary databases. The fragments that do not match any sequences in the database are isolated, sequenced and identified as novel.







TABLE 2







Zone 3 Necrosis in vitro













Zone 3




Concentration
Necrosis


Compound
Annotation
(microM)
Pathology













1,3-Dibromobenzene

63



1,3-Dibromobenzene

250
Present


2AAF

10


2AAF

50


2-nitrofluorene

30


2-nitrofluorene

150


Acetamide

500


Acetamide

2000


Acetaminophen

100


Acetaminophen

1000
Present


Aflatoxin

0.008


Aflatoxin

0.04


Allylalcohol

16


Allylalcohol

80


Allylformate

0.06


Allylformate

0.3


Amiodarone

5


Amiodarone

20


ANIT

3.1


ANIT

13


Aspirin
non-toxic
55.5
Absent


Aspirin
non-toxic
555


Atorvastatin
non-toxic
0.0095
Absent


Atorvastatin
non-toxic
0.095


Azaserine

100


Azaserine

500
Present


Azobenzene

1.6
Absent


Azobenzene

8


BCNU

50


BCNU

250


Butylhydroxytoluene

75


Butylhydroxytoluene

150


Bretylium
non-toxic
1.2
Absent


Bretylium
non-toxic
12


Bromobenzene

600


Bromobenzene

3000
Present


Carbamate

300


Carbamate

1500


Carbamazepine

200


Carbamazepine

1000


CCNU

8


CCNU

40


CdCl

0.1


CdCl

0.5


CeCl3

4


CeCl3

20
Present


Cefuroxime
non-toxic
224
Absent


Cefuroxime
non-toxic
2240


Chlordane

8
Absent


Chlordane

40


ClDirect

1


ClDirect

5


Ciprofibrate

100


Ciprofibrate

500


Clofibrate

100
Absent


Clofibrate

400


Clonidine
non-toxic
0.0165
Absent


Clonidine
non-toxic
0.165


Colchicine

500


Colchicine

2000


Concanavalin A

5000


Concanavalin A

20000


Coumarin

63


Coumarin

250
Present


4-chlorobenzotrifluoride

250


4-chlorobenzotrifluoride

1000


Cyclosporine

3.1
Absent


Cyclosporine

13


Cyproterone

10


Cyproterone

50


2,4-diaminotoluene

0.8


2,4-diaminotoluene

4


Di(2-Ethylhexyl)phthalate

500


Di(2-Ethylhexyl)phthalate

2000


Dehydroepiandosterone

1.2


Dehydroepiandosterone

6


Deoxycholate

6.3


Deoxycholate

25


Dexamethasone

100
Absent


Dexamethasone

500


Dibutyltin

0.2


Dibutyltin

1


Dichlorobenzene

100
Absent


Dichlorobenzene

500


Diethylnitrosamine

60


Diethylnitrosamine

300
Present


Diethylstillbestrol

5


Diethylstillbestrol

50


Dimethylnitrosamine

200


Dimethylnitrosamine

1000
Present


Disopyramide
non-toxic
3
Absent


Disopyramide
non-toxic
30


Dimethylformamide

1000


Dimethylformamide

5000
Present


DMSO
control
0
Absent


Doxorubicin
non-toxic
0.5
Absent


Doxorubicin
non-toxic
5


Endotoxin

30


Endotoxin

100


Erythromycin

20
Absent


Erythromycin

100


Ethanol

200
Absent


Ethanol

1000


Ethinylestradiol

25


Ethinylestradiol

100


Ethionine

200


Ethionine

1000
Present


Ethosuximide
non-toxic
1000
Absent


Ethosuximide
non-toxic
10000


Ethylenethiourea

200


Ethylenethiourea

1000


Fenarimol

20
Absent


Fenarimol

100


Fenbendazole

16


Fenbendazole

63


Fluconazole
non-toxic
0.816
Absent


Fluconazole
non-toxic
8.16


Gabapentin
non-toxic
2
Absent


Gabapentin
non-toxic
20


Galactosamine

12


Galactosamine

60


Hydrazine

20


Hydrazine

100


Ibuprofen
non-toxic
50
Absent


Ketoconazole

2
Absent


Ketoconazole

10


Mephenytoin
non-toxic
14.2
Absent


Mephenytoin
non-toxic
142


Methapyriline

30


Methapyriline

100


Methylcholanthrene

40
Absent


Methylcholanthrene

200


Methylenedianiline

1.4


Methylenedianiline

7.8


Methylleugenol

100


Methylleugenol

500


Microcystin

0.005


Microcystin

0.025


Minoxidil
non-toxic
0.166
Absent


Minoxidil
non-toxic
1.66


Mirex

50
Absent


Mirex

100


Molybdenum

20


Molybdenum

50
Present


Monocrotaline

30


Monocrotaline

100
Present


Methyl-tert-butyl ether

1000


Methyl-tert-butyl ether

4000


Nifedipine
non-toxic
0.335
Absent


Nifedipine
non-toxic
3.35


Nitrofurantoin

4
Absent


Nitrofurantoin

20


Nitrosodibutylamine

200
Absent


Nitrosodibutylamine

1000


Nitrosodiethanolamine

1000
Absent


Nitrosodiethanolamine

5000


Nitrosomethylethylamine

200


Nitrosomethylethylamine

1000
Present


Nitrosomorpholine

750


Nitrosomorpholine

3750
Present


Nitrosopiperidine

640


Nitrosopiperidine

3200
Present


NNK

200


NNK

1000


Norethindrone

40
Absent


Norethindrone

200


Pentachlorophenol

19
Absent


Pentachlorophenol

38


Piperonyl

20


Piperonyl

100


Prazosin
non-toxic
0.0148
Absent


Prazosin
non-toxic
0.148


Pregnenolone

38
Absent


Pregnenoione

150


Propranolol
non-toxic
0.125
Absent


Propranolol
non-toxic
1.25


Pyridine

800


Pyridine

4000
Present


Ranitidine
non-toxic
0.128
Absent


Ranitidine
non-toxic
1.28


Rifampicin

20
Absent


Rifampicin

100


Terfenadine
non-toxic
0.15
Absent


Terfenadine
non-toxic
1.5


Thioacetamide

500


Thioacetamide

2000
Present


Thiobenzamide

8


Thiobenzamide

40
Present


Untreated
control
0
Absent


Verapamil
non-toxic
0.1
Absent


Verapamil
non-toxic
1


WY14643

20


WY14643

100


Xylidine

13


Xylidine

50









Example 3
Methods of Analysis
Data Preparation:
We used GeneCalling® to estimate the activity of several thousand transcripts simultaneously. These data generally have  ̃5% missing data and are log normally distributed. The data are log transformed and missing values are filled using k-nearest neighbor (knn) replacement [14]. The knn algorithm was initially validated using a complete data set and randomly eliminating constant percentages of the data. It was determined that using correlation as a similarity index and imputing missing values with 6 nearest neighbors resulted in the smallest error of prediction.
Initially our data sets contained between 6000 and 8000 genes, which poses two problems. These large numbers of genes make most marker selection procedures computationally intractable with most computer algorithms. Second, the inclusion of markers with low variation, or low association with pathology results in a significant risk of choosing markers that over fit the models. To eliminate these problems we imposed an initial filter on the data, requiring that there be a significant difference between negative control samples and positive control samples for each pathology mode (Kruskal-Wallis test, p<0.001). Depending on mode, this process reduced our gene set to a more tractable number of genes (approximately 200–800 genes depending on mode).
Initial Marker Selection:
We define a marker as a gene that helps to explain some variation in pathology. In order to avoid selecting markers that particularly fit our current data set well at the expense of predictability outside our training set, we employed a leave one out cross-validation method to identify markers that contribute some explanatory power to the data set. Specifically, after the Kruskal-Wallis filter, a series of leave one out models are created leaving out all of the samples for each compound until all compounds have been left out once. This process results in a marker list and a count of the number of leave-one out models the marker was used in. This marker list contains all of the genes that explain some portion of the variation in pathology but is almost certain to over fit the data because of its size. In order to refine this marker list a second series of leave one compound out models is created for each different count of markers within the marker list. For example, the initial leave one out model may produce a gene list consisting of 5 genes that occur at frequencies of 20, 19, 19, 2, and 1 leave one out models respectively. The first step will use all genes that occur in 1 or more leave one out models, the second step 2 or more, the third 19 or more and finally 20 or more. The genes that are considered to be markers will have been used in a majority of models and result in a highly sensitive model. In most cases the first modeling step resulted in a sharp cutoff (e.g. 19 or more in the above example), which guides the marker selection process. In a few cases, the change in frequency was so gradual that no clear cutoff was available. Marker selection then proceeded with the most sensitive model first, and then the most specific model and in the case of ties the least number of markers.
Models:
The above process is a general strategy that is applied to all of our marker selection models. The models we used covered a range of statistical power and assumption stringency. The most powerful model with the strongest assumptions is a linear discriminant analysis, followed by logistic regression and finally by classification trees, which is virtually devoid of assumptions but does have a cost in terms of predictivity. These modeling methods are common statistical procedures that need not be developed here [15] [16] for a more detailed discussion). All three methods went through the algorithm outlined above with the exception of discriminant analysis, which did not utilize a Kruskal-Wallis filter. Both discriminant analysis and logistic regression create poor models when too many intercorrelated variables are used. To minimize this problem, these methods utilized a stepwise selection procedure (incorporating both forward and reverse selection) to select the best discriminating set of markers.
In Vivo Pathology Annotation:
In order to construct in vivo models, the pathology of each sample had to be determined. Pathology was assigned to each liver sample by the pathologists at Bayer's Stillwell, Kans. facility. Criteria for inclusion of a compound into a particular mode included the following:

    
    
        i. Zone-3 Necrosis: Necrosis or individual cell necrosis in centrilobular zone with an increase in some or all serum transaminases.
        ii. Cholestasis: Increased plasma billirubin with bile duct necrosis or hyperplasia.
        iii. Hypertrophy: Increase in cell size and liver weight.
        iv. Genotoxic Carcinogens: Some evidence of mild parenchymal damage in vivo which may be associated with an increase in mitosis. Compounds assigned to this group must have strong historical documentation. Compounds from other pathology modes cannot be added to this list.
        V. Non-genotoxic Carcinogens: Evidence of parenchymal changes and an increase in mitosis. Compounds assigned to this group must have strong historical documentation. Compounds from other pathology modes cannot be added to this list.
        vi. Steatosis: Increase in lipid accumulation or “vacuolar degeneration.”
        vii. Zone 1 Necrosis: Necrosis or individual cell necrosis in the periportal zone, with an increase in some or all serum transaminases.
        vii. Inflammation: Increase in inflammatory cells (e.g. Kupffer cells, neutrophils, macrophages, lymphocytes)
        viii. Apoptosis: Shrinking or fragmentation of the nucleus and increased “blebbing.”
    
    


In Vitro Pathology Annotation:
In order to construct in vitro models we need to make a decision about what pathology each sample represents. Two approaches were used. The first is to ascribe the in vivo compound annotation to the two highest concentrations in vitro (these are traditionally within five fold of each other). For example, clofibrate produced hypertrophy in vivo, so the two highest doses in vitro are used as a positive control for the model construction process. The second annotation strategy uses a nearest neighbor algorithm to assign annotation from in vivo samples to in vitro samples. Briefly, each in vitro sample was correlated, across in vivo markers, to all in vivo samples of the same compound. The annotation of the most correlated in vivo sample was used as the in vitro annotation.
Final Marker Selection:
This process resulted in 6 sets of models being generated for each mode of pathology (LDA, logistic, and classification trees for each annotation strategy, nearest neighbor and high dose). These markers are then correlated with the original data set (between 6000 and 8000 genes) and additional correlated markers (r>=0.60 across 329 samples, up to 3 per marker) were added back in to the final gene set for representation on a microarray.
Example 4
Identification of Zone 3 Necrosis Related Genes In Vivo


    
    
        SP=Secreted Protein
        NC=Novel Rat Composition
        NU=Novel Rat Utility
    
    
















TABLE 3








TOX








MARKER
SEQ ID



ACCNO
ASSIGNMENT
NO:
Definition
Description
Bin






















NU
scr_gb-
1
1
Rattus norvegicus
Betaine-homocysteine methyltransferase
Amino Acid



af03887


betaine homocysteine
(BHMT) catalyzes the transfer of an N-methyl
Metabolism



0_4


methyltransferase
group from betaine to homocysteine to produce






(BHMT) [AF038870].
dimethylglycine and methionine, respectively.







The enzyme is found in the pathway of choline







oxidation and is abundantly expressed in liver







and kidney. It has been known for at least 50







years that alterations in methionine metabolism







occur in human liver cirrhosis. Recently human







BHMT had been shown to be a zinc







metalloenzyme [14] [15].


NC
scr_gb-
2
2
Rat gene fragment -
Betaine-homocysteine methyltransferase
Amino Acid


NU
z83053—


1984 bp. 88% SI
(BHMT) catalyzes the transfer of an N-methyl
Metabolism



3


(1241/1396) to Mus
group from betaine to homocysteine to produce






musculus betaine-
dimethylglycine and methionine, respectively.






homocysteine
The enzyme is found in the pathway of choline






methyltransferase 2
oxidation and is abundantly expressed in liver






(Bhmt2) [AF257474].
and kidney. It has been known for at least 50







years that alterations in methionine metabolism







occur in human liver cirrhosis. Recently human







BHMT had been shown to be a zinc







metalloenzyme [14] [15].


NU
scr_gb-
3
3
Rattus norvegicus
Rat liver peroxisomes contain three acyl-CoA
Lipid Metabolism



x95189—


Trihydroxycoprostano-
oxidases:palmitoyl-CoA oxidase, pristanoyl-CoA



4


yl-CoA Oxidase
oxidase, and trihydroxycoprostanoyl-CoA






[X95189].
oxidase. Mammalian liver peroxisomes are







capable of beta-oxidizing a variety of substrates







including very long chain fatty acids and the







side chains of the bile acid intermediates di- and







trihydroxycoprostanic acid. The first enzyme of







peroxisomal beta-oxidation is acyl-CoA oxidase







[16].


NU
scr_gb-
4
4
Rattus norvegicus
Eph receptor tyrosine kinases and their
Cell Cycle



m59814


Ephrin type-B
membrane-bound ligands, ephrins, have thus
Regulation



_4


receptor 1 precursor
emerged as mediators of cell-contact-
(Regulation Of






(EphB1) [P09759]
dependent repulsion. The actin cytoskeleton is
Proliferation)







also a major target of the intracellular pathways







activated by Eph receptors [17]. More







specifically, activation of EphB1 by its ligand,







ephrin-B1/Fc has been shown to recruit Nck to







native receptor complexes and activate c-Jun







kinase (JNK/SAPK) [18]


NU
scr_gb-
5
5
Rattus norvegicus
It has been known for 20 years that the
Protein



m29358


ribosomal protein S6
ribosomal protein S6 is rapidly phosphorylated
Metabolism



_5


[M29358].
when cells are stimulated to grow or divide [19].







S6 is phosphorylated in response to mitogens







by activation of one or more protein kinase







cascades. Members of the 90 kDa S6 kinases







are activated in vitro by 42 kDa and 44 kDa







MAP kinases, which are in turn activated by







mitogen-dependent activators [20].


NU
aj29733
6
6
Rattus norvegicus
In addition to appearing in response to
Protein



6


heat shock protein 86
biological stresses, heat shock proteins are
Metabolism






(hsp86) [AJ428213].
expressed as ‘chaperones’ by some cells living







in physiological conditions. Among these







proteins, the Hsp90 family, consisting of







isoforms Hsp84 and Hsp86, seems to function







under normal growth conditions in the pathways







of numerous signal transducers, cell cycle and







developmental regulators. [21, 22]


NU
j00719
7
7
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification






cytochrome p-450
constituent proteins of the liver mixed function
Response/






isoform,
monooxygenases. They play a central role in
Biotransformation-






(phenobarbital-
the metabolism of steroids, the detoxification of
TOX






inducible or 2B1)
drugs and xenobiotics, and the activation of






[J00719] [P04167].
procarcinogens. In liver microsomes, this







enzyme is involved in an NADPH-dependent







electron transport pathway. It oxidizes a variety







of structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [23].


NU
j00720
8
8
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification






cytochrome p-450
constituent proteins of the liver mixed function
Response/






isoform,
monooxygenases. They play a central role in
Biotransformation-






(phenobarbital-
the metabolism of steroids, the detoxification of
TOX






inducible or 2B2)
drugs and xenobiotics, and the activation of






[P04167] [J00719].
procarcinogens. In liver microsomes, this







enzyme is involved in an NADPH-dependent







electron transport pathway. It oxidizes a variety







of structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [23].


NU
j00728
9
9
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification






cytochrome p-450
constituent proteins of the liver mixed function
Response/






isoform,
monooxygenases. They play a central role in
Biotransformation-






(phenobarbital-
the metabolism of steroids, the detoxification of
TOX






inducible, 2B1, or
drugs and xenobiotics, and the activation of






2B2) [P00176]
procarcinogens. In liver microsomes, this






[P04167] [Q64584].
enzyme is involved in an NADPH-dependent







electron transport pathway. It oxidizes a variety







of structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [23].


NU
l00320
10
10
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification






cytochrome p-450
constituent proteins of the liver mixed function
Response/






isoform
monooxygenases. They play a central role in
Biotransformation-






(phenobarbital-
the metabolism of steroids, the detoxification of
TOX






inducible, 2B1, or
drugs and xenobiotics, and the activation of






2B2) [J00719]
procarinogens. In liver microsomes, this






[P00176] [P04167].
enzyme is involved in an NADPH-dependent







electron transport pathway. It oxidizes a variety







of structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [23].


NU
m11251
11
11
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification






cytochrome p-450
constituent proteins of the liver mixed function
Response/






isoform
monooxygenases. They play a central role in
Biotransformation-






(phenobarbital-
the metabolism of steroids, the detoxification of
TOX






inducible, 2B1, or
drugs and xenobiotics, and the activation of






2B2) [Q64584]
procarcinogens. In liver microsomes, this






P00176] [P04167].
enzyme is involved in an NADPH-dependent 







electron transport pathway. It oxidizes a variety







of structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [23].


NU
m26125
12
12
Rattus norvegicus
Epoxide formation from drugs, chemicals, food
Detoxification






epoxide hydrolase
additives and environmental pollutants is
Response/






[M26125].
catalyzed by cytochrome P-450 dependent
Biotransformation-







monooxygenase(s). Epoxides are converted to
TOX







glycols or dihydrodiols by epoxide hydrolase.







These enzymes are known to be present in the







microsomes of different mammalian tissues and







in the hepatic nuclei from rats and humans. The







balance between the epoxide forming (AHH)







and metabolizing (EH) enzyme activities may







provide information about the “epoxide







exposure” of a tissue [Kuklin, 1976 #2].


NU
m34452
13
13
Rattus norvegicus
Epoxide formation from drugs, chemicals, food
Detoxification






cytochrome P450e-L
additives and environmental pollutants is
Response/






(P450IIB2) [M34452].
catalyzed by cytochrome P-450 dependent
Biotransformation-







monooxygenase(s). Epoxides are converted to
TOX







glycols or dihydrodiols by epoxide hydrolase.







These enzymes are known to be present in the







microsomes of different mammalian tissues and







in the hepatic nuclei from rats and humans. The







balance between the epoxide forming (AHH)







and metabolizing (EH) enzyme activities may







provide information about the “epoxide







exposure” of a tissue [Kuklin, 1976 #2].


NU
u33546
14
14
Rattus norvegicus
CYP2B16P is an apparent pseudogene in the
Detoxification






CYP2B16P
rat cytochrome P450 2B (CYP2B) subfamily
Response/






[CAB35441].
[24].
Biotransformation-








TOX


NU
x74673
15
15
Rattus norvegicus
Aflatoxin B1 aldehyde reductase/Succinic
Detoxification






aflatoxin B1 aldehyde
semialdehyde reductase is believed to be
Response/






reductase (AFAR)
involved in the detoxification of xenobiotic
Biotransformation-






[X74673].
carbonyl compounds [25].
TOX


SP
scr_gb-
16
16
Rattus norvegicus
MHC class II molecules at the surface of
Immunity And



x13044—


MHC-associated
antigen presenting cells present antigenic
Defense



4


invariant chain
peptides to CD4+ T helper cells. Ii plays a






gamma (Ia antigen-
critical role in MHC class II antigen processing






associated invariant
by stabilizing peptide-free class II alpha/beta






chain) (Ii) [X13044].
heterodimers [26].


SP
scr_gb-
17
17
Rattus norvegicus
MHC class II molecules at the surface of
Immunity And



x14254—


MHC-associated
antigen presenting cells present antigenic
Defense



5


invariant chain
peptides to CD4+ T helper cells. Ii plays a






gamma (Ia antigen-
critical role in MHC class II antigen processing






associated invariant
by stabilizing peptide-free class II alpha/beta






chain) (Ii) [X14254].
heterodimers [26].


NU
scr_gb-
18
18
Rattus norvegicus
Rab proteins form the largest branch of the Ras
Intracellular



bi27563


Ras-related protein
superfamily of GTPases. They are localized to
Transport



8_1


Rab-2 [P05712].
the cytoplasmic face of organelles and vesicles







involved in the biosynthetic/secretory and







endocytic pathways in eukaryotic cells [27].



scr_gb-
19
19
Rattus norvegicus
The annexins are a family of proteins that bind
Intracellular



x66871—


calpactin I heavy
acidic phospholipids in the presence of Ca2+.
Transport



3


chain (annexin II)
Because annexin II bridge secretory granules to






[X66871].
plasma membrane it has suggested that this







protein may play a role in Ca(2+)-dependent







exocytosis. Annexin II tetramer has also been







demonstrated on the extracellular face of some







metastatic cells where it mediates the binding of







certain metastatic cells to normal cells. Annexin







II tetramer is a major cellular substrate of







protein kinase C and pp60src [28].


NU
scr_gb-
20
20
Rattus norvegicus
cMOAT mediates the hepatobiliary excretion of
Oxidative Stress-



l49379—


canalicular
numerous organic anions. It has been shown
TOX



3


multispecific organic
that both multidrug resistance-associated






anion transporter
protein (MRP1) and canalicular multispecific






(cMOAT) [L49379].
organic anion transporter (cMOAT/MRP2) have







the ability to extrude glutathione conjugates







(GS-X pump activity) from cells [29] [30].


SP
scr_sc-
21
21
Rat gene fragment -
Inter-alpha-trypsin inhibitor (ITI) is a complex
Other



1326905


775 bp. 85% SI
protein containing two heavy polypeptide chains



01_1


(618/722) to Homo
(H1 and H2) and a light chain, which in the free






sapiens inter-alpha-
state is known as bikunin [31]. ITI is a 220 kDa






trypsin inhibitor heavy
serine proteinase inhibitor found in human






chain IIH1 [X63652].
serum [32].


NC
scr_gb-
22
22
Rat gene fragment -
Serine proteinase inhibitors (serpins) are
Other



aw1417


1561 bp. 98% SI
classically regulators of extracellular proteolysis.



35_3


(1002/1022) to Mus
Evidence suggests that some function






musculus serine
intracellularly as well [33].






proteinase inhibitor






mBM2A [U96701].


SP
af18498
23
23
Rattus norvegicus
Osteoactivin cDNA was recently isolated from
Unknown



3


osteoactivin
long bone and calvaria. In primary rat






[AF184983].
osteoblast cultures it was expressed at the







highest levels during the later stages of matrix







maturation and mineralization and correlated







with the expression of alkaline phosphatase and







osteocalcin. [34].


NU
scr_cg-
24
24
Unknown, 241 bp.

Novel



2251067



4_1


NU
scr_cg-
25
25
Unknown, 283 bp.

Novel



5721522



4_1


NU
scr_gb-
26
26
Unknown, 642 bp.

Novel



aa85076



7_2


NU
scr_gb-
27
27
Unknown, 866 bp.

Novel



ai01199



4_2


NU
scr_gb-
28
28
Unknown, 629 bp.

Novel



aw1422



93_1


NU
scr_gb-
29
29
Unknown, 1145 bp.

Novel



bm3833



27_1


NU
scr_gb-
30
30
Unknown, 3087 bp.

Novel



bm3866



25_1


NU
scr_sc-
31
31
Unknown, 434 bp.

Novel



1335569



69_1


NU
scr_sc-
32
32
Unknown, 221 bp.

Novel



1701427



36_1


NU
scr_sc-
33
33
Unknown, 581 bp.

Novel



2563586



_2


NU
scr_sc-
34
34
Unknown, 221 bp.

Novel



8761825



7_1









Example 5
Identification of Zone 3 Necrosis Related Genes In Vitro


    
    
        SP=Secreted Protein
        NC=Novel Rat Composition
        NU=Novel Rat Utility
    
    
















TABLE 4








TOX








MARKER




ASSIGN-
SEQ ID



ACCNO
MENT
NO
Definition
Description
Bin






















NU
cszr_96561
35
35
Rattus norvegicus
Mitochondrial protein involved in the urea acid cycle
Amino Acid



134_837604


Carbamoyl-
of ureotelic animals where the enzyme plays an
Metabolism



93


phosphate
important role in removing excess ammonia from






synthase
the cell. Catalytic Activity: 2 ATP + NH(3) + CO(2)






[ammonia]
+ H(2)O = 2 ADP + ORTHOPHOSPHATE +






(CPSASE I),
CARBAMOYL PHOSPHATE [17].






mitochondrial






precursor






[P07756].


NU
scr_gb-
36
36
Rattus norvegicus
EP3 receptors for Prostaglandin (PG) E(2) are
Carbohydrate



x83855_1


hepatocyte
primarily involved in inhibition of adenylyl cyclase
Metabolism






EP3alpha
via G(i) activation, and in Ca(2+)-mobilization






receptor [X83855].
through Gbetagamma from G(i). Along with G(i)







activation, the EP3 receptor can stimulate cAMP







production via G(s) activation [18].


NU
cszr_229800
37
37
Rattus norvegicus
Enolase is a vital enzyme of the glycolytic pathway.
Carbohydrate



465_190907


non-neuronal
It exists mainly in two forms, non-neuronal enolase
Metabolism



286


enolase (NNE)
(NNE) and neuron specific enolase (NSE).






(alpha-alpha
Catalytic Activity: 2-phospho-D-glycerate =






enolase, 2-
phosphoenolpyruvate + H(2)O [19].






phospho-D-






glycerate






hydrolase






[X02610].


SP
scr_gb-
38
38
Rat gene
Two genes, HEXA and HEXB, encode the alpha-
Carbohydrate



bi277612_1


fragment - 1381
and beta-subunits, respectively, of human beta-
Metabolism






bp. 89% SI
hexosaminidase. In the mouse, the corresponding






(816/910) to Mus
genes are termed Hexa and Hexb. The subunits






musculus for
have the capacity to degrade a variety of substrates






beta-
including oligosaccharides, glycosaminoglycans,






hexosaminidase
and glycolipids containing beta-linked N-






[Y00964].
acetylglucosaminyl or N-galactosaminyl residues







[20].


NU
scr_gb-
39
39
Rattus norvegicus
H+-ATP synthase catalyzes the synthesis and/or
Energy



j05266_3


mitochondrial H+-
hydrolysis of ATP [21].
Metabolism






ATP synthase






alpha subunit






[J05266].


NU
scr_gb-
40
40
Rattus norvegicus
Egfr is involved in the initiation of oncogenic effect
Cell Cycle



m37394_5


epidermal growth
such as DNA synthesis, enhanced cell growth,
Regulation






factor receptor
invasion, and metastasis. Specific abrogation of
(Regulation Of






(Egfr) [M37394].
EGFR results in cell cycle arrest, apoptosis, or
Proliferation)







dedifferentiation of cancer cells [22].


NU
scr_gb-
41
41
Rattus norvegicus
The Raf/MEK/ERK signaling was the first MAP
Cell Cycle



m64300_4


extracellular
kinase cascade to be characterized. It is probably
Regulation






signal-related
one of the most well known signal transduction
(Regulation Of






kinase (ERK2)
pathways among biologists because of its
Proliferation)






[M64300].
implication in a wide variety of cellular functions as







diverse- and occasionally contradictory- as cell







proliferation, cell-cycle arrest, terminal differentiation







and apoptosis [23].


SP
scr_gb-
42
42
Rat gene
IFNAR-2, is expressed ubiquitously, and exists as
Cell Cycle



bi294409_1


fragment - 526 bp.
both transmembrane and soluble forms. Recent
Regulation






88% SI (313/355)
evidence suggests murine IFNAR-2 as an efficient
(Regulation Of






to Mus musculus
regulator of IFN responses. Type I interferons are
Proliferation)






type I interferon
cytokines that are important in defense against viral






receptor soluble
infections well as in the control of cell proliferation






isoform precursor
[24] [25].






(IFNAR2)






[AF013486].


NU
scr_gb-
43
43
Rattus norvegicus
IL-4 is a pleiotropic cytokine which plays a pivotal
Cell Cycle



ab015747_3


interleukin-4
role in shaping immune responses. The effects of
Regulation






receptor
IL-4 are mediated after binding to high affinity
(Regulation Of






(membrane-bound
receptor complexes present on hematopoietic as
Proliferation)






form) [AB015747].
well as non-hematopoietic cells. There is also







evidence that IL-4 interaction with its receptor leads







to signal transduction mechanisms that result in







cellular proliferation and/or gene activation [26].


NU
scr_sc-
44
44
Rattus norvegicus
The Crk-associated substrate (Cas) is a unique
Cell Cycle



191879433—


Crk-associated
docking protein with a Src homology 3 (SH3)
Regulation



1


substrate, p130
domain. Aberrant CAS tyrosine phosphorylation
(Regulation Of






[D29766].
may contribute to cell transformation by certain
Proliferation)







oncoproteins, including v-Crk and v-Src, and to







tumor growth and metastasis [27] [28].


NC
scr_sr-
45
45
Rat gene
Diacylglycerol kinase (DGK) plays an important role
Cell Cycle/



140438096—


fragment - 383 bp.
in the signal transduction through modulating the
Proliferation



1


98% SI (125/127)
balance between two signaling lipids, diacylglycerol
(Basic






to Homo sapiens
and phosphatidic acid. Diacylglycerol is a protein
Machinery)






Diacylglycerol
kinase c activator. Thus, DGK is considered to






kinase, delta
regulate protein kinase C activity through the






[Q16760].
reduction of diacylglycerol [29] [30].


NU
scr_gb-
46
46
Rattus norvegicus
Neurotensin is a 13-amino acid hormonal peptide
Cellular



x87157_5


neurotensin
which was first isolated from bovine hypothalamus.
Communication






endopeptidase
It is present in the digestive tract as well as in the






[X87157].
central nervous system. It has a variety of biological







activities as a central neurotransmitter or







neuromodulator, and a peripheral hormone [20].


NU
scr_gb-
47
47
Rattus norvegicus
Densin-180 is a transmembrane protein that is
Cellular



u66707_2


densin-180
strongly associated with the postsynaptic density in
Communication






[U66707].
CNS neurons and is believed to function as a







synaptic adhesion molecule [31].


NU
scr_gb-
48
48
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification



af017393_2


cytochrome
constituent proteins of the liver mixed function
Response/






P4502F4
monooxygenases. They play a central role in the
Biotransformation-






(CYP4502F4)
metabolism of steroids, the detoxification of drugs
TOX






[AF017393].
and xenobiotics, and the activation of







procarcinogens. In liver microsomes, this enzyme







is involved in an NADPH-dependent electron







transport pathway. It oxidizes a variety of







structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [32].


NU
scr_sc-
49
49
Rattus norvegicus
The cytochromes P-450 are among the major
Detoxification



134241980—


cytochrome P450
constituent proteins of the liver mixed function
Response/



1


2B3 (CYP2B3)
monooxygenases. They play a central role in the
Biotransformation-






[U16214].
metabolism of steroids, the detoxification of drugs
TOX







and xenobiotics, and the activation of







procarcinogens. In liver microsomes, this enzyme







is involved in an NADPH-dependent electron







transport pathway. It oxidizes a variety of







structurally unrelated compounds, including







steroids, fatty acids, and xenobiotics [32].


NC
scr_sc-
50
50
Rat gene
DNA polymerase III is a replicative enzyme known
DNA Metabolism



191609675—


fragment - 217 bp.
to be essential in the DNA synthesis of Gram-



1


94% SI (67/71) to
positive bacteria [33].






Streptococcus






pneumoniae DNA






polymerase III,






alpha subunit R6






[AAK99055].


NU
scr_gb-
51
51
Rat OX40 antigen
OX40 is a member of the tumor necrosis factor
Immunity And



x17037_2


[X17037].
family which is expressed by activated T
Defense







lymphocytes [34].


SP
scr_gb-
52
52
Rat gene
Fc gamma BP is widely expressed on mucosal
Immunity And



bi291805_1


fragment - 528 bp.
surfaces and in external secretions lending support
Defense






76% SI (290/380)
to the concept that Fc gamma BP is an important






to Homo sapiens
component of mucosal immunological defenses






IgG Fc binding
[35].






protein [D84239].


NU
scr_gb-
53
53
Rattus norvegicus
The proteins of the kinesin superfamily (KIFs) are
Intracellular



aj000696_5


novel kinesin-
microtubule-based molecular motors whose
Transport






related protein,
functions include the transport of membrane-bound






KIF1D
organelles. The KIF1 subfamily members are






[AJ000696].
monomeric and contain a number of amino acid







inserts in surface loops [36].


NU
scr_gb-
54
54
Rattus norvegicus
r-sly1 is a mammalian homologue to yeast Sly1p
Intracellular



d79221_3


r-sly1 [U35364].
which plays a critical role in endoplasmic reticulum
Transport







to Golgi apparatus vesicle trafficking [37].


NU
m61937
55
55
Rattus norvegicus
Dihydrodiol dehydrogenase(s) (DD) have been
Oxidative Stress-






dihydrodiol
implicated in the detoxication of proximate (trans-
TOX






dehydrogenase
dihydrodiol) and ultimate carcinogenic (anti-diol-






[M61937].
epoxide) metabolites of polycyclic aromatic







hydrocarbons (PAHs). Although this pathway







suppresses the formation of the PAH anti- and syn-







diol epoxides (ultimate carcinogens), the process of







autoxidation is anticipated to yield reactive oxygen







species (ROS) [38].


NU
cszr_229602
56
56
Rattus norvegicus
Metallothionein (MT) is a small, cysteine-rich, metal-
Oxidative Stress-



935_183895


metallothionein-i
binding protein. MT synthesis is induced by various
TOX



355


(mt-1) [J00750].
stimuli such as heavy metals, oxidative stress,







anticancer drugs and fasting stress. MT is capable







of not only reducing metal toxicity but also







scavenging free radicals [39].


NU
scr_gb-
57
57
Rattus norvegicus
Peroxiredoxins are novel family of anti-oxidative
Oxidative Stress-



af106944_3


Peroxiredoxin III
proteins comprise six members in mammals. They
TOX






[AF106944].
share a common reactive Cys residue in the N-







terminal region, and are capable of serving as a







peroxidase and involve thioredoxin and/or







glutathione as the electron donor [40].


NU
scr_gb-
58
58
Rattus norvegicus
Metallothionein (MT) is a small, cysteine-rich, metal-
Oxidative Stress-



m11794_3


metallothionein-2
binding protein. MT synthesis is induced by various
TOX






and
stimuli such as heavy metals, oxidative stress,






metallothionein-1
anticancer drugs and fasting stress. MT is capable






genes [M11794].
of not only reducing metal toxicity but also







scavenging free radicals [39].


NU
scr_gb-
59
59
Rattus norvegicus
Transaldolase is a key enzyme of the reversible
Oxidative Stress-



af069306_1


transaldolase
nonoxidative branch of the pentose phosphate
TOX






[AF069306].
pathway (PPP) that is responsible for the generation







of NADPH to maintain glutathione at a reduced







state (GSH) and, thus, to protect cellular integrity







from reactive oxygen intermediates (ROIs) [41].


NU
scr_gb-
60
60
Rattus norvegicus
Steroid 3-alpha-dehydrogenase is an important
Oxidative Stress-



d17310_4


steroid 3-alpha-
multifunctional oxidoreductase capable of
TOX






dehydrogenase
metabolizing steroid hormones, polycyclic aromatic






[D17310].
hydrocarbons, and prostaglandins. It is also







required for bile acid synthesis and has been







suggested to play an important role in net bile acid







transport across the hepatocyte [42].


NC
scr_gb-
61
61
Rat gene
PRT1 is a component of translation initiation factor
Protein



bf281368_2


fragment - 1086
elF-3 and originally discovered in Saccharomyces
Metabolism






bp. 80% SI
cerevisiae [43].






(754/938) to






Human Prt1






homolog






[U62583].


NU
scr_gb-
62
62
Rattus norvegicus
Ubiquitin-conjugating enzymes (UBC) catalyze the
Protein



u56407_3


ubiquitin
covalent attachment of ubiquitin to target proteins
Metabolism






conjugating
and are distinguished by the presence of a UBC






enzyme
domain required for catalysis [44].






[U56407].


NC
scr_gb-
63
63
Rat gene
Heterogeneous nuclear ribonucleoprotein (hnRNP)
RNA metabolism



ai406674_1


fragment - 796 bp.
complexes, the structures that contain






96% SI (634/660)
heterogeneous nuclear RNA and its associated






Mus musculus
proteins, constitute one of the most abundant






heterogeneous
components of the eukaryotic nucleus. hnRNPs






nuclear
appear to play important roles in the processing,






ribonucleoprotein
and possibly also in the transport, of mRNA [45].






C, clone






MGC:5715






IMAGE:3499283






[BC004706].


NC
scr_gb-
64
64
Rat gene
Heterogeneous nuclear ribonucleoprotein (hnRNP)
RNA metabolism



bf290678_2


fragment - 716 bp.
complexes, the structures that contain






84% SI (542/643)
heterogeneous nuclear RNA and its associated






to Mus musculus
proteins, constitute one of the most abundant






heterogeneous
components of the eukaryotic nucleus. hnRNPs






nuclear
appear to play important roles in the processing,






ribonucleoprotein
and possibly also in the transport, of mRNA [45].






G, splice variant 1






[AJ237847].


NC
scr_gb-
65
65
Rat gene
Mouse Clp-1 is a potential cardiac transcriptional
RNA Metabolism



bi288503_1


fragment - 456 bp.
regulatory factor [46].






94% SI (430/456)






to Mus musculus






cardiac lineage






protein 1 (Clp1)






[AY090614].


NU
scr_gb-
66
66
Rattus norvegicus
Hex is a homeobox protein which is believed to
RNA metabolism



d86383_2


Hex [D86383].
function as a transcriptional repressor and may be







involved in the differentiation and/or maintenance of







the differentiated state in hepatocytes [47].


NU
scr_sc-
67
67
Rattus norvegicus
Thymosin beta 10 is one of a small family of
Structural Repair-



133366194—


thymosin beta-10
proteins closely related in sequence to thymosin
TOX



1


[M58405].
beta 4, recently identified as an actin-sequestering







protein [48].


NC
cszr_230290
68
68
Rat gene
FEM-1 is a signal-transducing regulator in the C.
Other



139_182026


fragment - 249 bp.
elegans sex-determination pathway. The existence



368


91% SI (227/248)
of FEM-1 homologs in the mouse raises the






to Mus musculus
possibility that evolutionary conservation of ancient






sex-determination
FEM-1 signaling interactions may play a role in






protein homolog
vertebrate cell-fate determination [49].






Fem1a






[AF064447].


NU
scr_gb-
69
69
Rattus norvegicus
VL30 elements constitute a family of
Other



ai013477_2


VL30 element
retrotransposons that are associated with cancer by






[M91234].
their overexpression in rodent malignancies, their







induction in a fibroblast response to anoxia which







shares features with the malignant phenotype, and







their presence recombined into Harvey murine







sarcoma virus (HaSV) and Kirsten murine sarcoma







virus [50].


NU
scr_gb-
70
70
Rattus norvegicus
VL30 elements constitute a family of
Other



m91235_3


VL30 element
retrotransposons that are associated with cancer by






[M91235].
their overexpression in rodent malignancies, their







induction in a fibroblast response to anoxia which







shares features with the malignant phenotype, and







their presence recombined into Harvey murine







sarcoma virus (HaSV) and Kirsten murine sarcoma







virus [50].


NU
cszr_204152
71
71
Unknown, 63 bp.

Novel



648_191521



095


NU
cszr_204152
72
72
Unknown, 133 bp.

Novel



792_191517



979


NU
cszr_204229
73
73
Unknown, 124 bp.

Novel



614_191891



958


NU
cszr_204229
74
74
Unknown, 124 bp.

Novel



615_191892



510


NU
scr_gb-
75
75
Unknown, 1252

Novel



aa801331_1


bp.


NU
scr_gb-
76
76
Unknown, 1241

Novel



aa899865_3


bp.


NU
scr_gb-
77
77
Unknown, 396 bp.

Novel



aa997629_1


NU 
scr_gb-
78
78
Unknown, 473 bp.

Novel



aa997691_1


NU
scr_gb-
79
79
Unknown, 1221

Novel



ai411514_4


bp.


NU
scr_gb-
80
80
Unknown, 695 bp.

Novel



aw142560_3





NU
scr_gb-
81
81
Unknown, 771 bp.

Novel



aw533305_2





NU
scr_gb-
82
82
Unknown, 2262

Novel



aw915573_2


bp.


NU
scr_gb-
83
83
Unknown, 422 bp.

Novel



be108509_1


NU
scr_gb-
84
84
Unknown, 445 bp.

Novel



be111483_1


NU
scr_gb-
85
85
Unknown, 482 bp.

Novel



be120910_1


NU
scr_gb-
86
86
Unknown, 784 bp.

Novel



bf285287_1


NU
scr_gb-
87
87
Unknown, 486 bp.

Novel



bf390383_1


NU
scr_gb-
88
88
Unknown, 921 bp.

Novel



bf558463_2


NU
scr_gb-
89
89
Unknown, 525 bp.

Novel



bf560709_1


NU
scr_gb-
90
90
Unknown, 930 bp.

Novel



bg662990_2


NU
scr_gb-
91
91
Unknown, 1060

Novel



bi278552_1


bp.


NU
scr_gb-
92
92
Unknown, 1158

Novel



bi278749_1


bp.


NU
scr_gb-
93
93
Unknown, 1241

Novel



bi295938_1


bp.


NU
scr_gb-
94
94
Unknown, 2695

Novel



bi296376_1


bp.


NU
scr_gb-
95
95
Unknown, 2423

Novel



bm384392_1


bp.


NU
scr_gb-
96
96
Unknown, 610 bp.

Novel



bm387477_1


NU
scr_gb-
97
97
Unknown, 1047

Novel



bm986259_1


bp.


NU
scr_gb-
98
98
Unknown, 1191

Novel



s69874_5


bp.


NU
scr_sc-
99
99
Unknown, 384 bp.

Novel



119263563—



1


NU
scr_sc-
100
100
Unknown, 181 bp.

Novel



132556005_



1


NU
scr_sc-
101
101
Unknown, 130 bp.

Novel



132570828_



1


NU
scr_sc-
102
102
Unknown, 50 bp.

Novel



132947646_



1


NU
scr_sc-
103
103
Unknown, 296 bp.

Novel



133387221_



1


NU
scr_sc-
104
104
Unknown, 321 bp.

Novel



133555783_



1


NU
scr_sc-
105
105
Unknown, 92 bp.

