Abstract:
Methods for detecting G-protein coupled receptor (GPCR) activity; methods for assaying GPCR activity; and methods for screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described. Included are methods for expanding ICAST technologies for assaying GPCR activity with applications for ligand fishing, and agonist or antagonist screening. These methods include: engineering seronine/threonine phosphorylation sites into known or orphan GPCR open reading frames in order to increase the affinity of arrestin for the activated form of the GPCR or to increase the reside time of arrestin on the activated GPCR; engineering mutant arrestin proteins that bind to activated GPCRs in the absence of G-protein coupled receptor kinases which may be limiting; and engineering mutant super arrestin proteins that have an increased affinity for activated GPCRs with or without phosphorylation. These methods are intended to increase the robustness of the GPCR/ICAST technology in situations in which G-protein coupled receptor kinases are absent or limiting, or in which the GPCR is not efficiently down-regulated or is rapidly resensitized (thus having a labile interaction with arrestin). Included are also more specific methods for using ICAST complementary enzyme fragments to monitor GPCR homo- and hetero-dimerization with applications for drug lead discovery and ligand and function discovery for orphan GPCRs.

Description:
This application is a continuation-in-part of U.S. application Ser. No. 09/654,499, filed Sep. 1, 2000, which claims the benefit from Provisional Application Ser. No. 60/180,669, filed Feb. 7, 2000. The entirety of U.S. application Ser. No. 09/654,499 and Provisional Application Ser. No. 60/180,669 are incorporated herein by reference. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The present invention relates to methods of detecting G-protein-coupled receptor (GPCR) activity, and provides methods of assaying GPCR activity, methods for screening for GPCR ligands, agonists and/or antagonists, methods for screening natural and surrogate ligands for orphan GPCRs, and methods for screening compounds that interact with components of the GPCR regulatory process. 
     2. Background of the Technology 
     The actions of many extracellular signals are mediated by the interaction of G-protein-coupled receptors (GPCRs) and guanine nucleotide-binding regulatory proteins (G-proteins). G-protein-mediated signaling systems have been identified in many divergent organisms, such as mammals and yeast. The GPCRs represent a large super family of proteins which have divergent amino acid sequences, but share common structural features, in particular, the presence of seven transmembrane helical domains. GPCRs respond to, among other extracellular signals, neurotransmitters, hormones, odorants and light. Individual GPCR types activate a particular signal transduction pathway; at least ten different signal transduction pathways are known to be activated via GPCRs. For example, the beta 2-adrenergic receptor (β2AR) is a prototype mammalian GPCR. In response to agonist binding, β2AR receptors activate a G-protein (Gs) which in turn stimulates adenylate cyclase activity and results in increased cyclic adenosine monophosphate (cAMP) production in the cell. 
     The signaling pathway and final cellular response that result from GPCR stimulation depends on the specific class of G-protein with which the particular receptor is coupled (Hamm, “The Many Faces of G-Protein Signaling.” J. Biol. Chem., 273:669-672 (1998)). For instance, coupling to the Gs class of G-proteins stimulates cAMP production and activation of the Protein Kinase A and C pathways, whereas coupling to the Gi class of G-proteins down regulates cAMP. Other second messenger systems such as calcium, phospholipase C, and phosphatidylinositol 3 may also be utilized. As a consequence, GPCR signaling events have predominantly been measured via quantification of these second messenger products. 
     The decrease of a response to a persistent stimulus is a widespread biological phenomenon. Signaling by diverse GPCRs is believed to be terminated by a uniform two-step mechanism. Activated receptor is first phosphorylated by a GPCR kinase (GRK). An arrestin protein binds to the activated and phosphorylated receptor, thus blocking G-protein interaction. This process is commonly referred to as desensitization, a general mechanism that has been demonstrated in a variety of functionally diverse GPCRs. Arrestin also plays a part in regulating GPCR internalization and resensitization, processes that are heterogenous among different GPCRs (Oakley, et al., J. Biol. Chem., 274:32248-32257 (1999)). The interaction between an arrestin and GPCR in processes of internalization and resensitization is dictated by the specific sequence motif in the carboxyl terminus of a given GPCR. Only a subset of GPCRs, which possess clusters of three serine or threonine residues at the carboxyl termini, were found to co-traffick with the arrestins into the endocytic vesicles after ligand stimulation. The number of receptor kinases and arrestins involved in desensitization of GPCRs is rather limited. 
     A common feature of GPCR physiology is desensitization and recycling of the receptor through the processes of receptor phosphorylation, endocytosis and dephosphorylation (Ferguson, et al., “G-protein-coupled receptor regulation: role of G-protein-coupled receptor kinases and arrestins.” Can. J. Physiol. Pharmacol., 74:1095-1110 (1996)). Ligand-occupied GPCRs can be phosphorylated by two families of serine/threonine kinases, the G-protein-coupled receptor kinases (GRKs) and the second messenger-dependent protein kinases such as protein kinase A and protein kinase C. Phosphorylation by either class of kinases serves to down-regulate the receptor by uncoupling it from its corresponding G-protein. GRK-phosphorylation also serves to down-regulate the receptor by recruitment of a class of proteins known as the arrestins that bind the cytoplasmic domain of the receptor and promote clustering of the receptor into endocytic vescicles. Once the receptor is endocytosed, it will either be degraded in lysosomes or dephosphorylated and recycled back to the plasma membrane as a fully-functional receptor. 
     Binding of an arrestin protein to an activated receptor has been documented as a common phenomenon of a variety of GPCRs ranging from rhodopsin to β2AR to the neurotensin receptor (Barak, et al., “A β-arrestin/Green Fluorescent Fusion Protein Biosensor for Detecting G-Protein-Coupled Receptor Activation,” J. Biol. Chem., 272:27497-500 (1997)). Consequently, monitoring arrestin interaction with a specific GPCR can be utilized as a generic tool for measuring GPCR activation. Similarly, a single G-protein and GRK also partner with a variety of receptors (Hamm, et al. (1998) and Pitcher et al., “G-Protein-Coupled Receptor Kinases,” Annu. Rev. Biochem., 67:653-92 (1998)), such that these protein/protein interactions may also be monitored to determine receptor activity. 
     Many therapeutic drugs in use today target GPCRs, as they regulate vital physiological responses, including vasodilation, heart rate, bronchodilation, endocrine secretion and gut peristalsis. See, e.g., Lefkowitz et al., Annu. Rev. Biochem., 52:159 (1983). Some of these drugs mimic the ligand for this receptor. Other drugs act to antagonize the receptor in cases when disease arises from spontaneous activity of the receptor. 
     Efforts such as the Human Genome Project are identifying new GPCRs (“orphan” receptors) whose physiological roles and ligands are unknown. It is estimated that several thousand GPCRs exist in the human genome. 
     Various approaches have been used to monitor intracellular activity in response to a stimulant, e.g., enzyme-linked immunosorbent assay (ELISA); Fluorescense Imaging Plate Reader assay (FLIPR™, Molecular Devices Corp., Sunnyvale, Calif.); EVOscreen™, EVOTEC™, Evotec Biosystems Gmbh, Hamburg, Germany; and techniques developed by CELLOMICS™, Cellomics, Inc., Pittsburgh, Pa. 
     Germino et al., “Screening for in vivo protein-protein interactions.” Proc. Natl. Acad. Sci., 90(3):933-937 (1993), discloses an in vivo approach for the isolation of proteins interacting with a protein of interest. 
     Phizicky et al., “Protein-protein interactions: methods for detection and analysis.” Microbiol. Rev., 59(1): 94-123 (1995), discloses a review of biochemical, molecular biological and genetic methods used to study protein-protein interactions. 
     Offermanns et al., “Gα 15  and Gα 16  Couple a Wide Variety of Receptors to Phospholipase C.” J. Biol. Chem., 270(25):15175-15180 (1995), discloses that Gα 15  and Gα 16  can be activated by a wide variety of G-protein-coupled receptors. The selective coupling of an activated receptor to a distinct pattern of G-proteins is regarded as an important requirement to achieve accurate signal transduction. Id. 
     Barak et al., “A β-arrestin/Green Fluorescent Protein Biosensor for Detecting G Protein-coupled Receptor Activation.” J. Biol. Chem., 272(44):27497-27500 (1997) and U.S. Pat. Nos. 5,891,646 and 6,110,693 disclose the use of a β-arrestin/green fluorescent fusion protein (GFP) for imaging protein translocation upon stimulation of GPCR with optical devices. 
     Each of the references described above has drawbacks. For example, 
     The prior art methodologies require over-expression of the proteins, which could cause artifact and tip the balance of cellular regulatory machineries. 
     The prior art visualization or imaging assays are low throughput and lack thorough quantification. Therefore, they are not suitable for high throughput pharmacological and kinetic assays. 
     In addition, many of the prior art assays require isolation of the GPCR rather than observation of the GPCR in a cell. There thus exists a need for improved methods for monitoring GPCR function. 
     SUMMARY OF THE INVENTION 
     The present invention provides modifications to the disclosure in U.S. application Ser. No. 09/654,499. In particular, the present invention is directed to modifications of the below aspects of the invention to further enhance assay sensitivity. The modifications include the use of genetically modified arrestins that exhibit enhanced binding to activated GPCR regardless of whether the GPCR is phosphorylated or non-phosphorylated; the use of a serine/threonine cluster strategy to facilitate screening assays for orphan receptors that do not possess this structural motif on their own; and the use of a combination of the above modifications to achieve even more enhanced detection. 
     A first aspect of the present invention is a method that monitors GPCR function proximally at the site of receptor activation, thus providing more information for drug discovery purposes due to fewer competing mechanisms. Activation of the GPCR is measured by a read-out for interaction of the receptor with a regulatory component such as arrestin, G-protein, GRK or other kinases, the binding of which to the receptor is dependent upon agonist occupation of the receptor. The present invention involves the detection of protein/protein interaction by complementation of mutant reporter enzymes. 
     Binding of arrestin to activated GPCR is a common process in the first step of desensitization that has been demonstrated for most, if not all, GPCRs studied so far. Measurement of GPCR interaction with arrestin via mutant enzyme complementation (i.e., ICAST) provides a more generic assay technology applicable for a wide variety of GPCRs and orphan receptors. 
     A further aspect of the present invention is a method of assessing GPCR pathway activity under test conditions by providing a test cell that expresses a GPCR, e.g., muscarinic, adrenergic, dopamine, angiotensin or endothelin, as a fusion protein to a mutant reporter enzyme and interacting a protein in the GPCR pathway, e.g., G-protein, arrestin or GRK, as a fusion protein with a complementing mutant reporter enzyme. When test cells are exposed to a known agonist to the target GPCR under test conditions, activation of the GPCR will be monitored by complementation of the reporter enzyme. Increased reporter enzyme activity reflects interaction of the GPCR with its interacting protein partner. 
     A further aspect of the present invention is a method of assessing GPCR pathway activity in the presence of a test arrestin, e.g., β-arrestin. 
     A further aspect of the present invention is a method of assessing GPCR pathway activity in the presence of a test G-protein. 
     A further aspect of the present invention is a method of assessing GPCR pathway activity upon exposure of the test cell to a test ligand. 
     A further aspect of the present invention is a method of assessing GPCR activity upon co-expression in the test cell of a second receptor. The second receptor could be the same GPCR or orphan receptor (i.e., homo-dimerization), a different GPCR or orphan receptor (i.e., hetero-dimerization) or could be a receptor of another type. 
     A further aspect of the present invention is a method for screening for a ligand or agonist to an orphan GPCR. The ligand or agonist could be contained in natural or synthetic libraries or mixtures or could be a physical stimulus. A test cell is provided that expresses the orphan GPCR as a fusion protein with a mutant reporter enzyme, e.g., a β-galactosidase mutant, and, for example, an arrestin or mutant form of arrestin as a fusion protein with a complementing mutant reporter enzyme, e.g., another β-galactosidase mutant. The interaction of the arrestin with the orphan GPCR upon receptor activation is measured by enzymatic activity of the complemented reporter enzyme. The test cell is exposed to a test compound, and an increase in reporter enzyme activity indicates the presence of a ligand or agonist. 