Novel



133678871_



1


NU
scr_sc-
106
106
Unknown, 94 bp.

Novel



133725675_



1


NU
scr_sc-
107
107
Unknown, 343 bp.

Novel



133955481—



1


NU
scr_sc-
108
108
Unknown, 238 bp.

Novel



134521597—



1


NU
scr_sc-
109
109
Unknown, 247 bp.

Novel



172126480_



1


NU
scr_sc-
110
110
Unknown, 196 bp.

Novel



172130231_



1


NU
scr_sc-
111
111
Unknown, 457 bp.

Novel



172755010—



1


NU
scr_sc-
112
112
Unknown, 85 bp.

Novel



188295137_



1


NU
scr_sc-
113
113
Unknown, 241 bp.

Novel



190079504—



1


NU
scr_sc-
114
114
Unknown, 388 bp.

Novel



191455923—



1


NU
scr_sc-
115
115
Unknown, 444 bp.

Novel



195460151—



1


NU
scr_sc-
116
116
Unknown, 135 bp.

Novel



198205946—



1


NU
scr_sc-
117
117
Unknown, 246 bp.

Novel



2573087_1


NU
scr_sc-
118
118
Unknown, 203 bp.

Novel



2585074_1


NU
scr_sc-
119
119
Unknown, 233 bp.

Novel



8571871_2


NU
scr_sc-
120
120
Unknown, 300 bp.

Novel



87731837_1


NU
scr_sc-
121
121
Unknown, 351 bp.

Novel



87869413_1


NU
scr_gb-
122
122
Unknown, 889 bp.

Novel



ai233262_2


NU
cgrrs0h0310.
123
123
Unknown, 310 bp.

Novel



9_13952-135


NU
scr_gb-
124
124
Unknown, 100 bp.

Novel



m13100.5_2


NU
scr_sc-
125
125
Unknown, 350 bp.

Novel



170396977—



1


NU
scr_sc-
126
126
Rat gene patent

Unknown



14059147_2


WO0210453, 254






bp.


NU
scr_sc-
127
127
Rat gene patent

Unknown



87750810_1


WO0210453,






1063 bp.


NU
cszr_202034
128
128
Rattus norvegicus

Unknown



260_190929


Tclone4 [U30788].



676









Using the TOXMARKER gene information listed in Table 4, zone 3 necrosis-related genes expressed in vitro were confirmed. Confirmed genes are listed in Table 5.












TABLE 5






TOX
SEQ ID





Gene ID
Number
NO
Definition
Human Ortholog
SEQ ID NO:















Amino Acid Metabolism












cszr_96561134—
35
39
Rattus
CPS1: Carbamyl phosphate synthetase I [D90282,
129


83760493


norvegicus
NM_001875]





Carbamoyl-





phosphate





synthase





[ammonia]





(CPSASE I),





mitochondrial





precursor





[P07756].







Carbohydrate Metabolism












scr_gb-
38
38
Rat gene
NM_000521
130


bi277612_1


fragment - 1381
>rshd:REFSEQHUMANDNA-





bp. 89% SI
ID:NM_000521|acc:NM_000521





(816/910) to Mus
/geneName=“HEXB” /definition=“Homo sapiens





musculus for
hexosaminidase B (beta polypeptide) (HEXB),





beta-
mRNA.” /protein_id=“NP_000512.1”





hexosaminidase
/organism=“Homo sapiens” /CDS=“76 . . . 1746”, 1857





[Y00964].
bp.






>gb:GENBANK-ID:AF378118|acc:AF378118.1






Homo sapiens cervical cancer proto-oncogene 7






mRNA, complete cds - Homo sapiens, 1892 bp.


cszr_229800465—
129
131
Rat non-neuronal


190907286


enolase (NNE)





(alpha-alpha





enolase, 2-





phospho-D-





glycerate





hydrolase)





[X02610].







Energy Metabolism












scr_gb-j05266_3
39
.39
Rattus
NM_004046
132





norvegicus
>rshd:REFSEQHUMANDNA-





mitochondrial H+-
ID:NM_004046|acc:NM_004046





ATP synthase
/geneName=“ATP5A1” /definition=“Homo sapiens





alpha subunit
ATP synthase, H+ transporting, mitochondrial F1





[J05266].
complex, alpha subunit, isoform 1, cardiac muscle






(ATP5A1), mRNA.” /protein_id=“NP_004037.1”






/organism=“Homo sapiens” /CDS=912 . . . 2573”,






2725 bp.







Cell Cycle Regulation (Regulation Of Proliferation)












scr_gb-
43
43
Rat interleukin-4
X52425.1 Human IL-4-R mRNA for the interleukin 4
133


ab015747_3


receptor
receptor





(membrane-





bound





form) (AB015747.1:





99%/3501,





p = 0.000000),





3520 bp.


scr_gb-
122
122
Rat gene
weakly similar to PUTATIVE IMPORTIN BETA-4
134


ai233262_2


fragment, 889 bp,
SUBUNIT [AK027871, NM_024658].





93% identical





over 679 bp to





mouse RANBP4





[AF123388].


scr_gb-
42
42
Rat gene
Ifnar2: interferon receptor ifnar2-1 [L41944,
135


bi294409_1


fragment, 526 bp,
NM_000874]





89% SI (327/365)





to mouse type I





interferon





receptor, IFNaR2





subunit, 1109 bp





(Y09813).


scr_gb-m37394_5
40
40
Rattus
EGFR: epidermal growth factor receptor [K03193,
136





norvegicus
NM_005228].





epidermal growth





factor receptor





(Egfr) [M37394].


scr_gb-m64300_4
41
41
Rattus
MAPK1: Mitogen-activated protein kinase 1
137





norvegicus
[BC017832, NM_002745].





extracellular





signal-related





kinase (ERK2)





[M64300].


scr_sc-
44
44
Rat Crk-
BCAR1: breast cancer anti-estrogen resistance 1
138


191879433_1


associated
[AK026121, NM_014567].





substrate, p130





[D29766].







Cellular Communication












scr_gb-
124
124
Rattus




m13100.5_2


norvegicus gene





for histamine N-





methyltransferase,





exon 1 and 2





[AB007833].


scr_gb-x87157_5
46
46
Rattus
NLN: neurolysin [AJ300837, NM_020726].
139





norvegicus





neurotensin





endopeptidase





[X87157].







Detoxification Response/Biotransformation-TOX












scr_gb-
48
48
Rattus
CYP2F1: cytochrome P450, subfamily IIF,
140


af017393_2


norvegicus
polypeptide 1[J02906, NM_000774].





cytochrome





P4502F4





(CYP4502F4)





[AF017393].


scr-sc-
49
49
Rat cytochrome
MSRA: methionine sulfoxide reductase A
141


134241980_1


P450IIB3
[AJ242973, NM_012331].





(P450IIB





subfamily)





mRNA, complete





cds (M20406.1:





100%/279,





p = 5.0e−156), 367





bp.







DNA Metabolism












scr_gb-
94
94
Rat ribosomal




bi296376_1


DNA external





transcribed





spacer 1 (ETS1)





[X16321], Rat





45S rDNA gene





transcription





initiation region





[X00677], and





Rat ribosomal





RNA 1.6 small





subunit (SS1.6)





gene, 3′ end





[M30843] - 2695





bp.







Immunity And Defense












scr_gb-x17037_2
51
51
Rat OX40 antigen
TNFRSF4: tumor necrosis factor receptor
142





[X17037].
superfamily, member 4






[AW293499, S76792, X75962, NM_003327].


cgrrs0h0310.9_13
123
123
Rat lipocortin-V


952-135


mRNA, complete





cds (M21730.1:





99%/1419,





p = 0.000000),





1744 bp.







Intracellular Transport












scr_gb-
53
53
Rattus
KIF1C: kinesin family member 1C [AB014606,
143


aj000696_5


norvegicus novel
NM_006612].





kinesin-related





protein, KIF1D





[AJ000696].


scr_gb-d79221_3
54
54
Rattus
Vesicle transport-related protein [AF110646,
144





norvegicus r-sly1
NM_016163].





[U35364].







Oxidative Stress-TOX












cszr_229602935—
56
56
Rat




183895355


metallothionein-i





(mt-1)


m61937
55
55
Rattus





norvegicus





dihydrodiol





dehydrogenase





[M61937].


scr_gb-
59
59
Rattus
clone MGC:2838 IMAGE:2966784 [BC001517].
145


af069306_1


norvegicus





transaldolase





[AF069306].


scr_gb-
57
57
Rattus


af106944_3


norvegicus





Peroxiredoxin III





[AF106944].


scr_gb-d17310_4
60
60
Rattus
MTCH2: mitochondrial carrier homolog 2
146





norvegicus
[AF085361, NM_014342]





steroid 3-alpha-





dehydrogenase





[D17310].


scr_gb-m11794_3
58
58
Rattus
MT2A: metallothionein 2A [S52379, NM_005953].
147





norvegicus





metallothionein-2





and





metallothionein-1





genes [M11794].







Protein Metabolism












scr_gb-
61
61
Rat gene
eIF3: eukaryotic translation initiation factor 3
148


bf281368_2


fragment - 1086
[U78525, NM_003751]





bp. 80% SI





(754/938) to





Human Prt1





homolog





[U62583].







RNA metabolism












scr_gb-
63
63
Rat gene
HNRPC: heterogeneous nuclear ribonucleoprotein
149


ai406674_1


fragment, 796 bp.
C [BC003394, NM_004500].





91% SI to human





heterogeneous





nuclear





ribonucleoprotein





C (C1/C2),





[XM_166936]


scr_gb-
65
65
Rat gene
HEMIX1 protein [AB021179, NM_006460].
150


bi288503_1


fragment 456 bp.





94% SI (430/456)





to Mus





musculus cardiac





lineage protein 1





(Clp1)





(AY090614.1)


scr_gb-d86383_2
66
66
Rattus
HHEX: hematopoietically expressed homeobox
151





norvegicus Hex
[BC015110, X67235, NM_002729].





[D86383].







Membrane Transport/Transporters












scr_sc-
119
119
Rattus
U90545.1|HSU90545 Human sodium phosphate
152


8571871_2


norvegicus Na/Pi
transporter (NPT4) mRNA, complete cds





[AB025224.1].







Other












scr_gb-
130
153
Rattus




af311311_2


norvegicus





P116RIP mRNA,





complete cds





(AF311311.1:





99%/3285,





p = 0.000000),





3905 bp.


scr_bg-
69
69
Rattus


ai013477_2


norvegicus VL30





element





[M91234].


scr_gb-
97
97
Rat gene
HSPC155: hypothetical protein HSPC155
154


bm986259_1


fragment, 1047
[AF161504, NM_016406]





bp, 86% SI





(647/748) to





human CGI-126





protein mRNA,





1091 bp





(AF151884).


cszr_230290139—
131
155
Similar to Mus


182026368


musculus,





feminization 1 a





homolog (C.





elegans), clone





MGC:6309





IMAGE:2811079,





mRNA, complete





cds





(BC009161.1:





91%/244, p = 3.4e−





085), 249 bp.


scr_gb-m91235_3
70
70
Rattus





norvegicus VL30





element





[M91235].







Novel












scr_gb-
75
75
Unknown, 1252




aa801331_1


bp.


scr_gb-
76
76
Unknown, 1241


aa899865_3


bp.


scr_gb-
79
79
Unknown, 1221
KIAA0652 gene product [BC006191, NM_014741].
156


ai411514_4


bp.


scr_gb-
88
88
Rat gene
Clone IMAGE:4052356, partial cds. [BC014348].
157


bf558463_2


fragment, 921 bp.


scr_gb-
91
91
Unknown, 1060
KIAA0427 gene product [AB007887, NM_014772].
158


bi278552_1


bp.


scr_gb-
83
83
Unknown, 1241
Hypothetical protein FLJ13409 [BC015897,
159


bi295938_1


bp.
NM_024617].


scr_gb-
85
85
Unknown, 2423
Hypothetical protein MGC3067 [BC002457,
160


bm364392_1


bp.
NM_024295].


scr_gb-s69874_5
98
98
Unknown, 1191
COL4A1: collagen, type IV, alpha 1 [AH002741,
161





bp.
NM_001845].


scr_sc-
102
102
Rat gene


132947646_1


fragment, 50 bp.


scr_sc-
109
109
Unknown, 247


172126480_1


bp.


scr_sc-
112
112
Unknown Gene,


188295137_1


85 bp.


scr_sc-
114
114
Unknown, 388
Heterogeneous nuclear ribonucleoprotein M
162


191455923_1


bp.
[NM_005968].


scr_sc-
115
115
Unknown, 444


195460151_1


bp.







Unknown












cszr_204152648—
71
71
UI-R-A1-ek-e-09-
XM_114110
163


191521095


0-UI.s1 UI-R-A1
>rshd:REFSEQHUMANDNA-





Rattus
ID:XM_114110|acc:XM_114110





norvegicus cDNA
/geneName = “LOC200081” /definition=“Homo





clone UI-R-A1-
sapiens similar to muscle-derived protein MDP77





ek-e-09-0-UI 3′,
isoform 2 (LOC200081), mRNA.”





63 bp.
/protein id = “XP_114110.1” /organism = “Homo






sapiens” /CDS = “94 . . . 873”, 3962 bp.


cszr_204152792—
72
72
Rat gene
AB032968.1 Homo sapiens mRNA for KIAA1142
164


191517979


fragment, 131 bp,
protein, partial cds





99% SI (130/131)





to mouse IMAGE





clone





(BC030389),





2072 bp.


scr_gb-
87
87
UI-R-CA1-bcg-d-


bf390383_1


03-0-UI.s1 UI-R-





CA1 Rattus





norvegicus cDNA





clone UI-R-CA1-





bcg-d-03-0-UI 3′,





486 bp.





[BF390383.1].


scr_gb-
89
89
UI-R-C1-kd-h-12-
CTNNA2: Catenin (cadherin-associated protein),
165


bf560709_1


0-UI.r2 UI-R-C1
alpha 2 [M94151, NM_004389].





Rattus





norvegicus cDNA





clone UI-R-C1-





kd-h-12-0-UI 5′,





mRNA





sequence (BF560





709.1: 99%/464,





p = 9.2e−264), 525





bp.


scr_sc-
126
126
Rat gene
>gb:GENBANK-ID:HSAJ2030|acc:AJ002030.1
166


14059147_2


fragment - 254 bp.
Homo sapiens mRNA for putative progesterone





75% SI (190/252)
binding protein - Homo sapiens, 1874 bp.





to Homo sapiens





mRNA for





putative





progesterone





binding protein





[AJ002030].


scr_sc-
110
110
Rat gene
AK056165.1|Homo sapiens cDNA FLJ31603 fis,
167


172130231_1


fragment - 196
clone NT2RI2002654





bp. 92% SI





(181/196) to Mus





musculus cDNA





clone





IMAGE:5051929





5′[BI146266.1].









Example 6
Genes Involved in Zone 3 Necrosis In Vivo
There is ample evidence to support the importance of ATP depletion as a mechanism that leads to cell necrosis [3]. Rat trihydroxycoprostanoyl-CoA oxidase was selected as an in vivo marker for zone 3 necrosis and was found to be downregulated in the dataset. This enzyme is one of the three acyl-CoA oxidases found in rat liver peroxisomes and is responsible for the beta-oxidation of fatty acids [16]. Downregulation of an enzyme involved in beta-oxidation represents an interruption in a pathway that can lead to ATP production. Consistent with this is the downregulation of an enzyme involved in amino acid metabolism. Betaine-homocysteine methyltransferase (BHMT) catalyzes the transfer of an N-methyl group from betaine to homocysteine to form dimethylglycine and methionine, respectively [14]. Interestingly, alterations in methionine metabolism have been known to be associated with human liver cirrhosis for many years [15]. As discussed earlier, severe necrosis is involved in the pathogenesis of cirrhosis [9]. Thus, necrosis itself can lead to an inflammation mediated injury. A marker involved in an inflammation and defense response was found to be upregulated in this in vivo marker set. Rat MHC-associated invariant chain gamma stabilizes MHC class II molecules that are at the surface of antigen presenting cells [26]. Rat MHC-associated invariant chain gamma is predicted to be a secreted protein based upon PSORT, SIGNALP, and Hydropathic Profile (HP) analysis
Rat ephrin type-B receptor 1 precursor (EphB1) was downregulated as compared to controls. Upon binding to its ligand, EphB1 has been shown to activate c-Jun kinase through recruitment of an intermediate protein Nck [18]. c-Jun is an oncogene involved in cell proliferation. Regulation of proliferation is an integral part of necrotic cell death, whether it results from compensatory liver regeneration of healthy cells or cell cycle arrest of unhealthy cells. Indeed, ribosomal protein S6 was selected as a marker and was found to be upregulated. It has been known for 20 years that the ribosomal protein S6 is quickly phosphorylated when cells are stimulated to grow or divide [19]. Phosphorylation of S6 occurs in response to mitogens by activation of one or more protein kinase cascades, including MAP kinases [20]. Rat annexin II belongs to a family of proteins that in the presence of Ca2+ bind to acidic phospholipids. They may also cross-link plasma membrane phospholipids with actin and the cytoskeleton, and possibly play a part in exocytosis, since they are also involved in granule aggregation and membrane fusion [28]. Annexin II was found to be an upregulated marker and may be indicative of loss of structural integrity within the cell. Though annexin I and annexin II have also been identified as major substrates for the tyrosine kinase activity associated with epidermal growth factor receptor (Egfr). Rat Egfr signals through a number of pathways, including the MAP kinase pathway, to regulate proliferation [35].
Canalicular multispecific organic anion transporter (cMOAT), an upregulated gene in this in vivo marker set, has been shown to arbitrate the hepatobiliary elimination of many organic ions [29]. It has also been shown to remove glutathione conjugates from cells [30]. Glutathione conjugation is a cellular adaptation to the generation of reactive oxygen species (ROS) [36]. ROS occurs when oxidative phosphorylation becomes uncoupled during ATP generation. When molecular oxygen is reduced in such a manner damaging amounts of O2—, H2O2 and OH are formed in the process. ROS are thought to participate in necrosis through their reaction with all forms of biological macromolecules including lipids, proteins, nucleic acids and carbohydrates [37–39].
Markers were chosen from other biochemical pathways as well. This includes the upregulation of rat heat shock protein 86 (hsp86) which may be indicative of cellular stress. Rat ras-related protein (Rab-2), a protein involved in intracellular transport, was also upregulated. Two protease inhibitors, rat homologue to human inter-alpha-trypsin inhibitor heavy chain (ITIH1) and rat homologue to mouse serine proteinase inhibitor mBM2A were down-regulated and upregulated, respectively. Human ITI is found in human serum and is predicted to be a secreted protein based upon PSORT, SIGNALP, and HP analysis [32]. Finally, numerous genes involved in xenobiotic metabolism were diminished after exposure to the zone 3 necrotic agents. However, two, rat epoxide hydrolase and rat aflatoxin B1 aldehyde reductase, were found to be upregulated.
There were 11 markers chosen for this PTS marker set that did not match any known genes in the database and have novel composition. One gene, rat osteoactivin has no known association to any of the histopathologically relevant biochemical or toxicological pathways but is predicted to be a secreted protein based upon PSORT, SIGNALP, and HP analysis.
Example 7
Genes Involved in Zone 3 Necrosis In Vitro
Several of the biochemical events consistent with necrosis are represented in the in vitro marker set that is predictive of zone 3 necrosis. There is ample evidence to support the importance of ATP depletion as a mechanism that leads to cell necrosis [3]. Included in this is documentation that the production of ATP via glycolysis can protect a cell from necrosis when oxidative phosphorylation is inhibited [51–53]. The alpha subunit of rat ATP synthase was found to be downregulated in the in vitro zone 3 necrosis marker set. ATP synthase is the final enzyme in the electron transport chain and is ultimately responsible for catalyzing the synthesis of ATP. Downregulation of such a key enzyme is indicative of loss of ATP within the cell. Rat nonneuronal enolase is another marker that was found to be downregulated. Enolase is a vital enzyme in the glycolysis pathway that converts glucose to pyruvate. Glucose is a preferred carbon source and generated the highest return of ATP per unit of expended energy. Downregulation of enolase may represent a depletion of glucose stores within the cell. The rat homologue to mouse beta-hexosaminidase is a protein involved in oligosaccharide and glycosaminoglycan degradation and was found to be upregulated in this marker set [20]. Upregulation of this marker may represent the cells attempt to maintain glucose supplies. The rat homologue of this gene was found to be a secreted protein based on protein based upon PSORT, SIGNALP, and HP analysis. Two markers related to ATP depletion were found to be upregulated in this marker set. Rat EP3 alpha receptors for prostaglandin has been found to be involved in the inhibition of adenylyl cyclase, which catalyzes the conversion of ATP to cAMP [18]. Inhibition of this process would be consistent with a lack of ATP within the cell. Carbamoylphosphate synthase is a mitochondrial protein that removes excess ammonia in the cell via the urea acid cycle. Upregulation of this rat enzyme may indicate a resort to utilize amino acids as a source of energy.
Uncoupling of electron transport during oxidative phosphorylation in the above process can lead to the formation of excessive amounts of Reactive Oxygen Species (ROS). When molecular oxygen is reduced in such a manner damaging amounts of O2—, H2O2 and OH are formed in the process. ROS are thought to participate in necrosis through their reaction with all forms of biological macromolecules including lipids, proteins, nucleic acids and carbohydrates [54–56]. Cells have adapted to the generation ROS through an elaborate antioxidant defense system. Two such mechanisms of defense are found to be upregulated in this marker set. A rat metallothionein isoform represents one of these mechanisms. Metallothionein is a small cysteine-rich metal binding protein that mediates heavy metal response and can play a role in ion homeostasis has the ability to scavenge free radicals and has been found to be induced under oxidative stress conditions [39]. Transaldolase is a key enzyme in the nonoxidative branch of the pentose phosphate pathway that can reduce the amount of reactive oxygen intermediates though the maintenance of glutathione at a reduced state [41]. Glutathione is critical for scavenging mitochondrial ROS through glutathione reductase and peroxidase systems. Interestingly rat peroxiredoxin III, a member of a novel family of anti-oxidative proteins, was found to be downregulated in this marker set. Peroxiredoxins have the ability to reduce H2O2 by using thioredoxin or glutathione as an electron donor [40]. Downregulation of this protein may represent a preference to remove H2O2 through one of the other defense mechanisms available to the cell. Similarly rat metallothionein 1, another isoform of metallothionein, was down-regulated. The fact that there are two rat metallothionein isoforms found in this marker set modulated in opposite directions may also indicate a preference of one form over the other or may represent a redundancy in the pathway. Dihydrodiol dehydrogenase is a marker, which was found downregulated and may play a role in the amount of ROS generated in the cell. This enzyme is believed to yield ROS upon detoxification of polycyclic aromatic hydrocarbons [38]. Thus downregulation of this process may be an attempt to limit the overall amount of ROS within the cell.
As discussed earlier, hepatic regeneration is a response to cellular necrosis. This process involves re-entry of surviving liver cells into the cell cycle to replace lost tissue mass [57]. Though this normal reaction to liver injury can, if uncontrolled, lead to the early onset of hepatic carcinogenesis. Several markers predictive of in vitro zone 3 necrosis were found to be involved in the regulation of cellular proliferation in the cell. These markers were consistently upregulated and are represented by growth factor receptors (rat Egfr), cytokine receptors (rat IL-4r & a rat gene homologue to Mus musculus Inar-2 receptor), MAPK signaling cascades (rat Erk2), as well as a gene involved in the regulation of protein kinase C activity (rat homologue to human DGK-delta). Rat epidermal growth factor receptor signals through a number of pathways, including the MAP kinase pathway, to regulate proliferation. However, under certain conditions stimulation of this pathway can lead to cell growth arrest and the induction of apoptosis [22]. Interestingly rat extracellular signal-related kinase 2 (Erk2) was also found to be upregulated. Erk2 is a member of the Raf/MEK/ERK signaling pathway that was the first MAP kinase cascade to be characterized [23]. Rat interleukin-4 receptor (IL-4r) is an upregulated marker for in vitro zone 3 necrosis. While IL-4 is a cytokine that has immunomodulatory effects, there is evidence that IL-4 interaction with its receptor can lead to such downstream effects as gene activation and cellular proliferation [26]. A rat homologue to mouse soluble isoform precursor type I interferon receptor (Ifnar-2) represents the upregulation of another cytokine receptor. Recent data shows that murine Ifnar-2 is an effective regulator of interferon responses [24]. It is known that type I interferons play a role in cell proliferation [25]. Murine Ifnar-2 soluble form is predicted to be a secreted protein based upon PSORT, SIGNALP, and Hydropathic Profile (HP) analysis. A rat homologue to human diacylglycerol kinase, delta (DGK-delta) was found to be upregulated in this marker set. DGK is a signal transduction enzyme that mediates protein kinase C activity by modulating intracellular concentrations of two signaling lipids, diacylglycerol and phosphatidic acid [29]. Protein kinase C is a family of serine-threonine kinases that is known to regulate proliferation and apoptosis [30]. The only downregulated marker involved in regulation of cell proliferation was rat Crk-associated substrate (Cas) p130, a unique docking protein with a Src homology 3 (SH3) domain. Tyrosine phosphorylation of Cas has been implicated in integrin mediated activities including cell proliferation and survival [27]. Additionally, Cas, upon interaction with Src, has been shown to be involved in a H2O2 activation of cJun NH(2) terminal kinase (Jnk) pathway [28].
Some markers may have a less clear association with a necrosis specific pathway. This includes the upregulation of rat VL30 element, a retrotransposon that has been found to be upregulated in rodent malignancies but which a specific role has not been identified [50]. Rat OX40 antigen was found to be downregulated. OX40 is a member of the tumor necrosis factor family that is expressed by activated T lymphocytes and may indicate the presence of inflammatory events [34]. Also relevant to an immunological response is the upregulation rat homologue to Homo sapiens IgG Fc binding protein. This protein is widely expressed on mucosal surfaces and in external secretions [35]. The rat homologue to human IgG Fc binding protein is predicted to be a secreted protein based upon PSORT, SIGNALP, and HP analysis. This marker set includes two markers involved with protein metabolism. This includes the upregulation of rat ubiquitin-conjugating enzyme (UBC), which catalyzes the covalent attachment of ubiquitin to a target protein. The ubiquitin/proteasome pathway is the main non-lysosomal route for intracellular protein degradation in eukaryotes. It is important to many cell processes including cell-cycle progression and more recently has been found to target regulatory molecules found in the apoptotic cell death pathway [44]. Another protein that is involved with protein metabolism and was found to be downregulated is a rat homologue to human Prt1. Little is known of this protein, except that it a component of the translation initiation factor elf-3 [43]. Rat thymosin beta-10 is a marker that was downregulated. It is a protein that is believed to be involved in the sequestering of actin and may be indicative of loss of structural integrity of the cell [48].
Markers were chosen from several other biochemical pathways as well. A couple of genes involved in xenobiotic metabolism were induced after exposure to the zone 3 necrotic agents. These are rat cytochrome P-450's 2F4 and 2B3, which were both upregulated. Several genes that are involved with the synthesis or transport of RNA were also chosen as markers. These include the upregulation of the rat homologue to mouse heterogeneous ribonucleoprotein C, rat homologue to mouse cardiac lineage protein 1 and rat Hex. The rat homologue to mouse heterogeneous ribonucleoprotein G was also chosen as a marker but was found to be downregulated. Two markers involved in the intracellular trafficking were also selected. This includes the upregulation of a rat novel related kinesin protein which may be involved in the transport of membrane bound organelles and the downregulation of rat r-sly1 which plays a role in ER to Golgi trafficking [36, 37]. Two markers in this set are involved with cellular communication. Rat neurotensin endopeptidase was downregulated. Neurotensin is a hormonal peptide that functions as a central neurotransmitter or neuromodulator as well as a peripheral hormone [20]. Rat densin-180 has a strong association with the postsynaptic density in CNS neurons and is believed to function as a synaptic adhesion molecule. Densin-180 was found to be upregulated in this in vitro marker set [31]. Rat homologue to mouse sex-determination protein homologue Fem1a was upregulated in this marker set but has no known association to any of the histopathologically relevant biochemical or toxicological pathways. Fem1 is a known signal transducing regulator in the C. elegans sex-determination pathway [49].
There were 55 markers chosen for this PTS marker set that did not match any known genes in the database and have novel composition. There were also two genes that blasted to rat gene patents and have novel utility. One additional marker, rat Tclone4, had some similarity to a known gene but has no known function.
Example 8
Prediction of the Toxicity of a Test Compound
The following example describes the application of the TOXMARKER expression profiles generated, as described above, to identify hepatotoxic compounds.
Hepatocyte Culture
Animals. Male Wistar Han rats (Crl:WI[Glx/BRL/Han]IGS BR) were obtained from Charles River Laboratories, Inc. (Raleigh, N.C.). The animals were housed for 6 or 7 days in a temperature-, humidity-, light-controlled facility prior to hepatocyte preparation and were at this time 200–250 g in weight.
Hepatocyte isolation Hepatocytes were prepared by in situ liver perfusion according to a protocol used at the Yale Liver Center (Yale University School of Medicine, Yale University, New Haven, Conn.). To minimize the risk of contamination all equipment and solutions used during the perfusion procedure were autoclaved. The animals were anesthesized by sodium phenobarbital (approx. 50 mg/kg) and the abdomen opened to expose the liver. To perform the perfusion a catheter was attached to vena porta and secured by a ligature. After disrupting vena cava inferior 37° C. Hanks A buffer (120 mM NaCl, 5 mM KCl, 0.4 mM KH2PO4, 25 mM NaHCO3, 0.5 mM EGTA, 0.1% glucose) was circulated through the liver a 40 ml/min for 10 min. The perfusion was continued with 37° C. Hanks B buffer (120 mM NaCl, 5 mM KCl, 0.4 mM KH2PO4, 25 mM NaHCO30.4 mM MgSO4, 0.5 mM MgCl2, 3 mM CaCl2, 0.1% glucose) supplemented with collagenase (Liberase Blendzyme 3, 120 mg/400 ml; Roche Diagnostics Corp., Indianapolis, Ind.) until clear signs of liver disintegration were visible (approx. 10 min). The liver was transfered into ice-cold Leibovitz L-15 media (Invitrogen, Carlsbad, Calif.) and the organ was disrupted mechanically with forceps. The cell suspension was filtered through a 80 μm mesh followed by a 45 μm mesh, and the medium replaced twice with fresh ice-cold L-15 medium following 5 min low speed centrifugations (30 G). Cell were transported on ice to Curagen's tissue culture facility and washed twice as above in William's E media (Sigma, St. Louis, Mo.) with supplements (10% fetal bovine serum (Gemini, Woodland, Calif.); 9.6 ug/ml prednisolon, glucagon 0.014 ug/ml, insulin 0.16 units/ml, glutamin and antibiotic-antimycotic solution (all purchased from Sigma, St. Louis, Mo.). Cells were counted and viability was measured in a hemacytometer after Trypan blue staining
Hepatocyte culturing Cells were cultured in 12 or 24 well tissue culture plates coated with rat tail collagen (Becton-Dickinson, Bedford, Mass.) according to a protocol provided by Dr. Grazyna Wasinska-Kempka at Bayer (Wuppertal, Germany). Cells were seeded at a density of approximately 80,000 cells/cm2 or 270,000 cells/well in 12 well plates and 140,000 cells/well in 24 well plates and incubated at 37° C., 5% CO2 for 2 h. To suppress contamination the amount of antibiotic-mycotic solution was increased (3-fold) and gentamicin (44 ug/ml; Invitrogen, Carlsbad,) was added during the seeding stage. After 2 h the media was replaced with fresh William's E media (as above) supplemented with 75 ug/ml rat tail collagen (Becton-Dickinson). The media was replaced approximately 16 h later with fresh collagen-containing media, with or without test compound, and thereafter every 24 h.
Test compounds Test compounds were solubilized at 200-fold final concentration in 100% DMSO and diluted into hepatocyte culture media to final concentrations ranging from approximately 200 pM to 10 mM depending on the compound. Compound stock solutions in DMSO were prepared at the initiation of the dosing regimen and stored at 4° C. Compounds were added to the culture media immediately prior to addition to the cells.
Control compounds (compounds that do not elicit a toxic histopathogy, see for example Table 1 and 2)) are selected from the training set and dosed on each batch of hepatocytes along with the unknown compounds subjected to toxicity prediction. The data from these controls are used to determine hepatocyte quality over time
RNA isolation Qiagen's (Valencia, Calif.) RNeasy 96 kit was used for isolation of RNA. For cell lysis the culture media was carefully removed and 400 ul/sample in 12 well plates and 200 ul/sample in 24 well plates of RLT lysis buffer supplemented with 10 mM DTT was added per well. The RLT buffer efficiently lysed cells and solubilized the collagen layer covering the cells. The lysates were homogenized by pipetting 12–15 times, snap frozen and stored at −80° C.
Cell viability In parallel with the compound dosing, a viability assay was performed to make sure that the concentrations used to treat the hepatocytes were not excessively toxic to the cells. Cell viability was monitored for each compound concentration using the CellTiter Assay (Promega, Madison, Wis.), a modified MTT assay. Cells were seeded in 96 well plates coated with rat-tail collagen (Becton-Dickinson) at a density of 27,000 cells/well. After 16 h in culture cells were treated in triplicate for each compound and concentration. Following 24, 48 or 72 h incubation in the presence of the drug, the MTS/PMS reagent of the CellTiter kit was diluted 1⁄6 in culture medium, added to the cells and after 60 min incubation of the cells optical density at 490 nm was measured using a PowerWaveX Select 96 well spectrophotometer (Bio-Tek Instruments, Winooski, Vt.). Viability was calculated in relation to no-drug control after subtraction of a no-cell background value. Only compound concentrations that showed 70% or more viability in this assay were used for gene expression profiling.
RNA Purification The hepatocytes are harvested in 200 μl of lysis buffer (RLT) provided with the Qiagen RNA isolation kit. Total RNA is isolated from the lysates using the Qiagen RNeasy 96® isolation kit following the manufacture's instructions with some modification. An equal volume of 70% ethanol is added to each of the lysates and the samples are added to the membrane on the 96 well plate. Membranes are then washed thoroughly (once with 800 μl of buffer RW1 and twice with 800 μl buffer RPE) to remove unbound material followed by DNase I treatment (50 units of DNase I from Promega (10 u/μl) in buffer RDD from Qiagen; total volume 60 ul) for 1 hr at room temperature to remove all traces of genomic DNA that might be co-purifying with the RNA. Following DNase I treatment, the membranes are again washed three times as before (once with 800 μl of buffer RW1 and twice with 800 μl buffer RPE), and dried with a centrifugation step (6000×g for 7 min; to remove all residual traces of ethanol from the washing buffers). RNA is then subsequently eluted from the columns with 40 μl of RNase/DNase-free water. This process has been automated using the Tecan Genesis Freedom robotic system.
Quantity of RNA is determined by fluorometry using Ribogreen dye from Molecular Probes and quantified using a fluorometer (Spectrafluor Plus instrument, Tecan). This procedure involves diluting each 2 μl sample ten-fold in Rnase-free water and then measuring fluorescence (after addition of dye). An average of triplicate subsamples is used to calculate the concentration and total RNA yield for each sample (by comparison to a standard curve generated from known amount of RNA standards). At this point the samples are evaluated as passed or failed based on a concentration criteria (60 ng/μl or more considered as “PASS”). The samples that have a concentration greater than 60 ng/μl are further diluted to 60 ng/μl with DEPC treated water.
cDNA Synthesis Double stranded cDNA is synthesized using the Roche cDNA synthesis kit, following the manufacturer's instructions, with some modifications. 600 ng of total RNA isolated from the hepatocytes (60 ng/μl), are spiked with 2 μl of reference mRNA (7×106 copies of hyaB and 2.5×107 copies of mhpR) and this mix is incubated in the presence of 2 ug of oligo [(dT)24 T7prom]65 primer at 70° C. for 10 min, immediately followed by quick chilling on ice. To each sample first strand synthesis mix is added such that the final mix contains 1× AMV RT buffer, 8.1 mM DTT, 25 units of AMV reverse transcriptase, 12.5 units of RNase-inhibitor and a dNTP-mix (1 mM of each nucleotide). This mix is incubated at 42° C. for 1 hour followed by chilling on ice. The second strand synthesis involves the addition of the second strand buffer to a final concentration of 1×, a dNTP mix (80 μM each) and the second strand enzyme blend provided in the Roche cDNA synthesis kit. The mix is incubated at 16° C. for 2 hours. Adding 10 units of T4 DNA polymerase to each reaction and incubating at 16° C. for a further 5 minutes to terminate the elongation. The reactions are stopped by adding 11 μl of 0.2 M EDTA pH 8.0. The double stranded cDNA is purified using Qiagen's QIAquickTM Multiwell PCR Purification kit, following the manufacturer's instructions. The cDNA is then quantified by fluorometry using the Picogreen® dsDNA Quantification Kit (Molecular Probes) following manufacturers instructions.
Microarrays
In Vitro Transcription, cRNA Purification, and cRNA Quantitation The complete yield of double stranded cDNA (minimum 50 ng) is placed in a Centrivap Concentrator (Labconco) for 2 hours at 45° C. or until liquid is completely evaporated. In vitro transcription is performed using Ambion's MEGAscriptTM T7 Kit, following the manufacturer's instructions, with the following modifications. Biotin labels are incorporated during cRNA synthesis by adding biotin 14-CTP (Invitrogen) and biotin 16-UTP (Roche Applied Science) to the in vitro transcription reaction to a final concentration of 1.5 mM. Transcription reactions are incubated at 37° C. during 16 to 18 hours. Reactions are stopped by adding 2 U of DNase 1, and incubating at 37° C. for 15 minutes. cRNA is purified using Qiagen's RNeasy 96TM kit, following the manufacturer's instructions. Purified labeled cRNA is recovered in 30 ul of DEPC-treated water (see STM TS-MAH-104). Labeled cRNA yield and quality are determined by measuring the 260/280 nm optical density ratio, using a Powerwave HT spectrophotometer (Bio-Tek). Labeled cRNA is then diluted to 0.225 ug/ul using DEPC-treated water.
Hybridization, and Fragmentation We use a format with two microarrays on each slide and dual hybridization chambers consisting of two 0.8 mm height, 22 mm2 square chambers. Dual hybridization chambers are installed on glass arrays using a Slide-Chamber Alignment Tool (SCAT). The SCAT is linked to a vacuum pump to create pressure that makes the chamber and the microarray hold together. Once assembled, microarrays are incubated at 37° C. for 10 minutes. Slides are then placed chamber side down on a lint free paper towel (Texwipe Company LLC) on a flat surface, and using a finger, gentle pressure is applied along the periphery of the chamber.
Synthetic cRNAs (1 ng each of bacterial araA and ybiw) are combined with 9 ug hepatocyte-derived labeled cRNA prior to fragmentation. The cRNA mixtures are fragmented at 94° C. for 20 minutes in a 50 ul reaction containing 40 mM Tris-Acetate, 100 mM Potassium-Acetate, 31.5 mM Magnesium-Acetate, pH 8.1. After the fragmentation is complete, 300 ul of cold hybridization buffer (Mergen Ltd., proprietary composition) is added to the fragmented RNA, and kept on ice until ready for loading onto a microarray. 320 ul of cRNA are then loaded onto a barcoded microarray. After air bubbles have been removed from the liquid, loading ports of the chamber are tightly sealed using Mergen adhesive dots. Slides are incubated for 16–18 hours at 30° C. in a hybridization rotisserie oven set at 18 rpm (Robbins Scientific, model 400)
Microarray Washing and Staining After hybridization for 16 hours, the hybridization chambers are removed from the microarrays slowly using forceps. The microarrays are placed in a reservoir containing TNT (0.1 M Tris-HCl, pH 7.6, 0.15 M NaCl; 0.05% Tween-20) and incubated at 39° C. for 1 hour. The following staining procedure is light sensitive so all incubations are done under foil taking care to minimize light exposure. Following the TNT wash, the arrays are incubated in Buffer B (Mergen LTD, proprietary composition) at 4° C. with gentle agitation for 30 minutes. The slides are then placed in a 1:500 dilution of 1 mg/ml streptavidin-alexa 647 (Molecular Probes) in Buffer B at 4° C. with gentle shaking for 30 minutes. Following a TNT wash of three times 5 minutes each, the arrays are incubated in a 1:500 dilution of 0.5 mg/ml biotinylated anti-streptavidin antibody (Vector Labs) in Buffer B at 4° C. with gentle agitation for 30 minutes. The microarrays are washed three times in TNT for 5 minutes each. A 1:1 mix of streptavidin-alexa 647 and biotinylated anti-streptavidin antibody is incubated at 25° C. for 1 hour to encourage complex formation. This complex is diluted 1:267 in Buffer B and incubated with the microarrays for 30 minutes at 4° C. with gentle shaking. The slides are washed again in TNT three times for 5 minutes each followed by 2 washes of 5 seconds each in redistilled H2O. The arrays are washed individually in redistilled H2O five times for 1 second each. The microarrays are placed in a slide holder and centrifuged for 15 minutes at 300 rpm to ensure complete dryness. The slides are placed in a clean dry box and stored at room temperature until being scanned.
Scanning and Analysis of Microarrays Each slide is individually scanned using a GenePix 4000B scanner (Axon Instruments Inc.), using the 630 nm laser at 100% power and a PMT setting of 600 volts.
Image QC and Data Export Tiff images from each slide are analyzed using the GenePix 4.0 software (Axon Instruments Inc.). The scanned image is first aligned with a grid consisting of an array of circular features, such that each spot on the image is contained within a feature. After the array is roughly aligned, the software conducts fine alignment of each feature with each spot on the array. Additionally the software calculates local median foreground and local median background for each feature. The microarray is then evaluated manually to determine whether the features found by the software were true data points and not false intensity due to a defect or contamination. If contamination affects more than 30% of a feature, the feature is flagged as bad. If contamination affects more than 30% of the local background area of a feature and the local foreground/local background intensity ratio is less than 3, the feature is flagged as bad. After flagging is complete, a GenePix output file is generated. The output file contains spot IDs, spot location information, median local foreground, median local background, and bad spot flag information.
A quality control log is kept for each array to document background levels, noise/contamination problems as well as the number of features flagged. A final Image QC pass or fail decision is made for each array and added to the log. An array must contain no more then 1% flagged spots in order to pass Image QC. If an array contains an area of contamination of >150 counts which covers more then 5% of the array then the array fails
PTS Microarray Data Processing The GenePix output files generated during the Image QC and Data Export are processed using Microsoft excel. A macro is used to split the data from each of the two arrays, calculate spot intensities, and filter out low quality data. The following procedure is used to calculate gene intensity and filter the data.
A. Calculate Spot Intensity


    
    
        spot-intensity=median local foreground−median local background
    
    


B. Calculate Threshold
There are 15 probes on the array that are either yeast or mouse negative controls.

    
    
        If (spot_intensity is from probe type “yeast_neg” or spot_intensity is from probe_type “mouse_neg”)
        then spot_intensity=thres_value
        mean_thres=average (thres_value[1], thres_value[n])
        stdev_thres=stdev (thres_value[1], thres_value[n])
        If (thres_value<(mean_thres+3* stdev_thres) and thres_value>(mean_thres−3*stdev_thres)
        then thres_value=good_thres_value
        else thres_value=bad_thres_value
        mean_good_thres=average (good_thres_value[1], good_thres_value[n])
        if mean_good_thres>0
        then threshold=mean_good_thres+5* stdev (local_bg_median[1], local_bg_median[n])
        else threshold=5* stdev (local_bg_median[1], local_bg_median[n])
    
    


C. Data Filtration

    
    
        If (spot_intensity has no problem spot flag then spot_intensity=good_spot_intensity
        Only data of type good_spot_intensity is passed to the Discovery department.
    
    


PTS Microarray Quantitative QC The array quality is assessed by calculating a set of descriptive statistics and testing if they pass set criteria. In order for array data to be accepted the following criteria must be met:
Toxicity marker spot intensity trim mean/threshold>8
Maximum spot intensity of blank spots<300
Number of marker spots above threshold>800
Labeling spike mean spot intensity/threshold>50
Hybridization spike mean spot intensity>50
Toxicity Prediction
The PTS presents a single overall likelihood of toxicity for unknown samples (i.e. the probability that a sample is toxic) that can be ranked to indicate severity of toxic insult. In order to compute the toxicity likelihood of an unknown we calculate a likelihood estimate from 3 different modeling types (Classification Trees, Discriminant Analysis, and Logistic Regression) using three independent gene lists for a total of 9 models per mode. These 9 models are then averaged to provide a likelihood (probability) of toxicity for each mode. The rationale behind combining the results of several models as opposed to relying on a single model that performs best for a given mode is to control for the risk of over-fit (a model that performs well on training data but inadequately on novel samples). The best performing model would be expected to be more over-fit than a weaker performer. In order to reconcile this we average the results of several models in order to determine the true likelihood of toxicity for an unknown. This section attempts to describe the methods we will use for model averaging, explain how compounds can be ranked, and how we can draw inferences about the severity of toxic insult.
Common Terms
Likelihood of Toxicity: A value between 0 and 1 indicating how confident we are that a given compound/dose combination is toxic.
Model: A statistical algorithm for prediction. This section focuses on Logistic Regression, Discriminant Analysis, and classification trees which are explained elsewhere.
Model Type: Logistic Regression, Discriminant Analysis, or classification trees
Mode: A specific type of hepatotoxicity (e.g. hypertrophy)
Present and Absent: Present means a compound produced a given toxicity in vivo absent means it did not.
Model Background
One obvious problem with model averaging is that different models provide different outputs. Discriminant Analysis produces a number without bounds, logistic regression produce a likelihood estimate with a value of 0 to 1, and classification trees a proportion of node impurity with a value between 0 and 1. Therefore an arithmetic mean of these three results may be misleading because the scale of discriminant analysis is so different than the other models. Our approach is to scale the results of all models to produce a single likelihood, PP, the probability that this sample belongs to the toxicity class as opposed to PA, the probability that the sample does not manifest itself as toxic for this mode. By definition:

Pjp+PjA=1  Equation 1

Where j is the jth model (the three models mentioned above). In order to proceed we first need to find PP for each model.
Equation 1 means that our classfications (present and absent) are mutually exclusive and collectively exhaustive. If a sample is absent for hypertrophy, it cannot also be present for hypertrophy (mutually exclusive) and if a sample is not hypertrophic it must be absent of hypertrophy (collectively exhaustive).
Logistic Regression: This model returns PP by design.
Discriminant Analysis: This model returns a linear discriminant that is a one-dimensional linear combination that establishes two separated normal distributions as follows:
where ‘Absent’ refers to the theoretical distribution of samples that did NOT produce pathology and ‘Present’ refers to the theoretical distribution of samples that did produce pathology. Let MP=the mean of the linear discriminant function for the training samples annotated as present, and MA=the mean of the linear discriminant function for the training samples annotated as absent. Let GP and GA denote the probability mass function for the Gaussian distributions of the linear discriminant values for the training samples annotated as present and absent, respectively. Having estimated these functions, calculations of percentiles is very straightforward. In the above figure, MP>MA, therefore, for an unknown sample with a linear discriminant value of ‘x’, we have:



  
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where Lower Tail Gp(x) refers to the area under Gp(y) for which y<x, and Upper Tail (GA(x)) refers to the area under GA(Z) where z>x. When MA>MP the above equation becomes:



  
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Equations 2 and 3 appear more complicated than they really are. They simply convert the percentile of (x) belonging to the “present” distribution to a conditional probability that it belongs to present and not absent. This is used to satisfy mutual exclusivity rule of equation 1.
Classification Trees. As mentioned above, classification trees return a probability of correct classification for each prediction. However, this probability is actually a proportion based on the node impurity of the classification tree (the fraction of training samples on that leaf that belong to the majority class, e.g. a leaf contains 9 samples with hypertrophy and 1 sample without, the probability returned is 0.9). This is not a good indication of the true probability of an unknown because the leaf may have very few members and because the confidences of branch decisions are not included in this calculation. In order to convert this proportion to likelihood, we simply construct many trees using a subset (n−1, where n=the number of compounds) of the compounds in the reference database until all compounds are NOT used once (this is identical to the leave one out cross validation described in the next section). This process is called a jack-knife estimate of confidence.
Summary
This section describes how we convery the results of the three different models to a likelihood estimate that satisfies the mutual exclusivity rule of equation 1. For logistic regression this is the result, for discriminant analysis we rely on the probability mass function of a normal distribution, and for classification trees we create a jack-knife estimate of node impurity. Having described these techniques the next section explains how we combine the results of the individual models.
Model Averaging
Each of the above models makes a decision as to whether a toxicity is “present” or “absent”, and some models are expected to perform better than others. What remains to be explained is how we determine model quality and how we combine the results of individual models.
a) Model Quality:
In order to determine how confident a given model is in its decision, we perform jack-knife estimates of each prediction. A jack-knife estimate computes n different models, with n−1 compounds in the training set (where n is the number of compounds used for training a particular toxicity mode). Each jack-knife casts a single vote for absent or present. The best models confidently (likelihood is much greater or less than 0.5) make the same decision consistently, while poorer performing models tend to have equivalent “present” and “absent” votes.
b) Combining Models:
The number of present and absent votes are tallied across all the jack knife estimates for each unknown compound. Models that are more confident in there decision are naturally weighted heavier by an ability to cast more votes. The vote totals can then be evaluated using the binomial distribution as follows:




  
    L
    Tox
  
  =
  
    
      ∑
      
        i
        =
        0
      
      P
    
    ⁢
    
      
        (
        
          
            
              N
            
          
          
            
              i
            
          
        
        )
      
      
        2
        N
      
    
  




Where LTox is the likelihood that the observed vote distribution is greater than 0.5, P is the number of votes for “present” and N is the total number of votes cast.
Summary
This section explains how we use three different marker sets for each model and then compute a weighted average based on how consistent the prediction are within a given step. At the completion of this exercise each sample will have three sets of votes: one each for logistic regression, discriminant analysis, and classification trees. The likelihood for each mode is calculated from a binomial distribution, under the null hypothesis that voting is random.
Example 9
TOXMARKER Nucleic Acid Sequences
This example provides exemplary TOXMARKER nucleic acid sequences, useful in methods of screening compounds for hepatoxicity according to the invention.








TABLE 9A







>scr_gb-af038870_4 (TOXMARKER Assignment: 1; SEQ ID NO: 1)



 


ttttttttttttttttttttgaaggttttcaaccggcatgtttttattaatgaaatggaa


 


tggaagcagtcagaacagagattacagaattacagaatggatcagttatctgttaagttt


 


tacagggctggtgtgtgttgtttctgcctaagggtcctgctcaaaagatcttggaatcca


 


cttgggaagcatcttagatatagatggttgctgtgtcacttatgatacggtccctgaatg


 


gttctatgtcactcgtggaggtggtgtcctatccccctatctgaaatgagattgacgtcg


 


ggtgactttctcttcgctgcagtgactcctgtgcgcctgtaatgcgacaggcacgtagga


 


aatgtgttcaggatttactgtggacttctcctttcttccttctaggtaaaattctaaagc


 


gtagttttgtaactgtgaaatgctatctgtgactccattttgtctaactagcaccaatca


 


caggtgtaagccggcatcaacacaaacgctggtttagagatgccttctccttccgggtgc


 


acactgtggcccggacctggaggaattcgccccgaaccgctggcctgtggctactgtgcg


 


gatttgaatttttgtttttcgaagagcgctctcagctgctgctcagtggtggcttccttc


 


tgctgcatcagctctgctgcccctttcgtcactccccaagcatccggcttggacatcgaa


 


ggattgtacggtctgccggaagctattcgaagattctgccagtattctttcctggccctt


 


gccctgatccagggtttggtgtgcatgtccaaaccacttccccagctgccatgtttttct


 


gaagctggtggtaaaaatcccctttctggggcgagctcctctgcaatggccctgatgtgg


 


tagggctcaaatccgcagcagccgccaatgtacctgacccccaggttgtaggcctctctg


 


gcgtatttttgaatatcccatctggtggcaactctgggttccaatccaaaggggaattct


 


gggagatcaataaatccctgtttgccacagtcaggggtgtggtaggccaggggctggctc


 


atcaagtaagccttcagccgagctgcttccagaccctccttcatgagctttattgtctgc


 


aagctggtgctggggtcgaagtggcagttcacaccgacaatggcggcacctgcttttacc


 


aaacgcactgcgcactctccaggagacacgccatgtagatctccttcaggtccgatgcac


 


atggtagccgctataggcttcccggatgtttttaaggcctcgactgcccacacggcttct


 


tcaacatgttcaaaatactctgcaatgaggaagtccacattcttcttcatgaagacctca


 


agctgttggtgaaatatctttttaacttccgtctcactcttgcagctgaggtaggaaggt


 


gtctgactcacacctcctgcaaccaatgcatccccttcgtcagcaacttgccgtgcaatg


 


tcacaagcagcttcattgaccttctgcccagatatcttctctgccacgtagttccctcgg


 


ttttccagcttgtcctcacttgcatagaaagtgaaggtctgcatgacgttcgatccagct


 


ctgaggaactcccgatgaagctgccgaactgcctcggggtgctccaccgcagcctctggg


 


gtccagggtccagcctttacgtagcccctcttttccagtgcaaagacaaatcccccatct


 


ccgatcacgacttcgccagcatttaagcgttctaagattcccctcttggccttcttgccg


 


gcaatcggtgccatctttccggtgtcctgagtggcgctgaacgcagctgcggactggaca


 


ggagcggtctccagcaaaggcttgactgctgagccgcttctggcctctttatatacagca


 


gctaggattccccagccttgaccgggtccaacacatggcctcaggcggggaacacgccca


 


ccagcctttgaaacaggcctggggctagctgggaattc


 


>scr_gb-z83053_3 (TOXMARKER Assignment: 2; SEQ ID NO: 2)


 


gacatggcaccagccggaggcccacgagtcaagaagggtatcttggagcgtctggacagc


 


ggggaggttgtggttggggacggcggctttctcttcactctggaaaagagaggctttgtg


 


aaggcaggactttggactccagaagcagtggtagagtatccaagtgcagttcgtcagctt


 


cacacagaattcttgagagcgggagccgatgtcttgcagacattcaccttttcggctgct


 


gaagacagaatggaaagcaagtgggaagctgtgaatgcagctgcctgtgacctggcccag


 


gaggtggctgatggaggggctgctttggtggcagggggcatctgccagacatcactgtac


 


aagtaccacaaggatgaaactagaattaaaaacattttccgactacagctaggtgttttt


 


gccaggaaaaatgtggacttcttgattgcagagtattttgagcatgtggaagaagccgtg


 


tgggctgtggaagtcttgagagaggtgggggcacctgtggctgtgaccatgtgcatcggc


 


ccagagggggacatgcacggcgtgacaccgggagagtgtgcggtgagactgtctcgtgca


 


ggggcgaacatcattggggtaaactgccggtttgggcctggaccagcttacaggaccatg


 


agctcatgaaggagggcctcagggattgcggcctactagctcaccttatggtccagtgct


 


tgggttttctcacactgggactgtggcaagggagggttgtggacttcctgatatcctttt


 


cgcctggggcaagagttgccaccagatgggatattcaaaaatacgccagagaggcctaca


 


acctgggggtcaggtacattggcggctgctgcggatttgagccctaccacatcaggggcc


 


attgcagaggagctcgccccagaaaggggatttttgccaccagcttcagaaaaacatggc


 


atctggggaagtggtttggacatgcacaccaaaccctggatcagagcaagggctagacgg


 


gaatactgggaaactctgttgccagcttcgggaagacctttctgtccttccctatcaaag


 


ccagatgcttgagaagccatgaaagagacctctgaagtgacagaaaggaggaaacagcct


 


caagccccatctggaatcttcctggctgctgtcctcagcccgttcttctggctgttgagc


 


atcgatgagctgtcgtcccttccaattgagtgacatatcactcctgagtatgcccactag


 


atgcggtggagatgcagaggcatccggaccccacgccccaccccctcccctcacacactt


 


actctctgcctagtaatgccacagagcttccatccccatccaaaggtcatcaggcatggc


 


tatcagttggctctcagggtggatttgacattctcagatgattagaagttggcaagaagc


 


aaccttggtgaataactctggtgtctaaactctgtacttgagttacagtctcagtagagg


 


agacgccaaagctgttgcgagtgacggcagaattattgaacagtcatgatgcttggcttt


 


caaaggcgattatcgctttaaggtcttagaattagtaagtgcatctttataaccaggcat


 


agctagatcataaactactgatggccaaggaccatagaacgtgcttcttaccttcctctc


 


tagttagcattacgacaaacataatcaccaacgctcagggaaacacttgctgattcaagt


 


aaaatgcatgaaccttggaagacctttctagaagtcagagatcaagttcatcttgttcta


 


gcactttccacattcatgtttggtttgtatgctgcgccctacttttgttttttgctacaa


 


tgtaacaaattagtgagtaaccattagtgaaattgcgaataattttccttttctaaattt


 


tgatttctttggaacattgatttaaaaaaaatagtgtgttgcttgtcaaaaaaaaaaaaa


 


aaaa


 


>scr_gb-x95189_4 (TOXMARKER Assignment: 3; SEQ ID NO: 3)


 


ccatagcgaagacttcatgaagactgtcccaggcatgctgtgacacaaactacagaaggt


 


gggaaaagatctttgtggtcaaaccatccggaccttggctaccgcagacagaacaatact


 


gaccgcattcactcatacacagttctcggcacctcccagtgctcagagcagaccctcaag


 


gagatgagcagatccaggatggggagcccaatgcaccgagtgtccctgggggacacctgg


 


agctggcaagtgcacccggacatagacagcgaaaggcactcaccgtccttcagtgtggag


 


cgactcaccaacatccttgatggaggcctcccaaacaccgtgctgcgaagaaaagtcgaa


 


agcatcatacaaagtgacccagtgtttaatttgaagaagctttacttcatgacccgagag


 


gagctatatgaggatgcgattcaaaagagattccatctcgagaagctagcctggagcctg


 


ggctggtcagaagatggtcctgaacgcatttatgctaacagagtccttgatggaaacgtc


 


aacttaagcttacatggtgttgccatgaatgctatccgaagcctgggctcagatgaacag


 


attgctaaatggggccaactctgcaaaaacttccaaatcatcacaacatacgcccagaca


 


gagctgggacacgggacatacctacagggcctggagactgaagccacctatgatgaagcc


 


aggcaggagcttgtgatacacagccctacgatgacttccaccaagtggtggcctggggac


 


ttgggatggtcggtcacccatgctgtggtcctagcccagttgacctgcttaggagtccgg


 


cacggcatgcacgccttcattgtgcccattcggagcctagaggatcacaccccactgcca


 


ggaatcacagttggggacataggccccaagatgggtttggaacacatagacaatggcttc


 


ctgcaactgaaccacgtgcgggttcccagagaaaacatgctcagtcgctttgcagaggtc


 


ttgccagatggtacctaccagaggcttgggacgccacagagcaattatcttggcatgttg


 


gtgacccgggtgcagctgctgtgtaaaggaatcctaccctccctccagaaggcttgcatc


 


attgccacgcgctactcagtaatccgccatcagtctcgacttcggcccagtgacccagag


 


gcaaaaatcctggaataccagacgcagcagcagaaactccttcctcagcttgctgtgagc


 


tatgccttccacttcacggccaccagcctctcagaattcttccacagctcctacagtgct


 


attctgaagagagacttcagcctcctgcctgagctccatgcattgagcactggtatgaag


 


gccacgtttgcagacttctgtgcccagggcgccgagatctgtcgcagagcttgcgggggc


 


catggctactcaaagctgagcggcctgccgacactggttgctcgagcaacagcctcttgc


 


acatatgagggtgagaatacggtgctctacctgcaagtggccaggtttctgatgaagagc


 


tatctgcaggctcaagcgtccccaggcgccacaccacagaagcctctccctcagtccgtc


 


atgtatattgccacacaaaggccagccaggtgctcagcccagactgcagctgacttccgc


 


tgcccagatgtctataccacagcctgggcatatgtgtctaccaggctcataagagatgca


 


gcacaccgtacacagaccctcatgaagtccggggttgaccagcatgatgcctggaatcaa


 


actactgtcatccaccttcaggctgctaaggctcactgctacttcatcactgtgaagaat


 


ttcaaggaagctgtggagaaactagacaaggaaccagagattcagcgtgtgctccaacgc


 


ctctgtgacctctatgccttacacggtgttctgactaactcaggggactttctgcatgat


 


ggcttcctgtctggggcccaggtggacatggccagagaagccttcctagacctgcttccc


 


ttgatccggaaggatgccatcttgttaaccgatgcttttgacttctcggaccattgttta


 


aactcggcacttggctgttatgatggacacgtctacgaacgcctgtttgagtgggctcag


 


aagtacccagccaatactcaggagaaccctgcctataagaagtatatccgaccactgatg


 


ctcggctggagacacaagatgtgaaaagtcaaaggatttgggaccgagaagcaccacggc


 


cttactatggcacatatacatagagaatttaaagcacggggggggggggggggggggtgc


 


tgctcggttaaatcaggtagtaaattggtacatgaatggatggtcatcctattagtctac


 


tattgagcatgtttgaaactttcccttgtccatctatagcatgtatttggctaaatgcta


 


aaatttttgttttacatacaggaaaagctaataaacttgtcagttacaaa


 


>scr_gb-m59814_4 (TOXMARKER Assignment: 4; SEQ ID NO: 4)


 


tttttttttttttttttttttttttttttttttttttttttaacaatgagacatatacag


 


ctttatttaacctgtaaaaagtcacactctgcagagtgacacctttcttatctcagcaga


 


aagcaaggagtgtgtgaaaaaccttttcctcaggttgggaaccgtatgaccctggctggg


 


ctcacatgtggatccttccagagtccttgtgtgtggcagcttcttcccagaggtctccct


 


ggctggtgtgacccctcaccaacaacagacaggggggcaaaatatttctacctggacaag


 


gctgccctgagattgtccctttccctcctattaagggacattacatgcttaagaccttcc


 


cagaaaagtcaccttcaaggtgacttggctttcatcatgtctgctgacacttaggctcca


 


cttatttaccatgatggtgtgtgctaacggtccttcctcttccaataacctcaccatcga


 


tggcattttaaatatcactctgttctctgggaccgagggatggagaaccgctctccctca


 


gaccaggttttgactcaggagctgggttttattttgaagaaacttccctacatgagtcat


 


gagcaagggaaatggatgtgggggagggaggaggggctctgagggaggagtacgaatgga


 


ggaaagaaaagaatgtcattggcgagggagagcatggcacagcccagggcttccctctct


 


tccctccacctccttcctttcttcctgcagacggggaactccagtccctctcagatggga


 


actgagttcaccctggttcccaacgcatacggtttcagcttcgcttctgtttagcatcac


 


ctttctctgtctttatcgtcaatcattacgcgtttggtttcccacggcttctacacactt


 


ccatggccgagaaatggcggttgcccatgggcagcaggtccagttcattcttcacaggtg


 


ggaagttgtttctcagccaagaagctgatctttctggcacattccaccgtggtcaacctc


 


tgtttcccctttgaccctggtccttttcattcctctcctcccctaggaacatcgagttct


 


catgccattaccgacggtgactggttcatctggaccctcatcgagtggatgctgctaaga


 


atcttcttctgatggcctgccaaggtgacccctattctcaggaggtcttctgatgtcatc


 


tgggtgaccagctggagggaggtgaagccagcggtgaggaagctgtccctgtactggacc


 


attttgatggcacttagccagtcatccacggtggtaaaggccgtgaagtctgggatagag


 


cggtcaagcaggggttgggaaggcacagcggtgatggttgccacagtcttgagactagct


 


gggttccggatcatcttgtccagggtgttgacgatctctgcaaaacggggccggctattt


 


cgatccttctgccaacagtccagcatgagctggtgcagggcagctgggcagtccatagga


 


gggggcagccggtagtcctgctcaatggcattgatgacatcttgattggacatatcccag


 


taaggtctctctccaaatgacattacttcccacatgacaatcccgtagctccagacatcg


 


ctggctgacgtaaacttgcggtaggcgatggcctctggagctgtccatctaacaggtatc


 


ttccctcccaaggagctggtgtaggtggggtctgaggtgtcatcctggaggtagcgagag


 


aggccaaagtcagacactttgcacaccaggttgctgttcaccagaatgttcctagcagcc


 


aggtcccggtgcacataattcatctcagataggtacttcatgccagcagcgatgcccctc


 


agcatccccacaagctggatcacggtgaactgtccgtcattttgccggaggaaagagtct


 


aaagcgccattctccatgaactccgtaatgatcatgacaggtcggctcttggtgacaaca


 


ccctctaggcgaatgatgttgggatggtcaaactggcccatgatgctcgcctcgctcaga


 


aaatcccgacgctgtttctctgagtacccagctttcagggtcttgatggccacatagatt


 


tccctcttgcctggcagcttcaatcggcccttgtacacttctccaaactcccctgctccg


 


atgacctcttcaattttcacaaaagacacatcaatctccttggcaaactcccggacagct


 


tcattagggtcctcataagtgaacgggtcaatgtagatcttcatccctggggagcctcgg


 


cctgtgctgtaatgctgaagtttatcactgtacacagcctctttgctgtaagctcgtttc


 


ctgctgcagacaatggagatagccaccagagacacaacaaatacaaccccagctgctgca


 


gagccagcgatcaggggtagctgctctctcagctccgacttgtaatcatcatctgtcaga


 


gtctggaagcacatcttgccactgaacttgccatagccagccacggttcgagctcgtacc


 


tggaccacatacaccatgccgggccgtagcccatcgatacgtgccgtgttggtctggctc


 


ctggccatggaagagttgaactcattgtgctccttctcatagtaccggatctcatagtcc


 


aggatgatgccattaggctgctccggctgaggccatgacaaggtgatgctcctcatggtg


 


gcactgacctggtgcatgataggaacagtggagggggcagcttggtttgtggtgatgttg


 


acagagacatgctgtggggggaagggactcttgctagagactccattgatggcctggata


 


tcaaaagtgtatggggtgtgggcccataggctactgatagagacacgacactcagtcaag


 


cccagctgtctgggtacaaactccacattgtcatcgcagcgggagcaactccggcggtct


 


gctctgcacttcttgcagatgatgttgtaggtcacatcatctcgcccaccggtctctctt


 


ggagggtgccactctagaatgatagatgtctcattcacaatggagatgacatttcgaggg


 


cctgatgggacactagtgcacgccacttctgggggatcaaagtctgctcggtaatagcca


 


gtccggcaggtgcagatgggagacgcctctgaaggggagcggctgttggaggggcagtgg


 


gagcagccttcagcttcctggctggccttgaaggttcccgcaggacaggccttgcaggcc


 


acgctgttctcaggttcatagccagccttacaggtgcagcgcccaatgggcaccatccac


 


tctccatctccattgcagtagagttttatgggcacatccacttcttctgcattagggatg


 


catgtgccccgagcaatcaccagagatgtgctctctgctcctgtcatggtttctgggaac


 


actgcaaaattttgcacaatgctgggacactttttgaagaagacacggacagaaagtaga


 


gacatacaggctccataatcctggaaagcgaggtaaaaaccattcctagtaagaggccca


 


aagctcctgacttctgtgttgaccttcatcaaccttcccccaaaatccacctgggagaag


 


ctctcatctgcagcaatggtgtcaactttgaggtagggggcttcagaccagaaggctgac


 


ttcttggtggcaatgacagagtcagtctcatagtagtataagttgaaggtctctttgcag


 


gagcctgggacatttggaaggctgctgcagtccctcacagtgaagcgcatctctgtatag


 


atgcgatgggcgccccgtctgttgataaaggtggtaagcagccagttgttctggttgggt


 


tcaaagacgttgcacacttggtaagtacggatggtgttcaggttttcatcgtagccactg


 


acttcttcccacccagaggcagggttggccgtccatcccaactctgcagtggcagtcctt


 


gtgtccatcaatgtttcttccatcgcggccactgcagatgccaggaggaacagcagcagg


 


caatccagggccatcgccggccagcggcccccaggccgagccccagcggagacgcgccgc


 


gtcccagggcgccgctgcgctcccggcgggtggcttctccgtgtcctttcgcgctctggc


 


cgggaccggactccccggagcgcggcgtgggcgtgggcgggagtgtgcgcgcgtggggcg


 


gtgcgggcgcgcgtggatgtgggtgtgcatgtgtgtgtgtgtgtttatgggagaggtggg


 


tgtgtgcgtgcgtgtgtgagagagggtgagggagagcgagccaaaccataaaaagatgga


 


gggggagttgtgggtgggcgaccctgctagtttcatagctggcattcttggggctggaaa


 


ccccatggcacaagacgttaggatggctggtctgctcaaccactgtgccgtgtgtgaggg


 


gtctctcggcttgtgtctctatcctgctctcattgagtcggatgacctgtacagctctgt


 


ctaccatggaggatgtattgtgaagtctctgtgctaaggactcacgtttgggtgctttgg


 


agatgaaatggatgacatgtacactggatatccccctcgtg


 


>scr_gb-m29358_5 (TOXMARKER Assignment: 5; SEQ ID NO: 5)


 


ccccccctcgaggtgttttctttcatttcattccttgtctttagggcttttttttttttc


 


aaggtctcattatttatttgttactctttaaagacttatttttgactggactcagattta


 


gaagtagaagctctcagcgaagacagcctacgtctcttggcaatctgttcctggcgcttc


 


tctttggcttccttcattctcttggccaaaagtttagcatattctgcagcctcctccttg


 


tttttcttagtgcgttgcttcttcagagcaatacgtcggcgtttgtgttgcaggacacgg


 


ggagtaacaagacgctgaatcttgggcgctttggtcctgggcttcttaccttctttgttt


 


aagggctttctgacaacatactggcggacatcatcttctttggagagattaaaaagcttt


 


cggattctactagctcttttaggtcccaaccgacgaggcacagtggtatctgtcagtcct


 


ggaatatccttctctcctttttttacaataaccaagttgagaacactcaggttggcatcc


 


acaatgcatcctcggacagacttgcgcttcctctctccagttctcctaggtctataacaa


 


gaatgccccttactcaaaagcaggcgcactctgccatgggtcaaaacgccttgcttcatg


 


ggaaaaccttgtttgtcattcccaccgctgatccggaccacataacccttccactcttca


 


ccaagagcatcagcagctacttctgtggccatgcgcttctcatagaacgtacgaagcttg


 


cgttcgtcatccacttctatgagtttctgacagccagtggcagggaaggagatattcagc


 


ttcatcttgacacagccgaccgcctaggaggcgtgttaccattctgatgttggagcggcc


 


gc


 


>aj297736 (TOXMARKER Assignment: 6; SEQ ID NO: 6)


 


agttgcttcagtgtcccggtgcggttagtcacgtttcgtgcgtgctcattctgccaagat


 


gcctgaggaaacccagacccaagaccaaccaatggaggaagaggaggtcgaaacctttgc


 


ctttcaggcagaaattgcccagttaatgtccttgatcatcaacactttctactcgaacaa


 


agagatctttctgagggagctcatttccaactcctcagacgctctggataagatcagata


 


cgagagcttgaccgaccctagtaaactggactcggggaaggagctgcacattaatctcat


 


tcccaacaagcaagaccgaaccctcactattgtggatactggcattggaatgaccaaggc


 


tgacttgatcaataaccttggcactattgccaagtcaggcaccaaagccttcatggaggc


 


tttgcaggctggtgcagatatctctatgattggccagtttggtgttggtttttactctgc


 


gtatttggttgctgagaaagtgactgtcatcaccaagcataatgatgacgagcagtacgc


 


ctgggagtcctcagctggaggatccttcactgtgaggacagacacaggtgaaccaatggg


 


tcgtggaacaaaggttatcttgcatctaaaagaagaccaaactgagtatttggaggaaag


 


gagaataaaagaaattgtgaagaaacattctcagtttattggctaccccattactctctt


 


tgtggagaaggaacgtgacaaggaagtcagtgatgatgaggctgaagaaaaggaagagaa


 


agaggaagagaaagaaaaagaagaaaaggagtctgatgacaagcctgaaatagaagatgt


 


tggttctgatgaagaagaagaagagaagaaggatggtgacaagaagaaaaagaagaagat


 


aaaggaaaagtacattgatcaagaagaactcaacaaaacaaagccgatctggaccagaaa


 


tcctgatgacattacgaatgaagaatacggagagttctacaagagcttaaccaacgactg


 


ggaagaacatttggcagtaaagcatttttctgttgaaggacaattagaattccgggctct


 


tctttttgtcccaagacgcgctccttttgatctatttgaaaacagaaagaaaaagaacaa


 


catcaagttgtatgttcgcagagtttttatcatggataactgtgaggagttaatccccga


 


gtatctgaatttcatcagaggggtggtggattctgaggatctccctctaaatatttcccg


 


tgaaatgctgcaacaaagcaaaattctgaaagttatcaggaagaatttggtcaagaaatg


 


cctagaactatttactgaactggctgaagataaagagaactacaaaaagttttatgagca


 


gttctcaaaaaatataaagcttggaattcatgaagactctcaaaatcggaagaagctttc


 


agagctgttgagatactacacatctgcttctggggatgagatggtttctctgaaggacta


 


ctgcaccagaatgaaggaaaaccagaagcacatctattttatcacaggtgagaccaagga


 


ccaggttgctaactcagcctttgtggaacgtctccgaaagcatggcttagaagtaatcta


 


tatgattgagcccattgatgagtattgtgtgcaacagctgaaggaatttgagggcaagac


 


cttggtgtcagttaccaaagaaggactggaacttccagaagatgaagaggaaaagaagaa


 


acaggaagagaaaaagacaaaatttgagaacctctgcaaaattatgaaggatattttaga


 


gaaaaaggttgaaaaggtggttgtgtcaaaccgattggtgacatccccatgctgtattgt


 


cacaagcacatatggctggacagcaaacatggagagaatcatgaaagctcaagccctcag


 


agacaactcaacaatgggttacatggcagcaaagaaacacctggagataaaccctgatca


 


ctccattattgaaaccttaaggcaaaaggcagaggctgacaagaatgacaagtctgtgaa


 


agatctggtcatcttgctgtacgaaacagcactcctgtcttccggcttcagtctggaaga


 


tccccagacccatgctaacaggatctacaggatgatcaagcttggtctaggtattgatga


 


ggatgatcctactgtggatgataccagtgctgctgtaactgaagaaatgccacccctgga


 


aggagatgatgacacatcacgcatggaagaagtagactaggcttcaccagaactatgtgt


 


ttgatgcttaccttcattccttctgataatatattttccatgatttttgtttatttttgt


 


taacatttaaaacatctgtgtggcatgaaaactaggggaaggtaaaaatttctacatgtg


 


atactgtgatactataggtttgactcaagaggttgatagaacgtttgttgtaagacgtaa


 


tgtaacctacggtacttgttaactatgggggtctgaaagtgtttagctgttgagctggat


 


tcctttagtagaccaaattaagatgacttaagtttcatct


 


>j00719 (TOXMARKER Assignment: 7; SEQ ID NO: 7)


 


ttgctcctccttgctctcctcgtgggcttcttgttactcttagtcaggggacacccaaag


 


tcccgtggcaacttcccaccaggacctcgtccccttcccctcttggggaacctcctgcag


 


ttggacagagggggcctcctcaattccttcatgcagcttcgagaaaaatatggagatgtg


 


ttcacagtacacctgggaccaaggcctgtggtcatgctatgtgggacagacaccataaag


 


gaggctctggtgggccaagctgaggatttctctggtcggggaacaatcgctgtgattgag


 


ccaatcttcaaggaatatggtgtgatctttgccaatggggaacgctggaaggcccttcgg


 


cgattctctctggctaccatgagagactttgggatgggaaagaggagtgtggaagaacgg


 


attcaggaggaagcccaatgtttggtggaggaactgcggaaatcccagggagccccactg


 


gatcccaccttcctcttccagtgcatcacagccaacatcatctgctccattgtgtttgga


 


gagcgctttgactacacagaccgccagttcctgcgcctgttggagctgttctaccggacc


 


ttttccctcctaagttcattctccagccaggtgtttgagttcttctctgggttcctgaaa


 


tactttcctggtgcccacagacaaatctccaaaaacctccaggaaatcctcgattacatt


 


ggccatattgtggagaagcacagggccaccttagacccaagcgctccacgagacttcatc


 


gacacttaccttctgcgcatggagaaggagaagtcgaaccaccacacagagttccatcat


 


gagaacctcatgatctccctgctctctctcttctttgctggcactgagaccagcagcacc


 


acactccgctatggtttcctgctgatgctcaagtacccccatgtcgcagagaaagtccaa


 


aaggagattgatcaggtgatcggctcacaccggctaccaacccttgatgaccgcagtaaa


 


atgccatacactgatgcagttatccacgagattcagaggttttcagatcttgtccctatt


 


ggagtaccacacagagtcaccaaagacaccatgttccgagggtacctgcttcccaagaac


 


actgaagtgtaccccatcctgagttcagctctccatgacccacagtactttgaccaccca


 


gacagcttcaatcctgaacacttcctggatgccaatggggcactgaaaaagagtgaagct


 


ttcatgcccttctccacaggaaagcgcatttgtcttggcgaaggcattgcccgaaatgaa


 


ttgttcctcttcttcaccaccatcctccagaacttctctgtgtcaagccatttggctccc


 


aaggacattgacctcacgcccaaggagagtggcattggaaaaatacctccaacgtaccag


 


atctgcttctcagctcggtgatccggctgaggcagccaggtgccccagttctgttgggaa


 


tggcctcatgtttctgcctctgggggacctgctgaaaaccaggctccaaggccactgctc


 


cacatct


 


>j00720 (TOXMARKER Assignment: 8; SEQ ID NO: 8)


 


cccagtgcccttttgtcctgtgtatctgtttcgtggtgtccttgccaacatctatggtgt


 


gggtaagggaatgaggagtgaatagccaaagcaggaggcgtgaacatctgaagttgcata


 


actgagtgtaggggcagattcagcataaaagatcctgctggagagcatgcactgaagtct


 


accgtggttacaccaggaccatggagcccagtatcttgctcctccttgctctccttgtgg


 


gcttcttgttactcttagtcaggggacacccaaagtcccgtggcaacttcccaccaggac


 


ctcgtccccttcccctcttggggaacctcctgcagttggacagaggaggcctcctcaatt


 


ccttcatgcagcttcgcgaaaaatatggagatgtgttcacagtacacctgggaccaaggc


 


ctgtggtcatgctatgtgggacagacaccataaaggaggctctggtgggccaagctgagg


 


atttctctggtcggggaacaatcgctgtgattgagccaatcttcaaggaatatggtgtga


 


tctttgccaatggggaacgctggaaggcccttcggcgattctctctggctaccatgagag


 


actttgggatgggaaagaggagtgtggaagaacggattcaggaggaagcccaatgtttgg


 


tggaggaactgcggaaatcccagggagccccactggatcccaccttcctcttccagtgca


 


tcacagccaacatcatctgctccattgtgtttggagagcgctttgactacacagaccgcc


 


agttcctgcgcctgttggagctgttctaccggaccttttccctcctaagttcattctcca


 


gccaggtgtttgagttcttctctgggttcctgaaatactttcctggtgcccacagacaaa


 


tctccaaaaacctccaggaaatcctcgattacattggccatattgtggagaagcacaggg


 


ccaccttagaccccagcgctccacgagacttcatcgacacttaccttctgcgcatggaga


 


aggagaagtcgaaccaccacacagagttccatcatgagaacctcatgatctccctgctct


 


ctctcttctttgctggcactgagaccggcagcaccacactccgctatggtttcctgctca


 


tgctcaagtacccccatgtcacagtgaaagtccaaaaggagattgatcaggtgattggct


 


ctcacaggccaccatcccttgatgatcgtaccaaaatgccatacactgatgcagtcatcc


 


acgagattcagaggtttgcagatcttgccccaattggtttaccacacagagtcaccaaag


 


acaccatgttccgagggtacctgctccccaagaacactgaggtgtatcccatcctgagtt


 


cagctctccatgacccacagtactttgaccatccagacaccttcaatcctgagcacttcc


 


tggatgccgatgggacactgaaaaagagtgaagcttttatgcccttctccacaggaaagc


 


gcatttgtcttggcgaaggcattgcccgaaatgaattgttcctcttcttcaccaccatcc


 


tccagaacttctctgtgtcaagccatttggctcccaaggacattgacctcacgcccatgg


 


agagtggcattgcaaaaatacctccaacgtaccagatctgcttctcagctcggtgatcgg


 


gctgag


 


>j00728 (TOXMARKER Assignment: 9; SEQ ID NO: 9)


 


atggagcccagtatcttgctcctccttgctctccttgtgggcttcttgttactcttagtc


 


aggggacacccaaagtcccgtggcaacttcccaccaggacctcgtccccttcccctcttg


 


gggaacctcctgcagttggacagaggaggcctcctcaattccttcatgcagcttcgcgaa


 


aaatatggagatgtgttcacagtacacctgggaccaaggcctgtggtcatgctatgtggg


 


acagacaccataaaggaggctctggtgggccaagctgaggatttctctggtcggggaaca


 


atcgctgtgattgagccaatcttcaaggaatatggtgtgatctttgccaatggggaacgc


 


tggaaggcccttcggcgattctctctggctaccatgagagactttgggatgggaaagagg


 


agtgtggaagaacggattcaggaggaagcccaatgtttggtggaggaactgcggaaatcc


 


cagggagccccactggatcccaccttcctcttccagtgcatcacagccaacatcatctgc


 


tccattgtgtttggagagcgctttgactacacagaccgccagttcctgcgcctgttggag


 


ctgttctaccggaccttttccctcctaagttcattctccagccaggtgtttgagttcttc


 


tctgggttcctgaaatactttcctggtgcccacagacaaatctccaaaaacctccaggaa


 


atcctcgattacattggccatattgtggagaagcacagggccaccttagaccccagcgct


 


ccacgagacttcatcgacacttaccttctgcgcatggagaaggagaagtcgaaccaccac


 


acagagttccatcatgagaacctcatgatctccctgctctctctcttctttgctggcact


 


gagaccggcagcaccacactccgctatggtttcctgctcatgctcaagtacccccatgtc


 


acagtgaaagtccaaaaggagattgatcaggtgattggctctcacaggccaccatccctt


 


gatgatcgtaccaaaatgccatacactgatgcagtcatccacgagattcagaggtttgca


 


gatcttgccccaattggtttaccacacagagtcaccaaagacaccatgttccgagggtac


 


ctgctccccaagaacactgaggtgtatcccatcctgagttcagctctccatgacccacag


 


tactttgaccatccagacaccttcaatcctgagcacttcctggatgccgatgggacactg


 


aaaaagagtgaagcttttatgcccttctccacaggaaagcgcatttgtcttggcgaaggc


 


attgcccgaaatgaattgttcctcttcttcaccaccatcctccagaacttctctgtgtca


 


agccatttggctcccaaggacattgacctcacgcccatggagagtggcattgcaaaaata


 


cctccaacgtaccagatctgcttctcagctcggtga


 


>l00320 (TOXMARKER Assignment: 10; SEQ ID NO: 10)


 


atggagcccagtatcttgctcctccttgctctccttgtgggcttcttgttactcttagtc


 


aggggacacccaaagtcccgtggcaacttcccaccaggacctcgtccccttcccctcttg


 


gggaacctcctgcagttggacagagggggcctcctcaattccttcatgcagcttcgagaa


 


aaatatggagatgtgttcacagtacacctgggaccaaggcctgtggtcatgctatgtggg


 


acagacaccataaaggaggctctggtgggccaacctgaggatttctctggtcggggaaca


 


atcgctgtgattgagccaatcttcaaggaatatggtgtgatctttgccaatggggaacgc


 


tggaaggcccttcggcgattctctctggctaccatgagagactttgggatgggaaagagg


 


agtgtggaagaacggattcaggaggaagcccaatgtttggtggaggaactgcggaaatcc


 


cagggagccccactggatcccaccttcctcttccagtgcatcacagccaacatcatctgc


 


tccattgtgtttggagagcgctttgactacacagaccgccagttcctgcgcctgttggag


 


ctgttctaccggaggttttccctcctaagttcattctccagccaggtgtttgagttcttc


 


tctgggttcctgaaatactttcctggtgcccacagacaaatctccaaaaacctccaggaa


 


atcctcgattacattggccatattgtggagaagcacagggccaccttagacccaagcgct


 


ccacgagacttcatcgacacttaccttctgcgcatggagaaggagaagtcgaaccaccac


 


acagagttccatcatgagaacctcatgatctccctgctctctctcttctttgctggcact


 


gagaccagcagcaccacactccgctatggtttcctgctgatgctcaagtacccccatgtc


 


gcagagaaagtccaaaaggaggttgatcaggtgatcggttcacaccggctaccaaccctt


 


gatgaccgcagtaaaatgccatacactgatgcagttatccatgagattcataggttttca


 


gatcttgtccctattggagtaccacacagagtcaccaaagacaccatgttccgagggtac


 


ctgcttcccaagaacactgaagtgtaccccatccggagttcagctctccatgacccacag


 


tactttgaccacccagacagcttcaatcctgaacacttcctggacgttaacggggcactg


 


aaaaagagtgaagctttcatgcccttctccacaggaaagcacatttgtcttggcgaaggc


 


attgcccgaaatgaattgttcctcttcttcaccaccatcctccagaacttctctgtgtca


 


agccatttggctcccaaggacattgacctcacgcccaaggagagtggcattggaaaaata


 


cctccaacgtaccagatctgcttctcagctcggtga


 


>m11251 (TOXMARKER Assignment: 11; SEQ ID NO: 11)