     A further aspect of the present invention is a method for screening a protein of interest, for example, an arrestin protein (or mutant form of the arrestin protein) for the ability to bind to a phosphorylated, or activated, GPCR. A test cell is provided that expresses a GPCR as a fusion protein with a mutant reporter enzyme, e.g., a β-galactosidase mutant, and contains arrestin (or a mutant form of arrestin) as a fusion protein with a complementing mutant reporter enzyme, e.g., another β-galactosidase mutant. The interaction of arrestin with the GPCR upon receptor activation is measured by enzymatic activity of the complemented reporter enzyme. The test cell is exposed to a known GPCR agonist and then reporter enzyme activity is detected. Increased reporter enzyme activity indicates that the β-arrestin molecule can bind to phosphorylated, or activated, GPCR in the test cell. 
     A further aspect of the present invention is a method to screen for an agonist to a specific GPCR. The agonist could be contained in natural or synthetic libraries or could be a physical stimulus. A test cell is provided that expresses a GPCR as a fusion protein with a mutant reporter enzyme, e.g., a β-galactosidase mutant, and, for example, an arrestin as a fusion protein with a complementing mutant reporter enzyme, e.g., another β-galactosidase mutant. The interaction of arrestin with the GPCR upon receptor activation is measured by enzymatic activity of the complemented reporter enzyme. The test cell is exposed to a test compound, and an increase in reporter enzyme activity indicates the presence of an agonist. The test cell may express a known GPCR or a variety of known GPCRs, or may express an unknown GPCR or a variety of unknown GPCRs. The GPCR may be, for example, an odorant GPCR or a βAR GPCR. 
     A further aspect of the present invention is a method for screening a test compound for GPCR antagonist activity. A test cell is provided that expresses a GPCR as a fusion protein with a mutant reporter enzyme, e.g., a β-galactosidase mutant, and, for example, an arrestin as a fusion protein with a complementing mutant reporter enzyme, e.g., another β-galactosidase mutant. The interaction of arrestin with the GPCR upon receptor activation is measured by enzymatic activity of the complemented reporter enzyme. The test cell is exposed to a test compound, and an increase in reporter enzyme activity indicates the presence of an agonist. The cell is exposed to a test compound and to a GPCR agonist, and reporter enzyme activity is detected. When exposure to the agonist occurs at the same time as or subsequent to exposure to the test compound, a decrease in reporter enzyme activity after exposure to the test compound indicates that the test compound has antagonist activity to the GPCR. 
     A further aspect of the present invention is a method of screening a sample solution for the presence of an agonist, antagonist or ligand to a GPCR. A test cell is provided that expresses GPCR as a fusion protein with a mutant reporter enzyme, e.g., a β-galactosidase mutant, and contains, for example, a β-arrestin as a fusion protein with a complementing reporter, e.g., another β-galactosidase mutant. The test cell is exposed to a sample solution, and reporter enzyme activity is assessed. Changed reporter enzyme activity after exposure to the sample solution indicates the sample solution contains an agonist, antagonist or ligand for a GPCR expressed in the cell. 
     A further aspect of the present invention is a method of screening a cell for the presence of a GPCR. According to this aspect, an arrestin fusion protein with a mutant reporter enzyme and a GPCR downstream signaling fusion protein with a mutant reporter enzyme are employed to detect GPCR action. A modification of this aspect of the invention can be employed to provide a method of screening a plurality of cells for those cells which contain a GPCR. According to this aspect, a plurality of cells containing a conjugate comprising a β-arrestin protein as a fusion protein with a reporter enzyme are provided; the plurality of cells are exposed to a GPCR agonist; and activity of reporter enzyme activity is detected. An increase in reporter enzymatic activity after exposure to the GPCR agonist indicates β-arrestin protein binding to a GPCR, thereby indicating that the cell contains a GPCR responsive to the GPCR agonist. 
     A further aspect of the invention is a method for mapping GPCR-mediated signaling pathways. For instance, the system could be utilized to monitor interaction of c-src with β-arrestin-1 upon GPCR activation. Additionally, the system could be used to monitor protein/protein interactions involved in cross-talk between GPCR signaling pathways and other pathways such as that of the receptor tyrosine kinases or Ras/Raf. According to this aspect, a test cell is provided that expresses a GPCR or other related protein with a mutant reporter enzyme, e.g., a β-galactosidase mutant, and contains a protein from another pathway as a fusion protein with a complementing mutant reporter enzyme, e.g., another β-galactosidase mutant. Increased reporter enzymatic activity indicates protein/protein interaction. 
     A further aspect of the invention is a method for monitoring homo- or hetero-dimerization of GPCRs upon agonist or antagonist stimulation. Increasing evidence indicates that GPCR dimerization is important for biological activity (AbdAlla, et al., “AT1-receptor heterodimers show enhanced G-protein activation and altered receptor sequestration.” Nature, 407:94-98 (2000); Bockaert, et al., “Molecular tinkering of G protein-coupled receptors: an evolutionary success.” EMBO J. 18:1723-29 (1999)). Jordan, et al., “G-protein-coupled receptor heterodimerization modulates receptor function.” Nature, 399:697-700 (1999), demonstrated that two non-functional opioid receptors, κ and δ, heterodimerize to form a functional receptor. Gordon et al., “Dopamine D2 receptor dimers and receptor blocking peptides.” Bioch. Biophys. Res. Commun. 227:200-204 (1996), showed different pharmacological properties associated with the monomeric and dimeric forms of Dopamine receptor D2. The D2 receptors exist either as monomers that are selective targets for spiperone or as dimer forms that are targets for nemonapride. Herbert, et al., “A peptide derived from a β2-adrenergic receptor transmembrane domain inhibits both receptor dimerization and activation.” J.B.C. 271:16384-92 (1996), demonstrated that the agonist stimulation was found to stabilize the dimeric state of the receptor, whereas inverse agonists favored the monomeric form. Indeed, the same study showed that a peptide corresponding to the sixth transmembrane domain of the β2-adrenergic receptor inhibited both receptor dimerization and activation. Further, Angers et al., Detection of beta-2-adrenergic receptor dimerization in living cells using bioluminescence resonance energy transfer, Proc. Natl. Acad. Sci. USA, 97(7):3684-3689, discloses the use of β2-adrenergic receptor fusion proteins (i.e., β2-adrenergic receptor fused to luciferase and β2-adrenergic receptor fused to an enhanced red-shifted green fluorescent protein) to study β2-adrenergic receptor dimerization. 
     GPCR dimerization in the context of cellular physiology and pharmacology can be monitored in accordance with the invention. For example, β-galactosidase complementation can be measured in test cells that co-express GPCR fusion proteins of β-galactosidase mutant enzymes, e.g., GPCR 1 Δα and GPCR 2 Δω (FIG.  27 ). According to this aspect, the interconversion between monomeric to dimeric forms of the GPCRs or orphan receptors can be measured by mutant reporter enzyme complementation. FIG. 27 illustrates a test cell co-expressing GPCR or an orphan receptor as a fusion protein with Δα form of β-galactosidase mutant (e.g., GPCR 1 Δα), and the same GPCR or orphan receptor as a fusion protein with Δω form of β-galactosidase mutant (e.g. GPCR 1 Δω). Formation of the GPCR homodimer is reflected by formation of an active enzyme, which can be measured by enzyme activity assays, such as the Gal-Screen™ assay. Similarly, hetero-dimerization between two distinct GPCRs, or two distinct orphan receptors, or between one known GPCR and one orphan receptor can be analyzed in test cells co-expressing two fusion proteins, e.g., GPCR 1 Δα and GPCR 2 Δω. The increased β-galactosidase activity indicates that the two receptors can form a heterodimer. 
     A further aspect of the invention is a method of monitoring the interconversion between the monomeric and dimeric form of GPCRs under the influence of agonist or antagonist treatment. The test receptor(s) can be between the same GPCR or orphan receptor (homodimer), or between two distinct GPCRs or orphan receptors (heterodimer). The increased β-galactosidase activity after treatment with a compound means that the compound binds to and/or stabilizes the dimeric form of the receptor. The decreased β-galactosidase activity after treatment with a compound means that the compound binds to and/or stabilizes the monomeric form of the receptor. 
     A further aspect of the invention is a method of screening a cell for the presence of a GPCR responsive to a GPCR agonist. A cell is provided that contains protein partners that interact downstream in the GPCR&#39;s pathway. The protein partners are expressed as fusion proteins to the mutant, complementing enzyme and are used to monitor activation of the GPCR. The cell is exposed to a GPCR agonist and then enzymatic activity of the reporter enzyme is detected. Increased reporter enzyme activity indicates that the cell contains a GPCR responsive to the agonist. 
     The present invention involves the use of a combination of proprietary technologies (including ICAST™, Intercistronic Complementation Analysis Screening Technology, Gal-Screen™, etc.) to monitor protein/protein interactions in GPCR signaling. As disclosed in U.S. application Ser. No. 09/654,499, the method of the invention in part involves using ICAST™, which in turn involves the use of two inactive β-galactosidase mutants, each of which is fused with one of two interacting target protein pairs, such as a GPCR and an arrestin. The formation of an active β-galactosidase complex is driven by interaction of the target proteins. In this system, β-galactosidase activity can be detected using, e.g., the Gal-Screen™ assay system, wherein direct cell lysis is combined with rapid ultrasensitive chemiluminescent detection of β-galactosidase reporter enzyme. This system uses, e.g., a Galacton-Star® chemiluminescent substrate for measurement in a luminometer as a read out of GPCR activity. 
     FIG. 23 is a schematic depicting the use of the complementation technology in the method of the present invention. FIG. 23 shows two inactive β-galactosidase mutants that become active when they are forced together by specific interactions between the fusion partners of an arrestin molecule and an activated GPCR or orphan receptor. This assay technology will be especially useful in high throughput screening assays for ligand fishing for orphan receptors, a process called de-orphaning. As illustrated in FIG. 28, a β-galactosidase fusion protein of an orphan receptor (e.g., GPCR orphan Δα) is co-expressed in the test cell with a fusion protein of β-arrestin (e.g., β-ArrΔω). When the test cell is subjected to compounds, which could be natural or synthetic, the increased β-galactosidase activity means the compound is either a natural or surrogate ligand for this GPCR. The same assay system can be used to find drug leads for the new GPCRs. The increased β-galactosidase activity in the test cell after treatment indicates the agonist activity of the compound. The decreased β-galactosidase activity in the test cell indicates antagonist activity or inverse agonist activity of the compound. In addition, the method of the invention could be used to monitor GPCR-mediated signaling pathways via other downstream signaling components such as G-proteins, GRKs or the proto-oncogene c-Src. 
     The invention is achieved in part by using ICAST™ protein/protein interaction screening to map signaling pathways. This technology is applicable to a variety of known and unknown GPCRs with diverse functions. They include, but are not limited to, the following sub-families of GPCRs: 
     (a) receptors that bind to amine-like ligands-Acetylcholine muscarinic receptor (M1 to M5), alpha and beta Adrenoceptors, Dopamine receptors (D1, D2, D3 and D4), Histamine receptors (H1 and H2), Octopamine receptor and Serotonin receptors (5HT1, 5HT2, 5HT4, 5HT5, 5HT6, 5HT7); 
     (b) receptors that bind to a peptide ligand-Angiotensin receptor, Bombesin receptor, Bradykinin receptor, C—C chemokine receptors (CCR1 to CCR8, and CCR10), C-X-C type Chemokine receptors (CXC-R5), Cholecystokinin type A receptor, CCK type receptors, Endothelin receptor, Neurotesin receptor, FMLP-related receptors, Somatostatin receptors (type 1 to type 5) and Opioid receptors (type D, K, M, X); 
     (c) receptors that bind to hormone proteins-Follic stimulating hormone receptor, Thyrotrophin receptor and Lutropin-choriogonadotropic hormone receptor; 
     (d) receptors that bind to neurotransmitters-substance P receptor, Substance K receptor and neuropeptide Y receptor; 
     (e) Olfactory receptors-Olfactory type 1 to type 11, Gustatory and odorant receptors; 
     (f) Prostanoid receptors-Prostaglandin E2 (EP1 to EP4 subtypes), Prostacyclin and Thromboxane; 
     (g) receptors that bind to metabotropic substances-Metabotropic glutamate group I to group III receptors; 