 


cccagtgcccttttgtcctgtgtatctgtttcgtggtgtccttgccaacatgtatggtgt


 


gggtaagggaatgaggagtgaatagctaaagcaggaggcgtgaacatctgaagttgcata


 


actgagtggaggggcggattcagcataaaagatcctgctggagagcatgcactgaagtct


 


accgtggttacaccaggaccatggagcccagtatcttgctcctccttgctctccttgtgg


 


gcttcttgttactcttagtcaggggacacccaaagtcccgtggcaacttcccaccaggac


 


ctcgtccccttcccctcttggggaacctcctgcagttggacagagggggcctcctcaatt


 


ccttcatgcagcttcgagaaaaatatggagatgtgttcacagtacacctgggaccaaggc


 


ctgtggtcatgctatgtgggacagacaccataaaggaggctctggtgggccaacctgagg


 


atttctctggtcggggaacaatcgctgtgattgagccaatcttcaaggaatatggtgtga


 


tctttgccaatggggaacgctggaaggcccttcggcgattctctctggctaccatgagag


 


actttgggatgggaaagaggagtgtggaagaacggattcaggaggaagcccaatgtttgg


 


tggaggaactgcggaaatcccagggagccccactggatcccaccttcctcttccagtgca


 


tcacagccaacatcatctgctccattgtgtttggagagcgctttgactacacagaccgcc


 


agttcctgcgcctgttggagctgttctaccggaggttttccctcctaagttcattctcca


 


gccaggtgtttgagttcttctctgggttcctgaaatactttcctggtgcccacagacaaa


 


tctccaaaaacctccaggaaatcctcgattacattggccatattgtggagaagcacaggg


 


ccaccttagacccaagcgctccacgagacttcatcgacacttaccttctgcgcatggaga


 


aggagaagtcgaaccaccacacagagttccatcatgagaacctcatgatctccctgctct


 


ctctcttctttgctggcactgagaccagcagcaccacactccgctatggtttcctgctga


 


tgctcaagtacccccatgtcgcagagaaagtccaaaaggaggttgatcaggtgatcggtt


 


cacaccggctaccaacccttgatgaccgcagtaaaatgccatacactgatgcagttatcc


 


atgagattcataggttttcagatcttgtccctattggagtaccacacagagtcaccaaag


 


acaccatgttccgagggtacctgcttcccaagaacactgaagtgtaccccatccggagtt


 


cagctctccatgacccacagtactttgaccacccagacagcttcaatcctgaacacttcc


 


tggacgttaacggggcactgaaaaagagtgaagctttcatgcccttctccacaggaaagc


 


acatttgtcttggcgaaggcattgcccgaaatgaattgttcctcttcttcaccaccatcc


 


tccagaacttctctgtgtcaagccatttggctcccaaggacattgacctcacgcccaagg


 


agagtggcattggaaaaatacctccaacgtaccagatctgcttctcagctcggtgatccg


 


gctgaggcagccatgtgccccagttctgttgggaatggcctcatgtttctgcctctgggg


 


gacctgctgaaaaccaggct


 


>m26125 (TOXMARKER Assignment: 12; SEQ ID NO: 12)


 


gacttgggaggaaccagggcctacacttagccctggtaaacagcagagcatgctgggata


 


attcttcccagaaaaggaaaagcaggcacttctgttcccagggaaaacaacaggagcact


 


ttggacctccctgctgcagtcaggagtcatgtggctggaacttgtcctggcttcccttct


 


gggctttgtcatctactggtttgtctcccgggacaaggaggaaaccttaccactaggaga


 


tggatggtgggggccagggtcaaagccatcagccaaagaagatgagagcatccggccctt


 


caaggtggaaacatcagatgaggagatcaaggacttacaccagaggatagataggttccg


 


ggcatccccacctttggagggcagccgcttccactatggcttcaactccaactacatgaa


 


gaaagtggtgtcctactggaggaacgagtttgactggaggaagcaggtggagatcctcaa


 


ccagtaccctcacttcaagaccaagatcgaagggcttgacatccacttcatccatgtgaa


 


gcctccccagctgccctcagggcgcaccccaaagcccttgctgatggtgcatggctggcc


 


tggatccttctatgagttttacaagatcatcccactactgactgaccccaagtcccacgg


 


tctgagtgacgagcacgtgtttgaagtcatctgtccctcgattcctggctatggctactc


 


agaggcatccagcaagaaaggtttaaattcggtggccactgcgaggattttctacaagct


 


gatgacacggctgggcttccagaaattctacattcaaggcggggactgggggtccctcat


 


ctgcaccaacatggcccagatggttcccaaccacgtgaaaggcctgcacttaaatatggc


 


tttcatttcgagaagtttttacaccatgactcctctcctgggccaacgcttcgggagatt


 


ccttggctacacagagaaggatatcgagctcttgtacccctataaggagaaggttttcta


 


cagcatcatgagggagagtggctacttacacatccaagccaccaagccagacactgtggg


 


ctgtgctctcaatgactctcccgtgggcctggctgcctacatcttagagaagttctccac


 


ctggaccaagtcagagtaccgtgaactggaggatggaggcctggagaggaagttctccct


 


ggatgatctgctggttaacatcatgatctactggacgacaggaaccattgtctcctccca


 


acgctactacaaggagaatttgggccagggcatcatggtccataaacatgaggggatgaa


 


ggtctttgtgcccactggcttttcagccttcccttccgagctactgcatgccccagaaaa


 


gtgggtgaaggtcaagtaccccaaactcatctcctattcctacatggaacgtgggggcca


 


ctttgctgcctttgaagagcccaagcttctggcccaggacatccgcaagttcgtgtccct


 


ggctgagctgcagtagtgacactggataccaactgtggctttagcagcagccctggttcc


 


tcccaagtcacacttatggaagatgacccctttctgaggaataagtttgttccctgacca


 


cactcgaggacccagacttaaactccacagagtcgtatgttacccccatatgcttcacct


 


cactacatagctgtgttagctacatggctttaatgataaatggatttatttct


 


>m34452 (TOXMARKER Assignment: 13; SEQ ID NO: 13)


 


tgagccaatcttcaaggaatatggtgtgttctttgccaatggggaacgctggaaggccct


 


tcggcgattctctctggctaccatgagagactttgggatgggaaagaggagtgtggaaga


 


acggattcaggaggaagcccaatgtttggtggaggaactgcggaaatcccagggagcccc


 


actggatcccaccttcctcttccagtgcatcacagccaacatcatctgctccattgtgtt


 


tggagagcgctttgactacacagaccgccagttcctgcgcctgttggagctgttctaccg


 


gaccttttccctcctaagttcattctccagccaggtgtttgagttcttctctgggttcct


 


gaaatactttcctggtgcccacagacaaatctccaaaaacctccaggaaatcctcgatta


 


cattggccatattgtggagaagcacagggccaccttagaccccagcgctccacgagactt


 


catcgacacttaccttctgcgcatggagaaagtgagtcctgcatggatgagagaggagaa


 


gtcgaaccaccacacagagttccatcatgagaacctcatgatctccctgctctctctctt


 


ctttgctggcactgagaccggcagcaccacactccgctatggtttcctgctcatgctcaa


 


gtacccccatgtcacagagaaagtccaaaaggagattgatcaggtgattggctctcacag


 


gccaccatcccttgatgatcgtaccaaaatgccatacactgatgcagtcatccacgagat


 


tcagagatttgcagatcttgccccaattggtttaccacacagagtcaccaaagacaccat


 


gttccgagggtacctgctccccaagaacactgaggtgtatcccatcctgagttcagctct


 


ccatgacccacagtactttgaccatccagacaccttcaatcctgagcacttcctggatgc


 


cgatgggacactgaaaaagagtgaagcttttatgcccttctccacaggaaagcgcatttg


 


tcttggcgaaggcattgcccgaaatgaattgttcctcttcttcaccaccatcctccagaa


 


cttctctgtgtcaagccatttggctcccaaggacattgacctcacgcccaaggagagtgg


 


cattgcaaaaatacctccaacataccagatctgcttctcagctcggtgatcgggctgagg


 


cagccaggtgccccagttctgttgggaatggcctcatgtttctgcctctgggggacctgc


 


tgaaaaccaggctcaaggccactgctcacatcttcctattgcagttctccaaagtcccaa


 


ggcttgttcttattcctgtgaatggcactgaagaagtcaatcgactgtcttattttgaca


 


tgtgaacagagatttcatgagtacacatctcatgctgagtcacttccctcttcctcctaa


 


tagcccacgtccccacttatcagccctccatggtctgtgatctgtgctaatggactctgt


 


atatggtctcagtgctatgtctacagacttacatagtatgtatggttcaggtaaacagaa


 


tcacagagtgtgtg


 


>u33546 (TOXMARKER Assignment: 14; SEQ ID NO: 14)


 


atggaacctagtgtcctacttctccttgctgtcctcctcagcttcttgctactcctggtc


 


aggggccatgcaaagatccatggtcgtcttccaccaggaccctgccctgtaccccttttg


 


ggaaatctcttgcagatggacagaagaggcctcctcaagtcttttattcagcttcaagaa


 


aaatatggagatgtgttcacagtgcacttaggactgaggccagtggtcgtgttatgtggg


 


acacagaccataagagaggctctggtggaccatgctgaggctttctctggccgggggaca


 


attgctgggcttgagccagttttccaggactatggtatattcttttccagtggagaacag


 


tggaagacccttcgacgattctctatggccaccatgagagactttgggatgagaaagaag


 


agtgtggaggagagaataaaggaagaatcccaatgtttggtggaggaactgaagaaatac


 


cagggagcccccctggatcccaccttccttttccagtgcatcacatccaacatcatctgc


 


tccattgtctttggagagtgctttgactacacagatcaccaattcctgcacctgctggat


 


ctgatgtatcagacgttttcactcttaagctcaatcttcagtcaggtatttgaactcttc


 


cctggtgtcctgaagtactttcctggtgcccacagacaaatctccagaaacctccatgaa


 


atcctggacttcattggccagagtgtggagaagcacagggccactttggacccaaatgct


 


ccacgagactttatatatacttaccttctgcacatggagaaaaagtcaaaccattataca


 


gagttccatcactggaacctactgtcgtctgtactctctctcttctttgctggcactgag


 


actagcagcaccacactccgctatggcttcctgatcatgctcaagtaccctcatatcaca


 


gagaaagtccaaaaagagattgattgtgtgattggctcacaccggctacctaccctggat


 


gaccgcagcaaaatgccatacaccgaggcagttatccatgagattcagagattttcagat


 


cttgcccctattggaacaccacacagagtcatcaaagacaccattttccgagggtacctg


 


ctccctaagaacactgaggtgttccccatcctgagttcagttctccatgatccacagtac


 


tttgaacaaccagacatcttcaatcttcagcactttctggatgccaatggggcactgaag


 


ataattgaagcttttctgcccttctccacaggaaagcgaatttgtcttggtgaaagcatt


 


gcccgcaatgaattgttccttttcttcactaccatcctccagaacttctccgtgtccagc


 


cctgtggctcctaaagacattgatctcactcccaaagagagtggtattggaagaataccc


 


caagtgtaccagatctgcttcttggcccactga


 


>x74673 (TOXMARKER Assignment: 15; SEQ ID NO: 15)


 


gaattccgcggccgccaacgtcctctcttacccgccaccttcttctgccacctctaccac


 


ggtcaccatgtcgcaagcccggcctgccactgtgctgggtgccatggagatgggtcgccg


 


catggatgtgacctccagctccgcgtcggtgcgcgccttcctgcagcgcggccacacgga


 


gatagacaccgccttcgtgtatgcgaacggtcagtctgagaccatcctaggagacctggg


 


gctcggactgggccgcagcggctgcaaagtaaaaattgccaccaaggctgccccaatgtt


 


tgggaagacactgaagccagccgatgttcggttccagctggagacgtcactgaagaggct


 


gcagtgtccccgggtggacctcttctatttacactttccagaccacggcactcctataga


 


ggagaccctgcaggcctgccaccacgtgcatcaggagggcaagtttgtggagcttggtct


 


gtccaactatgtctcctgggaagtggctgagatttgtaccctctgcaagaaaaatggctg


 


gatcatgccaactgtgtaccagggcatgtacaacgccatcaccaggcaggtggagactga


 


gctcttcccctgcctcagacacttcggactaaggttctacgccttcaaccctttggctgg


 


gggcctgctgactggcagatataaataccaggataaggatgggaagaatcctgagagccg


 


cttctttgggaatccattttctcaactgtacatggaccgctactggaaggaggaacactt


 


caatggcatcgccttggtggagaaggctctgaagactacctatggccccactgcccccag


 


tatgatctcagctgccgtacggtggatgtaccatcactcacagctcaagggcacccaagg


 


ggatgcagtcattctgggcatgtccagtctggaacaactggagcagaacttggccttggt


 


cgaggaagggcctctggagccagctgttgtggatgcctttgaccaagcctggaacctagt


 


tgcccacgagtgtcccaactatttccgctaagatacatctgccttggggatggcgcagct


 


tactgcctgccccgccttgtcctgggctcgatctgatctggttctttcctttttagacag


 


gtcactgtctttttcttccctgctttctatacagccagttgctttcaaagtgagagctgg


 


ctgagccccaatacctcctgctgaataaaactgttccctgtcacagcctgggctacaact


 


ggcggccga


 


>scr_gb-x13044_4 (TOXMARKER Assignment: 16; SEQ ID NO: 16)


 


ttttttttttttttttttctaccttctaccttttattgtcacgtgaaccatggtcctaca


 


ggctgctgacaagcttggctgagcagggatcccaggggcgtcggcaggacatgaggaagg


 


gttgctgggagggcttggcctcttccttgagaagacagcaaatgtatccagcctagatta


 


agggtagggcatcccctatccctgtcagtgggcctagatctcagagccccacattaaaga


 


ctgctaatgggtcagaaatgggggtcccttagatgggggtaggcagcaaggccctccctc


 


cagtgttctcattctgttccggtttcatttgttgtgtccagggacggtgaagcagatacc


 


agtctcaagccccagggtgcaggaagacgggaaatggggtgtgatgttagggagtgtaag


 


aagggctgaggagcaggggagctgccgccgtgcagagctggcttctgtcttcacaagaac


 


atttggcccatatcctgcttggtcactcccaggccagaagatgggtcttccatgtccagt


 


ggctctttaggtggagtctgggtgggctgcttctcctccagggagttcttgctcatttca


 


aacaacagccactgtttcatccagctctcaaagaccttccagtccagaccattcatagag


 


ttcttaaggtgcttcagattctccgggaagctccccttcagctgtgggtagttcacgggt


 


ccagacttcgtaagcaggtgcatcacgtggtcctgggtcatgttgccatacttggtaaca


 


ttcttcacgggcgcttggagcatgttatccatggacagtgggcgcatcagcaagggagta


 


gccatgcgcatcgggctcacaggtttggcagatttcggaagcttcatgcgaaggttctcc


 


agttgcaggttctgggaggtgacggtcagcttgtccaggcggccctgctgctggtacagg


 


aagtaagcagtggtggcctgcccagccaagagcagagccaccaggacagagacactggtg


 


tacaggactccacggttgcaattgctttctggggctctagcacgctggcccaggatgggc


 


agctgctcatggttagagatgaggtcgcgctggtcatccatgactctagcctctagcttt


 


tcccccaagtgctgctggtgctgctgctgctgctgct


 


>scr_gb-x14254_5 (TOXMARKER Assignment: 17; SEQ ID NO: 17)


 


tttttttttttttttgttctaccttctaccttttattgtcacgtgaaccatggtcctaca


 


ggctgctgacaagcttggctgagcagggatcccaggggcgtcggcaggacatgaggaagg


 


gttgctgggagggcttggcctcttccttgagaagacagcaaatgtatccagcctagatta


 


agggtagggcatcccctatccctgtcagtgggcctagatctcagagccccacattaaaga


 


ctgctaatgggtcagaaatgggggtcccttagatgggggtaggcagcaaggccctccctc


 


cagtgttctcattctgttccggtttcatttgttgtgtccagggacggtgaagcagatacc


 


agtctcaagccccagggtgcaggaagacgggaaatggggtgtgatgttagggagtgtaag


 


aagggctgaggagcaggggagctgccgccgtgcagagctggcttctgtcttcacaagaac


 


atttggcccatatcctgcttggtcactcccaggccagaagatgggtcttccatgtccagt


 


ggctcactgcagttatggcgcccgcggctcttggtgtgagggacctcagtgccgttgggg


 


aacacacaccagcagtagccagtgctcccatggcactggagtggcatatagttaccgttc


 


tcatcacacttgggacggaacgcccccgggtggacatcagggatgtggctgacttcttcc


 


tggcacttggtcaatactttaggtggagtctgggtgggctgcttctcctccagggagttc


 


ttgctcatttcaaacaacagccactgtttcatccagctctcaaagaccttccagtccaga


 


ccattcatagagttcttaaggtgcttcagattctccgggaagctccccttcagctgtggg


 


tagttcacgggtccagacttcgtaagcaggtgcatcacgtggtcctgggtcatgttgcca


 


tacttggtaacattcttcacgggcgcttggagcatgttatccatggacagtgggcgcatc


 


agcaagggagtagccatgcgcatcgggctcacaggtttggcagatttcggaagcttcatg


 


cgaaggttctccagttgcaggttctgggaggtgacggtcagcttgtccaggcggccctgc


 


tgctggtacaggaagtaagcagtggtggcctgcccagccaagagcagagccaccaggaca


 


gagacactggtgtacaggactccacggttgcaattgctttctggggctctagcacgctgg


 


cccaggatgggcagctgctcatggttagagatgaggtcgcgctggtcatccatgactcta


 


gcctctagcttttcccccaagtgctgctggtgctgctgctgctgctgctgctg


 


>scr_gb-bi275638_1 (TOXMARKER Assignment: 18; SEQ ID NO: 18)


 


cggcacgaggcgcgctcggcgctgtcagttcgtcccgctgcccctcggcccttgctgctg


 


gctctgacggcgaccgacggcgggcggggcccgggttcgcggccgagcggcgccggtgag


 


ggcgcggaggaggcgcacagcgggaggaggagccgtgagcctggcacggagcggccgcgg


 


ccatggcgtacgcctatctcttcaagtacatcatcatcggcgacacaggtgttggtaaat


 


cgtgcttattgctacagtttacagacaagaggtttcagccggtgcatgacctcacaattg


 


gtgtagagtttggtgctcgaatgataaccattgatgggaaacagataaaactccagatct


 


gggatacagcagggcaggagtcctttcgttctatcacaaggtcatattacagaggtgcag


 


cgggggctttactagtgtatgatattacaaggagagacacgttcaaccacttgacaacct


 


ggttagaagacgcccgtcagcattccaattccaacatggtcatcatgcttattggaaata


 


aaagtgacttagaatctaggagagaagtgaaaaaggaagaaggtgaagcttttgcacgag


 


agcatggacttatcttcatggaaacttctgccaagactgcttctaatgtagaggaggcat


 


ttattaacacagcaaaagaaatttatgaaaaaatccaagaaggggtctttgacattaata


 


atgaggcaaacggcatcaaaattggccctcagcatgctgctaccaatgcatctcacggag


 


gcaaccaaggagggcagcaggcagggggaggctgctgctgagtctgctgttgccggctag


 


ctgcccagtggagccacgcactctgtcaccctctctcctcatgctcagctgagacatgaa


 


actattgaaatggctttgtgtcacaggagactttaatccttcagattcttgtataacttt


 


gaataaatggttaatgttcacttaaaaagacagattttggagattgtattcatatctatt


 


tgcatttgatttctaggtcaattg


 


>scr_gb-x66871_3 (TOXMARKER Assignment: 19; SEQ ID NO: 19)


 


tttttttttttttttttttttttttttttttttttttttttttttttttttttttttcta


 


aagtaaaaatggtttattcacgacacatatgaggaagtgtctcatgtcacagacggtacg


 


tccaactccctggaatgttcatttctttggcataaaggagagaatgaggggaaagccagg


 


caaaggcagctaagatgggggatgggtcggcagctctgtcgtcatcttcacagggaggag


 


ttcaggggtccattagtggcaggctgattctctagaacattaggttggggcacaggtagg


 


gccacttctgggcaatccaccatgccaagcccttcagtcgtccccaccacacaggtacag


 


cagcgccttctggtagtcacccttagtgtcttgctggatgaagtagtacagggatttgcc


 


atatttcctcttgaattcagatctgattttcaacatgtccacttcactgcgagagaccat


 


gattctaatcaggaccttgtctcgagtccccttgcccttcatggagtcatacagccggtc


 


agcaaagtacaggggcttgttctgaatgcactgaaccaggttcaggaaggcgttctccag


 


gtctcctttgacctctttcctgatgctctccagcatgtcataaggactgtagctcttgta


 


cctttcgaacactttctggaggtggcacacactgcgctcagtcatgatgctgatccactt


 


ggggacatcggttcctttcctcttcaccccagcatcatagagctcccgggcatcctggtc


 


aatcagctcgtagtcaataacagaaccatcctctgcccgtttaccctttgcaagggcgac


 


caacagctttcggaattctccagatgtgtcagagatgatgtccttctccagatcggtctt


 


gtacatttccttatacactcggttaatctcctgcagctcctggttggttcttgagcagat


 


gatctcgatgagggagtcctcatcagtccccaggcccttcatggaggctttgagctcaga


 


ggcatcgtactgagcaggtgtcttcaacaggcctaacatcacggtctccaggtgaccaga


 


caaggccgacttcatcgccgatggcagttcctttttggtcctcctctggtaggcgaaggc


 


aatgtcctgcctctgtgcattgctgcggttagtcagaatgttgacaatggtgacctcgtc


 


cacgcctttggtcttgattgctgtttcaatgttcaaagcatccctctcagcgtcgaagtt


 


ggtgtagggtttgaccgacccataggcacttgggggtgtagaatgctgagaatcaccctc


 


caagctgagcttgcacaggatttcgtggacagtagacattttgaaaaaaaagctgggccg


 


ggcacctattgcagagagcctcc


 


>scr_gb-149379_3 (TOXMARKER Assignment: 20; SEQ ID NO: 20)


 


gggatgacatagagtacaacattcagagaagttaactattaagtcgtcaggatgaaaggt


 


caggaggcaggcctttaactgggctgtgagaatggagaaagcacggtgcactttaacatc


 


tgctttcccagaggaaaaagtaaaggagaaacagtacaatcatagaagagtcttcgtaac


 


agaagcgcgaggagagcattatggacaagttctgcaactctactttttgggatctctcat


 


tactggaaagtccagaggctgacctgcctctttgttttgagcaaactgttctggtgtgga


 


ttcccttgggctttctttggctcctggctccttggcaactttacagcgtgtacagatcca


 


ggaccaagagatcttctataaccaaattctaccttgccaagcaggtgttcgtcgtgtttc


 


ttcttattttagcagccatagacctgtctcttgcgctcacagaagatactggacaagcca


 


cagttcctcctgtcagatatacgaatccaatcctctacctgtgcacatggctcctggttt


 


tggcagtccagcacagcaggcaatggtgtgtacgaaagaactcttggttcctgtctctgt


 


tctggatcctctcggtcttatgcggcgtattccagtttcagactctgatacgagcactcc


 


tgaaggacagcaagtccaacatggcctactcctacctgttcttcgtctcctacggtttcc


 


agattgtcctcctgattcttacagccttttcaggaccaagtgactcaacacaaactccat


 


cagtcacggcttcctttctgagtagcattacatttagttggtatgacaggactgttctga


 


aaggttacaagcatccactgacactagaagatgtctgggatatcgatgaagggtttaaaa


 


caaggtcagtcaccagcaagtttgaggcggccatgacaaaggacctgcagaaagccaggc


 


aggcttttcagaggcggctgcagaagtcccagcggaaacctgaggccacactacacggac


 


tgaacaagaagcagagtcagagccaagacgttctcgtcctggaagaagcgaaaaagaagt


 


ctgagaagaccaccaaagactatcccaaatcgtggttgatcaagtctctcttcaaaacct


 


tccacgtagtgatcctgaaatcatttatactgaaattaatacatgaccttttggtgtttc


 


tgaatcctcagctgctgaagttgctgatcggtttcgtgaagagctctaactcatacgtgt


 


ggtttggctatatctgtgcaatcctaatgtttgctgtgactctcatccaatctttctgcc


 


ttcagtcttactttcaacattgttttgtgttgggaatgtgcgtacggacaaccgtcatgt


 


cttcgatatataagaaggcattgaccctatctaacttggctaggaagcagtacaccattg


 


gagagacggtgaacttgatgtctgtagattcccagaagctaatggatgcgaccaactaca


 


tgcagttggtgtggtcaagtgttatacagattactttgtccatcttcttcctgtggagag


 


agttgggaccgtccatcttagcaggtgttggggttatggttctcctaatcccagttaatg


 


gagttctggctaccaagatcagaaatattcaggtccaaaatatgaagaataaagacaaac


 


gtttaaaaatcatgaatgagattctcagtggaatcaagatcctgaaatactttgcctggg


 


agccttcatttcaagagcaagtccagggcattcggaagaaagaactcaagaacttgctgc


 


ggttcggccagctgcagagtctgctgatcttcattttacagataactccaatcctggtgt


 


ctgtggtcacattttctgtctatgtcctggtggatagcgccaatgttttgaatgcggaga


 


aggcatttacctccatcaccctcttcaatatcctacgcttccctctgtccatgcttccca


 


tggtgacctcatcgatcctccaggccagtgtttctgtggaccggctggagaggtatttgg


 


gaggagacgatttagacacatctgccattcgccgcgtcagcaattttgataaagctgtga


 


agttttcagaggcctcttttacttgggacccggacttggaagccacaatccaagatgtga


 


acctggacataaagccaggccaactggtggctgtggtgggcactgtaggctctgggaaat


 


cctctttggtatcagccatgctgggagaaatggaaaacgttcacgggcacatcaccatcc


 


agggatccacagcctatgtccctcagcagtcctggattcagaatggaaccatcaaagaca


 


acatcctgtttgggtccgaatacaatgaaaagaagtaccagcaagttctcaaagcatgcg


 


ctctcctcccagacttggaaatattgcctggaggagacatggctgagatcggagagaagg


 


ggataaatctcagtggtggtcagaagcagcgagtcagcctggccagagctgcctatcaag


 


atgctgacatctatattctggacgatcccctgtcggctgtggatgctcatgtgggaaaac


 


acattttcaacaaggttgtgggccccaacggcctgttggctggcaagacgagaatctttg


 


ttactcatggtattcacttccttccccaagtggatgagattgtagttctggggaaaggca


 


ccatcttagagaaaggatcctatcgtgacctgttggacaagaagggagtgtttgctagga


 


actggaagaccttcatgaagcattcagggcctgaaggagaggccacagtcaataatgaca


 


gtgaggcggaagacgacgatgatgggctgattcccaccatggaggaaatccctgaggatg


 


cagcttccttggccatgagaagagaaaatagtcttcgccgtacactgagccgcagctcta


 


ggtccagcagccgacgtgggaagtccctcaaaaactccttgaagattaaaaatgtgaatg


 


tcttgaaggagaaggaaaaagaagtggaaggacaaaaactaattaagaaagaatttgtgg


 


aaaccgggaaggtcaagttctccatctacctgaagtatctacaggcagtagggtggtggt


 


ccatacttttcatcatccttttctacggattgaataatgttgcttttatcggctctaacc


 


tctggctgagtgcttggaccagtgactctgacaacttgaatgggaccaacaattcgtctt


 


ctcatagggacatgagaattggggtctttggagctctgggattagcacaaggtatatgtt


 


tgcttatttcaactctgtggagcatatatgcttgcagaaatgcatcaaaagctttgcacg


 


ggcagctgttaaccaacatcctccgggcacccatgaggttttttgacacaactcccacag


 


gccggattgtgaacagattttctggtgatatttctactgtggacgacttgctcccccaga


 


cacttcgaagctggatgatgtgtttctttggcatcgctggcactcttgtcatgatctgca


 


tggccaccccagtcttcgctatcatcatcattcctctcagcattctttatatttcggtgc


 


aggttttttatgtggctacttcccgccagctgagacggttggattctgtcaccaaatctc


 


cgatctattctcacttcagtgagactgtcacaggtttgcccattatccgtgcctttgagc


 


accagcagcgatttctagcttggaatgagaagcagattgacatcaaccagaaatgtgtct


 


tttcctggattacctccaacaggtggcttgcaattcggctggagctggttggaaacttgg


 


tcgtcttctgttccgccttgctgctggttatttatagaaaaaccttaaccggggacgttg


 


tgggctttgttctgtccaacgccctcaatatcacacaaaccttgaactggctagtgagga


 


tgacgtcagaagcagagaccaacattgtggcagttgagcgaataagtgaatacataaatg


 


tagagaatgaggcgccctgggtgactgacaagaggcctccggcagactggcccagacatg


 


gtgagatccagtttaacaactatcaagtgcggtatcggccggagctggatctggtactga


 


aagggatcacttgtaacatcaagagcggagagaaggtcggcgtagtgggcaggactgggg


 


ctgggaaatcatccctcacaaactgcctcttcagaatcttagagtctgcggggggccaga


 


tcatcattgatgggatagatgttgcctccattggactgcacgaccttcgagagaggctga


 


ccatcattccccaggaccccattttgttctcggggagtctgaggatgaatctcgaccctt


 


tcaacaaatattcagatgaggaggtttggagggccctggagttggctcacctcagatcct


 


ttgtgtctggcctacagcttgggttgttatccgaagtgacagagggtggtgacaacctga


 


gcatagggcagaggcagctcctatgcctgggcagggctgtgcttcgaaaatccaaaatcc


 


tggtcctggatgaagccacggctgcagtggatctcgagacggatagcctcattcagacga


 


ccatccgaaaggagttctcccagtgcacggtcatcaccatcgctcacaggctgcacacca


 


tcatggacagtgacaagataatggtcctagacaacgggaagattgtcgagtatggcagtc


 


ctgaagaactgctgtccaacagaggttccttctatctgatggccaaggaagccggcattg


 


aaaatgtgaatcacacagagctctagcagctggttccgtggctggcggactataagaaca


 


gtttctattatttgctttggtttctgtgactgtgctctaggtgcaaagacacatattttg


 


ttcccgttgctcaggctggcctcaaactctaaggctccagcaatctctggtctcagccag


 


agacctgtaaaaatagacacttcaaagattatcatgaataaatatttaaataaatagtaa


 


aaaaaaaaaaaaaaaaaaaa


 


>scr_sc-132690501_1 (TOXMARKER Assignment: 21; SEQ ID NO: 21)


 


gaattctctgggcccatccgttgttctcaatggacatgacctccaggaagctaaagtcca


 


ggtcgtgaccaaagccaaggttgtagagcgggaatctgccccggatagcgttgcggacat


 


tcttgaggatctgggaacggtccgtctccccttcagtgggctctccgtcggtcaacatga


 


taagaattgaggcagggctgctgagttctgggtggcttccttgagctctgtttaagatct


 


cgattcctcggagcaagcctccattcaggtttgtggctccagccaaagaaaagcgcctca


 


caaagtcttgggctgcttgcaaattggcgtgagacgcgggtaccagtgagcccttccatg


 


actgcacttgagacccaaagaggaccaggtcaaagttgtctactggcttcatgtccccca


 


atatcttaaggagcgcctcctttgtctgcttcactttctggccttccatggacccactga


 


tatcaatcacaaaaaccaggttcttgctcatgttggtcaggttttggggggcaaagaaat


 


gtgtaaagtaattgttggccaccaggaggtcacagagcttgtctcggttcacatcgtagg


 


tcaccttgaagtctccattcagcaaggaggtagagcacgtggggcaggactgctgctggc


 


tcacagtggggcggaagagcacatgacccttcttccccgagaaagacttcttgatggttt


 


gagcacttgactggtgatgacgtagtgggcaaagcgagaggtgactttgcaattg


 


>scr_gb-aw141735_3 (TOXMARKER Assignment: 22; SEQ ID NO: 22)


 


ttttttttttttttttttactgtatatgtaatttaattcaaattggaacaatgacgtaga


 


tatataagccacaatccatgaaagtcttggaggaaaacataggagcagttatttctgtac


 


ttgattttagtggtgagattcttagctgtggcatggatacacatgatcagaacagtatta


 


aataaggagaacgtcactgaaaagagcaatctgtgtgcatcaaagaacattatcaagaaa


 


gcaaagaagcaatgtgtataaaacgtccctaataggtaaatctacatagataaagagaag


 


attggtggttagacaaccagagggaggaagaatggagagtcactgagtaatggttacagt


 


gtgtttgaaaggggataaagataagatcgtggcctgattttacccataaattgttgattc


 


tttacacaagaataatggttagaggaatgagccacaatagcagatattatccaaccatta


 


atgaaacttatgaccacttcttaaatttttatttatttttttaaaatttacttgtttctg


 


cataactttgagtgatgttacatgcttatacaggatgctggggccagtagtagccaaata


 


aaggcatcaagacatgggtggaaactggaatttccagaggttgtaagcagccatgtgggt


 


ggtgggaaatgtccctgtgtcctttgcaagatcagcaacttttcctagtatctgtccttc


 


tctccagcattcttacacattgattcagttctaccaggctgtaagttattggctataagt


 


tatgagtatcagcggcatagcaaaggctatatggcatcattagacataacctgcaaaagg


 


gcacaaatgcattcaggatagggagagctgaatgcaggcatcataagatcaggctggcag


 


gaagaaagtatcctcatcttggaacatggtttccccctacttgcccatcctgacagagct


 


ttggagtggtggagatactgaagagaggactctccccatgtagtaaatgtgtctttatgg


 


agatgagaacctgccacagaacagaatgctgctggttttgttgtgcttgatgaagaaaag


 


gaaggggtggtcagcacagaatgttgggacaaaagcagcacagcagtattctatgacagc


 


ggaggctgctgcagcctctgtgccttcctcattgacctccactacgctcttgtgaacaat


 


cttggacacacacaggtttctctctggagacattgctgataagtcagccttggcctcttg


 


gaagacatccactattcccaagcgctgaaacacagactccatgtcataatcctcttgcag


 


tttaaattttggaaggaaaacctcaacattagtgttcttcataaagtctgggttggtcca


 


ggctgttaacttctcaaaagtgagattgctttccaccttgctgaggtccccgtcattatc


 


tgggagtaggaccacgaagctcagctccattccttcatatggcatcatgagcacttgcgc


 


ctgcacctcgttcacatgggcaaggttatatgtgtcctcacaacacatcatctgcactag


 


t


 


>af184983 (TOXMARKER Assignment: 23; SEQ ID NO: 23)


 


gtatttcataaaacagagaggatcgcaggaggccggcactctgactcctggtggatggga


 


ctagggagtcagagtcaagccctgactggctgagggcgggcgctccgagtcagcatggaa


 


agtctctgcggggtcctggtatttctgctgctggctgcaggactgccgctccaggcggcc


 


aagcggttccgtgatgtgctgggccatgagcagtatccggatcacatgagggagaacaac


 


caattacgtggctggtcttcagatgaaaatgaatgggatgaacagctgtatccagtgtgg


 


aggaggggagagggcagatggaaggactcctgggaaggaggccgtgtgcaggcagcccta


 


accagtgattcaccggccttggtgggttccaatatcaccttcgtagtgaacctggtgttc


 


cccagatgccagaaggaagatgccaacggcaatatcgtctatgagaggaactgcagaagt


 


gatttggagctggcttctgacccgtatgtctacaactggaccacaggggcagacgatgag


 


gactgggaagacaacaccagccaaggccagcacctcaggttccccgacgggaagcccttc


 


cctcgcccccacggacggaagaaatggaacttcgtctacgtcttccacacacttggtcag


 


tattttcaaaagctgggtcagtgttcagcacgagtttctataaacacagtcaacttgaca


 


gttggccctcaggtcatggaagtgattgtctttcgaagacacggccgggcatacattccc


 


atctccaaagtgaaagacgtgtatgtgataacagatcagatccctatattcgtgaccatg


 


taccagaagaatgaccggaactcgtctgatgaaaccttcctcagagacctccccattttc


 


ttcgatgtcctcattcacgatcccagtcatttcctcaactactctgccatttcctacaag


 


tggaactttggggacaacactggcctgtttgtctccaacaatcacactttgaatcacacg


 


tatgtgctcaatggaaccttcaactttaacctcaccgtgcaaactgcagtgccgggacca


 


tgcccctcacccacaccttcgccttcttcttcgacttctccttcgcctgcatcttcgcct


 


tcacccacattatcaacacctagtccctctttaatgcctactggctacaaatccatggag


 


ctgagtgacatttccaatgaaaactgccgaataaacagatatggttacttcagagccacc


 


atcacaattgtagatggaatcctagaagtcaacatcatccaggtagcagatgtcccaatc


 


cccacactgcagcctgacaactcactgatggacttcattgtgacctgcaaaggggccact


 


cccacggaagcctgtacgatcatctctgaccccacctgccagatcgcccagaacagggtg


 


tgcagcccggtggctgtggatgagctgtgcctcctgtccgtgaggagagccttcaatggg


 


tccggcacgtactgtgtgaatttcactctgggagacgatgcaagcctggccctcaccagc


 


gccctgatctctatccctggcaaagacctaggctcccctctgagaacagtgaatggtgtc


 


ctgatctccattggctgcctggccatgtttgtcaccatggttaccatcttgctgtacaaa


 


aaacacaagacgtacaagccaataggaaactgcaccaggaacgtggtcaagggcaaaggc


 


ctgagtgtttttctcagccatgcaaaagccccgttctcccgaggagaccgggagaaggat


 


ccactgctccaggacaagccatggatgctctaagtcttcactctcacttctgactgggaa


 


cccactcttctgtgcatgtatgtgagctgtgcagaagtacatgactggtagctgttgttt


 


tctacggattattgtaaaatgtatatcatggtttagggagtgtagttaattggcatttta


 


gtgaagggatgggaagacagtatttcttcgcatctgtattgtggtttttatactgttaat


 


agggtgggcacattgtgtctgaagggggagggggaggtcactgctacttaaggtcctagg


 


ttaactgggagaggatgccccaggctccttagatttctacacaagatgtgcctgaaccca


 


gctagtcctgacctaaaggccatgcttcatcaactctatctcagctcattgaacatacct


 


gagcgcctgatggaattataatggaaccaagcttgttgtatggtgtgtgtgtgtacataa


 


gatactcattaaaaagacagtctattaaaaaaaaaaaaaa


 


>scr_cg-22510674_1 (TOXMARKER Assignment: 24; SEQ ID NO: 24)


 


gaattcttgcagttacagagtatggctgttgtctactcgggagctcccagatcctcataa


 


ctcagggacgtgtccctatttatggacaaaaaagtttgacgccaggtcgggcctacatga


 


gctcttctctaccctgcaagtccccagtgtatctgaggaaggtgtattctgtcagagaag


 


caaggaagatcaatgcacacctttagtctcagccccataggaggcagagtcaagcagatc


 


t


 


>scr_cg-57215224_1 (TOXMARKER Assignment: 25; SEQ ID NO: 25)


 


aagctttatagtcaggcacagctggctgttgccaggcaactgtggggcagagcatacctg


 


gctgttgccaagtagctgtggggtggagcttagacagaatcccaacagatagtatagttg


 


gagagggtttcagtctgtcacagtggggaggcaggggcagtagttgagttcatggtgacc


 


agatcttgtgatggaggaaatttacatcatcatcccaggctagaaagcagtgagcagggc


 


agagacaggagcaggttatcaccttggaagacctgacactagt


 


>scr_gb-aa850767_2 (TOXMARKER Assignment: 26; SEQ ID NO: 26)


 


ttgcggccgcccaagtctgccacttcaacactgtatctaaaacttgaaaggcactgtcaa


 


aaaccctggtgggttcctagctttagggatccatcgttagagtcagtaaacatggcaact


 


ctgcctccgggcatgtgatacgtcgccagcagaggcttgctagcccttgccacacaacgc


 


tcagcttactcaaagcactgccaagacatggctgccctgagacggttgtctgggctcctt


 


ccttcctataccttagggcgcccccttcacagcactgggtaagcaatcagcccctcccgg


 


agaggagaagggaaggtaaaagacaaaggtatgttttacactatgcaaaacgttccagag


 


ggggaagatgaacgaagtaacaagtatccaacacagggttttaaaaagcaacgacatttc


 


aaatgagcttgtatgggagaaagaaaagcaggttttcaggaaaaatccaaacacattcag


 


gtgtgtcttttaagtcatgagtttatcatttattctaagttcattgggaggaaaactgga


 


gactatcagcatagctgtcttactggggaaggcattcccagtgaataaacatctccctta


 


cctgagctcttggcgagagattctgcccagcttgactctctc


 


>scr_gb-ai011994_2 (TOXMARKER Assignment: 27; SEQ ID NO: 27)


 


tttttttttttttttttccagaaatttgcccattcttttatttgaaggcaaaaattccca


 


tggaagtctggatgaagagagagacaaaggcttatagaaaataaattgaataactagaga


 


ttctctggatccagacatagttggttgataaatttgttacctatttctcattgtatttca


 


cattatttagacatagttcttgacatctctgttttgcatactgtctctggccaagagttt


 


tggtcttcctttctaaatatcaagaggaaaaatggcagaacaaaccagtaatgttacatg


 


gcatgtggttcctgagtatataatcaagcattagcagcagttgtagttatctgaatataa


 


tgcatagatataatacatgaccgaagagacacaccgatttaaacaaccaatgtcaacact


 


gaaacaaagaattttaatgctaaggcacccaatcacggtgtctttcagttatttgttgtt


 


ttctttaggagactggccatacacagcagggattcaaaattgtggcttgcagtcatgaat


 


caacatttgcatttgagtaacttacccatcttctttatgcttccacaaacatagtttcag


 


ttgggataatcactgaggtgtgcacagccctttcttcctgtagtttaggcaatatccaag


 


gctgtagaacttggggtaaggtgtaatggtgtcacaggaggagacatctactcactgtta


 


aatgttgctctgatgtaggttggccatagctccccatacgatctcacagggaagccgatg


 


ggtaatagcagcaggaagatcatggtctacataactgactctggaacttcttgacttata


 


acttattactttttgggtttcttttc


 


>scr_gb-aw142293_1 (TOXMARKER Assignment: 28; SEQ ID NO: 28)


 


agggaacccggtttctgaggttaagaacctggtatgaggtagaaagcagaatcggacctt


 


aggcactcgagcgtcgtgtcgaagaaacattaaatagaatagaggagtaaaggggatgtt


 


tcggataagcgctaggtcgagtcaaagaagtcttgcaagaagagttaagggagcaagaat


 


ttctagaagcatctagataaggagtcgtagcatactgacgttactagtaataagtagggt


 


gagtcggagaatcatgcgctcgatggtcataagatagtatctatcgaggagtgtaggagg


 


cctcgtccttcggcggaaaagtaacgcgtagcggttaagaatcttgtcgttcattatctt


 


aagggtaaggagccatcagtttagaagtcgttcccgcggtagtaagttcgcgtcgatttt


 


aataagactttagattgcgtcgtttagtcgacgtagtagacggttaatagtaacggtctt


 


acttccttaagcgtttcgctagttcttaagcttaattcggctactctagattttaccttt


 


ggggttaagtttccgttagcgttgttggaatcggttttgcctgcggggtggacgcccgtc


 


taggagaacgcattcgctacgaacggtgc


 


>scr_gb-bm383327_1 (TOXMARKER Assignment: 29; SEQ ID NO: 29)


 


tttttttttttttttttgatggccagtgacagtttttgctttttttatatttataaacaa


 


aaccaacctcccccccaagtaactccccaaacaaacaaaaaaccagattaaataaaattt


 


acagtgaacccagcaaacatctgtatgtgcaattaaatactgtgtctgttactgtggtgg


 


cacgaacctcaaacaaacaatatacaagtgttctggggttggatcaggggtcgggggagt


 


cccaagttttaactctgtggggtttggggagacaaggtggggaattgaacgaatggggaa


 


atcaatttatttttcttaattctgtccatataaatatattcatgaagaccaaaagaggga


 


agggcagttgggctggtgatgaagtgggagaaggggagggcagagccctctcaactctac


 


tcagccaaaaatatgaaacaaattaatttcatggtgggagaagagatttaaaaaatgata


 


gaagatgggaaggagggggagacagaaggggaccaaccagggaaaagggggacccatggc


 


aagggagtcccatgtcaaggagtcctgtgccggtgtgagaatctgtctgcttctctcttc


 


agccataatgtggtaagctctggcccaatccgccttcggctcccggcttggcccttgctc


 


ctattgtgccagcccctcccgcctccagctattgagagctagctcgctccaggatcctca


 


ggtcgtagttctttttagctactcgaagtttgaagcgactcacagagttgttgaggcgaa


 


gggaggcattgtgggcagccaggggactggggaacacagccactatagtgtacaaggcag


 


cgaggtccgcatggcggccattctcagcagtcccactgttgtcccccccacctgcaccag


 


gcaacccctgagcatccttaagccactggatcttggcaccagacatggcaagctgtgtga


 


agagtttgtctgcctctgtgcgggtgattccttccgggagatcagtcacctccagtaccc


 


ttcccagcacaacatccgctgtccccaggtcagtggaggcagacttgagtgcttgtctct


 


tgcctcggtttccatgcttcaatccactctgtccctggtgcaccgtatacgttgactggc


 


catgg


 


>scr_gb-bm386625_1 (TOXMARKER Assignment: 30; SEQ ID NO: 30)


 


tttttttttttttttttcacatgtcaacaactgctagctactattaaaatactgtcaccc


 


aaggaggtggaatgtttaacagaaaataggctttaacaattcatactggtcctcaataac


 


tgcagatgactagttcaagccaactgcaaaactgagcaagaaatgcagcttgaagaacag


 


gacaataaaatttaatcttgcaacttgatagacttggaggcattccggtcaatgtagaag


 


accttgcgggcctcagagttaaagcccaggccagcccctaggctgtacttccagctcatg


 


gcccggtcgtagtcctgctgcagactctgctggagcgtatctgaagacttcttgtccagg


 


gccatgttggacctgacagtcatgctgggaggacggttgaatgacggggatagatgctta


 


aagccgcccataagtttcaggaaattttagtttctgtttcttcattttcaaagcccgcag


 


tgtcccactggccaaactgggttccctgatctatgcaggcctcatccatattgccttttt


 


tttccagtaccacctccaagtctgtgtctgactcctctttttcctcctgccaggggtctt


 


cctttactccgctctcttttctcctcttctttttcttccttttcacagccagcccttcac


 


caactggctgctccaccttttttttggatttcactttcttcttcccaggggccttcaggc


 


tgtctatcgggatgaaatccaacgcttcacttctgactgacttcttattccctttcttca


 


agctgttctccatggagatcttggagttgaccgggaggatgtctccctcctgatgagtct


 


ttttcttcttcttctttttcatgctgtgatctctggggctcccctgcttccttttgtgcc


 


ccaaggccgcctgctcttcggcctctgccccctctaagcatgagtgcaaagcatccccag


 


cctcagggatccaagagtcctggggaggaaaagcttccatgtccggaagcttcttctcct


 


tcctgtgctttttaggcttcctaccagctttgcttacctccttggcatgcttggagtctg


 


gggaagtcttcagccctgagccttgggaggcaacctttgataaggacttccgcctcttct


 


ttttcttctctctgatgaggccttctgctgactgctcaaggacctgccttctaaggctag


 


gtgactttatctgtcttgtccatgtaggctcatccttgcccaggtactcatccaagtgtg


 


tgctacaggacttttttctcttctttctcttgcccagtgacatctcagggacctgcacct


 


cacccacattgttaaaaggggatgtagcccttggaggagaaacatctatgaagtaatcat


 


tattgtttaagactgagtactgagtctctggttctgagacatttgccaccttctttttct


 


tcttcttctttttcttcttctctgggagccgtgggcccaggtcttctttctgagtcttgt


 


tgaccattactggtctattagcaggccaagcatccccacgtgagcacccgcgcagccgcg


 


acccggaagtcagcttcgaatttctggcccgccccctcgaaatcgttctccttccgggtc


 


gcagcttcgcggcgccctgggttgctgtagaaacggcgtccatggccgtgcctagacaag


 


catccagcctcagcgtgctgcgtgaggagacgggaggcgctgcggactcgccggtcacta


 


cacgaatgcccgggctcgcagggtcgcctgggtcccccgaagttctcgtgttacccgcgc


 


aggtcgccgagcctccggggaagaacctgtgggagcagatctgcgaggagtatgaagccg


 


agcagcctacctttccggaaggatataaagtgaagtttagtttcctgccttgcccggaat


 


gctacgctttcacgtggccatcttccccgcagttgttgacatgcctagtgaccgtgacct


 


ctgacacccgttttcccacttttgccaggatctgtattttaacttacttcagagtcctct


 


tagttgtcttggtttgggggtggtttgggggtgttgggataacagatggggcaaggctgt


 


agccctactgagctgtttccagaggccgttgtcaggaaggatttccagtgttacagcccc


 


agagtataacagcagcgccctgttagcttaatggtccccattggttctgtggctgcggct


 


caccaggattctcccattcaaaaggcccagacatggctgacagcctcctctgtaggtctg


 


actgacaagctaccacgcgtcttaggtaaatagtaaagcctttattttcttgttaagaac


 


agcattttgaaaataaaacctatctgcccatgcttaacaacctttaaagtctgtgatatt


 


ttatatacagccctgtacatactgattgtctggaaatttcttaaacagtttttgtttata


 


agtatgcaagtcagccaggatgaggggaagagtgagggtacattataaaatacacattaa


 


tacatttaataaatatatattatctatcaaaaacgagccatagctcttaatgaataaagc


 


acctgccaagggctctcatcagctcacagttgctacatccttggatgtgtaaatgccagt


 


gcccccttctactttgccatttggcaaattcaaaagacaactcttccaccaccctgcact


 


tgttccctggccttgacctcctctgtgtgggggtggggcagacaacaaccagatcttaac


 


tttagaaacagctgacacattggagcccctcccctctgccattgtcctgctaccttggca


 


actgactccagacctctatggagtcttcactcaggaggggacagagcggtggttatagtc


 


ccaatatggtattagtacccgggcatgccaagttgtgcttgcagtttggggttattcaca


 


gatgactttctagaccattttccccaaccaagtgttgggtgtatcaacacttaaacaggt


 


gccatgggattatgcatttcagccttgctctgtcagaagctggctgccacagtatctggg


 


tggagttgcctcgtggtcctcctcgtg


 


>scr_sc-133556969_1 (TOXMARKER Assignment: 31; SEQ ID NO: 31)


 


tgtacaatgggggataaaagtgtcaaatgagatgttgctatagtttcatttcttttgccg


 


tgatagagcaccctgacaaaaagcagcacgagaggaaatgtatctggcttacgattccat


 


gttaaagcccgtcattgatgaggtgggtcggggagtcaaggtaagactgtaaacagctag


 


tcaatcacatccacagtcagagacagaaggacacaaattcatggatacttgctcctttgc


 


actcagctcagtttctccactcttacacagttttaaatgccctgcctagggagtgatgcc


 


acccacagtgggctggatgttcccacatcagttatgacaatctcccacctcatgcccata


 


ggccaacccaatgtagacaatctctcattgagactctcttcccaggccatgtcaagctga


 


cagttatagctagc


 


>scr_sc-170142736_1 (TOXMARKER Assignment: 32; SEQ ID NO: 32)


 


agatctcttaagtgaaaatagaaaaatgattactaacgagaagatagacgcctacaacga


 


agctgcagtcagcattctgaacagcagcaccaggacatccaagtccaatgtcaagatgtt


 


cagtgtttccaaactcatcgcccaagaaaccatcatggagtctttgggtggcttacacct


 


tcctgaatcaagcagagaaactagtgcaatgattctcatga


 


>scr_sc-2563586_2 (TOXMARKER Assignment: 33; SEQ ID NO: 33)


 


tcatgactcccagcattgacattcccctacaatagggctttgagccttcacaaaaccaag


 


ggcctctcctgccattgttgctcaacaaggccatcctctgcttgatatgcctctcgagtc


 


atgggtcattccatgtgtaatctttggtggtttagtacctggcagctctgcatggttgat


 


attgttgttcttactatggagtgacaagcctgttctgcttgttcaattatttgtctaact


 


ccttagttgagtaccctgtttgcagtccaatggttgggtgtcagaatctgcctctgtatt


 


tgtcaggctctggcagaggctctcaggagacagctatatctggctcctttcagccagcac


 


ttcttggcattagcaataatgtctaggtttgatgactataaatgggatggatccctaggt


 


gtgatagtttctggatggcctttccttcagtcactgctccacattaggtcttgatatttc


 


ctccttattttgtttccctttctgccccatcgttgtgcccttttgatagattttgcagtt


 


tagaaatacaatttacgtgcaggtttattgcattcagatct


 


>scr_sc-87618257_1 (TOXMARKER Assignment: 34; SEQ ID NO: 34)


 


tcatgatgaagaaatgggttctcggcaataggcaaaggcaggatgagagcagaggggtcc


 


atgggggtcgaaggctgcccatgggggtggttctatgctctgaccattttgagatgaact


 


aataatgttccggcagtggctatcccctaacaaagatcacaagccgcctagtggagggaa


 


tggaatctgaactctggtaccagcctccaagatccagatct


 


>cszr——96561134_83760493 (TOXMARKER Assignment: 35; SEQ ID NO: 35)


 


gaattcactagaccagcatattgctctatgctgcctttccagcgctgtactgcctgtagt


 


ggaacagactcttggagtccacagtacgagctttctgcacagcctcagcaaaaagtttgg


 


tcacctggaaattggtgagcagagcaattccactgtccacagctgtcctccgaatcacat


 


aattatcatggacaaatttggtgttgttattggggaggttaatcactaggtcaatgcttc


 


cgtctcttatcaactttctgatggaagagaggctgggattctgtccttcctgagatggcc


 


aagccactggggtggcaggaacattgttggcgttgagccagtctgatgtggcttctgtgg


 


caaaaagctt


 


>scr_gb-x83855_1 (TOXMARKER Assignment: 36; SEQ ID NO: 36)


 


ctagtccccgcagcctagcgcgggcggcggcgggcgatggaggagagcagagccccgggc


 


cccgccgtcctccagcgcgctccgctgcaaccccgcagctgagcccagaggctccggccc


 


tgtgcgccctaccgcggccccgccactatggccggcgtgtgggcgccggagcactcggtt


 


gaagcgcacagcaaccagtcaagtgctgccgacggctgcggctctgtgtccgtggccttc


 


cccatcaccatgatggtcactggcttcgtgggcaacgcgctggccatgttgcttgtgtcg


 


cgcagctatagacgccgggagagcaaacgcaaaaagtctttcctgctgtgcattggctgg


 


ctggcgctcaccgacttggtggggcagctcctgaccagtccggtggtcatcctcgtgtac


 


ctgtcgcagcgacgctgggagcaactcgacccatcggggcgcctgtgcaccttcttcggg


 


ctgaccatgacagtgttcggactgtcctcgctcttggtggccagcgccatggccgtggag


 


cgcgccctggctatccgtgcgccgcactggtatgccagccacatgaagactcgcgccacg


 


cgcgcggtactgctgggtgtgtggctgtctgtgctcgccttcgcgctgctgcctgtgctg


 


ggcgtgggccgctacagcgtgcagtggcccggcacgtggtgcttcatcagcaccgggccg


 


gcgggcaacgagacggactctgcgcgggagccgggcagcgtggcctttgcctccgccttc


 


gcctgtctaggcttgctggctctggtggtgacctttgcctgcaacctggcgaccatcaaa


 


gccctggtgtcccgctgccgggccaaagccgccgcctcgcagtccagcgcccagtggggc


 


cggatcaccacggagacggctatccagcttatggggatcatgtgtgtactgtccgtctgc


 


tggtcgccgctattgataatgatgctgaaaatgatcttcaatcagatgtcagtagagcaa


 


tgcaagacgcagatgggaaaggagaaggagtgcaattccttcctaatcgccgttcgcctg


 


gcttcgctgaaccagatcttggatccctgggtttatctgctgctaagaaagatccttctt


 


cgaaagttctgccagatcagggaccacaccaactatgcttccagctctacctccttgccc


 


tgcccaggcttctcagtcctgatgtggagtgaccagctagaaagatgatgaacaacctga


 


agcggagtttcattgcaatacctgcttccctgagtatgagaatttcttcccccagggaag


 


gataactgaatcattttggattgtatcttctttcggcctcatattttaagttttccttgc


 


cattaaacacaccgagacaagctt


 


>cszr_229800465_190907286 (TOXMARKER Assignment: 37; SEQ ID NO: 37)


 


agatctctacaccgcaaaaggtctcttccgtgctgcggtgcccagcggtgcgtccactgg


 


catctacgaggccctagaactccgagacaatgataagacccgcttcatggggaagggtgt


 


ctcaaaggctgttgagcacatcaataaaactattgcacctgctctggttagcaagaaact


 


gaatgttgtggagcaggagaagattgaccagctgatgatcgagatggacggcacagagaa


 


taaatctaagtttggcgcacatgccatcctgggagtgtccctggctgtctgcaaggctgg


 


tgccgtggagaagggggtgcccctttaccgtcacattgccgacttggccggcaaccctga


 


agtcatcctgccggtcccagctttcaatgtgatcaacggcggttctcatgctggcgacaa


 


gttggccatgcaagagttcatga


 


>scr_gb-bi277612_1 (TOXMARKER Assignment: 38; SEQ ID NO: 38)


 


gggcccctcctgctcgctgctgctggaggcgtttcggcgatattacaactatatttttgg


 


tttctacaagagacatcatggccctgctaaatttcaagataaaccacagttagagaagct


 


tctggtcttcattaacctcgaaccgcagtgtgatgccttccctagtatgtcatcagatga


 


gtcctattctctacttgtacaagaaccagtagctctcctcaaggccaacgaagtttgggg


 


agcactaagaggtttggagacctttagccagttggtttaccaggacgcttatgggacttt


 


taccatcaatgaatccactattgctgattctccaagattccctcatagaggaattctaat


 


tgatacatccagacactacctgcctgtgaagacaatttttaaaactctggatgtcatggc


 


ttttaataagtttaacgtccttcactggcacatagtggacgaccagtctttcccttatca


 


gagtatcacttttcctgagctaagcaacaagggaagctattctttgtctcatgtctatac


 


accaaacgacatccatatggtacttgaatatgcccggctccgagggattcgagtcatacc


 


agaattcgatagccccggccatacacagtcttgggggaaaggtcagaaaaaccttctaac


 


tccatgtttcattcaaaaaattagaactcaaaaggttggacctgtagacccaagtctaaa


 


tacaacatacgtattctttgacacattcttcaaagaaatcagcagggtgtttccagacca


 


gtttatccacttgggaggagatgaagtggaatttgaatgttgggcatcaaatccaaacat


 


ccaaaatttcatgaagaaaaagggctttggcaacaattttagaagactagaatcctttta


 


tatcaaaaagtaagtcatctgaaagcctaatcaccactgttttcatacaagtccaagctg


 


cgacttagctctctgctttacttctcatcttccccactgcttgcaagagtggagccaaga


 


acacctaggaggcagtaagcattttgcagtaactactgaaatagagggagaagccatgcg


 


cccgctaggagctctggctgccctttgtcttttgcactatccaggggctggaactcactc


 


cctttgtcctgagtgacctggggcatctctgctccttacacagtgcagtgacatttccaa


 


cattccacagccagggaattggtactgaagtggtggctgccttgttagaaaacacagaca


 


gaccacttcccaaaagtttggtggacagtctgttctctaagaatcagcacatttttcccc


 


atagggaccagaccacacttaggcatcatgggccatgtggagttgcaaatctcttttana


 


a


 


>scr_gb-j05266_3 (TOXMARKER Assignment: 39; SEQ ID NO: 39)


 


ttttttttttttccagagcagaggtcttttttaatcaatcacaaagtactttaaaatctc


 


ataggggacagccttgaatcatctatccacgctgattgtaccggtaagtagaacaggata


 


agagcaattcgccagctgcagcacagtctggtacacgagcagcccggggccagccatgcc


 


tggcgttacaatgtgctctcacaaaagtaactcatggaactcaacgtgaagtcgcgcttt


 


tttttttttggttcttttttttccggagctggggaccgaacccagggccttgcgcttcct


 


aggcaagcgctctaccactgagctaaatccccaacccctgaagtcgagctttaaataata


 


acctgagttaaattcccagggaaaggagggcactgactcctacaggctgctctctgacct


 


ccacaagtcccaggatacatctgagcccgtcccacacaaactagcactcaatatggaact


 


tttattcatgtgatttctgtacatcagggagtacaagagtaaacctttacaaatggtgct


 


gattttaccacaataaatgacaaaaccaaagcagtgtctggtgacagtggcagggcttta


 


aggttcaaacccagccaagaagtttgttacgatttccttcagctttgcatccgactgttc


 


tgagattttcccatcagacctgatattgcccaagaggctctggtgctggctcacaacatg


 


agacaagaaagcactctcgaactttgtgatcttactgggctccagtttatcaagataacc


 


ccggacgcctgcatagatgacagccacctgttcttcaatagccatgggagagtactgtcc


 


ttgctttagcagctcggtcaggcgcacgccacggctcaagagctgctgagtggcagcatc


 


cagatcagaaccaaactgggcaaaagcagcgacctcccggtactgggccaactccagctt


 


catggtgcctgccacctgcttcatggctctggtctgggcggcagatccgacacgggacac


 


agacaagcccacattaatggcagggcggatgcctttatagaacaattctgtttccaagaa


 


gatctgtccatcggtgatggaaataacgtttgttggaatgtaggcggacacatcaccagc


 


ctgtgtttcaatgactggtaaggcagtcaaagagccaccaccaaaggaatcgttcatctt


 


ggctgctctctccagcaggcgagagtgtaggtaaaacacatcaccgggataggcctctcg


 


acccgggggtcggcggagcagcagagacatctggcggtaagcaacagcctgcttggataa


 


gtcgtcatagatgatcagagcgtgcttgccattatctcggaaatactctcccatggagca


 


gccggagtaaggagccaagtactgaagcggggcagcatcagaggcagtggctgacaccac


 


aatggtgtacttcatggcatctgcgtctgtcagtctcttcaccaactgagcaacggtgga


 


ccgtttctgaccaatagcaacgtagatgcagtacagtttcttcttctcgtcagtcccatc


 


attgaaacgcttctggttgatgattgtgtcaatagcaatcgaggttttcccggtctgtct


 


gtctccaataatcagctcacgctgacctcggccaatcggcaccaggctatccacagcctt


 


gatgcccgtctgcattggttcccgcacagagattcgggggataattccaggggctttcag


 


gcccactcgtctgcgaatcttggaaccaactggacccttcccatcaatggcatttcccag


 


ggcatcaactacacggcccaacagttcatcgccaactggaacgtccacgatggctcctgt


 


tctcttcacgatatcaccttctttaattagcttgtcattcccaaacacgacaactccaac


 


attgtcgggttccaagttcagggacataccctttaagccggaagaaaactctaccatctc


 


ctcagcttgaacgttcctcagtccatgcactcgggcaataccatcaccaatgcttaagac


 


acggccagtctcttcaaggtcaacagaagtatcagctccaaggatccgctcctcgagaat


 


ggaggacatctcggcagtgccagtcttctgaagtcgagtgttagaggcatggagatttct


 


tgtaccaacaaaagatgaccccaaggcatttttggagaccagtcccgcccgtcgagggag


 


ggcacggcg


 


>scr_gb-m37394_5 (TOXMARKER Assignment: 40; SEQ ID NO: 40)


 


ttttttttttttttttttgcttgtttgtttgtttgtttacttcatgaaatgaaaacagga


 


aagcatattaaaactcaaaacaatgaaacagaaaacataaaaggtagtctaatagtcaga


 


aaacactggtaaactagcgtgtgttaagtatcagggacatatttatacaaaaaagtaagt


 


ctgagggaaaattctacccagtcattcttctcccagtcccagtaagtaacaaagtggctt


 


atcctattgtacctgccatggtttaatgctgtacaagtgtggcctgctgagcacatccag


 


gacttcttgtgcatgtagttatcttgccatggaagtgtcttgatgcagagctgctagaac


 


caactgtctggtcagttggctccaggcaactctgtgtaatacacgctacgggcaagcttc


 


ttcctttatggaagagtgcatgaatcaaatcaataaagacaagaatcccagagttcccta


 


tgtcagcaagcgccataggtctgttttttttccccctatgtacctcaccatgaggcaacc


 


ttctgttccaaaaggacaatgttctcgatggatacctttcagtggaatcttcacagttcg


 


aagaccaatagatataccttcaacttcccaaagagcatcaggggaggggcccacttcttg


 


gctcagtgacaaagcccgtcagagttatgctttaaagccagtctgagggtttgacatttg


 


acacaatgtggacatggctgtcaggagcagaggtgctgccatggcttggtcctgggcctc


 


tggaaagtccggtttgtaactggtacaatgcctcttcaatgtcatgctccactaaactca


 


ctgcttggcggtgccacccgcaggtactctgcattttcagctgtggggcccttaaagatg


 


ccattcggcttggcttctttgggaaagaagtcctgctggtagtcagggttgtccaggctc


 


atttggtggctgcctttctggatccagagggcagagctgtcaaacccactactgaggcag


 


gtcggctgggcagtgttgagatactcagggttgctcaccgcattgctatggggattttga


 


taatgcaggtctcttccaggagctggatgcaggggctgattgtgatagactgggttctgc


 


acagagccagccggcctcttgggaacagattggtttatatattcaggcacgggaaggaat


 


gtgtcatctatgttgtcctctgtcaggacgctggtgggatcggagctataccgttgcaag


 


aaggcgtcttctttgacacggcagctcccatttctattaatgcaagccacagtggaactg


 


ttgctatttgcactcagagagctcaagagtggagtccgtgacgtggatgggctgttgaag


 


aagccttgctgtgggatgaggtattcatcagcatcaactacgtcttccatgtcctcctcc


 


tccatcagggctcggtaaaagttggagtctgtagggctcggcaaatgcatcctttcatcc


 


ccctggataacaaggtagcgctgtgggtctctggccattttggagaattcgagaatcaac


 


tctcggaactttgggtggctatcagcatctatcatccagcacttgaccatgatcatgtag


 


acgtcgatggtgcagataggtggctgtggaaggcgctctcctttctctaggatggatgag


 


atctcacttgcagggatcccatcataaggcttggacccaaaggtcatcagttcccacacg


 


gtgactccatagctccagacgtcgctttggtgtgtataaattcggtgtaaaattgattcc


 


aaagccatccacttgataggcactttgcccccctctgcatggtattctttctcctcagca


 


ccaagcagtttggccagtccaaaatctgtgatcttgacatgctgtggtgtctttaccagt


 


acattcctggctgccaagtcacggtgtaccaaacgccggtcttccaggtagttcatgccc


 


tttgcaatctgcacacaccagttgagtaggtactgggagccaatgttgtccttatgttct


 


cggacatagtccaggaggcaaccatagggcatgagttgtgtaatgagctggacagtggag


 


gtcagacagatgcccaggaggcggcatacatgagggttgtccacactggccatcacgtag


 


gcttcatcaaggatttccttgttggctttgggagatgtggcttctcttaactccttgatg


 


gccacagggattttcactttctcgccttctgggatccagagacccttatacactgtgcca


 


aatgctcctgaacccagaactttgatctttttgaattctgtttcctttaatatcctcaag


 


tgggcttggttcggagcttctccgctgggtgtgagaggttccacgagctctctctcttga


 


agcaggcggcgtagtgtacgttttcggacaagctgacgtcgacgcatgaagaggccgatc


 


ccaagggccaccactactatgaagaggaggccacccacaatcccagtggcgatggatggg


 


atctttggcccttctggttgttgacatcctttaaggcctggcccagcacatccataggta


 


cagtttgcatggcagaggtggcagacgttattggcatctgcaaacttccagaccagggtg


 


ttgttctcccccatgatgcccgaagggcaggtcttgacacagtggggaccatcaacatag


 


tgggcacacttgatgcagttgtctggcccccggcctgtacaggtgatgttcatggtctgg


 


ggcagacattctggatggcactggatgcattcagaattttccacaaactccctcggttcc


 


ccctccaggatgttgcacttgtccacgcactccctgcctctgctcacattctggcaggag


 


acacagtccgtgggctcagggccccagcagccttccgaggagcataaaggattacagacg


 


tggttcgtggccttgcagtccttttcagctctgttgttcatgattttggtcttttgattg


 


ggcgtcccgaagagttttttccagtttatagtgtttgcgtagcacaaatttcggttccca


 


gaaataatcacatccccatcactgatctccttgagggaacgcaaccccagcgatgttatg


 


ttcaggccgacaaccgccagagaaaactgaccatgttgctttgttctgccacgaattatt


 


tctaggttctcaaaagcatggaggtcagtccagttttcaggccaagcctgaatcagcaaa


 


aaccctgttatttccttcacagttttgagaatttctagttcccgtgggtctagaggagga


 


gtgcgggtgaaagaatcccccttaaaggccactggcaggatgtggaggtccccactgatg


 


gcagtgcagtacttgaagtgtttgatgtttgtagcatttatggagagtgtgtctttaaat


 


tcaccaatgcctatgccattgcaaactttgcggcagggcccgtcacattttttacacttg


 


ctgactccatcttcttctacttcatagtagtctggcccacaggcccggacacacgagccg


 


tgatctgtcaccacgtagtttctggggcatttcttcacacaggtggcaccaaagctgtac


 


ttcccctcagggttgacatccatctggtacgtggtggggttgtacagcatgagtggtggg


 


caggtgtctttgcacgtggcttcatctcggaacctgtggcagaccagacagtcactctct


 


ctgggccctgtacaccctgcggcacactggttgtggcagcagtcgctaggggacctgcca


 


cgacaacgccgggaacattgctgggcgcagatgattttggtcaatttctggcagttctcc


 


tctcctcttccccagcagcttccattgggacagctcggatcacatttcgggcagcccgtc


 


aggtggcgctgtacgtccattgacatgttgctcagaaagacatcttggacgatgtccctc


 


cactggatggtctccatattgcagaggatggggttgttgctaaatcgcacagcaccgatc


 


agaatttcctgtaagttccgcatgggcagttccctaagcccagttttgttggttccatag


 


ttggacaggacggctaaggcgtaggtgttttcgtagagagcatttcccctgatgatctgc


 


aggttctccaaagggattctctccacggtgttcagggcaatgagaacatagccagccacc


 


tcctggatggtctttaagaaggaaaggtcataattcctttgcacataggtgatttccaag


 


tttccaaggaccacttcacagttgttgaacatcctctggaggctcagaaagtggtcttca


 


aaggtgcctagttgggtgagcctgttacttgtgccttggcaaactttcttttcctccagc


 


gccccacctgcggcgcagagcgcagccagcagcagcagtagcttggttctcgcagtccct


 


gagggtcgcatcccggctcggcagtcgttggctctggctctccgggattaatccgagtca


 


gactgagtcccacggtcgtgcccggtgactgcgtcggcaacgacgacgggacccggactc


 


agactcgcgtccaggtgacccgtcgcctgtcttggtggcggtagcctccgggactggctc


 


cagacgctcgagcccaggaagagcgcacagt


 


>scr_gb-m64300_4 (TOXMARKER Assignment: 41; SEQ ID NO: 41)


 


tccgatctgagcagacagctacagccaacagatggcgtgtaagtttggagctgtcactga


 


cttaaggtgccttatgtcttagccttccctaatgtaaggtgggtgggcataactggaaca


 


agtctgttaagacttgctctgaggaggctgacagttcagtaggtgacatgtaggaaggat


 


tcagggcagggaggaaccactgcatctttcatccgacacagtagttactgactaaacaac


 


agtgagcacttgagtgcactgagtgcaactgtgcagggcctggtgcaggagaactctctg


 


gactgaagaattccgtgaaagtataaaagccactacgaccagaactgcccctcggaacgg


 


ctcaaaggagtcaagagtgggtaagctgagacgggctggagacaggaccagggtcaagaa


 


ctggggggacaccgacatctgaacgcgtccagtcctctgagcccttgtcctgaccaattt


 


aagatctgtatcctggctggaatcgagcagtctcttcaaaaatgagttctttgagcttct


 


ccttaggtaagtcgtccagctccatgtcaaacttgaatggtgcttcagcaatgggctcat


 


cacttgggtcataatactgctccaggtacgggtgggccagagcctgttcaacttcaatcc


 


tcttgtgagggttaaatgtcaacattttatccagtaaatccagagctttggagtcagcgt


 


ttgggaacaacctgttccacggcaccttatttttgtgcgggagagaaagcaaatagtttc


 


tagcttttaaatttattatacaattcagatcttcctgtgatggagatccaagaataccca


 


ggatgtgattcagctggtcaaggtaatgctttcctgggaagataggcctgttggatagca


 


tctctgccaggatgcagcccacagaccaaatatcaatggacttggtataacccttggaat


 


tcaacataatttctggagctctgtaccaacgcgtggctacatactctgtcaagaaccctg


 


tatgatcatggtctggatctgcaacacgggcaaggccaaagtcacagatcttgagatcac


 


aagtggtgttcagcaggaggttggaaggcttgaggtcacggtgcagaacattagctgaat


 


gtatatactttaatcctctcaggatctgataaagaaaatagcagatatgatacttgctga


 


ggtgctgtgtcttcaagagcttgtaaagatctgtctccatgaggtcctgtactatatata


 


catctttcatctgctcaatggttggtgcccggatgatgtcattgatgccgatgatgttct


 


catgtctgaagcgcagtaggatttttatctctctcagggttctctgacagtaggtctggt


 


gctcaaaaggactgattttcttgatagcaactcgaactttgttgagattatcataagcag


 


aacaaaccatgccgtaggcgccttctccgatgtacgagagattagtgtagcgcggcccca


 


cgtcgaacacctgcccgcggaccatctccgggcccgccgccgccgccgccgccatgttgg


 


ctgcacagcctccgccgcgttgggctcgacgcttcgcgttaccgctcgacttgtgctgcg


 


cttcccacaggaaccgcgccgccgcccgtgtagccggctggcggcgatcgggaacgagga


 


gggaggacaacacagaagagagaactaaccgccggtagaaccacgg


 


>scr_gb-bi294409_1 (TOXMARKER Assignment: 42; SEQ ID NO: 42)


 


gtgcacagaggggactcaacggtgtgccgctgctcagactacatctggcccacaaatgtt


 


cttctagagccaccagaatttaagattattggctttaaggaccacataaatgtgatgatg


 


gagtttccacctgccacttacaagctattcggggaaagcttatggaaaagactggagtct


 


acatccttcgtcatcgaggaacagacagaggacagcattagggtgcacaagccccaaatg


 


aataatgtcactgggaacttcacgtatgtccttagagacttacttccaaagacaaactac


 


tgtgtgtctgtttattttgatgatacacctgtaataaaatctcccttaaaatgcaccgtc


 


cttcagcctgaccaggaatcaggtatggctaggcttttaaaatttgcactgttgttttga


 


tggaaaacttgctgaaagaaaaaaaaaaactcaagttctggtacactaaatgtacttctt


 


ccaataaatgcacatcactgagctgtttaaaaaaaaaaaaaaaaaa


 


>scr_gb-ab015747_3 (TOXMARKER Assignment: 43; SEQ ID NO: 43)


 


tttttttttttttttttgacaagataaagagtctttattgacatagagctccacgtgacc


 


tcttctgtcctgccctccttgcaaacatactaggtgtcccaaaggtagggacacgagcag


 


acagtcctgagcctggtcccgtcctccagaatgcagtcagactgcagtctgccatctgcc


 


atccctatcatctggccaccaaccagaaccagccccacagttcccttgtggtctcgcctt


 


ggctgccagtggtggtgtccactgggacctgccactaggctgctgtgtttgtttactggg


 


atcccacttccacatcctgggagccctggcttctggccacatgtgggtaactggcagtga


 


ctttgggcaatcaagtttgcgttcttgttgctttccacaactgggccaagctgggacagc


 


aggctctgcttctagtctcagtccgagctgttcaatgaatagcctccttggggcagtatc


 


taccctcccttaactcaaaatttccactagttagggcctcccaagccactgccaggccag


 


ctgcgagtttctaggaccagcttccagctggagaacccgacagctatgccaggactgctg


 


tgagccttgggcaaacggtctattgggtggacagaatgggcctgagcaggtagggcaaca


 


agagctaggagagcccagggcttaagaatatcagcactgctgtgggagaaagcaaaatga


 


gtccctgaatcccttgtgagggaggagagcccaggccaacggtaggggagacagccaggc


 


tctgaacttctagggtcaggccaagttcacatcttcacttcaccattctttcgatttctg


 


ggaaacctgccagctgggctgtctctcaggaagcacttccctggcttggaggaaccccgg


 


ccttagcacagacctcagcaacaacagcacactcacctaagacacagtgacgcccagagt


 


gcccacaggtacctcagtagtctggctgggaacaggagagtggccagggcccttgcccac


 


ccctgacaaattggagggtgtcctgggtgctaaggtgaggttggcttcctgtgacatttc


 


cccaggacagctctccaaggtccccgagagattccccaaggatggtgatttttcatcata


 


gcaacagccgcagccagggctagcaacgacatggatctgaccatcttcctcctggctgtg


 


gtgttgcttcaggtggccacacagatggcaggtgagggacgagtacacaatgccaaggcc


 


caggtcatccccaaagggcttgggcacctgctctgaaggaggaggggccttcagccaggt


 


gcctcctttgagccccagctctagaccaaggcattctggggtgctattgggtggggctga


 


gttcaggggactgggtggcagctccatgtccagtccgaaggtgaataagggcatggagtt


 


gggggactggttaggaacagggttctggaagggcttgtaccctccacatccactgtcagt


 


ccctgctgctgctgtgtccgtgcagacgccactgctgctgagcaggctcgagaaagcctt


 


gtaaccagtgtctccagaaggcccgacaccaggcacccacctggcctgggacgcaccctg


 


cttcactgcctgcacaaactcttggtagccactggtaggggctggggtggagccagctgt


 


cccgtgctgcaggacactcatgtgaaggatctgctcccagctctccgcttgctgcattgg


 


tggccctgaagaatgggggtcaaccgggctcagaagatccccttcttccagatgtccagc


 


ctgcttctgttctgaagccagctctccaggatttggggcggggctggagaagtcactaaa


 


actccggtaggcaggattgtctgaaatgacaagggggacctgtgtgcaggctgtgccagt


 


tgctctctcagggtctgggtgtggaggctgctgccctgtgacctggcatgtggtctcact


 


gggccccgtggggaagcaggcccaggacgtagaagcttgcccactttctgaaggcagaag


 


ggaggatgactctgccatgctcgactggccaacgcctccattctcagcccccagcaagtc


 


tgaaaacaggttctcagtgagccgggccatgatgtctgcctgactctcctggaagccccc


 


tccgctgttctcaggtgacatgctcaggtcccctttgaccatctcatcctcttcctcctc


 


cacattctgtactggggcctcaaacagctccatacagcgcaccacactgacatgaacgtt


 


ctctggccagaggacggtcctgctgacctccgcaggataccagcctgctttttcaggact


 


ctggagaggcttggttttggcagccttcggggattctctctctttcttcactctatgctc


 


cagcaagcagggcagcagcttggttagacaagtcttccagtgccggctcttggttgactc


 


ctggcttcgggtctgcttctcccagagggacaccttcgtgtcctgaatgatgatggctgc


 


taagggactgcgtgctggagtgggaatctggtcccaccatatcttcttaatcttgataat


 


gctgaagtaacaggtcaggcaaaacaataggatgcagatgcaggagatgctgacacccag


 


cgggaggcgctgcagcaggggcagctggaagtggttgtaccacgtgatgctgggactcca


 


ctcactccagatgccagggaagctctgggacaagaccctcacacgtgccctatagcgcac


 


ccctgatgttagggtgttgactgggaagctcagcttgggttccgtgtaggtcacattata


 


gactttgaattccgccgggttgtcctctctggagatgttgaccatgcagatgaggccttt


 


gtgcaggaagttgttcgatgggtatgggttgctccacatcagcagcaggccattggagac


 


attggtgtggagtgtgaggttgtctggagctgggggcttcacattgtcactaggcttgaa


 


ggagccttgccacagctgtccccgctcagaccacagttccagccagtatgtgtctgcctg


 


gatcggctcctctatggccatttggcacacacacacggtgtcggcactgttcttgggggt


 


gcatgtgaggttttcagagaactcgaagagcagcctgtagtccaggaggagctgagaact


 


gcagtccacagtgctatccagctgccactcacacgtagaagtgcggatgtagtcagagaa


 


gcaggtggggtcacccaggaccttgatgcccccagagccagtcacccatagcaaaatcag


 


acagctcacggaggacaggaacttggtgcaaagccgccccattgcggacacaaaggtgcc


 


tgggctatacagggagagactggaatgcagctcagtggcagcgtacctggcccccagatc


 


ctgggctccctctccagcacctgtgtgttcaggctccacgcgccgtgcggggctttcctg


 


cgcgaaggacctcgcccggtttcctacgccgcccggacgc


 


>scr_sc-191879433_1 (TOXMARKER Assignment: 44; SEQ ID NO: 44)


 


gtgcactaagaatgacaaacttgctgtgtgccacaaagatcttgggtggctggttggtgg


 


ccagtggtcaggttggcctcacactgctccaagtagaagagcagcagctgtcggtctgaa


 


ggccccagtccccctgtccgccccggcacaaggggctgggctggtgtccagttggccagg


 


tcatggtctatgggacgagacacctcctgctccagtcgctcaaactgtttcagctgctgc


 


agctccagttggccttttccctgtcgcacgatgttgcccttttccagcagttccttctgg


 


gtcttctcaaattcctccttcccctgcagatgaacgtagtcatagtcctccatccaaccc


 


ccttcactgttctcatactggccatccgga


 


>scr_sc-140438096_1 (TOXMARKER Assignment: 45; SEQ ID NO: 45)


 


tctagactttaacaacaagcgtgatgaacacccagagaaatgcaggagtcggactaagaa


 


catgatgtggtacggtgtccttgggaccaaagaactgcttcacagaacctacaggaacct


 


ggaacaaaaggtcctgctggagtgtgatgggcgcccgattcccctcccaagtcttcaggg


 


aattgctgtcctcaacattcccagctatgctggagggaccaacttctgggggggcaccaa


 


ggaagatgatacttttgcagctccatcattcgatgataagattctggaggtggtcgctgt


 


gttcggcagcatgcagatggctgtgtctcgtgtaattaagctacaacatcatcgaattgc


 


ccagtgtcgcacagtgaagatct


 


>scr_gb-x87157_5 (TOXMARKER Assignment: 46; SEQ ID NO: 46)


 


ttttttttttttttttttaaccaagaggaggaatataattgtgataggaaactaagaatc


 


atgaagctcactacaaaagacaaacactactgaaacatgttgtgctggccttgacacacg


 


caggcagactgtcgcctagctctgaggcagagggtcaaggttgacacagggctcggagga


 


aatatttaccagagagaatgtggtgattcatttatcagtccagagatcgcaagtataaaa


 


cttcaagatataagaaggatcaaattatatcatgtatgtgattcaatttaaaatgtctta


 


gccctcttacattatattatctggattataactgtaaaaaaaatcaaattacattcatat


 


gaaacttttatcaaaagaaatcaaatccatttttatgaaactttatagtacaattatttt


 


tagttggtctttccttaggtcacagtatttataattccatttacatctgtataattttta


 


aaattaaaaaacaaaagcaaatcaatagaaatctaagttttcttttgtaaaactctcttc


 


agtctccaggccggcaccacatgacagtgttgacttgtcctccagacatggacaactccc


 


aggatccctggcttacgaaccattcaggcctcgactcattaggaatgctttttggtttgg


 


ctcacgttgcaagaaattctggagcatgtccatgccgtccagggaccccccaggcttcag


 


gattaagtttctgtatttcattccaacctctggattcatgatcccctcttttttaaaaca


 


gctgtgaaacatgtccatggaaaacacttcactccaaagatatccataatattggccatc


 


ataccctcctgccaagtgtccaaaagtagctggcatatttgtgcctggcgtagctgcaac


 


tcccagaatttctgtgcagtatttagcgtattcgctcgcggcatccagagtcgcattggt


 


atggagagattggtcaactttgctcaaaacaatttggcgcagcgtcagaagacctgtgtt


 


gaccagcctagaagcaacaagcttctcgagcagctcgtctgtgatagggtgtccatcttt


 


ataatgctttgacagttttcgcagggaatcaacgtcccacacccagttttcaagcatttg


 


tgatggcacctctacaaagtcagtttccacgtttgttccactgaatcgtgcaaagtcagt


 


ctgcgcacagatctgatgcatgacgtgaccgaactcgtggaagtaagtccgcacttcatc


 


atgtctcaggagagagggccgacctgctacaggctgagagaagttgaccaccagggcggc


 


cacagacatcatccgactgccatcagggagaaggcagcctggctggagaccgaagcaggc


 


tgcatggttgtattttccttcccttggatagaggtccaggtagaactgccccaggacctc


 


tcctgtagctttatccttcacagtgtaaagtgaaacgctcttattccaaacatgagcatc


 


gggcacttgttcaaatgaaagtcccagcagctcctggtagatgcttagcaagccttccgt


 


gaccacctcaatggggaagtactccttaagggactcctggtccaccgagtacttgagctc


 


ctctgtctgtgtcatgtagtaatggaggtcccatgcattgatcttcccgtcgtattcaaa


 


acctcgctcttcacattccttcttcttcaggctcaaaataaactcccgttctgcctcacc


 


caagggtttcaatttctggcttaaatcatctagaaaggcggccacgcggctggtgctctt


 


cgcagtgttcagttcaaggacaaagtcagcatgggtgttatagcccagcagcttggccac


 


ttgagctcgcagcgggaggagctgttgcagaattgcggtgttttcctgtttgcacctggt


 


atgaaaagccatttccatcttccttcgagtttcagggacacagcatttcttcatgacagg


 


gaagtagtgaggatactttaaggtaactttgtacttgtcttcatctgttttttctaaact


 


gtcaatgaagtcatcaggaagagcaccaagttcagccttggagaatacaagggaagtgtc


 


gtcctcattgaggttcttgttgaagtcaatgcatagctcactcattctcttcttcattga


 


tttgatttcatttcttatgtgttctgaaagatggagtccattcctttttcccattttaat


 


tgacttttccaagtatcgcctggcttcaggctttatcttctccaaatcgcatgtttcttg


 


taaatgaacaattctctgaaacacatcttctctcatgctcatctcaatatcaaaacgaga


 


aagctttttgtctgcttctgtgcttgcagcccgcacttctctgtcagatgacacgtgctg


 


agggaagtccagcatggtcctttccactatgtacgtcacttctatgtcagccagcacctg


 


cagacagttctcataagttacttctttcagggcgattgtccccacggtgtcgtacacctg


 


cttggtctgtgctatgagctgctctgtcctcgtcttgatctgctctggagaaaggtccca


 


tctgagaacattcctgccagccgcagtgtaggaagacatagcttgaagaggagaagccag


 


ctcctttcccagtgtcattgtcagctgaagcctggagccaccagctctgtggaggcctcg


 


cagagtcgaaaggcacagggtgatcatgggcacgccgggaggccggcagcagctggcgcg


 


tcgtcctcccgcttgtaggtgcaggaggcaggcggtggtgtctgcgggcccggaagccag


 


gagtgggccaagccgaggagaccagatctcgagacggaggccgtcagtcc


 