     (h) receptors that respond to physical stimuli, such as light, or to chemical stimuli, such as taste and smell; and 
     (i) orphan GPCRs-the natural ligand to the receptor is undefined. 
     Use of the ICAST™ technology in combination with the invention provides many benefits to the GPCR screening process, including the ability to monitor protein interactions in any sub-cellular compartment-membrane, cytosol and nucleus; the ability to achieve a more physiologically relevant model without requiring protein overexpression; and the ability to achieve a functional assay for receptor binding allowing high information content. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIG.  1 . Cellular expression levels of β2 adrenergic receptor (β2AR) and β-arrestin-2 (βArr2) in C2 clones. Quantification of β-galactosidase (β-gal) fusion protein was performed using antibodies against β-gal and purified β-gal protein in a titration curve by a standardized ELISA assay. 
     FIG. 1A shows expression levels of β2AR-βgalΔα clones (in expression vector pICAST ALC). 
     FIG. 1B shows expression levels of 3Arr2-βgalΔω in expression vector pICAST OMC4 for clones 9-3,-7,-9,-10,-19 and -24, or in expression vector pICAST OMN4 for clones 12-4,-9,-16,-18,-22 and -24. 
     FIG.  2 . Receptor β2AR activation was measured by agonist-stimulated cAMP production. C2 cells expressing pICAST ALC β2AR (clone 5) or parental cells were treated with increasing concentrations of (−)isoproterenol and 0.1 mM IBMX. The quantification of cAMP level was expressed as pmol/well. 
     FIG.  3 . Interaction of activated receptor β2AR and arrestin can be measured by β-galactosidase complementation. 
     FIG. 3A shows a time course of β-galactosidase activity in response to agonist (−)isoproterenol stimulation in C2 expressing β2AR-βgalΔα (β2AR alone, in expression vector pICAST ALC), or a pool of doubly transduced C2 co-expressing β2AR-βgalΔα and βArr2-βgalΔω (in expression vectors pICAST ALC and pICAST OMC and clones isolated from the same pod (43-1, 43-2, 43-7 and 43-8)). 
     FIG. 3B shows a time course of β-galactosidase activity in response to agonist (−)isoproterenol stimulation in C2 cells expressing β2AR-βgalΔα alone (in expression vector pICAST ALC) and C2 clones co-expressing β2AR-βgalΔα and βArr1-βgalΔω (in expression vectors ICAST ALC and pICAST OMC). 
     FIG.  4 . Agonist dose response for interaction of β2AR and arrestin can be measured by β-galactosidase complementation. 
     FIG. 4A shows a dose response to agonists (−)isoproterenol and procaterol in C2 cells co-expressing β2AR-βgalΔα and βArr2-βgalΔω fusion constructs. 
     FIG. 4B shows a dose response to agonists (−)isoproterenol and procaterol in C2 cells co-expressing β2AR-βgalΔα and βArr1-βgalΔω fusion constructs. 
     FIG.  5 . Antagonist mediated inhibition of receptor activity can be measured by β-galactosidase complementation in cells co-expressing β2AR-βgalΔα and βArr-βgalΔω. 
     FIG. 5A shows specific inhibition with adrenergic antagonists ICI-118,551 and propranolol of β-galactosidase activity in C2 clones co-expressing β2AR-βgalΔα and βArr2-βgalΔω fusion constructs after incubation with agonist (−)isoproterenol. 
     FIG. 5B shows specific inhibition of β-galactosidase activity with adrenergic antagonists ICI-118,551 and propranolol in C2 clones co-expressing β2AR-βgalΔα and βArr1-βgalΔω fusion constructs in the presence of agonist (−)isoproterenol. 
     FIG. 6. C2 cells expressing adenosine receptor A2a show cAMP induction in response to agonist (CGS-21680) treatment. C2 parental cells and C2 cells co-expressing A2aR-βgalΔα and βArr1-βgalΔω as a pool or as selected clones (47-2 and 47-13) were measured for agonist-induced cAMP response (pmol/well). 
     FIG.  7 . Agonist stimulated cAMP response in C2 cells co-expressing Dopamine receptor D1 (D1-βgalΔα) and β-arrestin-2 (βArr2-βgalΔω). The clone expressing βArr2-βgalΔω (Arr2 alone) was used as a negative control in the assay. Cells expressing D1-βgalΔα in addition to βArr2-βgalΔω responded agonist treatment (3-hydroxytyramine hydrochloride at 3 μM). D1(PIC2) or D1(PIC3) designate D1 in expression vector pICAST ALC2 or pICAST ALC4, respectively. 
     FIG.  8 . Variety of mammalian cell lines can be used to generate stable cells for monitoring GPCR and arrestin interactions. 
     FIG. 8A, FIG.  8 B and FIG. 8C show the examples of HEK 293, CHO and CHW cell lines co-expressing adrenergic receptor β2AR and arrestin fusion proteins of β-galactosidase mutants. The β-galactosidase activity was used to monitor agonist-induced interaction of β2AR and arrestin proteins. 
     FIG.  9 . Beta-gal complementation can be used to monitor β2 adrenergic receptor homo-dimerization. 
     FIG. 9A shows β-galactosidase activity in HEK 293 clones co-expressing β2AR-βgalΔα and β2AR-βgalΔω. 
     FIG. 9B shows a cAMP response to agonist (−)isoproterenol in HEK 293 clones co-expressing β2AR-βgalΔα and β2AR-βgalΔω. HEK293 parental cells were included in the assays as negative controls. 
     FIG.  10 A. pICAST ALC: Vector for expression of β-galΔα as a C-terminal fusion to the target protein. This construct contains the following features: MCS, multiple cloning site for cloning the target protein in frame with the β-galΔα; GS Linker, (GGGGS)n (SEQ ID NO:10); NeoR, neomycin resistance gene; IRES, internal ribosome entry site; ColE1ori, origin of replication for growth in  E. coli;  5′MoMuLV LTR and 3′MoMuLV LTR, viral promoter and polyadenylation signals from the Moloney Murine leukemia virus. 
     FIGS. 10B-10P. Nucleotide sequence for pICAST ALC (SEQ ID NO:01). 
     FIG.  11 A. pICAST ALN: Vector for expression of β-galΔα as an N-terminal fusion to the target protein. This construct contains the following features: MCS, multiple cloning site for cloning the target protein in frame with the β-galΔα; GS Linker, (GGGGS)n (SEQ ID NO:10); NeoR, neomycin resistance gene; IRES, internal ribosome entry site; ColE1ori, origin of replication for growth in  E. coli;  5′MoMuLV LTR and 3′MoMuLV LTR, viral promoter and polyadenylation signals from the Moloney Murine leukemia virus. 
     FIGS. 11B-11L. Nucleotide sequence for pICAST ALN (SEQ ID NO:02). 
     FIG.  12 A. pICAST OMC: Vector for expression of β-galΔω as a C-terminal fusion to the target protein. This construct contains the following features: MCS, multiple cloning site for cloning the target protein in frame with the β-galΔω; GS Linker, (GGGGS)n (SEQ ID NO:10); Hygro, hygromycin resistance gene; IRES, internal ribosome entry site; ColE1ori, origin of replication for growth in  E. coli;  5′MoMuLV LTR and 3′MoMuLV LTR, viral promoter and polyadenylation signals from the Moloney Murine leukemia virus. 
     FIGS. 12B-12L. Nucleotide sequence for pICAST OMC (SEQ ID NO:03). 
     FIG.  13 A. pICAST OMN: Vector for expression of β-galΔω as an N-terminal fusion to the target protein. This construct contains the following features: MCS, multiple cloning site for cloning the target protein in frame with the β-galΔω; GS Linker, (GGGGS)n (SEQ ID NO:10); Hygro, hygromycin resistance gene; IRES, internal ribosome entry site; ColE1ori, origin of replication for growth in  E. coli;  5′MoMuLV LTR and 3′MoMuLV LTR, viral promoter and polyadenylation signals from the Moloney Murine leukemia virus. 
     FIGS. 13B-13L. Nucleotide sequence for pICAST OMN (SEQ ID NO:04). 
     FIG.  14 . pICAST ALC βArr2: Vector for expression of β-galΔα as a C-terminal fusion to β-arrestin-2. The coding sequence of human β-arrestin-2 (Genebank Accession Number: NM — 004313) was cloned in frame to β-galΔα in a pICAST ALC vector. 
     FIG.  15 . pICAST OMC βArr2: Vector for expression of β-galΔω as a C-terminal fusion to β-arrestin-2. The coding sequence of human β-arrestin-2 (Genebank Accession Number: NM — 004313) was cloned in frame to β-galΔω in a pICAST OMC vector. 
     FIG.  16 . pICAST ALC βArr1: Vector for expression of β-galΔα as a C-terminal fusion to β-arrestin-1. The coding sequence of human β-arrestin-1 (Genebank Accession Number: NM — 004041) was cloned in frame to β-galΔα in a pICAST ALC vector. 
     FIG.  17 . pICAST OMC βArr1: Vector for expression of β-galΔω as a C-terminal fusion to β-arrestin-1. The coding sequence of human β-arrestin-1 (Genebank Accession Number: NM — 004041) was cloned in frame to β-galΔω in a pICAST OMC vector. 
     FIG.  18 . pICAST ALC β2AR: Vector for expression of β-galΔ(α as a C-terminal fusion to β2 Adrenergic Receptor. The coding sequence of human β2 Adrenergic Receptor (Genebank Accession Number: NM — 000024) was cloned in frame to β-galΔα in a pICAST ALC vector. 
     FIG.  19 . pICAST OMC β2AR: Vector for expression of β-galΔω as a C-terminal fusion β2 Adrenergic Receptor. The coding sequence of human β2 Adrenergic Receptor (Genebank Accession Number: NM — 000024) was cloned in frame to β-galΔω in a pICAST OMC vector. 
     FIG.  20 . pICAST ALC A2aR: Vector for expression of β-galΔα as a C-terminal fusion to Adenosine 2a Receptor. The coding sequence of human Adenosine 2a Receptor (Genebank Accession Number: NM — 000675) was cloned in frame to β-galΔα in a pICAST ALC vector. 
     FIG.  21 . pICAST OMC A2aR: Vector for expression of β-galΔω as a C-terminal fusion to Adenosine 2a Receptor. The coding sequence of human Adenosine 2a Receptor (Genebank Accession Number: NM — 000675) was cloned in frame to β-galΔω in a pICAST OMC vector. 
     FIG.  22 . pICAST ALC D1: Vector for expression of β-galΔα as a C-terminal fusion to Dopamine D 1  Receptor. The coding sequence of human Dopamine D1 Receptor (Genebank Accession Number: X58987) was cloned in frame to β-galΔα in a pICAST ALC vector. 
     FIG. 23. A schematic depicting use of the complementation technology in the method of the invention. FIG. 23 shows two inactive mutant reporter enzymes that become active when the corresponding fusion partners, GPCR and β-arrestin interact. 
     FIG.  24 . Vector for expression of a GPCR with inserted seronine/threonine amino acid sequences as a fusion with β-galΔα. The open reading frame of a known or orphan GPCR is engineered to contain additional seronine/threonine sequences, such as SSS (seronine, seronine, seronine), within the C-terminal tail. The engineered GPCR is cloned in frame with β-galΔα in a pICAST ALC vector. The pICAST ALC vector contains the following features: MCS, multiple cloning site for cloning the target protein in frame with the β-galΔα; GS Linker, (GGGGS)n (SEQ ID NO:10); NeoR, neomycin resistance gene; IRES, internal ribosome entry site; ColE1ori, origin of replication for growth in  E. coli;  5′MoMuLV LTR and 3′MoMuLV LTR, viral promotor and polyadenylation signals from the Moloney Murine leukemia virus. 
     FIG.  25 . Vector for expression of mutant (R170E) β-arrestin2 as a fusion with β-galΔω. The open reading frame of β-arrestin2 is engineered to contain a point mutation that converts arginine 170 to a glutamate. The mutant β-arrestin2 is cloned in frame with β-galΔω in a pICAST OMC vector. The pICAST OMC vector contains the following features: MCS, multiple cloning site for cloning the target protein in frame with the β-galΔα; GS Linker, (GGGGS)n (SEQ ID NO:10); Hygro, hygromycin resistance gene; IRES, internal ribosome entry site; ColE1 ori, origin of replication for growth in  E. coli;  5′MoMuLV LTR and 3′MoMuLV LTR, viral promotor and polyadenylation signals from the Moloney Murine leukemia virus. 
     FIG.  26 . Phosphorylation insensitive Mutant R170E β-Arrestin2Δω binds to β2ARΔα in Response to Agonist Activation. A parental β2ARΔα C2 cell line was tranduced with the Mutant R170E β-Arrestin2Δω construct. Clonal populations co-expressing the two constructions were plated at 10,000 cells/well in 96 well plates and treated with 10 μM (−)isoproterenol, 0.3 mM ascorbic acid for the indicated time period. β-galactosidase activity was measured by addition of Tropix Gal-Screen™ assay system substrate (Applied Biosystems) and luminescence was measured using a Tropix TR717™ luminometer (Applied Biosystems). Treatments were performed in triplicate. For comparison, a clonal cell line (43-8) co-expressing β2ARΔα and wild-type β-Arrestin2Δω was also plated at 10,000 cells/well and given the same agonist treatment regimen. Minutes of (−)isoproterenol treatment is shown on the X-axis and β-galactosidase activity indicated by relative light units (RLU) is shown on the Y-axis. 
     FIG.  27 . GPCR dimerization measured by β-galactosidase complementation. A schematic depicting the utilization of the invention for monitoring GPCR homo- or hetero-dimerization. One GPCR is fused to one complement enzyme fragment, while the second GPCR is fused to the second complement enzyme fragment. Interaction of the two GPCRs is monitored by complementation of the enzyme fragments to produce an active enzyme complex (i.e., β-galactosidase activity). GPCR homo- or hetero-dimerization can be monitored in the absence or presence of ligand, agonists, inverse agonists or antagonists. 
     FIG.  28 . Ligand fishing for orphan receptors by β-galactosidase mutant complementation in ICAST™ system. A schematic depicting the utilization of the invention for ligand fishing and agonist/antagonist screening for orphan GPCRs. As an example, a test cell expressing two β-gal fission proteins, GPCR orphan- Δα and Arrestin-Δω, is subjected to treatments with samples from natural or synthetic compound libraries, or from tissue extracts, or from conditioned media of cultured cells. An increased β-gal activity after treatment indicates the activation of the orphan receptor by a ligand in the testing sample. The readout of increased β-gal activity reflects the interaction of an activated GPCR orphan receptor with a β-arrestin. Therefore, a cognate or a surrogate ligand for the testing receptor is identified. 
    