>scr_gb-u66707_2 (TOXMARKER Assignment: 47; SEQ ID NO: 47)


 


tttttttttttttgtttatatgccaacatataccttgtgctagaaatactttatggggtt


 


acaactctttatatacaattttttttgaggcagtatctctgatggagagcataacttgta


 


aagagcttgtgtgtgcttccgtgctccaaaatgataggaaatccactttgagaagacaac


 


ttatttgattttaaaaaaacaaaaacaaaaacaaaaacagaaacaaaaccgcaccaatgc


 


acagccagaggctccgctggaactgatacagaaccgcgcaaacgccgtgattataagtaa


 


cattttccagggtggtcaaggctaacgtacaatattatacacctggcactgatgtttgcc


 


attggtcagcaactggcaaaatttgtttctatgtataaatttatttttaaacattatctc


 


tggcctgacatatcttcactatttataaaaacatttagacagtgagctcacgttgaataa


 


ctaggtctactgtgttctggaagctcttcagtagtaaaacagctttttcgtgttccatat


 


gcacaaaactgtgtccatttgcctgaaggattttatccccgggctgtagaaggttggatg


 


ctggtccatcaggctgaaccctagtaacaaagatacccttgtcggaaggtttgaaaggat


 


ttccttgcccactaattccaccgctgatactaaatccaagcccagggttcttttctattc


 


tcacacagaactgctcgggataaccgtccatactcctctgccctttcgtttgaattaagc


 


accgtccaggctggggtccccgggtggcctgtgatgaggggatctggatgggcagtggcg


 


actggaactgctgaatggtcactttgttgatgttcccttcatatggctgctgctcccggc


 


tgcggtgctgaaggctctgtgaccccatcagggtctgaatgtggctgcctgcctttttag


 


taatggtgtccggaggtacatcccgcctcccaagtgggtaaggattccattggccactag


 


gagagacatcttcttgtccattgtctaaaatgttgctttgctgggacggggtcctgtcta


 


accttctagcttctatatgtctaagcagctgctgtctccagtctgccggcattttgccac


 


agctctcctctcccttcacaggggtgggccttgtctttatatcactgttatctgatgtct


 


tgtcaccatagttacccaagttatagtcagatggtattttttccaggagggctgccatgg


 


taggcctagctgaaactggcctggtttgggaggccccgtaactctctgtgctgtagctcc


 


tggcagacagtggcctcctctgggtaaggtttttagcaggaaaacttcccgcctgcgctt


 


tcacttcttgatatgaagggtgctcatcttcgtacctgccattcctcttgaggaattggg


 


gatcagtcattgaaacgctgctctggccttccagccctcccctataggctgctcggccat


 


acctgtcaccagggggcagctcgtggggctcactgacccttctgaacatggccatctctg


 


tggagctggccagggagtcagccctccttaggaagcctgccctgggcccctggctggcga


 


actgagcattcaccatggcatcctcatttacagatggttgggaaaaggagaacatctgct


 


ccatgggtgggtatcctctataggctctggggctgacgagatccttggcgatgtttttac


 


cgggctgttgtacatactcagaattgtgtgcaaaagggggtgggatcctcttctctgctt


 


ttacttccaccgctccttgtggattgaagctttggtcaaactgatagaccttcttagtca


 


tagatgctttttgttggggtgggcctttactacttccgtacatgagcatttcgtcatcca


 


gcatgggtaccgactggctcctggacatactggacatgccgtgctctggtcccaagaact


 


tgtctggccgctcgtggcttcctaagtgatcgcttccagaggcatagttttccagtggaa


 


tgttatagaccttgtaggtaccgacgtcaatctcatcaatactctgggactttttgaact


 


tgttggactttatatctttcatgagcggggaaagcctctccgtgcttttgctaattgcaa


 


taacacctttagacgaatctgggcggcagtggatttgagaaaagacattaccgagactcc


 


ggttgggggtggggtcgtggtactcccatggcactcccggagaaaagggacctggtgtct


 


ctgtaggctccttcatgtggtcttttctttcaggcaagggactggtagtaggggttgttt


 


ctaatttggagggaaaagcagtcctgtcctcaaatggactgggggttctggtccaattct


 


gccagggattggaaggaggcacttctgtttctggtgtgtgtctgtgggtggactgctcca


 


gttccaggggaacaccgacaatcctttcttgcctgattaaaggcctgcgcccatgagcag


 


gtacgcttctagccttggagcttaagagagggttattgttggcattctcgcctgtggctt


 


cctcggagacaaaacctgtgttatcgtaatgggagccatcggtccagttgtcagggaaag


 


catcactcattggcagccgatcaggacgctgggggaggttccctggagggacagcctccc


 


gttgggtgagtaatggctttgcatctagaggctgtgggaaaggtggtgcaatcctgttac


 


cccacaaagagttatgcacggcatctttagttgttggctgcaaggaccccactttcaccc


 


gggtgttggaggatgctgaggaagcctgggagggcgagtagtctgagtaggtgcctgagg


 


agacactgttattcagacagtgagttttgtcaacttcagactcatcagttgattcttttt


 


tgtccttccctagcagaacaagtttgggtgggtacagaggggtctcagctaatgaagggt


 


gaagttccccaatcctcatttcattagctggatgaacaaaagaatcctccactgtaatct


 


cctttggggccaccggccacttgtgttcaaacttttctttcacagtttgctccgtgttag


 


ctgttgggtttgcgttctcaacgcgcactccatggcttggcttacccaccagattttgaa


 


cagattttaccatgttctttaaatcctccgggtaaggagttggatatcgttttaggttta


 


tttcaacctgtctgccacttagagagggaagagtggtggattgggctgccactggtaatg


 


gagcacacatgctcctctcctgctggaggccacttatacaaccccatgccagctgggggt


 


cactctgggggacgggcatatcttggatctgctgatcacacctggcccatggtgtgcagc


 


aatcgccagacagcctggcaggttggagagtaatcccacgctggcccctgtcccaggggg


 


cttggcaggagagagccttaactttcccagcactttcgtcatcttcttttttatcctcaa


 


attcaaaggcaacagtcatgcgctgttgtctctgctcctcccacagggtggggttgaagc


 


tgtcgctgtctgactggaaatcttcatcaccacggggctgctggggaaacatgtagttgg


 


tcagtaccctttgcttggtttctggatgggcttctgtttgcagagggatgagggccttgg


 


actgattgtcagaaagccacaatgctgcaagctctttgagtttggtgaaggagaatggca


 


agttcttcaacctattatcacttagatttaagactcgaagtctctgcatctgcccgattt


 


cttcaggaagaaattctagcttattggagcgtagagacataacggtgacgttcttacagc


 


ttccaatttctctgggcaactctgggaggaaattctcgtccacagctaaggttcgcaggc


 


tgtgcaggtaaccaatggtgggagggagggactccagctcattgcaactgcagtcgaatt


 


cttctaataaagataagtttccgattgtgttgggtagcattgtaagctgattgtcatcta


 


cttttagagttgttaactttttcagcaatcctatagagtccggcagctgttgcaacatat


 


tggatgatagtaagaggtcctcgagggcttcacatccagaaatatccatgtcaaccgttt


 


ctatcctgttttttgacatatccaggtataccaacatctttaacttccctatagacccag


 


gcagcacttgcaatgcgttgttatccatccacagctccctcaaattctgaatttgatcca


 


gaacttcaggcagctcgctgaattcattattgcctaggtcaagtctttccagctgggcca


 


gcttgtgcattgactttggtagagttttcaagtgattttctcttaactccaagattcgca


 


atttgacaagtcttccaaaattagctggaagaaattcgaggaaggcgtcattcaggtaga


 


gctgggtcaggttaagaagctgcgtgaagccatcgggtagtttagaaatgggattgacac


 


tggcttcaataatggttaaacacttacagcactttatgttttctggaaattcttgtacac


 


cgtttttactgatgtcgagttctttcagattaactaggctagcaatggaggtcggcagac


 


ttgagaggtcattatcaggaatgcttagtttccttagagcttgacagttgaacaattgct


 


tgggtagctcctcaatctgattggcatctagatagagctcttctagtgtacgttcgaagt


 


tgaagacctccttgggtacctgttgcaggctgcagtgggagtaatccaacaccgagatga


 


tctcttcctcgccacggaagcagcggcatggcaccaggcggccgatgagcttccgtttgg


 


tggtcatctccaggcactgcattgctagtcactcctgtctctgaagacttctaggctgtg


 


ggcactttgacttgcattcttttcatgtagcgggctcactcttcttcaggcctcttccga


 


agtgctgcacgggcctccttacaaggacttctctgatattgtgggggattccttccccgt


 


attaggttctccatcatcgcagaagca


 


>scr_gb-af017393_2 (TOXMARKER Assignment: 48; SEQ ID NO: 48)


 


ttttttttttttttttttttactagtaaggtatttactaggaaatgatacaaacagccag


 


gaaaagggtgcatcgcagaacagggtctgtgcgtataagatgggtatttcccctttgtca


 


cgtcattttttccatgaagatgcgcttaagataggaagggtaaagtaccgacacgtggca


 


ggccccgggtttagggaagggaacgtgagagagacgtcaatggaggcccacaacagtgaa


 


acccctggaagagggccagagcagtcccctggtgcagtactcagcgaatgcgcatacaca


 


actggaaaggccttggcaaattgcccagccctgagctgagcggggtcaggtcgatgtcct


 


caggctccaccagcggatgcaacgtgaagttctggagaatggaggtgaggtatatgaaca


 


gctccatgcgtgccagtggctctcccagacacagtcggcgtcccgccgaaaatggcatga


 


aggcggggctcttcttgaaggattgattggcatccagaaaatgctcaggattgaactcct


 


gaggggtcttgaattggtcggagtcatagtgcacggtgttaaggagcgtgatgacatctg


 


tgcccttgggtatcaggaagcccctgaaaggtgtgtcccgaatgacgcggtggggcaggt


 


tcatggggatgacgtctgcaaagcgctgcacttcgtggatcaccgcgtctgtgtaaggca


 


tggatgcacggtcctccagcgtgggcatccgcgaacgtcccaccacacaatcaatctctt


 


cctgcacacgggcttgcactttggggtacttcataagaatgaggaaggcatggcgtaaag


 


tggtgcccacagtctccgttccaccaaagagcaggttgtgtgtggtcatcagnagggtgt


 


ccatattgaagtggctcagtgggtcttgcttctcctgtaccatttttgtgaggaagcagt


 


cgatgaagtcccggggagagttggggtccagggagtcctggtgctcgcggacgctgcggg


 


cgatgagatctttcatgcccccaaagttccggaacacgcgtctgtgcggcccaggcaccc


 


agtccaggagactcgggaagatgttgtacatctcgccccaggggctgctcataatctgga


 


agttgtcattgataaagtggataatggtgagcagccgttcatcgtcataatcgaagcgac


 


tgccgaagatgacagagcaaataatgttggagaccgagcggctcaggataaacacggggt


 


caaagggcttgccttccgttttccgcagcacgtccagcaggaagctgccttcttccagga


 


tccgctcctcgatgcttctttttcccatgccaaagttcctcaggatttggacagagaacc


 


ttcggaggatcttccagcgttctccatcggagaaggcgatgccgttgcccttggtgaagt


 


tgaaaaagatggggtatgagcctcggccactgaactcctcccctttgtccacaagagcct


 


ccttcacagtttgatatccgctgaggacaatcacacgcctgggccccaggtacaccgtga


 


acactgacccatagtccttgctaagcttggtgagtgaggtcagcaagtcttgggagcgaa


 


gctgcagcaggtttcctaggattgggagaggcttgggtcctggagggagctggcccttgc


 


cccatgaggtgaaggtcagggacagagagatgacagccaggaggagaagcaagatggctg


 


tgctcacaccatccatagtgaaggcagc


 


>scr_sc-134241980_1 (TOXMARKER Assignment: 49; SEQ ID NO: 49)


 


actatatgatcctgtttacatgaaccatacatactaggcaaacctgtagacatagaattc


 


agaccttatacatagtccaatagcatagatcacagagcatggagacctgataaatgggga


 


ctgaggctgttgggaagaagtgaggaatgactcagcaaccttgggcctggtctccagcag


 


gtctcccagaatcagaaaaatggggccattttgaacagaagtgagtcggctgactgcctc


 


agcacaatcagcgggctacaaagcaaatcttgtacactgagtctacaagcaacactctct


 


gctatggattcctgctcatgctcaagtaccctcatgttgcagagaaagtccaaaaggaga


 


ttgatca


 


>scr_sc-191609675_1 (TOXMARKER Assignment: 50; SEQ ID NO: 50)


 


gccggctcaaaggtctctgcgagcgcattggtgttttcaatgacaatcttgcgtgccaag


 


tcttctcccaaaaaggcgaattcatccagcatttcattggtggttctgaaatgcgctttt


 


ggcagtggcgctggctgggcatcttctccgtgcccaatggtccggttgatcatagcccct


 


tgaccgagactacggacaatgatctcccgatagatct


 


>scr_gb-x17037_2 (TOXMARKER Assignment: 51; SEQ ID NO: 51)


 


gaacacagacaaggatgtatgtgtgggttcagcagcccacagcatttctgctcctgggac


 


tctcacttggagttacagtgaagctcaactgtgttaaagatacctaccccagtggtcaca


 


agtgctgtcgtgagtgccagccaggccatggtatggtgagccgctgtgatcacaccaggg


 


acactgtatgtcatccatgtgagcctggcttctacaatgaggctgtcaattacgacacct


 


gcaagcagtgtacacagtgcaaccaccgaagtggaagtgaactcaagcagaactgcacac


 


ctactgaggatactgtctgccagtgtagaccaggcacccaaccccggcaagacagcagcc


 


acaagcttggagttgactgtgttccctgcccccctggccacttttctccaggcagcaacc


 


aagcctgcaagccctggaccaattgtaccttatctggaaagcagatccgccacccagcca


 


gtaacagcttggacacagtctgtgaagacagaagcctcctggccacactgctctgggaga


 


cccagcgcactacattcaggccaaccactgtcccgtccaccacagtctggcccaggactt


 


ctcagttgccctctacacccaccttggtggctcctgagggccctgcatttgctgttatcc


 


taggcctaggcctgggcttgctggctcccttgactgtcctgctagccttgtacctgctcc


 


gaaaggcttggagatcgcccaacactcccaaaccttgttggggaaacagcttcaggaccc


 


ctatccaggaggagcagaccgacacacactttactctagccaagatctgagcaataccac


 


aggagtggattttatggggcacagacagcccatatcctgatgcctgcctgccagggccct


 


ccacaccgttctaggcgctgggctggctgtgcactctcccatgtatgctgtgcatactac


 


ctgcctggtggcactcctaataaacatgctcgcagctgtgagtctgtcactggccctaaa


 


aaaaaaaaaaaaaa


 


>scr_gb-bi291805_1 (TOXMARKER Assignment: 52; SEQ ID NO: 52)


 


ttttttttttttttttttccggggtcaagatatttactcgatgctttcaggtttgaattc


 


aggggctcagcaagggggaggggcagggaagggacacacagggcatcttccaatcactgt


 


gacttctggcaggtctcgatgtcttcattgccagtggtgactgatcagttgggacatggg


 


gagaagtcctgtgccctccacgtctccattgaaatcttcttctgatatttatgcacatca


 


ttgctccggtccccgtcaaagtttccacaggccccacacaacatggccgcataatgctca


 


tcaaccatcacattcagatgcccatcctttccaagccacacctggactccggccttctgg


 


tggacaaacatggatccgtctgagatcttcctcacagacacagatgttaacacagtagct


 


gggagatccaactcggagaccattcacccatgcacccttgcttgggatcacagtcaccat


 


gccatcctggaagaagatgtggaccttgctcacgatcttgtcattgtt


 


>scr_gb-aj000696_5 (TOXMARKER Assignment: 53; SEQ ID NO: 53)


 


ttttttttttttttttttgttggtttggtttgtttttggagacagggtttctctgtgtag


 


tcctggctaccctggaactaactctgtagatgagactggcctctgactcaagagatctgc


 


ctatttctgtgaggattcaaagtgttcatcgcaatgcccggcttagaaaatgagtcttga


 


aatggcactcagaagggtggatgtggccttttgaacgggcaagtaacacaggtaaaatga


 


aaacacaacaggtgcagaagcctgatcaacactcaccgcccagacacctttcaaacaagg


 


agctaagtcaatgaggtagaaccccaaatcctccacctaggcgctgacaggcttaaagac


 


cccattgccccacacagccctccctcctttgtaaggtcactgagggtacaggacctgggc


 


agagacccagagcaaacagaaatgaaagaacaggctttgtaccctgaagagaggaacagg


 


aggttttcaactcaaggtaactggatggcagcatttgccggcttcgagtgctgagtggac


 


acacgtgcagaaatgacgtgagatgacacgcttagtaaaacgatgatacactttactcgc


 


acaacctgaacctctactaaaacccagccagccacaagctgtttgctatcctttattaag


 


aggtcccacattcttgcgggactccagccaaaccagacaggtcccctaaatatagcagga


 


ggcctggaggggaagggaatgacttaggatcccaccacaccaccctggaaacagaactcc


 


accacagacagacggacagacggacggacaagagccggggaggagaacccacctcactct


 


tggttctctccccgttgcatccactcaaaaagaaagtcaaacactggctatgcagacccc


 


agcccacccacccacccatagcagcgtttgtgggactcccccctgaaacgggtagcccca


 


agacaacttcctatggttcttccctgactttggtttgctcctggcaactccgcgccctct


 


tccttccctcagcctccagctctctctcagcatcttctaccacctactcggaccttccct


 


ctctcttgctctctgctttctggtctccctgccacgggcttcttggggaagcagcgggca


 


cctttctcctagcaagggccccactaggccctgtctgcccagcgtgggactcacacagcc


 


gccccactctctttgaggtcaggggctgagcgctgccttcgcattcgtggaggggtagtg


 


tatggtgggtagcggggccctggccgctgggctgggtaaggttggggctgttggggataa


 


gagttgtgcttctggggccgtaagtgctggggttctggctgtgtagaaccccctccccga


 


gatcggctccctccatctagggaattcctgcgaggacggtggggccttcggggacttgga


 


ggtctgcggttagggggaggagggggtgcagtgacctcttcagggggttggggacctggg


 


gcctcaccactccccattcctggccaggactccctgtgctgagggttgctcttgaagggg


 


aagcgcagcttgcaatcatgcgggggtacaaagcgagctgggggcctgcgtagtccccca


 


ccccggcccccggagccctgcagtccctgcagccgcagctgctcctgcttgagccagcga


 


aggcgaccgcgacggaaggcagggtcctcttccatcagtcttgacacacgctcccagctg


 


gactggggtggtgaggagggccggacagctggtgagtggtcattggacactgcctcctct


 


actgcctctgacccttcgggtggggcccaggtgaccaaaccagattcttcattatcatcc


 


tcaagatcctgggtcaacgggataaccctctccatgcggagcatccggtcccgcagggcc


 


tgcagctcccggtccttgctgctgttttgcagcttcacttcctgcagaattcctgtcagc


 


ttgtcaatatgagcccggaggtcttctacctctgccccacgggctccttcctcaccacca


 


cctccaccacctccacatccttcttcttcacccacagtatcccagacatccctggccaca


 


gctctccaggcgtctccaggaccctcaggcttgccgtaggtccggcacagctccctcatc


 


ttgagagcagccagagcttcaatctctgcccgtccgtgacggaagtcggccagggccacc


 


tcatagcagatctccttcactgcttgcatcttcaggtcagccatggtggcccaccggggg


 


tctttgccctggagccgccgtcgctgagggatctggtaaactcttcgaggggccctgcgc


 


ttcccactgctgggcaggccacagcgcttgacgatggtctggactgtgttagggggcagc


 


tcgtcccgcaaggaggaaatcagccgccagctttcttcacaagagcgcttgtcagagtct


 


tccccactgtcagaatctgcatacagccgctgctgctccagcaaaaggtcagcctcttcc


 


ttttctttccggtactgattctccaagtcttgtagcctcttctccatctctagcttgatg


 


tctatgccttgctgctccagccagtccttctgagcaaagttccagtccacaggctcagag


 


ggaggtcctgggggtggggggacccctcgctctcgttccagccgtgcttgctccgggtga


 


ttgaagcggaacacatggttcttgcccattacaatcctgttgcctgacttcagcaccagc


 


ggctccgtcacaagcttcccattgacatatgtctcagctccttcacaaggttccaatgtg


 


accatcacttctccatcaggctgagggatgctgcggaagaggcagtgctgctcccggatg


 


aactggccagtcaacttgatgtccacatctacctggccaaccctggtgacgccatctttg


 


atgtggtagagaaggcattcagacatcaaggggtcctcattcagatttaccaggtgggga


 


gtcttttttggagagaagacacccacagtacgccatcctcccggagagcccatctcagcc


 


agcaatgcttctctctccatcctcagagcttctgtcttacggagcttctcctcccaagtc


 


tcattcagctcagctataattttctctgtttcctgcagcctctccatggcctcctcaggc


 


ccaatctggggctcagcactgggtgaaaatgacggctccagctcgccgttatgtggagga


 


ggagatgagggtgaagctggggcagggggagatgatgcagcaggcagaacacctccagga


 


ctcccctcttccacctttagacctcctagagcagaggctgaaagcccctgagccatcagc


 


agttcccgcaaccgggccacctcctcctgcagctcccggataagccgggcattggggtcc


 


tcattgatgacagcattgcatcggatctgtttggtgcggtctgcgtacctgagagtgctg


 


agtgtctcctcgtaattgatgtcagcgggactcagggctgcaatcattgctgtgcgtgag


 


ttcccacccaaattctccttgagtagccaggtaagcacagagtctctgtaagggatgaag


 


tccgacttccgcttctttgattgcaaatctgccagggctgagatcaccttccctagagta


 


gtcagggacttattgatgtttgcaccttccttcagacgcatgccccgagcccctgaggag


 


tcggcccgctcgctcccggcaaggttcaccaagctgatcttactgaccttttctgaatcc


 


agtccagtaagctggtcatgggagcgctgggtaaagacgatagtaaagacagcgtgggag


 


cggctgctggtttcgttcatgttggtggcagccacagttcttgccttatttccacagtcc


 


atgaggtcagcaatgtctgcataggaagtcacagccaacttagacaggtcttgtacatat


 


gggcccaggatggggtgctcccggacccgcagagagccccgactcttggggttcaagagg


 


tctcgtactcgttcgcaatagatctccatatagctcacctccacagagtaggaaagttga


 


gcactctggttcacattaactcgagagaagaggtcctcgcagagctgaggtacaatgccc


 


tgctgccccggttcctgccgccccatcatggtgtaggacttgccagcccccgtctgaccg


 


taagcaaagatgcacacgttgtagccttcaaaggcatgcagcagcatctcctctcctatg


 


tctcgatacacctgctgttgagatgcaaactgtgggtcctccaccgaagtatgtgaccag


 


taagaatagtcgaatgaagcttttaaaaacatcctgctctgtttgggattaatgatggag


 


gtggtgttgccctgcatgctgaccacacacttggcatcctggctggtctcacgggcatta


 


aagggccgaaccctcactgccactttcacggaggcaccagccatagcttcagaatctcct


 


gccctcctcagctggtgtcctggccccagatcagcggggctgtatcagttctggctgcca


 


ccggccctcgtatgggaagccccatcctacacttggggcctggccacaccagcaaggctc


 


ctcgcggcagactcccggcagagagcaaagggacaatactttgctggcgagtagtgctat


 


gaactctgcgctaccggtgtaagagacgcatcggggccagttcggggctgcccccgcccc


 


tcg


 


>scr_gb-d79221_3 (TOXMARKER Assignment: 54; SEQ ID NO: 54)


 


atgggaaaaaaagataacccagggtgtgagcattctcgtgccgaattcggcacgagcagc


 


attcgggaaaggcaaacagtggctctgaagcggatgttgaatttcaatgtgcctcatgtt


 


aaaaacagtcctggagaacccgtatggaaggtactcatctatgacagatttggccaagat


 


atcatctctcctctgctgtctgtgaaggagctgagagacatgggcatcaccctgcatctc


 


cttttgcactcagaccgagatccaattcgagatgttcctgcggtgtactttgtgatgcca


 


accgaagaaaatattgacagactgtgccaggatcttcgaaatcagctctatgaatcctat


 


tatttaaattttatttctgcgatttcaagaagtaaactggaagacattgcaaatgcagca


 


ttggccgctaatgcagtcacacaggttgccaaggtttttgaccagtatctcaattttatt


 


actttggaagaggacatgtttgtattatgtaatcaaaataaggaacttgtttcatatcgg


 


gccattaataggccagatatcacagacacagagatggagactgttatggacactattgtt


 


gacagcctcttctgcttttttgttacattaggtgctgttcccatcatccgatgctcaaga


 


ggaacggcagcagaaatggtggcagtgaaactagataaaaaactgcgggagaatctaaga


 


gatgcaagaaacagcctttttactggtgatccacttgggactggccagttcagcttccaa


 


aggcccttattagtccttgtggacagaaacattgacttggcaacgcctctgcaccatacg


 


tggacataccaagcgctggtacacgatgtcctggatttccacttaaacagagtaaatttg


 


gaagaatctacaggagtggaaaattctccaactggtgctagaccaaagaggaaaaacaag


 


aagtcttacgatttaactccagttgataaattttggcagaaacataaaggaagtccattc


 


ccagaagtcgcagaatcagtccaacaagaactagaatcttacagagcacaagaagatgag


 


gtcaaacgactgaagagcattatgggcctagaaggagaggacgaaggagccatcagcatg


 


ctttctgataacactgctaagctcacatcagctgtcagttctttgccagaactccttgaa


 


aaaaaaagacttatcgatctccatacaaatgtcgccactgctgttttagaacacataaag


 


gcaagaaaactggatgtatattttgaatatgaagaaaaaataatgagcaagactactctg


 


gataagtcccttctcgacgtcatatctgaccctgacgcagggactccggaagacaaaatg


 


aggctgtttcttatctactacataagcgctcagcaggcaccatctgaggttgatttggag


 


cagtataaaaaggctttaacagatgcaggatgcaaccttagccctttacagtatatcaaa


 


cagtggaaggcttttgccaagatggcctcaactcctgccagctacggaaacactaccact


 


aaaccaatgggtctcttgtcccgagtcatgaatacaggatcccagtttgtgatggaaggc


 


gtcaagaacctggtattgaagcagcagaatctacctgttactcggattttagacaatctc


 


atggagatgaagtcaaaccccgagactgatgattacagatattttgatcccaaaatgctg


 


cggagcaatgacagctcagttcctaggaacaaaagtccattccaagaggccattgtcttt


 


gtggtaggaggaggcaactatattgagtatcagaatcttgttgactacataaagggaaag


 


caaggcaagcatattttgtatggctgcagtgagatttttaatgctacacagttcataaaa


 


cagctgtcacagcttggacaaaagtaacacagaagagtcataatgggtgatcagtgtgga


 


cagatgtaaaaagccagacgtgtccttctccatagcagtgccctaacagtgcaacctgcg


 


gaatcagtcatttttaaagaaattctatacttcatatactgtacaatgattaaaataata


 


aaccatttcagaagtaaaaaaaaaaaaaaaaaaccc


 


>m61937 (TOXMARKER Assignment: 55; SEQ ID NO: 55)


 


ctcaggtttctcacactcctggtaatactgtaaaactttaccatggaccacagttccaag


 


gactcctgaacacagtcttggagttaagcctgtgaacagcccacgcttcccatcgatgcg


 


taacaagcgatggattccatatctctgcgtgtagcactaaatgatggtaacttcattcct


 


gtactggggtttggaaccactgtgcctgagaaggttgctaaggatgaagttatcaaggct


 


actaaaatagctatagataatggattccgccattttgactctgcttatttgtacgaagta


 


gaagaggaagtgggccaagccattagaagcaagattgaagacggcactgtgaagagagaa


 


gatatattctatacttcaaagctttggagcactttccatagaccagagctggtccgaact


 


tgcttggaaaagacactgaaaagcactcaactggactatgtggatctttatattattcat


 


ttcccaatggctttgcagcctggagatatatttttcccacgagatgagcatggaaaacta


 


ttgtttgaaacagtggatatctgtgacacatgggaggccatggaaaagtgtaaggatgca


 


ggattggccaagtctattggggtgtccaactttaactgcaggcagctggagaggattctg


 


aataagccagggctcaaatacaagcctgtgtgcaaccaggtggaatgtcacctttatctc


 


aaccagagcaaaatgctggactattgtaagtcaaaagacatcattctggtttcctactgc


 


acgctgggaagttcacgagacaaaacatgggtggatcagaaaagtccagttctcctagat


 


gatccagttctttgtgccatagcaaagaagtacaagcaaaccccagccctagttgccctt


 


cgctaccagctgcagcgtggggttgtgcccctgatcaggagtttcaacgcgaagcggatc


 


aaagagctaacacaggtttttgaattccagttggcttcagaggacatgaaagccctggat


 


ggcttgaacagaaatttcagatacaacaatgcaaaatattttgatgaccatcccaatcat


 


ccatttactgatgaatagtaacatggtggactttgtcagcatttctatcggaagatctgt


 


ttatgcattgtgatttgaaagatatcttggatactggtgactgaatgcatcagaccactg


 


tttctgttaattcacagtcagctggagcaatgtccacagtgctatgagggaagccatgtt


 


tttgtcacactctgaaatggaacatcacgttgcttttccttgtgtttttaaatattcatt


 


tattttgctttccatatatgaatattttccctacatgtatgtgtatctcatgaatgtcta


 


tgtccatgcagggttgaagagtgttgcaggtcacttggaaccggagttacattgattatg


 


gagttaccatgtgggtgctgggagccaaacctaggtcttctgtgagactagcaagtgcct


 


ttgaatgctgagccatctcattaggtccaaccctaaagatccttgcctgccactatttct


 


gtgatctcaatgttttgttttctcctgacttctgacaccaagctgatttgctagaagtct


 


tgggcatgaagtgggtgttgaggacagttattgcaaagggatttctgggtgggagttgaa


 


agaacgttcaacattcagggaattaattgttcgaggttattgattagtcaatattcccc


 


>cszr_229602935_183895355 (TOXMARKER Assignment: 56; SEQ ID NO: 56)


 


gtgcacttgtccgaggcacctttgcagacacagccctgggcacatttggagcagcccacg


 


gggcagcaggagcagcagctcttcttgcaggaggtgcatttgcagttcttgcagccgcag


 


gagctggagcaggtgcaggagccgccggtggagcaggagcagttggggtccattccgaga


 


tctggtgaatctggagcaacggtgtaagcgacaagaaggcagtttttttttttttttttt


 


taaaataaacaggcttttattttccacctgctcggtacaaaacggggtttattaaactgg


 


gtggaggtgtacggcaagactctgagttggtccgga


 


>scr_gb-af106944_3 (TOXMARKER Assignment: 57; SEQ ID NO: 57)


 


ttttttttttttttttttccaaaacaaattcttttataagttgtcttgtcatgttttgtc


 


acagcagaaagaaaagccactaagacacttgctaatcccccgttctgtttttttttctca


 


aaaccccaagatatatatatatatatatatatttacactcattttacatatgcaaaaata


 


gaaccagactcttctccctaaagacttccctgaaaaacctactcagaaccctgcaagtac


 


ctgatttctgtttattgagcttctcttccagaatcaagggaataaagacaaaggtttatt


 


tttcttcactccaatgcctccaggaccaacctggcatggttttcattccaggagctagca


 


aaataagggatgaaagtttaggtatcttgcctgctaatttcagtttcctaagggtggaga


 


cagctccgtgtaaatgcccagtaaacaggtacttgttgagctaagtcatcaaaggaggag


 


cagtgccccagaataaattgacagttaatgatgtcaagtatcttaatgtttatttttatt


 


ctttacatccagcacttgaagaaaagaaaatgacatagtgttttagaaacatagtccttc


 


atgattataactcatcaataccttagaacacacaaggacactgtgagttaatgactacac


 


taaaaaataatgggaaattcagcataattaacaaaaatccaagaggaaatttcaggacct


 


tgatcagaagctttcactaagtgctggcactatatgctacttcatttcactaagtgctgg


 


cgctatgtgctacttcatttcactaagtactggtgctatgtgctacttcactgtagacca


 


agcttcagggcaggctaagaaatcttaaccctctgaagacatgatctaagaaatggggac


 


caagcacttgtagagaattggtagccatcaagaagtccctagtaaggacagctatggaag


 


gagctggccacctttaacctgaacctgtcttaaaattacaaagcccatggagcagtactt


 


ataaacacaagcatggtgaggttttgccattctataaataatcttcaggattccagctgg


 


ggctctcttttggcatgagaagcttcaggtaaaccagcagacataggatgacctattatt


 


gatggaccttctcaaagtactcttttgaagctgttggacttggcttgatcgtaggggact


 


ctggtgtccagttgggtgggcagacttctccatgggtctccacaaactggaacgccttta


 


ccaaacggagtggttcttccacacttcggcccaccggaaggtcattgacactcaggtgct


 


tgatgacaccattagggtcaataatgaagagacctctgagcgcaatgccagcactttcca


 


acagtactccgtagtctcgggatatctgcttagttaagtccgacaacagcgtgatgttca


 


tgtggcccaaaccaccattctttcttggcgtgttgatccaggcaagatgactgaagtggg


 


aatccacagaaaccgcaactacttcacagtttacgtcatgaaactcattggctttgtcac


 


tgaaagcaacaatttctgtaggacacacaaatgtgaaatccaaagggtagaagaaaagca


 


ccaagtatttccccttaaagtcgtcgagactcagctctttgaactctccattgacaacag


 


cagtacctttaaaatggggcgcatgctgggtgacagcaggggtgtggaatgaagaactgg


 


tgctaaaggcaaactttgcttggggacaggcagaccacagcatgtctgtcaagcaggttc


 


ttctagaagcaacaggcctaagaactgttgaggcagaaatactccggaaaatagtgctcg


 


caggccgagccaccgaggaccagagcaaccttcccgcagctgccgccatcttcagagaac


 


gcaagagccacgatagc


 


>scr_gb-m11794_3 (TOXMARKER Assignment: 58; SEQ ID NO: 58)


 


atgcccacatttgtgaccagtacatgtttctgcccaccatgttcgagactatcaaagtcc


 


agaggggtcatcaatccacttatcccaaatcaaggtgcaccaatcccatttcaacgcctc


 


tgccagccccttatttccaatgaacacagacaaagctgggttaatcaagtcaagtttttt


 


tattttattgtcagttacatgctttatagaaaaaagtgtggagaaccggtcagggttgta


 


caaaaaaaaggctaggttcctacgttgttttatttacaccattgtgaggacgcccccact


 


tcaggcgcagcagctgcacttgtccgaagcctctttgcagatgcagccctgggagcactt


 


cgcacagcccacggggcagcaggaacagcagcttttcttgcaggaggtgcatttgcattg


 


tttgcatttgcaggagccagcgcaggagcaggatccatctgtggcacaggagcagttggg


 


gtccatggcgaatggaggcggcagttggagatcaacgagagatcgctgtagagttctagg


 


agcgtgatggagagaagcacgcggagcgcgacctttatagcccagagtattgggtcgcgc


 


gcaaaagctccgcccgggtggcggggcgccacctgccctcctccccactgcctgcacacg


 


cccttcttctggctcaagggaaatgg


 


>scr_gb-af069306_1 (TOXMARKER Assignment: 59; SEQ ID NO: 59)


 


ttttttttttttttttttaggaaaagcgactgctttaatgaattagacaaaatttcacat


 


gaaatcagaatcctataatccttcccttctgatcactaaaaaatgcaagattcattcgtt


 


acaagccatgtgcgattcggacccctcgaaggcagtgcaggtctgcggtccagcctcagg


 


tgctgcactatttcccattctcagcgctgaacattcgttctgtgagcatccgctccaact


 


ttatggcatcagcagcaaacttgcggatcccatcagagagcttctccacagccatttggt


 


cctcattgtgcagccaacggaaggccttctcgtccagatgtatcttctccaagtcactgg


 


tctgggctgctttgacggaaagcgtgggtgccagcttgctgctgtccttgagcagctccc


 


ccagaagcttgggtgagatggtgaggaaatcacagcctgccagcgctttgatctcacccg


 


tgttacggaaggaagcacccatgacaatggtcttgtagccaaactttttgtagtagttgt


 


agatttttgtgacactcttcaccccagggtcctcctggggttcgtaggatttcttgtctg


 


tgtttgccacatgccagtcaaggatgcgccccacaaagggagagatgagcgtcacgcccg


 


cttcagcgcaggccacggcctgggcgaaggagaaaagcagtgtcatgttgcagtggatgc


 


catgctgctcctccagctcctttccggcctggattccctcccaggttgatgataacttga


 


tgagaattctgtccttgctgatcccagcttctttgtaaagctctatgatgcgcctggctc


 


gggccaccatggcatccttatcaaaggaaagccttgcatcgacttctgtggatacacggc


 


ctagaatcttctttagtatttctgccccaaacagcacaaaaagtttatcaatggcatttt


 


taatctgctcctcttgtggcccacccagcttcttgccgtaggcaatggcctcctccacca


 


gctcttggtaggcaggcatctgtgctgcagccaggatcagggatgggttggtggtggcat


 


cctggggcttgtactcatcgatggcgttgaaatcacccgtgtcagccaccacggtggtga


 


actgcttgagctggtccaaggcggactccatcctctggcgctttaccggggaccccgaca


 


tggcgaaacgcgcacagctgaggcggtagctggt


 


>scr_gb-d17310_4 (TOXMARKER Assignment: 60; SEQ ID NO: 60)


 


gcactctccagcctctcaccgacttttttttcaaggagacaattttatttttttaccaag


 


gctgaatttataccataacatgggtaacagagggaggggggaagtgtgaaacatttacac


 


aggccaagggcacagtatacatgtagtcagctgatgtcaacaggatgttggtttttcaga


 


aagcttacaggtcatcacattgggtatcttgatgtcagatgtatttctcagcaaggtcag


 


aactttatcatatcattattcatcctgaccaccagatttgtattagtcttctgcagctgg


 


ctggggattttccatgaacccagtcatacttaattctaaccataacatcaataatggagg


 


gtttcaagggcattgctcccaacatgtaattacaaaaagaaaaaagatgatatatttccc


 


aaaaagagagacacattcaaatttcctctcaaactccccacatctgaatcatgatgatgc


 


ttttaaattggttctcttcttaccaacattccaaccttcccacaagaacttgctctccag


 


gttcttggagctctggttcttgggctgttggagagaaccctgggtctcttggtcactcct


 


gccacaggtgccctacctcaaaactaagaaaaagggaaaatctatggagtactttcttct


 


tcctcaaagaatatggggaatattgactaatcaataacctcgaacaattaattccctgaa


 


tgttgaacgttctttcaactcccacccagaaatccctttgcaataactgtcctcaacacc


 


cacttcatgcccaagacttctagcaaatcagcttggtgtcagaagtcaggagaaaacaaa


 


acattgagatcacagaaatagtggcaggcaaggatctttagggttggacctaatgagatg


 


gctcagcattcaaaggcacttgctagtctcacagaagacctaggtttggctcccagcacc


 


cacatggtaactccataatcaatgtaactccggttccaagtgacctgcaacactcttcaa


 


ccctgcatggacatagacattcatgagatacacatacatgtagggaaaatattcatatat


 


ggaaagcaaaataaatgaatatttaaaaacacaaggaaaagcaacgtgatgttccatttc


 


agagtgtgacaaaaacatggcttccctcatagcactgtggacattgctccagctgactgt


 


gaattaacagaaacagtggtctgatgcattcagtcaccagtatccaagatatctttcaaa


 


tcacaatgcataaacagatcttccgatagaaatgctgacaaagtccaccatgttactatt


 


catcagtaaatggatgattgggatggtcatcaaaatattttgcattgttgtatctgaaat


 


ttctgttcaagccatccagggctttcatgtcctctgaagccaactggaattcaaaaacct


 


gtgttagctctttgatccgcttcgcgttgaaactcctgatcaggggcacaaccccacgct


 


gcagctggtagcgaagggcaactagggctggggtttgcttgtacttctttgctatggcac


 


aaagaactggatcatctaggagaactggacttttctgatccacccatgttttgtctcgtg


 


aacttcccagcgtgcagtaggaaaccagaatgatgtcttttgacttacaatagtccagca


 


ttttgctctggttgagataaaggtgacattccacctggttgcacacaggcttgtatttga


 


gccctggcttattcagaatcttctccagctgcctgcggttaaagttggacaccccgatgg


 


acttggccaatcctgcatccttacacttctccatggcctcccatgtgtcacagatatcca


 


ctgtttcaaacaatagttttccatgctcatctcgtgggaaaaatatatctccaggctgca


 


aagccattgggaaatgaataatataaagatccacatagtccagttgagtgcttttcagtg


 


tcttttccaagcaagttcggaccagctctggtctatggaaagtgctccaaagctttgaag


 


tatagaatatatcttctctcttcacagtgccgtcttcaatcttgcttctaatggcttggc


 


ccacttcctcttctacttcgtacaaataagcagagtcaaaatggcggaatccattatcta


 


tagctattttagtagccttgataacttcatccttagcaaccttctcaggcacagtggttc


 


caaaccccagtacaggaatgaagttaccatcatttagtgctacacgcagagatatggaat


 


ccatcgcttgttactcatgcaaccaagcaggtcttgggtctggcgagggtcttctgactg


 


ttctgagacagccctgtgtgaggaatgcactttcacagggttggaggtacttccaagacg


 


ccataggaaccacacgtgggtcacagctatcagttcactgtgggcaagaaacctctttat


 


ggccacctggtaacaaaaatttttctgtctgtgaattttttcttactatttaaa


 


>scr_gb-bf281368_2 (TOXMARKER Assignment: 61; SEQ ID NO: 61)


 


ttttttttttttttttttcacacagggttgcttttatttccacatccaacttgagcagag


 


gccctgccacaacctgaacagctgtgaggtgctgggtgcctccagagtttctggcacagt


 


aagtgttgggtgtgcagacttcctgatggccacatgacactggcccacacaggaacagca


 


agtccatgaatggaaatcccactgagctggaagtggaggctctggaaaccccatgggcag


 


cagcaggagttaaaggagccaccaggaacactgcagtgaggctccaatgcagacagggct


 


gataaaaacccaaacagggcattgtgagagcagaggctcgagtgtccccgctgaggaccc


 


ggggctgaaggcacagagctgtgtcgggatggaagaaccctgggtgcactcgcagtccag


 


agcacgaaagcacaggtgagaacccagcccgaggctctctgtgaagagtgtggccttgga


 


tcttgggcacggcacagtgacacacagtgctgaggtcactcctgacttcccagaggaatg


 


acctcttcagtgacaaaaaactcaatggtctcttcctcccagtcatccacgttgctgtcc


 


agctcgtcagtgtccacccctccccgtagctctagacgctcgttcttctgcttcatatag


 


agttcctgggccatttttcggtattgcctgaagtcctccatcatggtccgccttctttcc


 


accagttcctttgaagctttggactggctcaagcgatccttctgctcaaagatcttagag


 


tatttcttcagatcctttttaatttgctttatctgatcctgactgaggagtgttgggggc


 


cttggtctccagagcagctggcagaagcggtccttgttgttcttctggagaagacgacct


 


tggaaggtccacagccaataagcattgtccaccttatggctccaccacgacacagaggta


 


accacatagcggccagttgggtcccattcgacgtcggaggccatgtagtgctctgcaatg


 


ttcatgacggtgcagtctgaagtgtcgacaaacgccaaggcgccattcatgctcctcagc


 


ctcgtg


 