    
     DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS 
     The present invention provides a method to interrogate GPCR function and pathways. The G-protein-coupled superfamily continues to expand rapidly as new receptors are discovered through automated sequencing of cDNA libraries or genomic DNA. It is estimated that several thousand GPCRs may exist in the human genome. Only a portion have been cloned and even fewer have been associated with ligands. The means by which these, or newly discovered orphan receptors, will be associated with their cognate ligands and physiological functions represents a major challenge to biological and biomedical research. The identification of an orphan receptor generally requires an individualized assay and a guess as to its function. The present invention involves the interrogation of GPCR function by monitoring the activation of the receptor using activation dependent protein-protein interactions between the test GPCR or orphan receptor and a β-arrestin. The specific protein-protein interactions are measured using the mutant enzyme complementation technology disclosed herein. This assay system eliminates the prerequisite guessing because it can be performed with and without prior knowledge of other signaling events. It is sensitive, rapid and easily performed and is applicable to nearly all GPCRs because the majority of these receptors desensitize by a common mechanism. 
     The present invention provides a complete assay system for monitoring protein-protein interactions in GPCR pathways. The invention employs the complementation technology, ICAST™ (Intercistronic Complementation Analysis Screening Technology as disclosed in pending U.S. patent application Ser. No. 053,614, filed Apr. 1, 1998, the entire contents of which are incorporated herein by reference). The ICAST™ technology involves the use of two mutant forms of a reporter enzyme fused to proteins of interest. When the proteins of interest do not interact, the reporter enzyme remains inactive. When the proteins of interest do interact, the reporter enzyme mutants come together and form an active enzyme. According to an embodiment of the invention, the activity of β-galactosidase may be detected with the Gal-Screen™ assay system developed by Advanced Discovery Sciences™, which involves the use of Galacton-Star®, an ultrasensitive chemiluminescent substrate. The Gal-Screen™ assay system and the Galacton-Star® chemiluminescent substrate are disclosed in U.S. Pat. Nos. 5,851,771; 5,538,847; 5,326,882; 5,145,772; 4,978,614; and 4,931,569, the contents of which are incorporated herein by reference in their entirety. The invention provides an array of assays, including GPCR binding assays, that can be achieved directly within the cellular environment in a rapid, non-radioactive assay format. The methods of the invention are an advancement over the invention disclosed in U.S. Pat. Nos. 5,891,646 and 6,110,693 and the method disclosed in Angers et al., supra., which rely on microscopic imaging or spectrometry of GPCR components as fusion with Green-fluorescent-protein. The imaging technique disclosed in U.S. Pat. Nos. 5,891,646 and 6,110,693 and spectrometry-based technique in Angers et al. are limited by low-throughput and lack of thorough quantification. 
     The assay system of the invention combined with Advanced Discovery Sciences™ technologies provide highly sensitive cell-based methods for interrogating GPCR pathways which are amenable to high-throughput screening (HTS). Among some of the technologies developed by Advanced Discovery Sciences™ that may be used with the present invention are the Gal-Screen™ assay system (discussed above) and the cAMP-Screen™ immunoassay system. The cAMP-Screen™ immunoassay system provides ultrasensitive determination of cAMP levels in cell lysates. The cAMP-Screen™ assay utilizes the high-sensitivity chemiluminescent alkaline phosphatase (AP) substrate CSPD® (disodium 3-(4-methoxyspiro {1,2-dioxetane-3,2′-(5′-chloro) tricyclo 3.3.1.1. 3,7 } decan-4-yl phenyl phosphate) with Sapphire-II™ luminescence enhancer. 
     Unlike yeast-based-two-hybrid assays used to monitor protein/protein interactions in high-throughput assays, the present invention (1) is applicable to a variety of cells including mammalian cells, plant cells, protozoa cells such as  E. coli  and cells of invertebrate origin such as yeast, slime mold (Dictyostelium) and insects; (2) detects interactions at the membrane at the site of the receptor target or in the cytosol at the site of downstream target proteins rather than a limited cellular localization, i.e., nucleus; and (3) does not rely on indirect read-outs such as transcriptional activation. The present invention thus provides assays with greater physiological relevance and fewer false positives. 
     The present inventors have developed modifications to the embodiment disclosed in U.S. patent application Ser. No. 053,614 described above in order to enhance the sensitivity of the inventive GPCR assay. According to an embodiment, the invention incorporates the use of serine/threonine clusters to enhance and prolong the interaction of GPCR with arrestin in order to make the detection more robust. The clusters can be utilized for orphan receptors or known GPCRs, which do not have this sequence motif. By adding this sequence to the C-terminal tail of the receptor, the activation of the receptor can be detected more readily by readouts of arrestin binding to GPCR, i.e., β-galactosidase complementation from fusion proteins of target proteins with β-galactosidase mutants. 
     According to another embodiment, the invention incorporates the use of arrestin point mutations to bypass the requirement of phosphorylation, by the action of specific GRK, on the C-terminal tail or intracellular loops of GPCR upon activation. The applications include i) wherein the cognate GRK for a particular GPCR or orphan receptor is unknown; and ii) wherein the specific GRK for the receptor of interest (or under test) may not be present or may have low activity in the host cell that is used for receptor activation assay. 
     According to another embodiment, the invention incorporates the use of a super arrestin to increase the binding efficiency of arrestin to an activated GPCR and to stabilize the GPCR/arrestin complex during GPCR desensitization. This application can be used to increase the robustness of ICAST/GPCR applications in cases where the GPCR is normally resensitized rapidly post desensitization. 
     Each of these methodologies is discussed below. 
     The invention will now be described in the following non-limiting examples. 
     EXAMPLE 
     According to an embodiment of the invention, GPCR activation is measured through monitoring the binding of arrestin to ligand-activated GPCR. In this assay system, a GPCR, e.g., β-adrenergic receptor (β2AR), and an arrestin, e.g., β-arrestin, are co-expressed in the same cell as fusion proteins with mutant forms of a reporter enzyme, e.g., β-galactosidase (β-gal). As illustrated in FIG. 23, the β2AR is expressed as a fusion protein with Δα form of β-gal mutant (β2ARΔα) and the β-arrestin as a fusion protein with the Δω form of β-gal mutant (β-ArrΔω). The two fusion proteins, which at first exist in a resting (or un-stimulated) cell in separate compartments, i.e., the membrane for GPCR and the cytosol for arrestin, cannot form an active β-galactosidase enzyme. When such a cell is treated with an agonist or a ligand, the ligand-occupied and activated receptor becomes a high affinity binding site for arrestin. The interaction between an activated GPCR, β2ARΔα, and arrestin, β-ArrΔω, drives the β-gal mutant complementation. The enzyme activity can be measured by using an enzyme substrate, which upon cleavage releases a product measurable by colorimetry, fluorescence, or chemiluminescence (e.g., the Gal-Screen™ assay system). 
     Experiment Protocol 
     1. In the first step, the expression vectors for β2ARΔα and βArr2Δω were engineered in selectable retroviral vectors pICAST ALC, as described in FIG.  18  and pICAST OMC, as described in FIG.  15 . 
     2. In the second step, the two expression constructs were transduced into either C2C12 myoblast cells, or other mammalian cell lines, such as COS-7, CHO, A431, HEK 293, and CHW. Following selection with antibiotic drugs, stable clones expressing both fusion proteins at appropriate levels were selected. 
     3. In the last step, the cells expressing both β2ARΔα and βArr2Δω were tested for response by agonist/ligand stimulated β-galactosidase activity. Triplicate samples of cells were plated at 10,000 cells in 100 microliter volume into a well of 96-well culture plate. Cells were cultured for 24 hours before assay. For agonist assay (FIGS.  3  and  4 ), cells were treated with variable concentrations of agonist, for example, (−)isoproterenol, procaterol, dobutamine, terbutaline or L-L-phenylephrine for 60 min at 37° C. The induced β-galactosidase activity was measured by addition of Tropix Gal-Screen™ assay system substrate (Applied Biosystems) and luminescence measured in a Tropix TR717™ luminometer (Applied Biosystems). For antagonist assay (FIG.  5 ), cells were pre-incubated for 10 min in fresh medium without serum in the presence of ICI-118,551 or propranolol followed by addition of 10 micro molar (−)isoproterenol. 
     Serine/Threonine Cluster Strategy 
     Background 
     Based on structure-function relationship studies on β-arrestins, a large region within the amino-terminal half of β-arrestins (termed the activation-recognition domain) recognizes the agonist-activated state of GPCRs. This region of β-arrestin also contains a small positively charged domain (approximately 20 amino acids with net charge +7) called the phosphorylation-recognition domain, which appears to interact with the GRK-phosphorylated carboxyl termini of GPCRs. 
     GPCRs can be divided into two classes based on their affinities for β-arrestins. Oakley et al., “Association of β-Arrestin with G Protein-Coupled Receptors During Clathrin-Mediated Endocytosis Dictates the Profile of Receptor Resensitization.” J. Biol. Chem., 274(45):32248-32257 (1999). The molecular determinants underlying this classification appear to reside in specific serine or threonine residues located in the carboxyl-terminal tail of the receptor. The receptor class that contains serine/threonine clusters (defined as serine or threonine residues occupying three consecutive or three out of four positions) in the carboxyl-termini binds β-arrestin with high affinity upon activation and phosphorylation and remains bound with β-arrestin even after receptor internalization, whereas the receptor class that contains only scattered serine and threonine residues in the carboxy-terminal tail binds β-arrestins with less affinity and disassociates from the β-arrestin upon internalization. Several known GPCRs, such as vasopressin V2 receptor (Oakley, et al.), neurotensin receptor 1 and angiotensin II receptor type 1A (Zhang, et al., “Cellular Trafficking of G Protein-Coupled Receptor/β-Arrestin Endocytic Complexes.” J. Biol. Chem., 274(16):10999-11006 (1999)), which possess one or more of such serine/threonine clusters in their carboxyl-termini, were shown to bind β-arrestins with high affinity. 
     EXAMPLE 
     According to an embodiment of the invention, a serine/threonine cluster strategy is used to facilitate screening assays for orphan receptors that do not possess this structural motif of their own. The orphan receptors are easily classified by sequence alignment. Orphan receptors lacking the serine/threonine clusters are each cloned into an expression vector that is modified to introduce one or more serine/threonine cluster(s) to the carboxyl-terminal tail of the receptor (FIG.  24 ). The serine/threonine clusters enhance the receptor activation dependent interaction between the activated and phosphorylated receptor (negative charges) and β-arrestin (positive charges in the phosphorylation-recognition domain) through strong ionic interactions, thus prolonging interaction between the receptor and arrestin. The modification of the orphan receptor tail thus makes detection of receptor activation more robust. 
     Experiment Protocol 
     1. In a first step, the open-reading-frame (ORF) of an orphan receptor, which lacks the serine/threonine clusters, is cloned into a modified expression vector such as pICAST ALC described in FIG.  10 A. The modified pICAST ALC includes coding sequences for one or more sets of serine/threonine clusters (for example, SSS or SST) located downstream from the insert of the ORF of an orphan receptor (FIG.  24 ). 
     2. In a second step, chimeric orphan receptor, ORF orphan R -(SSS) n -Δα, is co-expressed in a mammalian cell with a β-arrestin chimera, such as βArr2Δω described in FIG.  15 . 
     3. In a third step, the cell is treated with an agonist or a ligand and the activated receptor with phosphorylated serine cluster(s) binds the β-arrestin with high affinity producing strong signals in readouts of β-gal complementation. 
     This assay, which provides a means for sensitive measurement of functional activation of the orphan receptors, can be used to screen for natural or surrogate ligands for orphan receptors, a process called de-orphaning or target discovery for new GPCRs (FIG.  28 ). Furthermore, this assay is also useful in screening for potential agonists and antagonists for lead discovery of GPCRs. 
     Enhanced Binding of Arrestin in the Presence and in the Absence of GPCR Phosphorylation 
     Background 
     Six different classes of G-protein coupled receptor kinases (GRKs) have been identified and each of these has been reported to be expressed as multiple splice variants. Krupnick et al., “The role of receptor kinases and arrestins in G protein-coupled receptor regulation.” Ann. Rev. Pharmacol. Toxicol., 38:289-319 (1998). Although many cell lines express a variety of GRKs, the specific GRK required for phosphorylation of a given GPCR may not always be present in the cell line used for recombinant GPCR and arrestin expression. This is particularly an issue for applications using orphan receptors, in which case the cognate GRK will likely be unknown. In other cases, the cell line used for recombinant expression work may have the required GRK, but may express the GRK at low levels. In order to bypass such caveats, genetically modified arrestins that bind specifically to activated GPCRs, but without the requirement of GRK phosphorylation are employed. 
     Mutagenesis studies on arrestins demonstrate that point mutations in the phosphorylation-recognition domain, particularly mutations converting Arg175 (of visual arrestin) to an oppositely charged residue such as glutamate (R175E mutation), result in an arrestin which specifically binds to activated GPCRs, but does so without the requirement for phosphorylation. 
     Numerous observations have led to the hypothesis that arrestin exists in an inactive state that has a low affinity for GPCRs. Once a GPCR is both activated and phosphorylated, the phosphorylated region of the GPCR C-terminus interacts with the phosphorylation-recognition domain of arrestin causing the arrestin to change conformations allowing the activation-recognition region to be exposed for binding to the activated/phosphorylated receptor. Vishnivetskiy et al., “How does arrestin respond to the phosphorylated state of rhodopsin?” J. Biol. Chem., 274(17):11451-11454 (1999); Gurevich et al., “Arrestin interactions with G protein-coupled receptors. Direct binding studies of wild-type and mutant arrestins with rhodopsin, beta 2-adrenergic and m2 muscarinic cholinergic receptors.” J. Biol. Chem., 270(2):720-731, (1995); Gurevich et al., “Mechanism of phosphorylation-recognition by visual arrestin and the transition of arrestin into a high affinity binding site.” Mol. Pharnacol., 51(1):161-169 (1997); Kovoor et al., “Targeted construction of phosphorylation-independent beta-arrestin mutants with constitutive activity in cells.” J. Biol. Chem., 274(11):6831-6834 (1999). In summary, binding studies of single mutation, double mutation, deletion, and chimerical arrestins with inactive, inactive and phosphorylated, activated but not phosphorylated, or activated and phosphorylated visual or non-visual GPCRs all support this model. 
     EXAMPLE 
     A phosphorylation insensitive mutant of arrestin fused to mutant reporter protein can be produced that will bind to activated GPCRs in a phosphorylation independent manner. As proof of concept, a point mutation for β-arrestin2, R170E β-arrestin2, has been produced and its interaction with β2AR has been analyzed in accordance with the invention. 
     Experimental Protocol: 
     1) In the first step, β-arrestin2 was mutated such that Arg170 was converted to Glu. This mutation is equivalent to the R175E mutation of visual arrestin. The mutant β-arrestin2 open reading frame was cloned in frame with Δω-β-galactosidase in the pICAST OMC expression vector to produce a modified expression vector R170E β-arrestin2 (FIG.  25 ). 
     2) In the second step, the R170E β-arrestin2 expression construct was transduced into a C2C12 myoblast cell line that had been engineered to express β2AR as a fusion to Δα-β-galactosidase as described in FIG. 18 of U.S. application Ser. No. 09/654,499. Following selection with antibiotic drugs, a population of clones expressing both fusion proteins was obtained. 
     3) In the last step, this population of cells expressing both R170E β-arrestin2Δω and β2AR Δα were tested for response by agonist/ligand stimulated β-galactosidase activity as demonstrated in FIG.  26 . The C2C12 clone 43-8 co-expressing β2ARΔα and wild-type 1β-arrestin2Δω (FIG. 26) was used as reference control. Triplicate samples of cells were plated at 10,000 cells in 100 microliter volume into wells of a 96-well culture plate. Cells were cultured for 24 hours before assay. For agonist assay as in FIG. 26, cells were treated with 10 μm (−)isoproterenol stabilized with 0.3 mM ascorbic acid 37° C. for 0, 5, 10, 15, 30, 45 or 60 minutes. The induced β-galactosidase activity was measured by addition of Tropix Gal-Screen™ assay system substrate (Applied Biosystems) and luminescence measured in a Tropix TR717™ luminometer (Applied Biosystems). As shown in FIG. 26, the mutant arrestin interacts with β2AR in an agonist-dependent manner and was comparable with that of wild-type arrestin. 
     4) To expand the application of phosphorylation-insensitive arrestin, cell lines such as C2C12, CHO or HEK 293, are developed that express the R170E β-arrestin2Δω construction. These cell lines can be used to transduce orphan or known GPCRs as fusions with Δα-β-galactosidase in order to develop cell lines for agonist and antagonist screening and 
     Development of Super Arrestins: 
     Background 
     Attenuation of GPCR signaling by the arrestin pathway serves to ensure that a cell or organism does not over-react to a stimulus. At the same time, the arrestin pathway often serves to recycle the GPCR such that it can be temporarily inactivated but then quickly resensitized to allow for sensitivity to new stimuli. The down-regulation process involves phosphorylation of the receptor, binding to arrestin and endocytosis. Following endocytosis of the desensitized receptor, the receptor is either degraded in lysosomes or resensitized and sent back to the membrane. Resensitization involves release of arrestin from the receptor, dephosphorylation and cycling back to the membrane. The actual route a GPCR follows upon activation depends on its biological function and the needs of the organism. Because of these diverse pathways that may be required of the down-regulation pathway, arrestin affinities for activated GPCRs vary from receptor to receptor. It would thus be very advantageous to engineer super arrestins that have a higher affinity and avidity for activated GPCRs than what nature has provided. 
     Although mutational, deletion and chimerical studies of arrestins have focused on understanding regulatory switches in the molecule that respond to GPCR phosphorylation states, several of these altered recombinant forms of arrestin have resulted in molecules with enhanced binding to activated, phosphorylated GPCRs. Conversion of Arg175 to histidine, tyrosine, phenylalanine or threonine results in significantly higher amounts of binding to phosphorylated, activated rhodopsin than wild-type arrestin or R175E arrestin, although these mutations result in less binding to activated, non-phosphorylated receptor. Gurevich et al. (1997). In addition, conversion of Valine 170 to alanine increased the constitutive affect of the R175E mutation, but also nearly doubled the amount of interaction of wild-type arrestin with activated, phosphorylated rhodopsin. Gurevich et al. (1997). 
     Truncation of β-arrestin1 at amino acid 382 has been reported to enhance binding of both R169E (equivalent to arrestin R175E) and wild-type β-arrestin1 to activated or activated and phosphorylated receptor, respectively. Kovoor et al. Chimerical arrestins in which functional regions of visual arrestin were swapped with those of β-arrestin1 have been reported to be altered in binding affinity to activated, phosphorylated GPCRs. Gurevich et al. (1995). Several of these chimeras, such as β-arrestin1 containing the visual arrestin extreme N-terminus, show increased specific binding to phosphorylated activated GPCRs compared to wild-type β-arrestin1 (Gurevich et al. (1995)). Modifications that enhance arrestin affinity for the activated GPCR such as described above, whether phosphorylated or non-phosphorylated, could also enhance signal to noise of β-galactosidase activity since the arrestin/GPCR complex is stabilized and/or more long-lived. The use of mutant arrestins with higher activated-GPCR affinity would improve the inventive technology for GPCR targets, without compromising receptor/ligand biology. 
     In addition, this “super arrestin” approach can be combined with the use of arrestin point mutations to provide a stronger signal to noise with or without GRK requirements. 
     EXAMPLE 
     An arrestin mutant fused to mutant reporter protein can be produced to enhance binding of the arrestin to an activated GPCR to enhance sensitivity of detection. 
     Experiment Protocol 
     1) In the first step, mutant β-arrestin2 constructions will be generated which include R170E/T/Y/or H, V165A, substitution of a.a. 1-43 with a.a. 1-47 of visual arrestin, or deletion of the C-terminal and combinations of these alterations. The mutant β-arrestin2 open reading frames will be cloned in frame with Δω-β-galactosidase in the pICAST OMC expression vector similar to cloning of the R170E β-arrestin2 mutation shown in FIG.  25 . 
     2) In the second step, mutant expression constructs will be transduced into a C2C12 myoblast cell line that has been engineered to express β2AR as a fusion to Δα-β-galactosidase. Following selection with antibiotic drugs, a population of clones expressing both fusion proteins will be obtained. Wild type and R170E β-arrestin2 constructions will be transduced to generate control, reference clonal populations. 
     3) In the third step, populations of cells expressing both β-arrestin2Δω (mutant or wild type) and β2ARΔα will be tested for response by agonist/ligand stimulated β-galactosidase activity. 
     4) In the next step, mutant (super) β-arrestin2Δω constructions that show a significantly higher signal to noise ratio in the agonist assay compared with wild-type β-arrestin2Δω will be chosen. These constructions will be used to develop stable cell lines expressing the “super” β-arrestin2Δω that can be used for transducing in known or orphan GPCRs. Use of a super β-arrestin2Δω could increase the signal to noise of ICAST/GPCR applications allowing improved screening capabilities for lead and ligand discovery. 
     Super Arrestin is used to increase the binding efficiency of arrestin to an activated GPCR and to stabilize the GPCR/arrestin complex during GPCR desensitization. This application can be used to increase the robustness of ICAST/GPCR applications in cases where the GPCR is normally resensitized rapidly post desensitization. 
     The assays of this invention, and their application and preparation have been described both generically, and by specific example. The examples are not intended as limiting. Other substituent identities, characteristics and assays will occur to those of ordinary skill in the art, without the exercise of inventive faculty. Such modifications remain within the scope of the invention, unless excluded by the express recitation of the claims advanced below. 
     
       
         
           
             10 
           
           
             1 
             6700 
             DNA 
             Unknown 
             
               pICAST ALC. 
             
           
            1
ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca     60
gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt    120
tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag    180
tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc    240
ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga    300
gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc    360
gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc    420
tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt    480
gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg    540
caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta    600
tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa    660
ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc    720
gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg    780
tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt    840
cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt    900
ccgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt    960
ttgaccttag gtaactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc   1020
aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg   1080
ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca   1140
cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct   1200
tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct   1260
ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt   1320
tatccagccc tcactccttc tctaggcgcc ggccgctcta gcccattaat acgactcact   1380
atagggcgat tcgaatcagg ccttggcgcg ccggatcctt aattaagcgc aattgggagg   1440
tggcggtagc ctcgag atg ggc gtg att acg gat tca ctg gcc gtc gtg gcc   1492
                  Met Gly Val Ile Thr Asp Ser Leu Ala Val Val Ala
                  1               5                   10
cgc acc gat cgc cct tcc caa cag tta cgc agc ctg aat ggc gaa tgg     1540
Arg Thr Asp Arg Pro Ser Gln Gln Leu Arg Ser Leu Asn Gly Glu Trp
        15                  20                  25
cgc ttt gcc tgg ttt ccg gca cca gaa gcg gtg ccg gaa agc tgg ctg     1588
Arg Phe Ala Trp Phe Pro Ala Pro Glu Ala Val Pro Glu Ser Trp Leu
    30                  35                  40
gag tgc gat ctt cct gag gcc gat act gtc gtc gtc ccc tca aac tgg     1636
Glu Cys Asp Leu Pro Glu Ala Asp Thr Val Val Val Pro Ser Asn Trp
45                  50                  55                  60
cag atg cac ggt tac gat gcg ccc atc tac acc aac gtg acc tat ccc     1684
Gln Met His Gly Tyr Asp Ala Pro Ile Tyr Thr Asn Val Thr Tyr Pro
                65                  70                  75
att acg gtc aat ccg ccg ttt gtt ccc acg gag aat ccg acg ggt tgt     1732
Ile Thr Val Asn Pro Pro Phe Val Pro Thr Glu Asn Pro Thr Gly Cys
            80                  85                  90
tac tcg ctc aca ttt aat gtt gat gaa agc tgg cta cag gaa ggc cag     1780
Tyr Ser Leu Thr Phe Asn Val Asp Glu Ser Trp Leu Gln Glu Gly Gln
        95                  100                 105
acg cga att att ttt gat ggc gtt aac tcg gcg ttt cat ctg tgg tgc     1828
Thr Arg Ile Ile Phe Asp Gly Val Asn Ser Ala Phe His Leu Trp Cys
    110                 115                 120
aac ggg cgc tgg gtc ggt tac ggc cag gac agt cgt ttg ccg tct gaa     1876
Asn Gly Arg Trp Val Gly Tyr Gly Gln Asp Ser Arg Leu Pro Ser Glu
125                 130                 135                 140
ttt gac ctg agc gca ttt tta cgc gcc gga gaa aac cgc ctc gcg gtg     1924
Phe Asp Leu Ser Ala Phe Leu Arg Ala Gly Glu Asn Arg Leu Ala Val
                145                 150                 155
atg gtg ctg cgc tgg agt gac ggc agt tat ctg gaa gat cag gat atg     1972
Met Val Leu Arg Trp Ser Asp Gly Ser Tyr Leu Glu Asp Gln Asp Met
            160                 165                 170
tgg cgg atg agc ggc att ttc cgt gac gtc tcg ttg ctg cat aaa ccg     2020
Trp Arg Met Ser Gly Ile Phe Arg Asp Val Ser Leu Leu His Lys Pro
        175                 180                 185
act aca caa atc agc gat ttc cat gtt gcc act cgc ttt aat gat gat     2068
Thr Thr Gln Ile Ser Asp Phe His Val Ala Thr Arg Phe Asn Asp Asp
    190                 195                 200
ttc agc cgc gct gta ctg gag gct gaa gtt cag atg tgc ggc gag ttg     2116
Phe Ser Arg Ala Val Leu Glu Ala Glu Val Gln Met Cys Gly Glu Leu
205                 210                 215                 220
cgt gac tac cta cgg gta aca gtt tct tta tgg cag ggt gaa acg cag     2164
Arg Asp Tyr Leu Arg Val Thr Val Ser Leu Trp Gln Gly Glu Thr Gln
                225                 230                 235
gtc gcc agc ggc acc gcg cct ttc ggc ggt gaa att atc gat gag cgt     2212
Val Ala Ser Gly Thr Ala Pro Phe Gly Gly Glu Ile Ile Asp Glu Arg
            240                 245                 250
ggt ggt tat gcc gat cgc gtc aca cta cgt ctg aac gtc gaa aac ccg     2260
Gly Gly Tyr Ala Asp Arg Val Thr Leu Arg Leu Asn Val Glu Asn Pro
        255                 260                 265
aaa ctg tgg agc gcc gaa atc ccg aat ctc tat cgt gcg gtg gtt gaa     2308
Lys Leu Trp Ser Ala Glu Ile Pro Asn Leu Tyr Arg Ala Val Val Glu
    270                 275                 280
ctg cac acc gcc gac ggc acg ctg att gaa gca gaa gcc tgc gat gtc     2356
Leu His Thr Ala Asp Gly Thr Leu Ile Glu Ala Glu Ala Cys Asp Val
285                 290                 295                 300
ggt ttc cgc gag gtg cgg att gaa aat ggt ctg ctg ctg ctg aac ggc     2404
Gly Phe Arg Glu Val Arg Ile Glu Asn Gly Leu Leu Leu Leu Asn Gly
                305                 310                 315
aag ccg ttg ctg att cga ggc gtt aac cgt cac gag cat cat cct ctg     2452
Lys Pro Leu Leu Ile Arg Gly Val Asn Arg His Glu His His Pro Leu
            320                 325                 330
cat ggt cag gtc atg gat gag cag acg atg gtg cag gat atc ctg ctg     2500
His Gly Gln Val Met Asp Glu Gln Thr Met Val Gln Asp Ile Leu Leu
        335                 340                 345
atg aag cag aac aac ttt aac gcc gtg cgc tgt tcg cat tat ccg aac     2548
Met Lys Gln Asn Asn Phe Asn Ala Val Arg Cys Ser His Tyr Pro Asn
    350                 355                 360
cat ccg ctg tgg tac acg ctg tgc gac cgc tac ggc ctg tat gtg gtg     2596
His Pro Leu Trp Tyr Thr Leu Cys Asp Arg Tyr Gly Leu Tyr Val Val
365                 370                 375                 380
gat gaa gcc aat att gaa acc cac ggc atg gtg cca atg aat cgt ctg     2644
Asp Glu Ala Asn Ile Glu Thr His Gly Met Val Pro Met Asn Arg Leu
                385                 390                 395
acc gat gat ccg cgc tgg cta ccg gcg atg agc gaa cgc gta acg cga     2692
Thr Asp Asp Pro Arg Trp Leu Pro Ala Met Ser Glu Arg Val Thr Arg
            400                 405                 410
atg gtg cag cgc gat cgt aat cac ccg agt gtg atc atc tgg tcg ctg     2740
Met Val Gln Arg Asp Arg Asn His Pro Ser Val Ile Ile Trp Ser Leu
        415                 420                 425
ggg aat gaa tca ggc cac ggc gct aat cac gac gcg ctg tat cgc tgg     2788
Gly Asn Glu Ser Gly His Gly Ala Asn His Asp Ala Leu Tyr Arg Trp
    430                 435                 440
atc aaa tct gtc gat cct tcc cgc ccg gtg cag tat gaa ggc ggc gga     2836
Ile Lys Ser Val Asp Pro Ser Arg Pro Val Gln Tyr Glu Gly Gly Gly
445                 450                 455                 460
gcc gac acc acg gcc acc gat att att tgc ccg atg tac gcg cgc gtg     2884
Ala Asp Thr Thr Ala Thr Asp Ile Ile Cys Pro Met Tyr Ala Arg Val
                465                 470                 475
gat gaa gac cag ccc ttc ccg gct gtg ccg aaa tgg tcc atc aaa aaa     2932
Asp Glu Asp Gln Pro Phe Pro Ala Val Pro Lys Trp Ser Ile Lys Lys
            480                 485                 490
tgg ctt tcg cta cct gga gag acg cgc ccg ctg atc ctt tgc gaa tac     2980
Trp Leu Ser Leu Pro Gly Glu Thr Arg Pro Leu Ile Leu Cys Glu Tyr
        495                 500                 505
gcc cac gcg atg ggt aac agt ctt ggc ggt ttc gct aaa tac tgg cag     3028
Ala His Ala Met Gly Asn Ser Leu Gly Gly Phe Ala Lys Tyr Trp Gln
    510                 515                 520
gcg ttt cgt cag tat ccc cgt tta cag ggc ggc ttc gtc tgg gac tgg     3076
Ala Phe Arg Gln Tyr Pro Arg Leu Gln Gly Gly Phe Val Trp Asp Trp
525                 530                 535                 540
gtg gat cag tcg ctg att aaa tat gat gaa aac ggc aac ccg tgg tcg     3124
Val Asp Gln Ser Leu Ile Lys Tyr Asp Glu Asn Gly Asn Pro Trp Ser
                545                 550                 555
gct tac ggc ggt gat ttt ggc gat acg ccg aac gat cgc cag ttc tgt     3172
Ala Tyr Gly Gly Asp Phe Gly Asp Thr Pro Asn Asp Arg Gln Phe Cys
            560                 565                 570
atg aac ggt ctg gtc ttt gcc gac cgc acg ccg cat cca gcg ctg acg     3220
Met Asn Gly Leu Val Phe Ala Asp Arg Thr Pro His Pro Ala Leu Thr
        575                 580                 585
gaa gca aaa cac cag cag cag ttt ttc cag ttc cgt tta tcc ggg caa     3268
Glu Ala Lys His Gln Gln Gln Phe Phe Gln Phe Arg Leu Ser Gly Gln
    590                 595                 600
acc atc gaa gtg acc agc gaa tac ctg ttc cgt cat agc gat aac gag     3316
Thr Ile Glu Val Thr Ser Glu Tyr Leu Phe Arg His Ser Asp Asn Glu
605                 610                 615                 620
ctc ctg cac tgg atg gtg gcg ctg gat ggt aag ccg ctg gca agc ggt     3364
Leu Leu His Trp Met Val Ala Leu Asp Gly Lys Pro Leu Ala Ser Gly
                625                 630                 635
gaa gtg cct ctg gat gtc gct cca caa ggt aaa cag ttg att gaa ctg     3412
Glu Val Pro Leu Asp Val Ala Pro Gln Gly Lys Gln Leu Ile Glu Leu
            640                 645                 650
cct gaa cta ccg cag ccg gag agc gcc ggg caa ctc tgg ctc aca gta     3460
Pro Glu Leu Pro Gln Pro Glu Ser Ala Gly Gln Leu Trp Leu Thr Val
        655                 660                 665
cgc gta gtg caa ccg aac gcg acc gca tgg tca gaa gcc ggg cac atc     3508
Arg Val Val Gln Pro Asn Ala Thr Ala Trp Ser Glu Ala Gly His Ile
    670                 675                 680
agc gcc tgg cag cag tgg cgt ctg gcg gaa aac ctc agt gtg acg ctc     3556
Ser Ala Trp Gln Gln Trp Arg Leu Ala Glu Asn Leu Ser Val Thr Leu
685                 690                 695                 700
ccc gcc gcg tcc cac gcc atc ccg cat ctg acc acc agc gaa atg gat     3604
Pro Ala Ala Ser His Ala Ile Pro His Leu Thr Thr Ser Glu Met Asp
                705                 710                 715
ttt tgc atc gag ctg ggt aat aag cgt tgg caa ttt aac cgc cag tca     3652
Phe Cys Ile Glu Leu Gly Asn Lys Arg Trp Gln Phe Asn Arg Gln Ser
            720                 725                 730
ggc ttt ctt tca cag atg tgg att ggc gat aaa aaa caa ctg ctg acg     3700
Gly Phe Leu Ser Gln Met Trp Ile Gly Asp Lys Lys Gln Leu Leu Thr
        735                 740                 745
ccg ctg cgc gat cag ttc acc cgt gca ccg ctg gat aac gac att ggc     3748
Pro Leu Arg Asp Gln Phe Thr Arg Ala Pro Leu Asp Asn Asp Ile Gly
    750                 755                 760
gta agt gaa gcg acc cgc att gac cct aac gcc tgg gtc gaa cgc tgg     3796
Val Ser Glu Ala Thr Arg Ile Asp Pro Asn Ala Trp Val Glu Arg Trp
765                 770                 775                 780
aag gcg gcg ggc cat tac cag gcc gaa gca gcg ttg ttg cag tgc acg     3844
Lys Ala Ala Gly His Tyr Gln Ala Glu Ala Ala Leu Leu Gln Cys Thr
                785                 790                 795
gca gat aca ctt gct gat gcg gtg ctg att acg acc gct cac gcg tgg     3892
Ala Asp Thr Leu Ala Asp Ala Val Leu Ile Thr Thr Ala His Ala Trp
            800                 805                 810
cag cat cag ggg aaa acc tta ttt atc agc cgg aaa acc tac cgg att     3940
Gln His Gln Gly Lys Thr Leu Phe Ile Ser Arg Lys Thr Tyr Arg Ile
        815                 820                 825
gat ggt agt ggt caa atg gcg att acc gtt gat gtt gaa gtg gcg agc     3988
Asp Gly Ser Gly Gln Met Ala Ile Thr Val Asp Val Glu Val Ala Ser
    830                 835                 840
gat aca ccg cat ccg gcg cgg att ggc ctg aac tgc cag ctg gcg cag     4036
Asp Thr Pro His Pro Ala Arg Ile Gly Leu Asn Cys Gln Leu Ala Gln
845                 850                 855                 860
gta gca gag cgg gta aac tgg ctc gga tta ggg ccg caa gaa aac tat     4084
Val Ala Glu Arg Val Asn Trp Leu Gly Leu Gly Pro Gln Glu Asn Tyr
                865                 870                 875
ccc gac cgc ctt act gcc gcc tgt ttt gac cgc tgg gat ctg cca ttg     4132
Pro Asp Arg Leu Thr Ala Ala Cys Phe Asp Arg Trp Asp Leu Pro Leu
            880                 885                 890
tca gac atg tat acc ccg tac gtc ttc ccg agc gaa aac ggt ctg cgc     4180
Ser Asp Met Tyr Thr Pro Tyr Val Phe Pro Ser Glu Asn Gly Leu Arg
        895                 900                 905
tgc ggg acg cgc gaa ttg aat tat ggc cca cac cag tgg cgc ggc gac     4228
Cys Gly Thr Arg Glu Leu Asn Tyr Gly Pro His Gln Trp Arg Gly Asp
    910                 915                 920
ttc cag ttc aac atc agc cgc tac agt caa cag caa ctg atg gaa acc     4276
Phe Gln Phe Asn Ile Ser Arg Tyr Ser Gln Gln Gln Leu Met Glu Thr
925                 930                 935                 940
agc cat cgc cat ctg ctg cac gcg gaa gaa ggc aca tgg ctg aat atc     4324
Ser His Arg His Leu Leu His Ala Glu Glu Gly Thr Trp Leu Asn Ile
                945                 950                 955
gac ggt ttc cat atg ggg att ggt ggc gac gac tcc tgg agc ccg tca     4372
Asp Gly Phe His Met Gly Ile Gly Gly Asp Asp Ser Trp Ser Pro Ser
            960                 965                 970
gta tcg gcg gaa ttc cag ctg agc gcc ggt cgc tac cat tac cag ttg     4420
Val Ser Ala Glu Phe Gln Leu Ser Ala Gly Arg Tyr His Tyr Gln Leu
        975                 980                 985
gtc tgg tgt caa aaa aga tct gac tat aaa gat gag  gac ctc gac cat    4468
Val Trp Cys Gln Lys Arg Ser Asp Tyr Lys Asp Glu  Asp Leu Asp His
    990                 995                 1000
cat  cat cat cat cac cgg  taataatagg tagataagtg actgattaga          4516
His  His His His His Arg
1005                 1010
tgcattgatc cctcgaccaa ttccggttat tttccaccat attgccgtct tttggcaatg   4576
tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc   4636
tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt   4696
cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg   4756
acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac   4816
cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg   4876
tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg   4936
ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc   4996
gaaccacggg gacgtggttt tcctttgaaa aacacgatga taataccatg attgaacaag   5056
atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg   5116
cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc   5176
cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag   5236
cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca   5296
ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat   5356
ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata   5416
cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac   5476
gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc   5536
tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg   5596
tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg   5656
gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta   5716
cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg   5776
gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct   5836
gagcgggact ctggggttcg catcgataaa ataaaagatt ttatttagtc tccagaaaaa   5896
ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg   5956
caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga   6016
tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc   6076
agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag   6136
ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca   6196
gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc   6256
cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg   6316
agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt   6376
cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg   6436
ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt   6496
catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc   6556
atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggcgctct   6616
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca   6676
gctcactcaa aggcggtaat acgg                                          6700
 