>scr_gb-u56407_3 (TOXMARKER Assignment: 62; SEQ ID NO: 62)


 


ccaaaccaacaaggcagccacaggccgtcggtgcctgccgccttccaccaggggcccgcc


 


aagacaaccttccaccatggctttgaagagaatccacaaggaactgaacgacctggcgca


 


ggatcccccagcacagtgttcagcaggtcctgtcggggaagatatgttccattggcaagc


 


tacaatcatggggccaaatgacagtccctaccagggtggagcatttttcttgacaattga


 


tttcccaacagagtaccccttcaaaccacctaaggttgaatttacaacaagaatttatca


 


tccaaatgttaacagtaatggcagcatttgtcttgatattcttcggtcacagtggtctcc


 


agcactaactatttcaaaagtacttttgtccatcagttctctgttgtgtgatcccaatcc


 


cgatgatcccttagtgcctgagattgctcagatctacaaaacagatagagacaagtacaa


 


cagaacagctcgggaatggactcagaagtatgccatgtgactaaagagattattggatcc


 


tctgcgaataaaagctaggggaactctgaaagagaaagtccttttgattcccacttgact


 


gtttgctgtgaacccacgatgtaccggcctcgtcctccctggtgcacggtcttcatctga


 


tacagtactgttgcatgttgcacgcaccaaaaatactgtgtttctgtaccaacactgtct


 


cctagcagacgagccttctccaggcataacctaggtgtgagattaaaagttttccttatt


 


gacttaaatctggataacaaggtgtgagtgagggtggtgggtacaagatactgctcagaa


 


ggggtaaaggtccccaacctataagacaatgagatggcttttcagtggaagccatttaca


 


gctaaatgtttaaatgaatgaaaagctaggtgaagaacatgaatgttcctgtactcattt


 


tattccaaaagacctagagcttaaatgaacattaaagccaaccagactaagccaacccac


 


ctcctgtattttaaagtctaattggtcaacaaaaatagatcggcactatcggtccataaa


 


gtgtgcctggctttgttcccaaatcctttatacacggatgactcaacctattttctttca


 


cactttctctccatattctttggtttacttgcggtttctcagttgattcatcactaatag


 


ctcttatttttattatattaactgcttaatctatttggatgtaaaggtagacattcaact


 


tgatgaaaaaagcttgtgtatagagacctaattgctcctcttggagcttgtacagtcaag


 


aatgatgcatctgtgtaataaaccaattattctagccattat


 


>scr_gb-ai406674_1 (TOXMARKER Assignment: 63; SEQ ID NO: 63)


 


tgtacactacccctcacaaaccacaagccgcagcaacatggatgcccagtctggagcagc


 


aacagccaggatgacctggagccaggggggcttcggaacagatgtgcacccttcctgggt


 


gatgttttcagctttgtgagaaaccttactatcagaggagatggctagcaatgttaccaa


 


caagacagatcctcgatccatgaattcccgtgtattcattgggaatctcaacactctggt


 


ggtcaagaagtctgatgtggaggccatcttttcaaagtatggcaaaattgtgggttgctc


 


tgtgcataagggctttgcctttgtccagtatgttaatgaaagaaatgcccgagctgctgt


 


agctggagaggatggcagaatgattgctggccaggttttagatattaacctggctgcaga


 


gccaaaagtgaaccgaggaaaagcgggtgtgaaacgatctgcagcggagatgtacggttc


 


ctcatttgacttggactatgactttcaacgcgattattatgacaggatgtacagttaccc


 


agcacgtgttcctcctcctcctcccattgctcgagctgtggtgccttccaaacgccagcg


 


tgtgtcggggaacacctcacgaaggggcaaaagtggattcaattcaaagagtggacaacg


 


gggatcttcttccaaatctggaaagttgaaaggtgatgaccttcaggccattaaaaagga


 


gctgactcagataaaacaaaaagtggattctctgctggaaagcctggaaaaaaaaaaaaa


 


aaaaacccctcgtgcc


 


>scr_gb-bf290678_2 (TOXMARKER Assignment: 64; SEQ ID NO: 64)


 


ttttttttttttttttttgattttggccaaactttttatttagtattttgtagttgttta


 


acacacacttaaatggtcttactcggggagggggaagggaggttcttgtanattcccaag


 


gaaaggtcagaaaagcaaaatatggccagcatccatttgctttttttgaggggggggggt


 


ttctgggtaaatagtacatgcctaggcatctgatctcagcttggtttgtttgtttgaata


 


tatatatactgcgaacattgagatttcagttggaagacaccctgaaatcctcacacccca


 


ccaaccctctctaatggctagcttgtctgcacaggcagggtgattcaactctcaatggag


 


accaaaggacatctagatggctaaatgtttgtggaagatcttggggttgcttgcctcatt


 


tgctgggaaaaatcaggaagtggccttcagggacacttttacttggaaaattacaacact


 


agttacaagtcacgggttacacatctaacatttgcttgttgaaagcaactcataatagca


 


aataaaattaaacatgtcttactttttccctcacaagaacataaaaattattaaggggaa


 


caggaaattttaaaaaggtaacacaatttttcctttagtagtccttgggtagtttatgac


 


agaaagtttccatttttttgtttgtttctttgaatggggattgttggtccctcgtg


 


>scr_gb-bi288503_1 (TOXMARKER Assignment: 65; SEQ ID NO: 65)


 


tgtacagttgctagtttgaggctggtgttgatgttctgacaagagtggctcagccatggc


 


tcagtagagtcctcttctggaagtttgagaaattctggcttacgggaaaaggtttttctt


 


tcttttcaagatatgtccaacaaagtcctcttcggtcagtaatttctgcagtgacgcctt


 


tcgtccgtcctgtcagcaaactccaatcgcaacttgggagtccagtcaataaagggttaa


 


gcgcacacaagcgtggccaactagtaggtccgagaggttcaccggcaggcaccgtactta


 


atatgcagaggggtgggcttcacgcctccccgccgagcgctcccacggtcgaggagttgg


 


tgggcaaggagatgaggtttaagtccaatgggttaaacccaaccccgagagggttaaaac


 


tacccgatgacgctgccacggaggggccgaatccac


 


>scr_gb-d86383_2 (TOXMARKER Assignment: 66; SEQ ID NO: 66)


 


ttttttttttttttttttcacaccagatgacgaatgtatatgaaagttaattcattaaat


 


taaaaaaaaaaatcaaacatttggggagggttttttttacaacgaataattctatacaca


 


tgctatagacacggtttctataaaacacactatctacaatctacttacatttaattgtcc


 


tgctatttctagttcatgtgagatcagtcacaagtgagtcagtttccctgcctgtagaga


 


ctgcgtcatcccttaataccagggtcagaggcactggccgagcaaaacaagattgtaaga


 


atcttatcaactatcttgcttatgagaacagacaccaggggccaagtgctctgaaccggc


 


tttggagttaaggcagcaatgtaaggtgtcacgtaaaaaccaagtgtgctctttgaaagc


 


attccatggatccccaaatgctggccccctttctaagtgcacctctgaagtcgagggaac


 


agctacacatttgggaaaagtcattcgagaacagccgcccaaaacctttaaagttatagt


 


ttaagcttcaggcaaaagttcaaattacttctcacaaatagaaagaattcactttttaaa


 


aacgaagtcacatttagccactttatcaaaacaacttaacaccggtacggaaaacgtacg


 


ctaaaccaaaagtatggtttcaatgcacgccgtgccaaatattttcaaaacgctagaaga


 


atggtacttctttctctcagaatttcccagtttgtctgtagcagaacggtattctaaagt


 


ccagtctctgaacatggtcacggccgatgactgtcatccagcattaaaatagcctttatc


 


accctcgatgtccacttcctggtcggaatcctctgagatctctgattcagggtcttccgg


 


agaggctggggagggtgaacactgagaactgtccaaagaggcacctttattctgttcact


 


gggcaagtcttggccctggtcacaggaagtgtccaaactgtccaactcatcctttttatt


 


gctttgaggattctcctgcttcagtcgtctccatttagctctgcgattctgaaaccaggt


 


tttgacctgtctctcgctgagctgcaacatcttggccagacgctttctctcaggtgggga


 


gaggtatttctgagtctcgaacttcttctccagctcgatggtctggtcgttggaaaacct


 


cacttgaccgcctttccttttgtgcagaggtcgctgtaggaaggggttccagagcaaggg


 


cttgcccagggggtcgtggcggagtagggcgtgcgtgtagtcgttcaccgtccgcgggaa


 


cgggtacagagggcctccgaagccaccggggccataggcagcggccagcgcggcggcggg


 


gtgatgcgagaaggcggggtggaccggcgtgggctcgtacaccggggtccggtaggagga


 


cacgaggctggtgaaggaggagttgggggacggcagcgtgggagtgggcgtgggagcggc


 


gggcccgcgacccaggatgtcgtcgatgtagaaaggcgtcgggtgagcgggctgcagcag


 


cggcgtgggcgcgtacagcgggactccgacggcgggcgcagccgcgggccccgggtgcgg


 


gaactgcatggctgctccgc


 


>scr_sc-133366194_1 (TOXMARKER Assignment: 67; SEQ ID NO: 67)


 


gctagcatcttttttctgccacgaggtgcgttttattttcatcaatcatacaaatgattt


 


tccatatcacagggcaagctgagtgcctgggtgtgttcacagtgtagcttgtcgcttgtg


 


tctgtccatcttccccgtcagaatggggtctcagaaatgatgaggtgaggtggagaaatc


 


ctcctaggcttgtaggaaattttactcctcttttcctgttgaatggtcttttggttggct


 


ggtgttcttctcatgctctttggttttctccagtgtggctttattgaagcttgtgatttc


 


ccccatggataacttgcctgccattttcttagaactcttggaatcttgctctgagctcat


 


gctccaattg


 


>cszr_230290139_182026368 (TOXMARKER Assignment: 68; SEQ ID NO: 68)


 


aagctttggagctgctaggtgctacctatgtcgataagaaaagggatctgcttggagccc


 


tgaagcattggagacgggcaatggaactccgccaccagggtggggactaccttcctaagc


 


ctgaaccccagcaactggttctagcctatgactattccagggaggtgagcacgccccaag


 


agttggaagccctcatcacagatcctgatgagatgcggatgcaggcactgctgatacggg


 


agaggatcc


 


>scr_gb-ai013477_2 (TOXMARKER Assignment: 69; SEQ ID NO: 69)


 


tttttttttttttttctaagaagctgttctcatctatgaaccagatggcatctaccccat


 


ctgttggctgatcagtccgatctttatgccactcctgtgctttagtgagcacctggtgac


 


agtcatgatggggggtgtctaggtcagggtccgggagcagggttgtagggtttagactcg


 


taggggcagtctgggatcacaaggaacaagtgggatacccggcccacgccaaggtccacc


 


gttcttcgggtagtccatgagtatcatttgttgtcagtagccccttgtactcaaggtctt


 


ttgcttgacactagcccatttggacgtaggagcacagagtgttgggcccccgtattcaca


 


caacaactgggcgggcttcccttctatctttttgcatagccagcactctaggaccaagag


 


gcttgccttccaggctgctggagaggcccctcttgttcttcctggggcagtccctgaccc


 


agtgtcctttttctttgcattaggcacactgatctttagccaggaattctcttctgttgc


 


caggtactgtcttcctaggttccctaactactgtggccagtatatgttcctctcttgtct


 


tttatctctctttagctctctagcttcctcttctttttgtctcttttcttccctagcttc


 


ctgctctttttaccttcttttctctttctctttgtttaaccttactttctctgtaactta


 


tactaactctcagcaacttagcttaacccttcaaatttctgtaactttctcttcataccc


 


tttccttatcttagccagattggtggggcattttccagcccctaggagacccaccctcgg


 


agcctgggggcagacctggagcactccctaccttcaggggcattgaagtcaacagtcagg


 


agccttccatccatgtctggaacattctttctggcctctagcaggattctgtctttcctc


 


agtggtaaagaagatctgtaacagttactaacaagcatctcacgtgggatggtgagaaaa


 


caagaagggaatctagaggagagaggtccactgaagaggacaaatagcatttagtcacac


 


agctaaaccaggaggcctttttttggacaaaaaggccactgtaaatataagcacaagctt


 


tgtctatgaaacagaaaggcgagcagagaggcagcctagctgttaccggctgtctctctg


 


ggcttagattttcccttaaggagtacctacctcccttcagtgtcagcttggtggctttgc


 


ctctcaagagaaccagcctccaaatgacactaggcttctagtaacaactaataacaaaag


 


gatggagagatggttagaacctgggtgctagatactaagcagctgacaaaagaattgtaa


 


ccagttcacctggggctttcaggactttagtaacagccctttaccaaactgtctcagtgg


 


gctataggcccatggaaaagaaaacattaatcctgaccttgtccaccaccaaagcctgaa


 


ttctaacctcgtgccg


 


>scr_gb-m91235_3 (TOXMARKER Assignment: 70; SEQ ID NO: 70)


 


aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaacagtctctctgcatcttcttctacagcta


 


ttaggtgctgtccacttttctgcacagaccctgaaccacgcatcaacttattttctctgc


 


aacttacaataactctctcagtgacttagctttacccttcaagtttctgtaactttctct


 


tcatatcctttccttatcttagccagatccagattggcgggggattttccagcccctagg


 


agacccaccctcggagcctgggggagacctggagcactccctaccttcaggggcattgaa


 


gtcaacagtcaggagccttccatccatgtctggaacattctttctggcctctagcaggat


 


ctgtctttcctcagtggtaaagaagatctgtaacagttactaacaagcatctcacgtggg


 


atggtgagaaaacaagaagggaatctagaggagagaggtccactgaagaggacaaatagc


 


atttagtcacacagctaaaccaggaggccttttttttggacaaaaaggccactgtaaata


 


taagcacaagctttgtctatgaaacagaaaggcgagcagagaggcagcctagctgttacc


 


ggctgtctctctggacttagattttcccttaaggagtacctacctcccttcagtgtcagc


 


ttggtggctttgcctctcaagagaaccagcctccaaatgacactaggcttctagtaacaa


 


ctaataacaaaaggatggagagatggttagaacctgggtgctagatactaagcagctgac


 


aaaagaattgtaaccagttcacctggggctttcaggactttagtaacagccctttaccaa


 


actgtctcagtgggctataggcccatggaaaagaaaacattaatcctgactggcaaaaca


 


aagttcttcacagttgtagattctttgaaactattttaggggctctttttgtcccccaac


 


ctggggcattttaaccataggggcaggaactggctgctgtggggataggaccaaaggcac


 


tctccatgttaatgatgatcagtggagaaaagtaattttgatgttggagactactcctcc


 


ttggataggacagcagataaggaggcttcttaagactcttaatgagcgctctcctacttg


 


agcgaaattcctttcctgttctgttttcctatagccccactagctctccagcctttttag


 


tcattcttccttgacgatttctaacacagcctgtcctttttttatagcctgttaacagca


 


tttctgatcttttaggcagctatcgactaagtgccataccgggtgaaactccgcctttaa


 


gattccttactcccaaggaaaatttaaatctttcccagttcatcacagctggctgcgagc


 


ataagcacagaataaaacactatatgtttttgttttgtttttctttccttttttcactag


 


gctggggcccgaacccagggccttgcgcttgctaggcaagagctctaccactgagctaaa


 


tccccaacccccaaaacactatgttttaaaaattaactttggctatcaaccaacacactg


 


ccactagagcggggtctctacaaaattaagtttcttactcactaagcgttaaggggacca


 


agtaaaactcttcgacgaacaaagcaaacagtttcatgatttcaaacacagtcgtcggtc


 


caagattttaaacacagtcgtcagtccaaattcaaacacgaaacaaaagtcaaaaagaca


 


ctaacagacacaacacgtccagaaaaccacagtcaggtcacaaagaagacaaacaattcc


 


aacagtcaaacaagtaacaagcagacgcgccgcgcagcttcggtaccaaactgaaaccaa


 


aaaattcagacggagtcatcaagggtgcggatccctccgaaaacggacggaggtgccacg


 


gatccggatctccctctcctccaaccacccttggaacgtcttccagggctgcgggggaga


 


agtccgagctcgtcagctccttctctggcccgcccagatagtccccagatctgagcctat


 


tgatcgatcgttcacaggacaagacaccctcgtgcc


 


>cszr_204152648_191521095 (TOXMARKER Assignment: 71; SEQ ID NO: 71)


 


tcatgacctcattttaggaccaagagctgtgttggtttcttagattgttagctttttctc


 


taga


 


>cszr_204152792_191517979 (TOXMARKER Assignment: 72; SEQ ID NO: 72)


 


tctagaaaacggaggctgtctggatgcagtagtcatttgctgcagaggttggggaagggg


 


aggccccatgtttctcctgtggaaagagggtgtggggctctgggaaaaggccactcttca


 


aacattcatga


 


>cszr_204229614_191891958 (TOXMARKER Assignment: 73; SEQ ID NO: 73)


 


gctagccttatgccagcctgccactgtcaacatattctgttcccattggttacatgcttg


 


atacatacactcttgtgtttttggctaattgagctttttaattctattgtaatattttca


 


attg


 


>cszr_204229615_191892510 (TOXMARKER Assignment: 74; SEQ ID NO: 74)


 


caattgaaaatattacaatagaattaaaaatctcaattagccaaaaacacaagagtgtat


 


gtatcaagcatgtaaccaatgggaacagaatatgttgacagtggcaggctggcataaggc


 


tagc


 


>scr_gb-aa801331_1 (TOXMARKER Assignment: 75; SEQ ID NO: 75)


 


tttttttttttttttatgaagacacgaaatgcatttattcacataacaaaaaacaaaaac


 


aaaaacgaaaaaaacactcactccctcttcacttgaaatgtgtcagtaatgactcaaagt


 


gtcatgatttaccaggtggtgaattcttctgacaaccaggtgaagaattaggaaaacata


 


cagttccagtctttatattctgaccctagaaatcggttcatttgtagctcttgggggtac


 


acagtaaagcaggcaagcaactgtccacactgtttcattccacatacttagtgagtgccc


 


ttattcagggcctaacttcactccaggcacaaaaacaaggcaggattgcctggtaagtct


 


gaacatgagaaaagaaaacgatttattacacaacagatatatccatttatgtgagtgttg


 


acatctaggaattctctgctttatagacaattagaagcagcatcctttctttagaatatt


 


tctatgccctcactaaacccatgagtaagtatcttgcttgggagtcatacccagagctaa


 


ttacaattcaatattctccctgtacatgcaatccttgaaaaacgttatatgtattttatc


 


tcattttcataaaagaattacaaagaccccaaaaaggtttagtgtttgtttgcatattaa


 


ggttgcaattctccagaaacccaaagttcggatagtatgtgacgttgtgcagacaatagt


 


ttacctcatgctacaggctataaatgtcagaacagagcttaaacactcacattagtgaac


 


gcattggcactacttgtactctttattttaagggctaagaaaaagcacacttctactcag


 


ccctatggaagttatcagtgagcacattctctatcgctcactgtacagtaaactatgtac


 


aacaggcactataacaaacagaattttagagtcaggtatgacatgaaactttttcaattt


 


tttatatttacactgtgggtttatcctcatcttaagatcagtttttcattttgttttgtt


 


cttctgtttttttggttttttttctgcctaaacggtatgctcaagtagcatggataaatc


 


ttccagaatatgcactgagtaactccttggctcttcccagagccttgccttcagcacagc


 


atgatgttaaaagatggtctcattgtagacatcaaagtaggtagaagaacaattgtgtct


 


gtatcagaggctctatgaagagacctggagtctcgaagttccttcttactac


 


>scr_gb-aa899865_3 (TOXMARKER Assignment: 76; SEQ ID NO: 76)


 


aaattttggctggatccacagcaagagtcctcagtattatttatttgttttgttttgtgt


 


tctgttttgtttttactgcaacttgacaataaaagatgtttggcattggaagagaataga


 


acattaggtctgggcccagcgctctgactccgtcttgtttaatagtttaacctgaagtcg


 


caagactgggataaacaggagagctgacatgaaggacatcatcgcacatgtttcggctta


 


ctgtgtcagaactacacgtgcttggccttatttctttgagcctgtggcagaagagtgtat


 


cgaggcagaaagcagaaaggtccaacctccctttctagaaaatgtcccttgatgtcctga


 


ttctcttccactagtcctcactactaaaggtcctgtcacctctcagtaacactgtgggcc


 


gggaaccaagcctcagggcacaggcctttgggagctgtattagagttctcgagaagaata


 


aacagcacttgcagaaggaattcccagaagaagaatgacttacaggcttctgtccagcta


 


atccaacagtgagcagaaagtccaaaaatccagcagttcgggccatgaggctgggtgtct


 


cggctggtcttcagtagactctggaatcccaatgacgtaggctctaacgccagtgaagga


 


atggacttgccaacaaggtgaggccaagcaggcaaagagcaaaagctcccttcgtcctgt


 


cctcaagtagacttctagcagaaggcgtggcccagactagaggtgtgtcttcccacctca


 


agatcaggattaaagaagatctactgacttcaaattaagcaaaactccctcccaggtgtg


 


ccctctgtcattagattttagttcattcaagatggagtcaagttgacaaccaagaatggc


 


catcaccggggacactccacatataaactgtataccaagcttcatatttcagacatgttt


 


cttaatgtcatccacgtctccagcccctgtagtgtgtatgtgttgtattctctgcagaat


 


ttagcatgcccgtgtttcctgtccttcacataaacgcctttgtgtgaagcttgcttgatc


 


ctccactcccctctccagcccccacccctgtgacactgcccagtaataactgttcgttgt


 


ttacctgttgcttgtaagtgcaagtattaaagcaatttgaaagctaaactcacctgtaag


 


actataataaatacctgtaatccaataaaaaaaaaaaaaaa


 


>scr_gb-aa997629_1 (TOXMARKER Assignment: 77; SEQ ID NO: 77)


 


tttttttttttttttttaaataataatgttactgtcgtgttggctgtatatcattgcata


 


tacttcaggaaaagttttcttgttcttgctaaataacaaagcacaattggtaagttccat


 


ggacagcaggctccctcagaacgtagccagttctgtgaggcaccccatatcccaaggaca


 


agcttgtggcatgccagatgaacagcagccttggcttacacgcacacctgtacataaaag


 


ctcatctttccaaccacgtgcagccaagagattaccacagacttgacacagggaccctaa


 


caggctcctatagacagtcctgccgctccatgaagtggggaaggaacaaatgcagtgacc


 


gcatctaatgcacttcctttgaaaatgtttgcttat


 


>scr_gb-aa997691_1 (TOXMARKER Assignment: 78; SEQ ID NO: 78)


 


agatctgagcggccgcccacggtcctgatgacagaagagctctcctccccgaaaggggca


 


gtccggagcccaccagtggataccgccaggaaggagataaaggcagctgagcacaatggt


 


gctccagaacgcacagaggagatgaggacaccggagcccctggaggagggtctagcagag


 


gaagctggcagggctgagcgcagtgacagcaggggcagcccacagggtggccggcgctat


 


gtgcaggtgatgggcagcgggctgctggcggagatgaaggctaaacaggagcggagagca


 


gcatgtgcgcagaagaagcttggcaacgatgtcatctcccaggatccctccagcccagtc


 


atgagcaacacagagcgattagatggaggggcaacagtgcctaaactgcaaccaggtctt


 


ccagaggcccgctttggtttgggaacaccagaaaagaatgccaaagctgaacc


 


>scr_gb-ai411514_4 (TOXMARKER Assignment: 79; SEQ ID NO: 79)


 


ttttttttttttttgtttgtgaaagtacagaaaactttattggaaatctcttgattatat


 


ttccaagtgtagctctcatttcctaacaaagcactggaggaggggcttcacagccacctg


 


gtcccagcctgagcttggctgcgggagttgtctagagcccgtttcttccattgtgtagga


 


ctgaggggcacaggccaccttgaaggatgcttcgctcagcttccctggcctctttcttaa


 


gaatctgggacataaaggctgctgtctagaggccactggctgagccctgaaaagaatccg


 


tgccctcaccccccttttagtgctggccctggggggtaaatcctgttcagtaggctatga


 


atgtgcccctgacccaaaggctgcaatggcacttggccaccactgctgggcacatttctc


 


tgtggcagcaaaagcatgcacaggggaaaggctccagtgttacatgcagattactaacag


 


cagttgagagccacctgctccaatgcgtaacggctgctgccagtgaggatccagggacaa


 


gaacaggacaggctggcagaggcacttgactgactcaagcaacaatacctgaaggtttaa


 


gtcaaccataggctcagctttggtttctcaaaagggaaccaatccagcttgtaagcccag


 


ggccatgtacagactctggaattagagggagggagagagggaggaacagctccctagtcc


 


tgctccagctcaggggctggagcagcaggttatacagtgctcctctgggcaccatgggca


 


acacacctctgaggagtcctcacactgaacacacctgagacctcctgggctgctagaaca


 


gagctagtcacattacagatgctgtgtcaacagagtatgctcggcaggagcacgcagcat


 


gccgggaagcctgatgcctgctcagttccatacacacagtttgagggggctactttgcct


 


ttgccagacccattgctgatctctccttaggtgtgacaggaagatcctcagagcagtagc


 


acaggttctgagtaatcttcaccggaggcctacagcccagagaaaccctcctccttcccc


 


agcagaactgctaaccccaaacatacttcttttataaaatatctgatttctctgacagta


 


ataaatatttaccatgttctatatccacgcagcagcgatcgagggaaaacgaggaggaaa


 


aaagatcctacaggcggccgc


 


>scr_gb-aw142560_3 (TOXMARKER Assignment: 80; SEQ ID NO: 80)


 


ttttttttttttttttttgattgaaaatgtttaatttgtaaggcacacagtttatgatca


 


ttttaatatctaaaagaacgaaattaacaggactaaaacctgattgtcgaatcatttacc


 


aagtttggatgtcacgttgtaaaagcaggcttaaaaagatgactccttacaaaggagtga


 


ggtggacctgggtgggacaggctagacatggccctgaaaaccttcttgggtgacaaagaa


 


acagactactggactgaagccacagcttccaagaaacaagaaaatgtagtggccaccaca


 


ttgggctttgtttccttatgagacattttccacctcatctcgggatcttactgttaccct


 


tgcccaaactgcttatggcatgagggttccagagcccagcgccccagccaagtgtacaaa


 


agacgtttcctgtagagtgtgcctgtgagggacaagcttgaggagtcctgtagagcgtcc


 


agacaagctcacatttcctcattcatggatgatgaaggggatgtcacaagcagaccagaa


 


actcctcaatgtctcaggaaaggaccgttttccagagcggcttacaagtgggactttctg


 


ggtttccatctggagtttggttttcctgcttggcctcagactgagatagaagagcagtga


 


gacagaaagtagacagagaatgagctagcctccgg


 


>scr_gb-aw533305_2 (TOXMARKER Assignment: 81; SEQ ID NO: 81)


 


tttttttttttttttttagaggttaaaggtgttcatttgccaaccggacagcctgagttg


 


gatcccctgagcccccatggtggaaggaaaggattggctcctgtaagttgtccactcttc


 


tgaagtatgtgcactgcggtgtgtacctgcccacatacacaaacaggctaggtagagaag


 


aaaagggaaaccattaatagtcaacactgatacttatcaaaaatggcactagatggtgat


 


ggtttaaaagcttcacttagaagccaacagtgacagcagagacagacctctgttaaccat


 


tgcaggcaaaactgaaagacatgctcacacaggaagcaagcacaggcggctttgttgacg


 


gcttagctgaaacagactcaagacaaagcgtgttaacagacagacgcacttcacggtgac


 


acgaggggccagctaccaagaagacattgaccccaaaacatgtatacacgccaacagaat


 


cccaaaaggcacagtgagaaaggacagaaggaaagttcgaaatagaactttgtgctgccg


 


aggtaggagattaacttcccctggagatttccacagtggccaaaacttcggtgaggatat


 


ggaagacggaggtaccatctgagcttgatcagactctctaaggtgtgatattgcaaatag


 


tgcaagccaaacgactcagcgggcacatcacaggttcaagaccagcctgagaaacttagc


 


agggccctgtctcaaaattaaaagaggttgtttttaaggaccagcctcgtg


 


>scr_gb-aw915573_2 (TOXMARKER Assignment: 82; SEQ ID NO: 82)


 


cacgagggatcaccagatgctgccaggtgctggttgccaaggttgaaatgagaagtttct


 


gttaactgggtacagagtttcagttttacaaggtgaagaagttgggcagacagatggtag


 


ggatggtcacacaaagatatgaatgtatttactgccactgaagcaacactaaggtggtta


 


atctgagaagttatgtttattatttaaaggactaaattgtcaagctaacttcaataactg


 


ttttattttgtacaactgacatattcatatagatgacatctctaaagatgtctttatcag


 


tatttaaaactgtgttacatctcttaggaatttgacacacagtttcacttgtaaggccag


 


ataaccaattgtagggtgcgttattacccagaatgtggtgggtccaagagcttgaactca


 


cgatcaagtttggtgacacttgcctttacccactgagccttctcatcaacccaagtttcc


 


caggaattaagtaatctgtttccctaattccccttaagcaaacatggcagtcaagtgtac


 


agcaggagacaggttatgatttgcatgatgtaatttaataatgtaaccatctttggggaa


 


tctaattagtaccaaaagagaaaaaaaaaaccaacaggaaacagctgtctctctcacaca


 


gtgttgagagctttccctcccactcattgccaatcagtgtcctggtgtcccctcaccctg


 


cctctgtctctgcaacctgccagcctccaactgaacagacttccattcctgtgcaatcta


 


agtcagtctctccagtctcttcctccctccctccctcgctctccctctctcttataaagg


 


aaagaaagcactcactgggtataattgatgtctatatgcaggtgagggcaggtacaagat


 


aaggcaagacctgtgattgggcagtgaaaaaagaaaggcgggggcagaggttttgtaagg


 


caggagagatgaggaggtagaagaaccaagaaaaaggcagagaaggacgacccagatctg


 


cgtggctttaaccgggcaaaggtagctatgaatatttcataagggacagatttatatagg


 


acaatttgtcttacctaggtgggcagtttacatcaataccaattggttgtgactttattg


 


tgtggacgttttgtggactgagaatttgctgatatgaatctgactgctaaattacaagct


 


ttgggttttgattttaactggctactgggagttgtgactgtagccacaggggcagatgct


 


gggattgtgagcagggttcacagcacagtcccaggatggcagctgctgctgggcccagag


 


aggagccagtgccaacatggggctagccatggaggtggagagatcgctggggacagagaa


 


gagcaggaggcagtgtggcttggtgcctggtgccccacccacccctgcatccattttaat


 


tatttactgctacaactgggtgcttgcttttagtttcagagggttagtccattagcatcc


 


tgaggagaagcatgcaggcaggcagacaggcatggtgctagaagggtagctgagagcttt


 


aaatcgtgatccgcacgctgcagagagagaaaaaggaaacagagatggagggatgactgt


 


ccctggcaaggactttcaaaccttaaaagccacctctagggacacacctcttccaacaag


 


gccacacccctactccttcccaacagtccaccaactgtgaacaaagcatccaaatgtatg


 


ggccgatggggccattcctattcaagccacctcactgaaggaataaattaacatgtccca


 


aagtattaaatgtagtcatttttctcagtactgagacaaaatatctcaagaaataaaaaa


 


acactgaaggacgtatttcgtttggctccccctttaaaagaaacagtccaccatggccgg


 


gaaggcatgtggctggtcagagtgcaccctcatgcaggaagcagagagtgggggagtgct


 


cctcgaagccttttcctttttatttagcatgcaccccaagcccacaggagctggctaacc


 


cagcaagccttgctggcctggaagccacccccaacaaccatcatcaccccagtgcctccc


 


tacagtggggattatgagttgccaccatgctgtttttcacatgggtgcaggggatttgaa


 


accacagcctcctgcttgtacagaaagcatcctgaggagccatctctctggattcaccct


 


tcacttttggctgactgggcctgagctggagtcacctgggct


 


>scr_gb-be108509_1 (TOXMARKER Assignment: 83; SEQ ID NO: 83)


 


tttttttttttttttttggtttgttttgttttgtttttctttgcttttctaaggatagtt


 


taaaatacaaacaaattaaagtatgtgatatgtcaacatgatcatgcccctcccagacac


 


agcctttaactgtccagctcaaataagagaaatgctgaagcttaagatgtctttgtcctc


 


aggaagacatcacatgtgtggttgtcctgacactgcacatggcagcttccccacaacatg


 


ggcccttcgccttcacactgacaagaagtgtatgcccttcacactgacaagaactgtgtg


 


ctcactacaacttgtattggttgtaccttccccaaaagcagtaatgtatttctcaagatg


 


tcctaaatcaagtggagactctcctctggaaggaactggactcagcctcgtgccgaattc


 


tt


 


>scr_gb-be111483_1 (TOXMARKER Assignment: 84; SEQ ID NO: 84)


 


tttttttttttttttttcagacaaggatggtttattgaatggaccccctgagactgatca


 


atcagggccagggccgcagcctcagaattcaggggctgagccatgactctgaccatttct


 


cagggccggcttataaagggaaaaccccacaaagccacaatgagctcgcatgcaggtgct


 


gccggatggttggctctgactcaagccatttcagacagaacagctcatatttacctttaa


 


tgtggtgggccatatgtaaagctttgtgtaatttattaagttgaacaaacctcacagcat


 


gaccttgctctgagtcgagtcattttctgtatcaatgatggcaggcatggaacaaaatgg


 


ctatagctatgctaggtggggtagacctcaacaggataagaaactaaaaagtaacaaaga


 


tgagaagacaattgggcatcctggt


 


>scr_gb-be120910_1 (TOXMARKER Assignment: 85; SEQ ID NO: 85)


 


tcttttttttcggagctggggaccgaacccagggccttgcgcttcctaggcaagcgctct


 


accactgagctaaatccccagccccaggaacaagccttcttaaacaaccaccccatctct


 


ccagtccctgatcaatattttatgactacgtttactctgtaaaacaaaggattaaaatct


 


aatccgattaccagtcttactagacaaaccttccaaatctgagttttctcaagtataaac


 


acttcacaacaccttctgagaaatgtccacatcactcaaagacaacacatttgggaggtt


 


tttatgggcttcttttcatacagaacctttcaaagcttgtaaaacttcgaacctagggac


 


atttgggagttcttctcggtctcacacaaaacggacttgctttcaaagatcccttcggat


 


tctatttgacttagcaaaaacacagcgcaaaacacacccctgtaagaacaaaggtgcaat


 


tg


 


>scr_gb-bf285287_1 (TOXMARKER Assignment: 86; SEQ ID NO: 86)


 


agttcatgtgcattggtgtttgctcacatgcatgtctgtgacgtatgcctgtaggagggc


 


atcagatccctgcaactggagttattgacagttgtgagctaccatgtgggctgtgggaat


 


taaacctcgaaaagcagccagtgctcttaaccactgagccatctttccagccacctcaac


 


tcattcttaaatccacttaagacatagaggaaacactattccttctattctgtttgctga


 


tatctgtaaaagtagacagacttgcagagtggtggtggcagacacctttaatgtcagcac


 


tcaagaggcagaggaagacagatctgagttcaaggctaccctgatctacagatagagttt


 


caggtcagtcagagctttatagagagacctgtctcaaaatacaaaaaacaaaaccaaatt


 


aagtagacagactcccacttacacgaaacgtaaacactgtttcacacacttcagaatcac


 


atttaaactaccaatcaacaagaactgacagaaccaatatcaggaaacctcatccatata


 


aagcaacgtcacagcaccaagcagttaacagcttttggctcgctctaatcgaggatccca


 


aacacaaatcttacacagacatggggaggtacatcctacatctcatctcggtcgcagctc


 


atcgtcagtcctagggatcttttgggtccccacaaagatggaggcatagccttgctcttc


 


ttgcccgacaaggaggccagcaggccaggaagttaaactgccaatacctgccaatgctgg


 


tctc


 


>scr_gb-bf390383_1 (TOXMARKER Assignment: 87; SEQ ID NO: 87)


 


tttttttttttttttttanagnagctgtattttctttatattctgcatgggatatgaaat


 


aggggttttgctccacagggagcctggtcaatatagacaggatgtantcagggtgtgtct


 


tccaaggtcatctccatttccaggcagatggaaaaaaaatcatgaacaatcatgttgatg


 


attttgaaagatgagtataggcaatagcatgtgtcctctgtcctgagcaacagatctcag


 


ggatgtgagggtgtgcgctttctggatggttcaccatacgcatcttcagcaccaaggcta


 


tgcaagctttgttcagtaaggcagaacatcaggaactcaggagagtggctcccggaaggt


 


gatcatgtggcttgacccctgattatccatcttcctcaccaatggtttgcttacattcga


 


agcttaaagccttaaagttaacttcgtcttgtgatgctgttaaatgttttcaattacagc


 


acgatc


 


>scr_gb-bf558463_2 (TOXMARKER Assignment: 88; SEQ ID NO: 88)


 


ttttttttttttttttttaagagaaacattttaatatctgcaggctcacgcaggattcaa


 


ctgtgtgtggtacagtctagagtgacttgcttctatttacttccacacacggtgactttc


 


gatgagatggttaagctgagcagtatacattcctgaacagtgccaaggatcctgttttca


 


aacagctttatcaatcgaaacatcctcaaagagccattggaggcagtgtggctgggccat


 


ctgcactaaaatcgcttattcagaaggtgtcaaagcagccgagggccttgagccacaggt


 


tgctggtgttcacatctcagctgggacgtgataaagactgcatgagctgcagatccgcaa


 


acagccttgcaggctggctctgctcctgcaaagtcaatggagccacaaggtacttcttaa


 


tggtgtcatctgttcaggttctccagggagttaagggaagcactgtctttgcacacagtc


 


tctatcacaagggctctggctagcagcatgagagttccctctcagccaggctgccacagt


 


gagccatctattgtcctcactgcagagtgcacaggatgaagatgtccactttcctcatca


 


gacttgctgacagcctcatttcctgccaaacggatcagaccacactttcaaccctggtgg


 


ctgcacatcttcctggacgataccagctcgatttacagcctgctccttctggtattcttc


 


cagccgcagaaggggccggaagtagatagggtagaaggcggctccgaccatagagatgaa


 


gcctccgaatatgagcgcggtgcgcaggttccgggccgcggccatggtgagaaagggggc


 


tgcagggcgggcgaaggcccggcacgctccgaaacccgactcccagccttaaggtcgcga


 


cccggctcggaagaggcggag


 


>scr_gb-bf560709_1 (TOXMARKER Assignment: 89; SEQ ID NO: 89)


 


tttttttttttttttttacatttatgaatttttaatactcctgtcaagatcttacaagga


 


gaaattactttgggaggtgggtatggaggttagaggtaggttggaaagtggatcatgatc


 


tcaaaatagtaaatgctagctgagtggctttcccagagagaagcgacatgccctgacgag


 


actggagaacatgtgtaaaggagagcttattttcaggtctccgctggcctccatcctctt


 


caaaaacctcagctcctgggttctgctcaacccacattctgtaatacttgctcaagtagg


 


cctgtagcaccttgtaggatacagacagttccaaatggatgtccactccagtctctggct


 


gctctattctgtacttctcttgaatcacagcttttatccatgtaagtagatgcctttacc


 


tgggcacttgaagttcagaggagacaggtctttagatagaaatgtgcaaattacttatgt


 


ggttattgacaatcaatgactgttctcccgtagtctcccctcgtg


 


>scr_gb-bg662990_2 (TOXMARKER Assignment: 90; SEQ ID NO: 90)


 


ttttttttttttttttttacataaactattttatttaaataaaaccaggactgaccctct


 


cccacacgcaccagcacatgcactcgcacaatcatgtcctccgtttctgttcctcctgaa


 


cagccacctcaaaccccacaggttttcattgtgaccatccttgaaacctgaaaattggga


 


gatcccatgcgaaacactggcactcttcccccaaccctgggcaagcattctcctcatcct


 


cctggtgggacaggagctcagctcttccaaggcacccagatctggtgtggtttcccttca


 


cacaacccgggaacaccaatacccagagctgctctttgaggctgggacccctcgcttcag


 


gtcaactcctctcacacaacagaggaggctttgtaaccatgcttaagcgctctccaaagg


 


ttcctggcataggtaccgtctggtatgaggaagagcgacagagagcaattgagcaccaag


 


ttccctaatgccaccctgaaggagggtgccaagctccagttcagtctgtaccaagaaaaa


 


gcaagcctagcgccacacatggggaaggtggggatggcaaggtctcagccttgagaatct


 


cacatctctaccctccagcatagatcccatgagggacccactagcaccttggcgattgta


 


agggctcagcccaactggagacacaccacacaaacagtggccatttggagttggcccaaa


 


tgcctgtgtcggtaacagggtttgactcccgcatctaacactgactgaaggacacacagc


 


acagcagctaaggtcacgagaggtgcactgacagaaggtgttgtcttccagaggcacatg


 


gacatttcacacactgctcacaggcaagctgggacaggagaagagcacaggctgccaggg


 


actcagcagcgtatctagggcatgccctct


 


>scr_gb-bi278552_1 (TOXMARKER Assignment: 91; SEQ ID NO: 91)


 


ttttttttttttttttttggggtttggtatcatttattttttttcttaaacccacttgta


 


gtttgggttcagctgggaagcaggatatacgggtagagggaaggggacggtgcgagcagg


 


attggcccatagctttgggggcaatctccaaaccctgctccagggaggtaggtcctgttg


 


tcaggctcccagctggctcaggtgaggctccaaatggatcttctggagcaagtgtcctga


 


gcagaggagaatttccattctctccaacccacctcctcaaagacccagtcagaaggtttt


 


ccaacacagtgccaggcagttgaggggacatcaggccacgggcaggcctgagtgggtggg


 


acaaggaacactgtctggcttctggttccaggtaacaacctaggatgtggctacccagag


 


gctgccatctagagtgacctccgggagctgcttctcttgcttcctgggctgcctgggatc


 


caaacctgcagctgccctggttgcaaccagtggtatacttcccaccccccacccctcaga


 


caaaataaaataaaataaaatacaataaaaattagaataaataccaatcgggtcaacatt


 


tacatttacacaaatggacaagatgatcccccaaaccgtagaagtttacagactggatgg


 


gaaggatacgcagatgaagatggttttggggaggaagaggttcgccgtggtggttgatgg


 


tggggggtcctggccctgtccaggggagggccagagccctgcaggaactgtggtctcaga


 


gcttaggcaatacggccagttcatgaggagaacagtgacctgcaggccacttgagtagaa


 


aacaaggaccaacttgtcctgacaggtaggggagcctaaaaaggctcaatatgagatcgc


 


catggccagcaggacaccacagtttgggagaggcttcgcctcctgttcatccattcagag


 


gcggctttgataggccgtccctctggcagcgggagagcctctggcctggggaggtcaggg


 


tctgtgggtacctgcaacgcccctacttcccctcgtgccg


 


>scr_gb-bi278749_1 (TOXMARKER Assignment: 92; SEQ ID NO: 92)