           
             2 
             1010 
             PRT 
             Unknown 
             
               pICAST ALC. 
             
           
            2
Met Gly Val Ile Thr Asp Ser Leu Ala Val Val Ala Arg Thr Asp Arg
1               5                   10                  15
Pro Ser Gln Gln Leu Arg Ser Leu Asn Gly Glu Trp Arg Phe Ala Trp
            20                  25                  30
Phe Pro Ala Pro Glu Ala Val Pro Glu Ser Trp Leu Glu Cys Asp Leu
        35                  40                  45
Pro Glu Ala Asp Thr Val Val Val Pro Ser Asn Trp Gln Met His Gly
    50                  55                  60
Tyr Asp Ala Pro Ile Tyr Thr Asn Val Thr Tyr Pro Ile Thr Val Asn
65                  70                  75                  80
Pro Pro Phe Val Pro Thr Glu Asn Pro Thr Gly Cys Tyr Ser Leu Thr
                85                  90                  95
Phe Asn Val Asp Glu Ser Trp Leu Gln Glu Gly Gln Thr Arg Ile Ile
            100                 105                 110
Phe Asp Gly Val Asn Ser Ala Phe His Leu Trp Cys Asn Gly Arg Trp
        115                 120                 125
Val Gly Tyr Gly Gln Asp Ser Arg Leu Pro Ser Glu Phe Asp Leu Ser
    130                 135                 140
Ala Phe Leu Arg Ala Gly Glu Asn Arg Leu Ala Val Met Val Leu Arg
145                 150                 155                 160
Trp Ser Asp Gly Ser Tyr Leu Glu Asp Gln Asp Met Trp Arg Met Ser
                165                 170                 175
Gly Ile Phe Arg Asp Val Ser Leu Leu His Lys Pro Thr Thr Gln Ile
            180                 185                 190
Ser Asp Phe His Val Ala Thr Arg Phe Asn Asp Asp Phe Ser Arg Ala
        195                 200                 205
Val Leu Glu Ala Glu Val Gln Met Cys Gly Glu Leu Arg Asp Tyr Leu
    210                 215                 220
Arg Val Thr Val Ser Leu Trp Gln Gly Glu Thr Gln Val Ala Ser Gly
225                 230                 235                 240
Thr Ala Pro Phe Gly Gly Glu Ile Ile Asp Glu Arg Gly Gly Tyr Ala
                245                 250                 255
Asp Arg Val Thr Leu Arg Leu Asn Val Glu Asn Pro Lys Leu Trp Ser
            260                 265                 270
Ala Glu Ile Pro Asn Leu Tyr Arg Ala Val Val Glu Leu His Thr Ala
        275                 280                 285
Asp Gly Thr Leu Ile Glu Ala Glu Ala Cys Asp Val Gly Phe Arg Glu
    290                 295                 300
Val Arg Ile Glu Asn Gly Leu Leu Leu Leu Asn Gly Lys Pro Leu Leu
305                 310                 315                 320
Ile Arg Gly Val Asn Arg His Glu His His Pro Leu His Gly Gln Val
                325                 330                 335
Met Asp Glu Gln Thr Met Val Gln Asp Ile Leu Leu Met Lys Gln Asn
            340                 345                 350
Asn Phe Asn Ala Val Arg Cys Ser His Tyr Pro Asn His Pro Leu Trp
        355                 360                 365
Tyr Thr Leu Cys Asp Arg Tyr Gly Leu Tyr Val Val Asp Glu Ala Asn
    370                 375                 380
Ile Glu Thr His Gly Met Val Pro Met Asn Arg Leu Thr Asp Asp Pro
385                 390                 395                 400
Arg Trp Leu Pro Ala Met Ser Glu Arg Val Thr Arg Met Val Gln Arg
                405                 410                 415
Asp Arg Asn His Pro Ser Val Ile Ile Trp Ser Leu Gly Asn Glu Ser
            420                 425                 430
Gly His Gly Ala Asn His Asp Ala Leu Tyr Arg Trp Ile Lys Ser Val
        435                 440                 445
Asp Pro Ser Arg Pro Val Gln Tyr Glu Gly Gly Gly Ala Asp Thr Thr
    450                 455                 460
Ala Thr Asp Ile Ile Cys Pro Met Tyr Ala Arg Val Asp Glu Asp Gln
465                 470                 475                 480
Pro Phe Pro Ala Val Pro Lys Trp Ser Ile Lys Lys Trp Leu Ser Leu
                485                 490                 495
Pro Gly Glu Thr Arg Pro Leu Ile Leu Cys Glu Tyr Ala His Ala Met
            500                 505                 510
Gly Asn Ser Leu Gly Gly Phe Ala Lys Tyr Trp Gln Ala Phe Arg Gln
        515                 520                 525
Tyr Pro Arg Leu Gln Gly Gly Phe Val Trp Asp Trp Val Asp Gln Ser
    530                 535                 540
Leu Ile Lys Tyr Asp Glu Asn Gly Asn Pro Trp Ser Ala Tyr Gly Gly
545                 550                 555                 560
Asp Phe Gly Asp Thr Pro Asn Asp Arg Gln Phe Cys Met Asn Gly Leu
                565                 570                 575
Val Phe Ala Asp Arg Thr Pro His Pro Ala Leu Thr Glu Ala Lys His
            580                 585                 590
Gln Gln Gln Phe Phe Gln Phe Arg Leu Ser Gly Gln Thr Ile Glu Val
        595                 600                 605
Thr Ser Glu Tyr Leu Phe Arg His Ser Asp Asn Glu Leu Leu His Trp
    610                 615                 620
Met Val Ala Leu Asp Gly Lys Pro Leu Ala Ser Gly Glu Val Pro Leu
625                 630                 635                 640
Asp Val Ala Pro Gln Gly Lys Gln Leu Ile Glu Leu Pro Glu Leu Pro
                645                 650                 655
Gln Pro Glu Ser Ala Gly Gln Leu Trp Leu Thr Val Arg Val Val Gln
            660                 665                 670
Pro Asn Ala Thr Ala Trp Ser Glu Ala Gly His Ile Ser Ala Trp Gln
        675                 680                 685
Gln Trp Arg Leu Ala Glu Asn Leu Ser Val Thr Leu Pro Ala Ala Ser
    690                 695                 700
His Ala Ile Pro His Leu Thr Thr Ser Glu Met Asp Phe Cys Ile Glu
705                 710                 715                 720
Leu Gly Asn Lys Arg Trp Gln Phe Asn Arg Gln Ser Gly Phe Leu Ser
                725                 730                 735
Gln Met Trp Ile Gly Asp Lys Lys Gln Leu Leu Thr Pro Leu Arg Asp
            740                 745                 750
Gln Phe Thr Arg Ala Pro Leu Asp Asn Asp Ile Gly Val Ser Glu Ala
        755                 760                 765
Thr Arg Ile Asp Pro Asn Ala Trp Val Glu Arg Trp Lys Ala Ala Gly
    770                 775                 780
His Tyr Gln Ala Glu Ala Ala Leu Leu Gln Cys Thr Ala Asp Thr Leu
785                 790                 795                 800
Ala Asp Ala Val Leu Ile Thr Thr Ala His Ala Trp Gln His Gln Gly
                805                 810                 815
Lys Thr Leu Phe Ile Ser Arg Lys Thr Tyr Arg Ile Asp Gly Ser Gly
            820                 825                 830
Gln Met Ala Ile Thr Val Asp Val Glu Val Ala Ser Asp Thr Pro His
        835                 840                 845
Pro Ala Arg Ile Gly Leu Asn Cys Gln Leu Ala Gln Val Ala Glu Arg
    850                 855                 860
Val Asn Trp Leu Gly Leu Gly Pro Gln Glu Asn Tyr Pro Asp Arg Leu
865                 870                 875                 880
Thr Ala Ala Cys Phe Asp Arg Trp Asp Leu Pro Leu Ser Asp Met Tyr
                885                 890                 895
Thr Pro Tyr Val Phe Pro Ser Glu Asn Gly Leu Arg Cys Gly Thr Arg
            900                 905                 910
Glu Leu Asn Tyr Gly Pro His Gln Trp Arg Gly Asp Phe Gln Phe Asn
        915                 920                 925
Ile Ser Arg Tyr Ser Gln Gln Gln Leu Met Glu Thr Ser His Arg His
    930                 935                 940
Leu Leu His Ala Glu Glu Gly Thr Trp Leu Asn Ile Asp Gly Phe His
945                 950                 955                 960
Met Gly Ile Gly Gly Asp Asp Ser Trp Ser Pro Ser Val Ser Ala Glu
                965                 970                 975
Phe Gln Leu Ser Ala Gly Arg Tyr His Tyr Gln Leu Val Trp Cys Gln
            980                 985                 990
Lys Arg Ser Asp Tyr Lys Asp Glu  Asp Leu Asp His His  His His His
        995                 1000                 1005
His Arg
    1010
 
           
             3 
             6700 
             DNA 
             Unknown 
             
               pICAST ALC. 
             
           
            3
gacgtcggac ttatacccgg tttgtcctat agacaccatt cgtcaaggac ggggccgagt     60
cccggttctt gtctaccttg tcgacttata cccggtttgt cctatagaca ccattcgtca    120
aggacggggc cgagtcccgg ttcttgtcta ccaggggtct acgccaggtc gggagtcgtc    180
aaagatctct tggtagtcta caaaggtccc acggggttcc tggactttac tgggacacgg    240
aataaacttg attggttagt caagcgaaga gcgaagacaa gcgcgcgaag acgaggggct    300
cgagttattt tctcgggtgt tggggagtga gccccgcggt caggaggcta actgactcag    360
cgggcccatg ggcacatagg ttatttggga gaacgtcaac gtaggctgaa caccagagcg    420
acaaggaacc ctcccagagg agactcacta actgatgggc agtcgccccc agaaagtaaa    480
cccccgagca ggccctagcc ctctggggac gggtccctgg tggctgggtg gtggccctcc    540
gttcgaccgg tcgttgaata gacacagaca ggctaacaga tcacagatac tgactaaaat    600
acgcggacgc agccatgatc aatcgattga tcgagacata gaccgcctgg gcaccacctt    660
gactgctcaa gacttgtggg ccggcgttgg gaccctctgc agggtccctg aaacccccgg    720
caaaaacacc gggctggact ccttccctca gctacacctt aggctggggc agtcctatac    780
accaagacca tcctctgctc ttggattttg tcaagggcgg aggcagactt aaaaacgaaa    840
gccaaacctt ggcttcggcg cgcagaacag acgacgtcgt agcaagacac aacagagaca    900
gactgacaca aagacataaa cagactttta atcccggtct gacaatggtg agggaattca    960
aactggaatc cattgacctt tctacagctc gccgagcgag tgttggtcag ccatctacag   1020
ttcttctctg caacccaatg gaagacgaga cgtcttaccg gttggaaatt gcagcctacc   1080
ggcgctctgc cgtggaaatt ggctctggag tagtgggtcc aattctagtt ccagaaaagt   1140
ggaccgggcg tacctgtggg tctggtccag gggatgtagc actggaccct tcggaaccga   1200
aaactggggg gagggaccca gttcgggaaa catgtgggat tcggaggcgg aggagaagga   1260
ggtaggcggg gcagagaggg ggaacttgga ggagcaagct ggggcggagc taggagggaa   1320
ataggtcggg agtgaggaag agatccgcgg ccggcgagat cgggtaatta tgctgagtga   1380
tatcccgcta agcttagtcc ggaaccgcgc ggcctaggaa ttaattcgcg ttaaccctcc   1440
accgccatcg gagctctacc cgcactaatg cctaagtgac cggcagcacc gggcgtggct   1500
agcgggaagg gttgtcaatg cgtcggactt accgcttacc gcgaaacgga ccaaaggccg   1560
tggtcttcgc cacggccttt cgaccgacct cacgctagaa ggactccggc tatgacagca   1620
gcaggggagt ttgaccgtct acgtgccaat gctacgcggg tagatgtggt tgcactggat   1680
agggtaatgc cagttaggcg gcaaacaagg gtgcctctta ggctgcccaa caatgagcga   1740
gtgtaaatta caactacttt cgaccgatgt ccttccggtc tgcgcttaat aaaaactacc   1800
gcaattgagc cgcaaagtag acaccacgtt gcccgcgacc cagccaatgc cggtcctgtc   1860
agcaaacggc agacttaaac tggactcgcg taaaaatgcg cggcctcttt tggcggagcg   1920
ccactaccac gacgcgacct cactgccgtc aatagacctt ctagtcctat acaccgccta   1980
ctcgccgtaa aaggcactgc agagcaacga cgtatttggc tgatgtgttt agtcgctaaa   2040
ggtacaacgg tgagcgaaat tactactaaa gtcggcgcga catgacctcc gacttcaagt   2100
ctacacgccg ctcaacgcac tgatggatgc ccattgtcaa agaaataccg tcccactttg   2160
cgtccagcgg tcgccgtggc gcggaaagcc gccactttaa tagctactcg caccaccaat   2220
acggctagcg cagtgtgatg cagacttgca gcttttgggc tttgacacct cgcggcttta   2280
gggcttagag atagcacgcc accaacttga cgtgtggcgg ctgccgtgcg actaacttcg   2340
tcttcggacg ctacagccaa aggcgctcca cgcctaactt ttaccagacg acgacgactt   2400
gccgttcggc aacgactaag ctccgcaatt ggcagtgctc gtagtaggag acgtaccagt   2460
ccagtaccta ctcgtctgct accacgtcct ataggacgac tacttcgtct tgttgaaatt   2520
gcggcacgcg acaagcgtaa taggcttggt aggcgacacc atgtgcgaca cgctggcgat   2580
gccggacata caccacctac ttcggttata actttgggtg ccgtaccacg gttacttagc   2640
agactggcta ctaggcgcga ccgatggccg ctactcgctt gcgcattgcg cttaccacgt   2700
cgcgctagca ttagtgggct cacactagta gaccagcgac cccttactta gtccggtgcc   2760
gcgattagtg ctgcgcgaca tagcgaccta gtttagacag ctaggaaggg cgggccacgt   2820
catacttccg ccgcctcggc tgtggtgccg gtggctataa taaacgggct acatgcgcgc   2880
gcacctactt ctggtcggga agggccgaca cggctttacc aggtagtttt ttaccgaaag   2940
cgatggacct ctctgcgcgg gcgactagga aacgcttatg cgggtgcgct acccattgtc   3000
agaaccgcca aagcgattta tgaccgtccg caaagcagtc ataggggcaa atgtcccgcc   3060
gaagcagacc ctgacccacc tagtcagcga ctaatttata ctacttttgc cgttgggcac   3120
cagccgaatg ccgccactaa aaccgctatg cggcttgcta gcggtcaaga catacttgcc   3180
agaccagaaa cggctggcgt gcggcgtagg tcgcgactgc cttcgttttg tggtcgtcgt   3240
caaaaaggtc aaggcaaata ggcccgtttg gtagcttcac tggtcgctta tggacaaggc   3300
agtatcgcta ttgctcgagg acgtgaccta ccaccgcgac ctaccattcg gcgaccgttc   3360
gccacttcac ggagacctac agcgaggtgt tccatttgtc aactaacttg acggacttga   3420
tggcgtcggc ctctcgcggc ccgttgagac cgagtgtcat gcgcatcacg ttggcttgcg   3480
ctggcgtacc agtcttcggc ccgtgtagtc gcggaccgtc gtcaccgcag accgcctttt   3540
ggagtcacac tgcgaggggc ggcgcagggt gcggtagggc gtagactggt ggtcgcttta   3600
cctaaaaacg tagctcgacc cattattcgc aaccgttaaa ttggcggtca gtccgaaaga   3660
aagtgtctac acctaaccgc tattttttgt tgacgactgc ggcgacgcgc tagtcaagtg   3720
ggcacgtggc gacctattgc tgtaaccgca ttcacttcgc tgggcgtaac tgggattgcg   3780
gacccagctt gcgaccttcc gccgcccggt aatggtccgg cttcgtcgca acaacgtcac   3840
gtgccgtcta tgtgaacgac tacgccacga ctaatgctgg cgagtgcgca ccgtcgtagt   3900
ccccttttgg aataaatagt cggccttttg gatggcctaa ctaccatcac cagtttaccg   3960
ctaatggcaa ctacaacttc accgctcgct atgtggcgta ggccgcgcct aaccggactt   4020
gacggtcgac cgcgtccatc gtctcgccca tttgaccgag cctaatcccg gcgttctttt   4080
gatagggctg gcggaatgac ggcggacaaa actggcgacc ctagacggta acagtctgta   4140
catatggggc atgcagaagg gctcgctttt gccagacgcg acgccctgcg cgcttaactt   4200
aataccgggt gtggtcaccg cgccgctgaa ggtcaagttg tagtcggcga tgtcagttgt   4260
cgttgactac ctttggtcgg tagcggtaga cgacgtgcgc cttcttccgt gtaccgactt   4320
atagctggca aaggtatacc cctaaccacc gctgctgagg acctcgggca gtcatagccg   4380
ccttaaggtc gactcgcggc cagcgatggt aatggtcaac cagaccacag ttttttctag   4440
actgatattt ctactcctgg agctggtagt agtagtagta gtggccatta ttatccatct   4500
attcactgac taatctacgt aactagggag ctggttaagg ccaataaaag gtggtataac   4560
ggcagaaaac cgttacactc ccgggccttt ggaccgggac agaagaactg ctcgtaagga   4620
tccccagaaa ggggagagcg gtttccttac gttccagaca acttacagca cttccttcgt   4680
caaggagacc ttcgaagaac ttctgtttgt tgcagacatc gctgggaaac gtccgtcgcc   4740
ttggggggtg gaccgctgtc cacggagacg ccggttttcg gtgcacatat tctatgtgga   4800
cgtttccgcc gtgttggggt cacggtgcaa cactcaacct atcaacacct ttctcagttt   4860
accgagagga gttcgcataa gttgttcccc gacttcctac gggtcttcca tggggtaaca   4920
taccctagac tagaccccgg agccacgtgt acgaaatgta cacaaatcag ctccaatttt   4980
ttgcagatcc ggggggcttg gtgcccctgc accaaaagga aactttttgt gctactatta   5040
tggtactaac ttgttctacc taacgtgcgt ccaagaggcc ggcgaaccca cctctccgat   5100
aagccgatac tgacccgtgt tgtctgttag ccgacgagac tacggcggca caaggccgac   5160
agtcgcgtcc ccgcgggcca agaaaaacag ttctggctgg acaggccacg ggacttactt   5220
gacgtcctgc tccgtcgcgc cgatagcacc gaccggtgct gcccgcaagg aacgcgtcga   5280
cacgagctgc aacagtgact tcgcccttcc ctgaccgacg ataacccgct tcacggcccc   5340
gtcctagagg acagtagagt ggaacgagga cggctctttc ataggtagta ccgactacgt   5400
tacgccgccg acgtatgcga actaggccga tggacgggta agctggtggt tcgctttgta   5460
gcgtagctcg ctcgtgcatg agcctacctt cggccagaac agctagtcct actagacctg   5520
cttctcgtag tccccgagcg cggtcggctt gacaagcggt ccgagttccg cgcgtacggg   5580
ctgccgctcc tagagcagca ctgggtaccg ctacggacga acggcttata gtaccacctt   5640
ttaccggcga aaagacctaa gtagctgaca ccggccgacc cacaccgcct ggcgatagtc   5700
ctgtatcgca accgatgggc actataacga cttctcgaac cgccgcttac ccgactggcg   5760
aaggagcacg aaatgccata gcggcgaggg ctaagcgtcg cgtagcggaa gatagcggaa   5820
gaactgctca agaagactcg ccctgagacc ccaagcgtag ctattttatt ttctaaaata   5880
aatcagaggt ctttttcccc ccttactttc tggggtggac atccaaaccg ttcgatcgaa   5940
ttcattgcgg taaaacgttc cgtacctttt tatgtattga ctcttatctc ttcaagtcta   6000
gttccagtcc ttgtctacct tgtcgactta tacccggttt gtcctataga caccattcgt   6060
caaggacggg gccgagtccc ggttcttgtc taccttgtcg acttataccc ggtttgtcct   6120
atagacacca ttcgtcaagg acggggccga gtcccggttc ttgtctacca ggggtctacg   6180
ccaggtcggg agtcgtcaaa gatctcttgg tagtctacaa aggtcccacg gggttcctgg   6240
actttactgg gacacggaat aaacttgatt ggttagtcaa gcgaagagcg aagacaagcg   6300
cgcgaagacg aggggctcga gttattttct cgggtgttgg ggagtgagcc ccgcggtcag   6360
gaggctaact gactcagcgg gcccatgggc acataggtta tttgggagaa cgtcaacgta   6420
ggctgaacac cagagcgaca aggaaccctc ccagaggaga ctcactaact gatgggcagt   6480
cgcccccaga aagtaagtac gtcgtacata gttttaatta aaccaaaaaa aagaattcat   6540
aaatgtaatt taccggtatc aacgtaatta cttagccggt tgcgcgcccc tctccgccaa   6600
acgcataacc gcgagaaggc gaaggagcga gtgactgagc gacgcgagcc agcaagccga   6660
cgccgctcgc catagtcgag tgagtttccg ccattatgcc                         6700
 
           
             4 
             8518 
             DNA 
             Unknown 
             
               pICAST ALN. 
             