 


tggctcagtggtcgagcacagtaacaacatggagattctaaaaacagagaaagagaaaag


 


caagaagatagtggagggagacaagaagaaaggctggggggccagttttgttatttttgt


 


tttggttcagctatatgctccacacttccaaagcagcaaatgtgttgcatcaccacccaa


 


acctgagaaagctacagcatcactggcaaggacaagctagcgcacgggtgacatcctcta


 


accctgccattgtaaattatacaactgcagtttccagcacacaccattgcctccgacact


 


attggagagcccgtgacactccaaaaactgctaaggcctttacagtatctgaccttcaat


 


ggccccgaaaactggtaggccgcttctccccattccaacccaaaaattacatgcgagcaa


 


cggaagagaaaagcttttaagcccgcgcggacgaagagaccagcggacgctgctgaagac


 


cacagaccaggtaagccagctgaggctggagtttattgccgatgagcgctgagtcctggg


 


gaggagcggggaaggataaggtcgggcaggatcaggaccttggctaggagaggcggcgcc


 


acgaaggcgaggccgggaggtgcagacagacaggcgcaggccacggtgggggcgggccag


 


gctatccaggcactcggtgagcggtctccggcgtcgctcccggagctgggtggcggctgt


 


ggcggcggctccgcggcagtcctggctgcggtcgtggcccaccggaggccccaagcaagc


 


aggacgcggcgggaggcggggcgggtggtgctgctcgagcacacggagcagctgcagcgc


 


tgggcaaggggtcggcggggcccgcaggcggccgcgtggggacccagatgagcccgtagt


 


ataccgcaagcaacacagcagccaaggatacacacaggaagtaggcgcagacaggggcga


 


gccgcagccatcgggcgcggggcccctcgctcagccccgtaccacctgggctctcgccac


 


cactgcccacgcagctcgagcccccgcatgcgctgcccactcagcctgtaccgaccccgc


 


ccccaccccgccgcttctagcaagccacgccccttctagagtcacgccctatcagaccgc


 


cacccccctcgtgccgaa


 


>scr_gb-bi295938_1 (TOXMARKER Assignment: 93; SEQ ID NO: 93)


 


aaaaatctcgatgccctcaactgttaggttaaagcctgacctgtgtcactatgtgctgtg


 


acacgaacctaattcccaagtggacagggacacctgagtggcatttcgtgcttcagttcc


 


ttccctcatgattcttgctgggtcctcttcactgaggctctcccctgagtcatatattta


 


ctggaaaggctacctggagagcctttgaattgtgggcattcctttttaatgtgtccctct


 


cttccacagatgaaacagcgcttttctcttgagtctctgtcatcctgtctcttccacttt


 


tcggctggtgtcctgacaagtttctcccggcccaggtcaacagctgccctcattggcttg


 


gctttggcagctgtgcacggtgcagccttgtcttcttttgctgacacttccttttctgtg


 


tacttgttctgaatttctttgtcctctttgcttcttttttctttgctctctgtgtacctt


 


tggtttggggtatcttcctggtctcgccgccgcctcacttttctcctcatgggacagtcc


 


ttcatgaagtggccaattttcccacagatccggcagcacctgtcatttggggccagttct


 


ccctcagtcaggacatccggatcaaagaagtatgccaggatgtcctttggaaatcctttg


 


actggaattccaaatactcttctaccattgataaaagctttcattataaaatttgtcatt


 


ttccttgataatccagcaccaagattgtggttcaaatcaaagggatcttcaatgacgatg


 


tattttgaggtccactgtttcttaaaagttgtaagcagactttttcttctgatgctgatt


 


acgtgttccttaaagtcaaactcctcagtgtagaagcgtagaagtcccaaccacagctgc


 


ccaacagattctgtattttttccatattctggccaacaagtgggcagttcatttatttga


 


tcgaaaaagtagatattccagccatcaacaagtatttctggtttcttttcacctttgtat


 


atctcctgaagcacagggatgacagggggggaccgctgctggaggaagtacagcaccata


 


agagtgtaagcgtatgatgacaagctgcctctggacgcgtcaccgatgtcacacatcttt


 


gtgaacactttcatggtgtagcacaggtatttcactctggggtcaatggctgagtatgca


 


aacaggagccgcgtgttgtgaagagccagtgtgtcctcgtg


 


>scr_gb-bi296376_1 (TOXMARKER Assignment: 94; SEQ ID NO: 94)


 


ttttttttttttttttttccaggagtcccttcggtccctgatagcgggagcctggacctc


 


tgaggccgagagggtgctgtgtccccggcctccgagccgaggtggcccggctagggggcg


 


ccacggagtttttttttttttttctttttcttttccaggagtcccttcggtcccagccag


 


cgggaccatagacacttttgaggccgagagggtgctgtgtccccggcctccgagccgagg


 


tggcccggctaggtggcgccacggattttttttttcttttccaggagtcccttcggtccc


 


tgatagcgggagcctggacctctgaggccgagagggtgctgtgtccccggcctccgagcc


 


gaggtggcccggctagggggcgcctccgaggctttattttttccaggatcctccccggtc


 


cctgccagcgggagcatggacttctgaggccgaggggaagctgtgttccaggctatctac


 


catggcctcctcggtctgtgagcactcagggttctaaggtcgaccagttgttcctttgcg


 


gtccggttctctttctacatggggacctcttggggacacgtcaccgaacatgacttccag


 


acgttccgtgtggcctgtcatgtttatccctgtgtcttttacacttttcatctttgctat


 


ctgtccttattgtacctggagatatatgctgacacgctgtccttttgactctttttgtca


 


ttaaaggacgttggaagaggcttgcaccaaggctgtttgcttgtccagccctagctcttt


 


tcttctgcgcatgggcctcttcgatgcttgaagcttagcgtccccccatgagtacgcgct


 


tcctgctttcccgtgcttgcttgcctgtgctctgtggggcagctttatgacaaccgtccc


 


gcgtgtcaggcgttcccgatttccccgtggtggttgtcgtccgttaccggtaggagtcgt


 


tggtgccgagtgcgactgaaagggttttcccgtttggtgctagtgaccccctggcgtgct


 


cctctgcggccgaccggtttttttatttgttttttttttttttgttttttttttgttttt


 


ttttttgttttttggaaggagttcccgaacctccgctgcttggtggtgtgtccctttctt


 


tcctgctgtgtgcctcccgagttgcaccttttctccttcgaaggggattttattttttta


 


tttttattttttttttatttttattttttttgaaggagttcccgaacctccgctgcccgt


 


tgagtcccgttcttccacgccacgtgcctcccgagtgcaacgcttccttttttttctcgc


 


cctcgagaagggtaaattttttttttgtgtgtgtgtgtggcagtgttagcgacttcttcc


 


cgtgctctctctcgctcttctcgctcgtattcccgtccagtgcgtgttagaaagctctca


 


cgcccgttgttcccgatgcatggcgtgtctcgctcccgttggatcgatgtggtgctgccg


 


cgttctcttcgggccggggcctaagccgcgccaggcgagggacggacattcatggcgaat


 


ggtcattcagcgcgaatggcgaccgctcttctcgttctgccagcgggcccctcgtctctc


 


ctccccattcctttgcagggtggtgtgtggaagtcaggggtgcggctgtccggcacgagc


 


gctgacccgcgcacacttgctgctgtggttcgcggtgtccctgtggacgtgtcgggggcg


 


cttgcccccacgccgttcactgcttcgcggccctcttcccccgtgccgggggaaggtggt


 


agacccgctgcggtgcatacccttcccgaatggtgtgtgcacgcgccctgctttgtgtga


 


gccttgcggtgctcctggagcgttccgggctttgaccaccaaggtgcccgcttctgagtt


 


ggcggtggcgcttcccgctccccggcgtgcctcctgtgctccatggtgcttgtgccttta


 


cgctttcccttgtcctagttgccggctttctgcacggtgacagaaagggggggggtcgag


 


gagttgagtgtgcggttaaaaggctccttccgttgggtgagcgcccaccccgtgcctatg


 


tttttggtgccttcacccgcgggccctgcgcggttagggtggtgctgagcgatcgcggct


 


ggccctttttaaagaccggactccctcaagtcaaggctcctcctttgtgtgcgccttgaa


 


gaggcctggccctcggcggggacctgtcgcaggtccccccggtccgcgaatgctcaagaa


 


gaccccggagaaagagacctttgccgataccgcagaccccccaccagctggcgcgtggtc


 


cttcccgttctgtcccgcgcctgttgctcgtttcccgttgcgtgcacggagcccttggct


 


gctcgtcggtgttgggttcgtcccgccctcagtgaggaatttgccttctctagctatctt


 


cggaaagggctttacgatctccgaggggcttctcccggatggtcccctcggctgcccgcc


 


ctgacctcagccttctgcgcgcagcgtttgctctctcgcctaccgcgacccgcgcctccc


 


cgctccgagtacgaggagggatcacgcgggacggggctctgtcgacctgccgctgtgcgg


 


agcttgtgggggagattgggtttctggtggcaggtggcggggaagggccgtgcac


 


>scr_gb-bm384392_1 (TOXMARKER Assignment: 95; SEQ ID NO: 95)


 


ttttttttttttgtttttcaagttgcacattttaatttacaatgtttaccagtaaaaagg


 


attagttacaaaaaggaaagctgtctgtacaaaataagggtttttttttttcacattcat


 


aaagagaacccactgtgaattcttaccttgtgaagtcaatactcaaacagctcactttgg


 


taaaactatcttggaaggactagtaatccaggcaagataataaaattatcagcttcccaa


 


tcatgtccaggagaaagaattttctgaacattttccctgtacagaaaagctctctgtact


 


tgcagatccttagaaaagccagtgctctcaggagacagcctggtaccaggacgaagcata


 


atctcctgctcactcaaatggcaatccttcctgaatctgacagacacacatttatcatag


 


cctcaggtcagcaggagaaccagatggttcaggatcagcctctctccactcaatagttta


 


tcatataaattaaatatggagaggtacacatgagaaagggggagctctttttcaaactcc


 


cacttcctaatataatacacatcacagttttaatgagcagagaagggtaagtcaccctgg


 


tttgggcacatttcctcaagggaaaaaccaaagtatcaaaagccttcaaagcatactggc


 


ccgtcccactgcagccagcagcctgattccagaatgaaagcatacagtagctgtaaagcc


 


ctggagccttcagaaagctttatttagtgataagctgagctctgctggcaaaagcccacc


 


tataaaaagggagcaggtctgattcacaaagtgtatacatgcatgacccaaggtaatgaa


 


gaccttcaaatgcaaatgatcctaaagctattggaacctctaattacgagtgacccgttc


 


agatgtgcctccattagccttaaaaactgaccaacacacatctgaagaggcacttccctt


 


agcattaacataaacacttgaccagaaaaggcatggtccaaaaaacagttaactaaaaat


 


ttagagtctaaacctctcttctccaccgactgaatgaacacacccgcaatgaggaccaaa


 


cagaatcagtgcctccagggacgtgtgtctgtctggccatgtgatcaggaacctcctaac


 


atagcacagcacagcacagctgctctgggcacacaaagccagttcaccccatgaagaaac


 


acaagggattgtgattaaacccatcccctgtgtcaggagcaactccactatggttttgat


 


cactcagctcagagggataggagtgcctagcaacaagtcctaatcctcgttactcccagt


 


ccgggccctcactgactcagaggtgcctttgtgtataaatatgtgagaggcagcaaatgg


 


cagcactgctgacaggctaatgcaggccccacagcggagaaagttcttcctctgctgctc


 


caatcttctccctacagttacagtcctgccagtgatggccaaggaccatgtgtgagccag


 


ctctttgtgaccaagctttggcaagtcagtaagtttgtcaaaggcaaaatccttctgtgg


 


acaatgctagctgcagctctggggacgtgtgagagaggagagggtcctctgacgggattg


 


gggacgtgtgagagaggagagggtcctctgagaggatttgactcatcagcccctcttgcc


 


cagttcattaatcagaaggaaggggagaggagaagacagcagaacatgagtcagttgtga


 


aatctgcacagctgacatttgctcttcacagcagaaaggacttgaatgagaatcatgaaa


 


cttgaggaacacttgtattttccttcgggatttaaaaatgtgtcttgtaccaaaagacta


 


cattcagtgtgggtcaggtccaagagcggcagcaagagctcggccattaagcgtgcccag


 


cactgggaggagactgtcatctgcttagcatggctggtgagcaggccagggctgctcctc


 


actggtctccaagtcggaagccctggccccagttgtgtctcccacctccgccattctgat


 


cagcagctcgcctcatgcttgcagggggcacaccgaagcccgacacccctcctctcctgc


 


tgggtagccagcggtacaaaaactgaggtgtggacagaaaattccttcctcccaaatcca


 


ttgggtatctgaacatcaggaagaaataaagatgtccgacaaggtttccaatgagctcat


 


tgatgaccgagcctccaatgatatagttgaatccgaggataacccaaggtaagtaacagg


 


ccttaaatcgtgttccaaaccaaaatgatacaatcaggtctctgttcagctgggcccaga


 


cgtaaagtactgacatgattagaggaatcatcagcaactgcatatccatggctaagccag


 


taataacaatgcagatccagttg


 


>scr_gb-bm387477_1 (TOXMARKER Assignment: 96; SEQ ID NO: 96)


 


aaatttcaagaggtcagagtggggcttagattaagtaactaatgcacagcaaaacgctgt


 


gagattaggtgtgaaggagctggctgccctcctgtctcttcccttctctatcccacagga


 


gctacagagagagcacagcagccagacgctggccaaacagggaacactctttatgccaag


 


tcgcaaagatgacaagcggcatgaggaggacccagggccctcctttgtgtggaaggacgg


 


agaggttctgggagggctgggaagggtatgggaggatcctttgtgtgggaggattgagga


 


aggcctgggcaggctgggaagggctaggaccgctctcctttgtgttagaggtctgggaaa


 


gtctgggaggatcctcctttgtgtgggaggactgaggggctctgggagggctgggagggc


 


cctcctttgcttcacagttttagatgttgttccatctgctctcggagtttgaatttctgg


 


atctttcctgagacagtgagaggatagccttccacaaacacgatgtatcggggaatctta


 


aaatgggaaatctttcctttgcagaaagctttgatctcctcctccgtggtggtctctccg


 


cctcgtgcca


 


>scr_gb-bm986259_1 (TOXMARKER Assignment: 97; SEQ ID NO: 97)


 


gtaacccacctccattctgttcttcggacgcttgcgccagtgggtcaattttattttctt


 


tcaaaaataaaagtcgagtgcattcagagacggccttaaggcaatacgcctcatcttccc


 


acagtaaagatggcgacgccgtgagtaagttacaagtaactccacttccgcaattttctt


 


gagccctggtccaagatggcggacgaggccacccggcgggtcgtgtctgagatcccggtg


 


ctgaagactaacgccggaccccgagatcgggaattgtgggtgcagcgactaaaggaggaa


 


tatcagtcccttatccggtatgtcgaaaacaacaagaatgcggacaatgattggttccga


 


ctggagtccaacaaggaagggacccggtggtttggaaaatgctggtacatccacgacttc


 


ctcaaatacgagtttgacatcgagtttgaaattcctatcacatatcccactactgctcca


 


gaaattgcagtccctgagctggatgggaaaacggcaaagatgtacaggggtggcaaaata


 


tgtctaactgatcatttcaaacctttgtgggccaggaatgtgcccaagtttggactagct


 


cacctcatggccctggggctgggtccttggctggcagtggaagtccctgatctgattcag


 


aagggtgtgatccagcacaaagaaaaatgcaaccaatgaaggatgaagcttctgaggcag


 


gacagagggactgttgctagactctgattctgtttcctcctttctcatgattccttcaag


 


ggtcacctctggccattacaaagtagctggagggacaaataacaaaacccaacaaaaggg


 


caaggtcacaaagttgctaaattaagctgtacagagaggtgaaagatttgggccttgaaa


 


gaggcggtttgtatcccttctccaagcagagccctggaggcattttggagacctggggtg


 


taactgacagcatatagctttttgatttctggagacaacctgtcaataaaagctgcttcc


 


catggtgtgaaaaaaaaaaaaaaaaaa


 


>scr_gb-s69874_5 (TOXMARKER Assignment: 98; SEQ ID NO: 98)


 


tttgctatctgcacagcccatcgagggacctgaggtggcaaaccctggacagtgggtcag


 


gcggcgctcacgtctggggtgacaggatgaagcgggctgtgggctgtgtggagcaccgtg


 


cacccctagcacctttgggtttcttgtggagttctcgccccagacatcagtgcactggat


 


tgcaaaaggcaattcatcttttattggatcaggagcgccatttggagtgtgccattatgg


 


gaggctcgtagctgtctgtccctcgtgccgaattcggcacgagccccccttttttttttt


 


tttttttttttttttttttttttttttttttgaattagcacaaacgcatttatttactaa


 


ccaaaggaatgatcctgggtaaaccaacggtctgacatgggtttcgggtaaagtgtctat


 


gatgaaaagtcatgaaaaataaaaccaaagaagtgaagcagtgtggttctgtacgacctg


 


ctcattgaattgagcttattccctcagccagctgactgctgtccaggatgacgagttagc


 


cagtcctcattgtaccttctcatagacccgagtacagatggcattgttcatgacgcactc


 


caccaccatcttcccgtccttcagttttctcgttatcgtgctttctttcccttcccactt


 


ctggtgctggaccagggcaccgtctgtgaaggtgcagaccgtctcagttttcctgccatc


 


agctgtggtttcatcaaacttctctcccaaggtgcaagaaaacacggtcgtcttcaccgt


 


gctctcagttttgacggtgaggttgttgccgtcgagggtaatgatgcagtctggtttggc


 


catggcacccatcttcctaagagccagccctactcctagttccttcatgtagtcctcaaa


 


cccgtggctttccaccagacgccacttcccttccaggtccttaaggctggccatggcgag


 


cgggagagcacaaaagcagcaaggagacgcggtggcgggggcgctgagggaataagctca


 


attcaatgagcaggtcgtacagaaccacactgcttcacttctttggttttatttttcatg


 


acttttcatcatagacactttacccgaaacccatgtcagaccgttggtttacccaggatc


 


attcctttggttagtaaataaatgcgtttgtgctaaaaaaaaaaaaaaaaa


 


>scr_sc-119263563_1 (TOXMARKER Assignment: 99; SEQ ID NO: 99)


 


ncctagcagaacgcttgttaggagtctgtgggacaagatagcctctgataaaataaactc


 


taaacatgaactccttcaagaaaaaggactggactccaccactgttcaataaagtcacag


 


cgagggatgctagaggcggtagacagaaattaagacattctagatacggggagtggccac


 


ttggttgggccaccacttgccttagcataggtaccataggctaagcatggaaggcagtaa


 


gggtggatgtcattttaatgagagcagcaaatttagtacatggtttatcaaataaaaggt


 


aaaggagtccaagatcaatctgacaaatagatctatcagctgaattgtaatcttggggtg


 


gaggggtcagaggtccggcaattg


 


>scr_sc-132556005_1 (TOXMARKER Assignment: 100; SEQ ID NO: 100)


 


caattgctgctctaggatagtcagagtgtgttctctgtctcctgggaaacagtggaccag


 


gaatgaaagcttcaacctggtacccagattttagatgttttagggacaatcagtcaaatt


 


tttgtgtgaatgtatgggtttatatgactataactgtgtaagacagagaaatggatgtac


 


a


 


>scr_sc-132570828_1 (TOXMARKER Assignment: 101; SEQ ID NO: 101)


 


ccatggacataactacctcctgattaagtccgttaattgagacctaatcagtctgttaga


 


ttattgaaacaggtcctgttagcagactgcagggagaaaacacggtcatgaaccaaagag


 


tgagtccgga


 


>scr_sc-132947646_1 (TOXMARKER Assignment: 102; SEQ ID NO: 102)


 


aagcttcctccatttcccagtagtgccatacgctggcaaccataggatcc


 


>scr_sc-133387221_1 (TOXMARKER Assignment: 103; SEQ ID NO: 103)


 


aagcttcaactgtctatttattcacagtcacactggctgagatgtcctacactgtgtcca


 


gtgcaagtgctgacactggacattgatgtcttcttctgtatcttagaggaaaggtcggta


 


gaggtagagcctggcttccggcttgtcatacatgacccctaagtgattatttctactgta


 


ccttattctcagaggaattttatcatgaaaggggtccaggagtctccccacaaaccttag


 


gaacaccaatctcagtcagacagggatgttttgaatgcacacctaaagtctgatca


 


>scr_sc-133555783_1 (TOXMARKER Assignment: 104; SEQ ID NO: 104)


 


gctagccatttggtatttattagataacaagttagggaactcatgccttggaaaggtgtt


 


gttggttgcttgtagttctttgtctggcacagggaagctacagctattatctcaataaaa


 


tagctgtcccttggattttttttttttaaataattgcttattcgagccaacatctaaata


 


aggtgcatgcattgtatttgcttgatacgtttgttgtgtctctttttcttcttctgtaag


 


tttcttcccctccttatttttctttcctcgtattgtatttactggaaaaaccagatcgcg


 


cgccctgcaggcttctgtaca


 


>scr_sc-133678871_1 (TOXMARKER Assignment: 105; SEQ ID NO: 105)


 


agatctgaaagttaggcaaaatataagagcagccctctgaagaggggacctgccagctca


 


cttgggactcaacattctactgtagagctagc


 


>scr_sc-133725675_1 (TOXMARKER Assignment: x106; SEQ ID NO: 106)


 


agatcttggggtttcaggcttgtttggcattcaattttaccttctgagcccaggagcgag


 


aatcttgaactaaagagggcttgacagtgctagc


 


>scr_sc-133955481_1 (TOXMARKER Assignment: 107; SEQ ID NO: 107)


 


caattgaacagtagtctgtaagtagtgcaacactgtaaaatgttctctttagttcagaga


 


gaaaattcccaagcattattccaactgctgctaaaatagatgttataattatcagtttaa


 


tgccagttccaaacccctaaataagcaaatattactgttattgccagcaacttcctgaaa


 


ctacacaaattcagtgtatccctccctccctcttttcctttcagtcatgaagggagcaga


 


tacaacccagggtccaagataggtaagtgatccttagatgattttagatagcaggtggtg


 


caaacttttaatcccagcacttgggaggtaaacaggtggatcc


 


>scr_sc-134521597_1 (TOXMARKER Assignment: 108; SEQ ID NO: 108)


 


nctaacaaagatggtttagagatccaggtcaccaatcctcttctcagacagacccatttc


 


tggggtcaacagccattactgcatgtagagtaaagggaagtaagacagagagagttcatg


 


ggcagtcctaactggctgtgtggaaacagctttccaattgttctgggaatgaatgtagag


 


tcagtgtccctgcatgggtcatgataagagtgcctgcaagtgaggcgctcacaagctt


 


>scr_sc-172126480_1 (TOXMARKER Assignment: 109; SEQ ID NO: 109)


 


ctcaggttggccttaaactcactatatactcaaggatgaggttgaacctatcttcctatc


 


tctgtctcctgagtgtactgggattgtacacatgtgccaccatacctggcttacgtgatg


 


ttgtggatcaaacccatggctttatgtatgctaagcaagcactttatcaactcaaccaca


 


attcatctctatattttaaatgtaatattcctaatatgtctttacattttccagctacat


 


tcctagg


 


>scr_sc-172130231_1 (TOXMARKER Assignment: 110; SEQ ID NO: 110)


 


tgatcaagagtcccaaacccagagagtctggggtgctgacatctgaatgtggctggcctg


 


ccctggctgactgctttcagtgccagccacactgatgccccttagccctctggggttaat


 


ttaggaacttgggctcaggccaccgtcaccagcaatgaactcacaaagaatgagatgtgg


 


ctgttgatttcctagg


 


>scr_sc-172755010_1 (TOXMARKER Assignment: 111; SEQ ID NO: 111)


 


agatcttccggagcaatggggttcagcttttgcagcgcctactggacacgggagagactg


 


acctcatgctggcagccctgcgcacactggtcggcatttgctctgagcaccagtctcgga


 


cagtggcgaccctgagtgtcctaggaactcggagagtcgtctccatcctgggtgtggaaa


 


accaggctgtgtcgctggcagcctgccacctgctgcaggttatgtttgatgccctcaagg


 


aaggtgtcaagaaaggcttccgaggcaaagaaggtgccattatcgtggatcctgcccggg


 


agctgaaggttctcatcagtaacctcttggagcttctgactgagatgggggtctctggcc


 


aaggccgggacaatgccctgaccctcctcattaaaatggtacctcggaagtcaccgaaag


 


atcccaacaacagcctcacactctgggtcattgatca


 


>scr_sc-188295137_1 (TOXMARKER Assignment: 112; SEQ ID NO: 112)


 


gctagcttaagggttcttctgtaggccgcctcatttcctggtttaattttactttatgta


 


tatgatgttgcctggatgtagatct


 


>scr_sc-190079504_1 (TOXMARKER Assignment: 113; SEQ ID NO: 113)


 


agatcttttttgcttcccttccttttattgatccttaggaataaatcctcccaaactctg


 


ttgtttttaaagttttttgaaagacctgattttttttccattttctttgcccttgcaaat


 


aaccatcagtgtaattagttgtccatgctgcaagggaatactttgtgagggaaataagca


 


agaattgagtgttgtttactaagaggtcacgcggatggtttttgggtaattatttactag


 


t


 


>scr_sc-191455923_1 (TOXMARKER Assignment: 114; SEQ ID NO: 114)


 


tccggagctggggactgaacccagggccttgtgcttcctaggcaagcgctctaccactga


 


gctaaatccccaaccccgtcaaaggccatttttatcctcatcaaacaattataccttact


 


ttttgagttggaaatgtaattcagtaatagtctgttttcctagtatgtacaaagtcttgg


 


gctccctcactaacaccaaaggaaaggggaaaaaagagctcacttctttgactttcagtg


 


gccttccactcagactatgcttgtttagaacttcggcagcttttttcatgctctcctcca


 


tcttgaactcaacaacactataaaaaagaaaagccaaaaacaaatgaataaaaccagtct


 


tacttggaaaattgaacttggaaaattt


 


>scr_sc-195460151_1 (TOXMARKER Assignment: 115; SEQ ID NO: 115)


 


tctagagaaatatacatagacagcaaggctggagttgagccaggcaacctaagctgggcc


 


accggagtcaggcagctgcagaaggtcacgtgagcaggcccagtgctagcctgtgacgga


 


gtgatgtagacactcagccacaccagggagccaatctccaagttgtcttggctagactgt


 


ggactctgcccttcatgggtctgccacacaggcattctggaactgtctagctagctcttg


 


gggaaacagctaaaaggactttggcttttctggggtttgcagggagggtaacagtgtctg


 


cgcccttgttctctacttctgaatgtagtaacctcaccctctggggtagcatatgacagg


 


tacccaactccttttcgtgggcaagcctctggcaggggagctctttctgttgcaatgtaa


 


cagaggcattgcctctttcaattg


 


>scr_sc-198205946_1 (TOXMARKER Assignment: 116; SEQ ID NO: 116)


 


gtgcacagaagtatgtgttctgggtcggaggaaagatggtaggtgtttgtcccaacacag


 


tgaaaaggaacagacatgtgaagtcttcagactgtgggcctttgatttacccctcagttg


 


gtctatgtgtgtaca


 


>scr_sc-2573087_1 (TOXMARKER Assignment: 117; SEQ ID NO: 117)


 


caattgcattgcaaaattttaaaggttacattgaaaacacttgaaaataagccaccaata


 


aatgagatgacgataataagagcccctaaataaagaggctaagaaggagttaagtgtaaa


 


ggaagagggaagaaatagttaaggcatttataagacactagaaagtctagaagagagaat


 


gttagcagtacggagtcacagctaaaaatctgcatcttgccctttaaaacccaagagaga


 


aagctt


 


>scr_sc-2585074_1 (TOXMARKER Assignment: 118; SEQ ID NO: 118)


 


agatctgctggtgtttgcctccacagtggtgaggttgcatgtacatgccgaccatgctcc


 


tatctttcacatgagtgctgtggaatgctcaggtcttagtgcttgtacaagcaccttact


 


caactgaaccattgtcttagcccaatagtgaaacactgaaaagttattttacccatgatc


 


agaagctttaacaatcaactagt


 


>scr_sc-8571871_2 (TOXMARKER Assignment: 119; SEQ ID NO: 119)


 


cctaggtctgccagtgaataagaagacccctccccggaaagtcccgagtttatgttccat


 


gcgctattcaatagccttcatcgcacatatctgcaacttcacattgatagcacagaattc


 


catcataagcatcaccatggtagccatggtcaacaacacggaccagccatcccacctcaa


 


tagctctactgaatggtttcctgatggtttaaacggtgatcaacatgaagctt


 


>scr_sc-87731837_1 (TOXMARKER Assignment: 120; SEQ ID NO: 120)


 


tgtacacaggtagtcttaggatttctgttgctgaaaccgtgggaagggaacagttcaatg


 


agtaaaaccaagacagaagtcaacctggttagaagctggaggcaggagaagatgcagagg


 


ctgtggaggggtgctgcttactggcttgctccccatggcttattcctgctttcttataga


 


acccaggaccaccggcccaagggttacaccatctgtggtgatctgggccctcctccatca


 


accactaattaagaaagtgtccaagtttggctatatcttacagagatgttttctcaattg


 


>scr_sc-87869413_1 (TOXMARKER Assignment: 121; SEQ ID NO: 121)


 


cctagggaatttgccattgtttagtttaagctaacactccaaaggtaatctcctatttcc


 


tcttttcctttctgtcctccatgtggctgtcatgggcatgcagcataccagttctcaggt


 


gcctggaacactggccagtgctctagcccagccactgtgccctgaaatccttccctgtgt


 


tcaatgctacagcacatcctccagactgcctccccacccccagcaaccgaattgagcagg


 


gacactaagacagtcctttggagacttccactggtctgttgaaactttggctgctctcac


 


agcatagctcctcttagcctgtaacttagtgctgctcaggctgactgatca


 


>scr_gb-ai233262_2 (TOXMARKER Assignment: 122; SEQ ID NO: 122)


 


tttttttttttttttttaaggggccaagcagaagacaagctgcctttattatagttgatg


 


tcacagctctgcttgtaatagattcagccccagaaacaccccggttaaaacagcacggtt


 


gacttcaatggatagagtctttggtaaggtgaaccagaccagggctgaccgacaatcttc


 


gggcccctggcccaggggtagcctgtagtcttacgtgaggcccagcatggcctgaagttc


 


ccgagctttatcatctggcagagagcccagggctgtgtggaagctgtcgctgtgctgctt


 


ggccaggaacgtcagtagtagtagcagtgcggccttggtgtctggggggatcctgttgtc


 


tggcaggatcaggctgcagatgcgcaggagctctgaagccacacccacaacctggtcagg


 


gttgttctggtgcaggaagctgaagaggtgacctatagtgacccattcctccatgtcttc


 


cttcaggggcagggcatgtagcagggtagctagcacctggggctctgtttttcctgccgg


 


actggccatcagcagacgggcaagagccccacagatgttatcacggactcgatcatgccg


 


ctcccttgccaggaggggcaaaaggaggcccagtagcttagggaagtggtcctgagcagg


 


gcagcccccatgctctgcaagtacgcccagcccaaagatggcattgctccgcacctcggg


 


gtctgcttcccgggcattgtttaacagcacaggaaacagccgggacacaaattgggctga


 


ggcagcacctagaccctgaatggattctgccagtgtccccactgcaaaggacttctctgc


 


cactgtacagctctgtttcgtcttacacagcaataatggcaacctcgtg


 


>cgrrs0h0310.9_13952-135 (TOXMARKER Assignment: 123; SEQ ID NO: 123)


 


tgatcaagggcgacacatctggagactataagaaggccctgctgctcctctgtggaggcg


 


aggatgactgaggagctgcctggagtgccctgggcccgcctgctgcccaccatcagcttc


 


cttcagcaccacgcctacttacgttcaatgcctgcctgcctgccacgctgccttactcac


 


acgagtgtgtgctaatgaccaaagctgtctcgaatgaaagcagtgttctgctgttctgtc


 


tgacatagaccttcccacgtctctcagtctagtatctctaagttgcgttttctatcctct


 


tctaaagctt


 


>scr_gb-m13100.5_2 (TOXMARKER Assignment: 124; SEQ ID NO: 124)


 


aagctctggttgcttgacattgttgtacatatagggtctcgagccccttagagctcgtcc


 


agttctttctctgattccttcaacgggggtcctattctcagttcagtggtttgctgctgg


 


cattcacctctgtatttgctgtattctggctgtgtctctcaggagagatctacatccggc


 


tcctgttggtctgcacttctttgcttcatccatcttgtctaattgggtggctgtatatgt


 


atgggccacatgtggggcaggctctgaatgggtgttccttctgcctctgttttaatcttt


 


gcctctctcttccctgccaagggtattcttgttccccttttaaagaaggagtgaagcatt


 


cacattttgatcatccgtcttgagtttcatttgttctgtgcatctagggtaattcaagca


 


tttgggctaatagccacttatcaatgagtgcataccatgtatgtctttctgtgattgggt


 


tagctcactcaggatgatattttccagttccaaccatttgcctacgaatttcataaactc


 


gttgtttttgatagctgagtaatattccattgtgtagatgtaccacattttctgtatcca


 


ttcctctgttgaagggcatctgggttctttccagcttctggctattataaataaggctgc


 


aatgaacatagtggagcacgtgtctcttttatatgttggggcatcttttgggtatatgcc


 


caagagaggtatagctggatcctcaggcagttcaatgtccaattttctgaggaacctcca


 


gactgatttccagaatggttgtaccagtttgcaatcccaccaacaatggaggagtgttcc


 


tctttctccacatcctcgccagcatctgttgtcccctgagtttttgatcatagccattct


 


cactggtgtgaggtgaaatctcacggttgttttgatttgcatttcccttatgactaaaga


 


tgttgaacatttctttaggtgtttctcagccatttggcattcctcagctgtgaattcttt


 


gtttagctctgaaccccattttttaatagggttatttgtttccctgcggtctaacttctt


 


gagttctttgtatattttggatataaggcctctatctgttgtaggattggtaaagatatt


 


ttcccaatctgttggttgccgttttgtcctaaccacagtgtcctttgccttacagaagct


 


ttgcagttttatgagatcccatttgtcgattcttgatcttagagcataagccattggtgt


 


tttgttcaggaaattttttccagtgcccatgtgttccagatgcttccctagtttttcttc


 


tattagtttgagtgtgtctggtttgatgtggaggtccttgatccacttggacttaagctt


 


tgtacagggtgataagcatggatcgatctgcattcttctacatgttgccctccagttgaa


 


ccagcaccatttgctgaaaatgctatcttttttccattggatggttttggctcctttgtc


 


aaaaatcaagtgaccataggtgtgtgggttcatttctgggtcttcagttctattccattg


 


gtctatctgtctgtctctgtaccaatcaccatgcagtttttatcactattgctctgtaat


 


actgcttgagttcagggatagtgattccccctgaagtccttttattgttgaggatagctt


 


tagctatcctgggttttttgttattccagatgaatttgcaaattgttctgtct


 


>scr_sc-170396977_1 (TOXMARKER Assignment: 125; SEQ ID NO: 125)


 


tgatcacgctcagcccttggtaggacattctacagagtctcttgctgcccctccgtctgt


 


gccagtggtaccacacggggcagcctccgtggaagtttctagttcacagtatgcagctca


 


gagtgaaagtgtggtgcatcaagactccagtgtccctggaatgccagtacaaactccagg


 


cccagtccaaggacagaattacagtgtctgggattcaaaccaacagtctgtcagtgtaca


 


gccccagtattctcctgcccaatctcaagcaaccatatattaccaaggacagacatgttc


 


aactgtctacggtgtgacctctccttattcacagacaactcctccaattg


 


>scr_sc-14059147_2 (TOXMARKER Assignment: 126; SEQ ID NO: 126)


 


gctagcatcgtgatggccaagtgcatccctgtgcttttttcttttctaagaaagattgaa


 


aaccaacagttcttccccaacagctgcctaaattttaaggggtctgacccttacatttca


 


attgggggaatgaagggggcccaaccggcttaattgctgtgggagagtgagtctggatgt


 


ctgagagagcaccttgggagggactcttcctgcaatgctgtaaatacgagtaccgtttta


 


ataaagcatgtaca


 


>scr_sc-87750810_1 (TOXMARKER Assignment: 127; SEQ ID NO: 127)


 


tttttttttttttttttggctcctgccatcttttttattggtctgggctgtgggctgggg


 


gaggcaggtgggctcacatctttatgcaagcagcaaggagacggttcacatgctcaggag


 


actccaggaaggccttgagcttgggtcgggctttgagacgcgctacataggcggagagca


 


gggggaagtctttcaagtaaccagggaacaggagctctaggttcagaagtaaatccagta


 


ggcggtagtcggcgaaggagatctggtcaccaacaatgaagcattggccacccttgttct


 


gggccagaagagtttcaaatggcttcaggtgtcctggaagctccttcctatattggccct


 


tgtcctccttacagatatggagatagtgccatgcaatgcgcctgaacacgtcttccagtc


 


cgtcgttcaccatgtccaccagtgctgcctcttgctggtctttgccgtagagcccgaagg


 


agtggcccaggtgccgtaggatggcattcgattggtacagagtgagctttccatcctgga


 


acttggggatctgcccaaacagacaggaagccttgaatgtgccttgctcccaaacatcca


 


aggtcaccacctcctccttccaactctggccctggtcggctagcagcatgcgcataacct


 


cacagcgcccagtgttggggtgcaggatggggatgaggccacagcgaagagacccaccct


 


cagagcatcctgggagagtttgggagactggaaagctgacaagtggactaaactagcttg


 


ggagcctcgaagggagggaaaaaatgtggtggtagaggccatgtcctaacattatcttgg


 


caagccaagacccagccccaccggcacagggaaggaggaaaagtgacagacagtgtagct


 


gcctatggaggctaagaggtcagtcctggccccaccaaccacaattgtagtcccgcccca


 


agtctcggtcttgcccccaacgtggtcttggccacatccctccagcaccagtgttgaggg


 


ggccccaggagtgactatggcttgtgcccttcatcttgaaaac


 


>cszr_2 02034260_190929676 (TOXMARKER Assignment: 128; SEQ ID NO: 128)


 


gtgcaccagtacctgatgctgggagatgaatggcttagcgctgttctacttggaacatat


 


cactcctgccagccgggcactaacaattatcacccaatccaggacttaaactgtgataga


 


ctggctgatgtttgcctttgaatagagtgtcccaaaagatgggaccactggtcagctgcc


 


atggactagattctccacctgttgggggcaatctggtcaccttgctgcccaatccgacct


 


ggagccaccacagcacgagtgtcaagcactggcagaagcccatgggtggaggaaagacct


 


ctgcgactggctgattgacccctgctgaaagccgaggctaccttgtccacagacgggaac


 


agttctcttcatga









Example 10
Identification of a TOXMARKER 76, 135, 147, 151, 152, 154, and 162 for Accurate Prediction of Hepatotoxcity
In order to determine the minimal number of markers required for prediction, backward selection from a larger set of putative markers was used. Each set of smaller markers was evaluated on cross-validation by linear regression and the smallest set that was significantly accurate (p<0.001, Fisher's exact test) was selected. The TOXMARKER genes that make up the TOXMARKER 76, 135, 147, 151, 152, 154, and 162 is listed in Table 6 below







TABLE 6







TOXMARKER 76, 135, 147, 151, 152, 154, and 162
















SEQ ID
Gene




PTS Code
Rat ID No.
TOXMARKER
NO
Name
Accuracy
P. Value





pts2.3014511.1
scr_gb-bi294409_1
42
42
IFNAR-2
0.771552
4.12E−13


pts2.3013420.1
scr_gb-af069306_1
59
59
Transaldolase


pts2.3015170.1
scr_gb-bi288503_1
65
65
Clp-1


pts2.3011880.1
scr_gb-d86383_2
66
66
Hex


pts2.3015871.2
scr_gb-bm986259_1
97
97
Novel


pts2.3012511.2
scr_gb-aa899865_3
76
76
Novel


pts2.3017180.2
cszr_204152648_1915
71
71
Novel



21095









“P.value” is the probability level that the observed classification is random.
“Accuracy” is the number of correct predictions divided by the number of samples (total number of predictions). It is a proportion of how often the disclosed TOXMARKER 76, 135, 147, 151, 152, 154, and 162s are accurate in screening for toxicity.
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OTHER EMBODIMENTS
It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

## Claims

1. An in vitro method of predicting the association of a test agent with zone 3 necrosis, comprising:
a) determining the level of expression of each of the genes listed in Table 5 in a cell exposed to said test agent;
b) comparing said level of expression to the level of expression of said genes in a control population exposed to at least one control agent;
c) identifying a statistically significant alteration in the level of expression of said genes in the presence of the test agent;

wherein, if present, said alteration indicates that said test agent is predicted to be associated with zone 3 necrosis.

2. The method of claim 1, wherein said level of expression is determined by detecting a gene transcript.

3. An in vitro method of predicting the association of a test agent with zone 3 necrosis, comprising:
a) determining the level of expression of each of the TOXMARKER 42, 59, 65, 66, 71, 76, and 97 genes in a cell exposed to said test agent;
b) comparing said level of expression to the level of expression of said genes in a control population exposed to at least one control agent;
c) identifying a statistically significant alteration in the level of expression of said genes in the presence of the test agent;

wherein, if present, said alteration indicates that said test agent is predicted to be associated with zone 3 necrosis.

4. The method of claim 3, wherein said level of expression is determined by detecting a gene transcript.

5. An in vivo method of predicting the association of a test agent with zone 3 necrosis, comprising:
a) providing a cell from a subject exposed to said test agent;
b) determining the level of expression of each of the TOXMARKER 42, 59, 65, 66, 71, 76, and 97 genes in said cell;
c) comparing said level of expression to the level of expression of said genes in a control population exposed to at least one control agent;
d) identifying a statistically significant alteration in the level of expression of said genes in the presence of the test agent;

wherein, if present, said alteration indicates that said test agent is predicted to be associated with zone 3 necrosis.

6. The method of claim 5, wherein said level of expression is determined by detecting a gene transcript.

7. A method for screening for changes in gene expression associated with a toxic agent, comprising:
a) determining the level of expression of each of the genes listed in Table 5 in a cell exposed to a test agent;
b) comparing said level of expression to the level of expression of said genes in a control population exposed to at least one control agent;
c) identifying a statistically significant alteration in the level of expression of said genes in the presence of the test agent

thereby screening for changes in gene expression associated with a toxic agent.

8. A method of predicting the association of a test agent with zone 3 necrosis, comprising:
a) determining the level of expression of TOXMARKER 71 in a cell exposed to said test agent;
b) determining the level of expression of at least one TOXMARKER selected from the group consisting of TOXMARKERS 1–70 and 72–129 in said cell;
c) comparing the level of expression in said cell of TOXMARKER 71 and the TOXMARKER(s) selected in step (b) to the level of expression in a control population exposed to at least one control agent; and
d) identifying a statistically significant alteration in the level of expression of said genes in the presence of the test agent,

wherein, if present, said alteration indicates that said test agent is predicted to be associated with zone 3 necrosis.

9. The method of claim 8, wherein the TOXMARKERs selected in step (b) are TOXMARKERs 42, 59, 65, 66, 76 and 97.

10. The method of claim 8, wherein said level of expression is determined by detecting a gene transcript.