           
            4
ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca     60
gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt    120
tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag    180
tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc    240
ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga    300
gctcaataaa agagcccaca acccgtcact cggggcgcca gtcctccgat tgactgagtc    360
gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc    420
tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt    480
gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg    540
caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta    600
tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa    660
ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc    720
gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg    780
tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt    840
cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt    900
ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt    960
ttgaccttag gtaactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc   1020
aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg   1080
ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca   1140
cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct   1200
tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct   1260
ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt   1320
tatccagccc tcactccttc tctaggcgcc ggccgctcta gcccattaat acgactcact   1380
atagggcgat tcgaacacca tgcaccatca tcatcatcac gtcgactata aagatgagga   1440
cctcgagatg ggcgtgatta cggattcact ggccgtcgtg gcccgcaccg atcgcccttc   1500
ccaacagtta cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc   1560
ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc   1620
aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac   1680
ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa   1740
tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttaactc   1800
ggcgtttcat ctgtggtgca acgggcgctg ggtcggttac ggccaggaca gtcgtttgcc   1860
gtctgaattt gacctgagcg catttttacg cgccggagaa aaccgcctcg cggtgatggt   1920
gctgggctgg agtgacggca gttatctgga agatcaggat atgtggcgga tgagcggcat   1980
tttccgtgac gtctcgttgc tgcataaacc gactacacaa atcagcgatt tccatgttgc   2040
cactcgcttt aatgatgatt rcagccgcgc tgtactggag gctgaagttc agatgtgcgg   2100
cgagttgcgt gactacctac gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc   2160
cagcggcacc gcgcctttcg gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg   2220
cgtcacacta cgtctgaacg tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct   2280
ctatcgtgcg gtggttgaac tgcacaccgc cgacggcacg ctgattgaag cagaagcctg   2340
cgatgtcggt ttccgcgagg tgcggattga aaatggtctg ctgctgctga acggcaagcc   2400
gttgctgatt cgaggcgtta accgtcacga gcatcatcct ctgcatggtc aggtcatgga   2460
tgagcagacg atggtgcagg atatcctgct gatgaagcag aacaacttta acgccgtgcg   2520
ctgttcgcat tatccgaacc atccgctgtg gtacacgctg tgcgaccgct acggcctgta   2580
tgtggtggat gaagccaata ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga   2640
tgatccgcgc tggctaccgg cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg   2700
taatcacccg agtgtgatca tctggtcgct ggggaatgaa tcaggccacg gcgctaatca   2760
cgacgcgctg tatcgctgga tcaaatctgt cgatccttcc cgcccggtgc agtatgaagg   2820
cggcggagcc gacaccacgg ccaccgatat tatttgcccg atgtacgcgc gcgtggatga   2880
agaccagccc ttcccggctg tgccgaaatg gtccatcaaa aaatggcttt cgctacctgg   2940
agagacgcgc ccgctgatcc tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg   3000
tttcgctaaa tactggcagg cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg   3060
ggactgggtg gatcagtcgc tgattaaata tgatgaaaac ggcaacccgt ggtcggctta   3120
cggcggtgat tttggcgata cgccgaacga tcgccagttc tgtatgaacg gtctggtctt   3180
tgccgaccgc acgccgcatc cagcgctgac ggaagcaaaa caccagcagc agtttttcca   3240
gttccgttta tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga   3300
taacgagctc ctgcactgga tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt   3360
gcctctggat gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc   3420
ggagagcgcc gggcaactct ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg   3480
gtcagaagcc gggcacatca gcgcctggca gcagtggcgt ctggcggaaa acctcagtgt   3540
gacgctcccc gccgcgtccc acgccatccc gcatctgacc accagcgaaa tggatttttg   3600
catcgagctg ggtaataagc gttggcaatt taaccgccag tcaggctttc tttcacagat   3660
gtggattggc gataaaaaac aactgctgac gccgctgcgc gatcagttca cccgtgcacc   3720
gctggataac gacattggcg taagtgaagc gacccgcatt gaccctaacg cctgggtcga   3780
acgctggaag gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt gcacggcaga   3840
tacacttgct gatgcggtgc tgattacgac cgctcacgcg tggcagcatc aggggaaaac   3900
cttatttatc agccggaaaa cctaccggat tgatggtagt ggtcaaatgg cgattaccgt   3960
tgatgttgaa gtggcgagcg atacaccgca tccggcgcgg attggcctga actgccagct   4020
ggcgcaggta gcagagcggg taaactggct cggattaggg ccgcaagaaa actatcccga   4080
ccgccttact gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca tgtatacccc   4140
gtacgtcttc ccgagcgaaa acggtctgcg ctgcgggacg cgcgaattga attatggccc   4200
acaccagtgg cgcggcgact tccagttcaa catcagccgc tacagtcaac agcaactgat   4260
ggaaaccagc catcgccatc tgctgcacgc ggaagaaggc acatggctga atatcgacgg   4320
tttccatatg gggattggtg gcgacgactc ctggagcccg tcagtatcgg cggaattcca   4380
gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaaagat ctggaggtgg   4440
tggcagcagg ccttggcgcg ccggatcctt aattaacaat tgaccggtaa taataggtag   4500
ataagtgact gattagatgc attgatccct cgaccaattc cggttatttt ccaccatatt   4560
gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc   4620
taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc   4680
agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg   4740
gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc   4800
tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa   4860
atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg   4920
tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa   4980
aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa   5040
taccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct   5100
attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct   5160
gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga   5220
actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc   5280
tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg   5340
gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc   5400
aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca   5460
tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga   5520
cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc   5580
cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga   5640
aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca   5700
ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg   5760
cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct   5820
tcttgacgag ttcttctgag cgggactctg gggttcgcat cgataaaata aaagatttta   5880
tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct   5940
taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga   6000
tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc   6060
agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg   6120
atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg   6180
cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac   6240
ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg   6300
cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt   6360
cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca   6420
tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc   6480
agcgggggtc tttcattcat gcagcatgta tcaaaattaa tttggttttt tttcttaagt   6540
atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt   6600
ttgcgtattg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc   6660
tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg   6720
ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg   6780
ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac   6840
gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg   6900
gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct   6960
ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg   7020
tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct   7080
gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac   7140
tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt   7200
tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc   7260
tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca   7320
ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat   7380
ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac   7440
gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc   7500
cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc   7560
agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc   7620
gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata   7680
ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg   7740
gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc   7800
cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct   7860
acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa   7920
cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt   7980
cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca   8040
ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac   8100
tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca   8160
atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt   8220
tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc   8280
actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca   8340
aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata   8400
ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc   8460
ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttc     8518
 
           
             5 
             8518 
             DNA 
             Unknown 
             
               pICAST ALN. 
             
           
            5
gacgtcggac ttatacccgg tttgtcctat agacaccatt cgtcaaggac ggggccgagt     60
cccggttctt gtctaccttg tcgacttata cccggtttgt cctatagaca ccattcgtca    120
aggacggggc cgagtcccgg ttcttgtcta ccaggggtct acgccaggtc gggagtcgtc    180
aaagatctct tggtagtcta caaaggtccc acggggttcc tggactttac tgggacacgg    240
aataaacttg attggttagt caagcgaaga gcgaagacaa gcgcgcgaag acgaggggct    300
cgagttattt tctcgggtgt tggggagtga gccccgcggt caggaggcta actgactcag    360
cgggcccatg ggcacatagg ttatttggga gaacgtcaac gtaggctgaa caccagagcg    420
acaaggaacc ctcccagagg agactcacta actgatgggc agtcgccccc agaaagtaaa    480
cccccgagca ggccctagcc ctctggggac gggtccctgg tggctgggtg gtggccctcc    540
gttcgaccgg tcgttgaata gacacagaca ggctaacaga tcacagatac tgactaaaat    600
acgcggacgc agccatgatc aatcgattga tcgagacata gaccgcctgg gcaccacctt    660
gactgctcaa gacttgtggg ccggcgttgg gaccctctgc agggtccctg aaacccccgg    720
caaaaacacc gggctggact ccttccctca gctacacctt aggctggggc agtcctatac    780
accaagacca tcctctgctc ttggattttg tcaagggcgg aggcagactt aaaaacgaaa    840
gccaaacctt ggcttcggcg cgcagaacag acgacgtcgt agcaagacac aacagagaca    900
gactgacaca aagacataaa cagactttta atcccggtct gacaatggtg agggaattca    960
aactggaatc cattgacctt tctacagctc gccgagcgag tgttggtcag ccatctacag   1020
ttcttctctg caacccaatg gaagacgaga cgtcttaccg gttggaaatt gcagcctacc   1080
ggcgctctgc cgtggaaatt ggctctggag tagtgggtcc aattctagtt ccagaaaagt   1140
ggaccgggcg tacctgtggg tctggtccag gggatgtagc actggaccct tcggaaccga   1200
aaactggggg gagggaccca gttcgggaaa catgtgggat tcggaggcgg aggagaagga   1260
ggtaggcggg gcagagaggg ggaacttgga ggagcaagct ggggcggagc taggagggaa   1320
ataggtcggg agtgaggaag agatccgcgg ccggcgagat cgggtaatta tgctgagtga   1380
tatcccgcta agcttgtggt acgtggtagt agtagtagtg cagctgatat ttctactcct   1440
ggagctctac ccgcactaat gcctaagtga ccggcagcac cgggcgtggc tagcgggaag   1500
ggttgtcaat gcgtcggact taccgcttac cgcgaaacgg accaaaggcc gtggtcttcg   1560
ccacggcctt tcgaccgacc tcacgctaga aggactccgg ctatgacagc agcaggggag   1620
tttgaccgtc tacgtgccaa tgctacgcgg gtagatgtgg ttgcactgga tagggtaatg   1680
ccagttaggc ggcaaacaag ggtgcctctt aggctgccca acaatgagcg agtgtaaatt   1740
acaactactt tcgaccgatg tccttccggt ctgcgcttaa taaaaactac cgcaattgag   1800
ccgcaaagta gacaccacgt tgcccgcgac ccagccaatg ccggtcctgt cagcaaacgg   1860
cagacttaaa ctggactcgc gtaaaaatgc gcggcctctt ttggcggagc gccactacca   1920
cgacgcgacc tcactgccgt caatagacct tctagtccta tacaccgcct actcgccgta   1980
aaaggcactg cagagcaacg acgtatttgg ctgatgtgtt tagtcgctaa aggtacaacg   2040
gtgagcgaaa ttactactaa agtcggcgcg acatgacctc cgacttcaag tctacacgcc   2100
gctcaacgca ctgatggatg cccattgtca aagaaatacc gtcccacttt gcgtccagcg   2160
gtcgccgtgg cgcggaaagc cgccacttta atagctactc gcaccaccaa tacggctagc   2220
gcagtgtgat gcagacttgc agcttttggg ctttgacacc tcgcggcttt agggcttaga   2280
gatagcacgc caccaacttg acgtgtggcg gctgccgtgc gactaacttc gtcttcggac   2340
gctacagcca aaggcgctcc acgcctaact tttaccagac gacgacgact tgccgttcgg   2400
caacgactaa gctccgcaat tggcagtgct cgtagtagga gacgtaccag tccagtacct   2460
actcgtctgc taccacgtcc tataggacga ctacttcgtc ttgttgaaat tgcggcacgc   2520
gacaagcgta ataggcttgg taggcgacac catgtgcgac acgctggcga tgccggacat   2580
acaccaccta cttcggttat aactttgggt gccgtaccac ggttacttag cagactggct   2640
actaggcgcg accgatggcc gctactcgct tgcgcattgc gcttaccacg tcgcgctagc   2700
attagtgggc tcacactagt agaccagcga ccccttactt agtccggtgc cgcgattagt   2760
gctgcgcgac atagcgacct agtttagaca gctaggaagg gcgggccacg tcatacttcc   2820
gccgcctcgg ctgtggtgcc ggtggctata ataaacgggc tacatgcgcg cgcacctact   2880
tctggtcggg aagggccgac acggctttac caggtagttt tttaccgaaa gcgatggacc   2940
tctctgcgcg ggcgactagg aaacgcttat gcgggtgcgc tacccattgt cagaaccgcc   3000
aaagcgattt atgaccgtcc gcaaagcagt cataggggca aatgtcccgc cgaagcagac   3060
cctgacccac ctagtcagcg actaatttat actacttttg ccgttgggca ccagccgaat   3120
gccgccacta aaaccgctat gcggcttgct agcggtcaag acatacttgc cagaccagaa   3180
acggctggcg tgcggcgtag gtcgcgactg ccttcgtttt gtggtcgtcg tcaaaaaggt   3240
caaggcaaat aggcccgttt ggtagcttca ctggtcgctt atggacaagg cagtatcgct   3300
attgctcgag gacgtgacct accaccgcga cctaccattc ggcgaccgtt cgccacttca   3360
cggagaccta cagcgaggtg ttccatttgt caactaactt gacggacttg atggcgtcgg   3420
cctctcgcgg cccgttgaga ccgagtgtca tgcgcatcac gttggcttgc gctggcgtac   3480
cagtcttcgg cccgtgtagt cgcggaccgt cgtcaccgca gaccgccttt tggagtcaca   3540
ctgcgagggg cggcgcaggg tgcggtaggg cgtagactgg tggtcgcttt acctaaaaac   3600
gtagctcgac ccattattcg caaccgttaa attggcggtc agtccgaaag aaagtgtcta   3660
cacctaaccg ctattttttg ttgacgactg cggcgacgcg ctagtcaagt gggcacgtgg   3720
cgacctattg ctgtaaccgc attcacttcg ctgggcgtaa ctgggattgc ggacccagct   3780
tgcgaccttc cgccgcccgg taatggtccg gcttcgtcgc aacaacgtca cgtgccgtct   3840
atgtgaacga ctacgccacg actaatgctg gcgagtgcgc accgtcgtag tccccttttg   3900
gaataaatag tcggcctttt ggatggccta actaccatca ccagtttacc gctaatggca   3960
actacaactt caccgctcgc tatgtggcgt aggccgcgcc taaccggact tgacggtcga   4020
ccgcgtccat cgtctcgccc atttgaccga gcctaatccc ggcgttcttt tgatagggct   4080
ggcggaatga cggcggacaa aactggcgac cctagacggt aacagtctgt acatatgggg   4140
catgcagaag ggctcgcttt tgccagacgc gacgccctgc gcgcttaact taataccggg   4200
tgtggtcacc gcgccgctga aggtcaagtt gtagtcggcg atgtcagttg tcgttgacta   4260
cctttggtcg gtagcggtag acgacgtgcg ccttcttccg tgtaccgact tatagctgcc   4320
aaaggtatac ccctaaccac cgctgctgag gacctcgggc agtcatagcc gccttaaggt   4380
cgactcgcgg ccagcgatgg taatggtcaa ccagaccaca gttttttcta gacctccacc   4440
accgtcgtcc ggaaccgcgc ggcctaggaa ttaattgtta actggccatt attatccatc   4500
tattcactga ctaatctacg taactaggga gctggttaag gccaataaaa ggtggtataa   4560
cggcagaaaa ccgttacact cccgggcctt tggaccggga cagaagaact gctcgtaagg   4620
atccccagaa aggggagagc ggtttcctta cgttccagac aacttacagc acttccttcg   4680
tcaaggagac cttcgaagaa cttctgtttg ttgcagacat cgctgggaaa cgtccgtcgc   4740
cttggggggt ggaccgctgt ccacggagac gccggttttc ggtgcacata ttctatgtgg   4800
acgtttccgc cgtgttgggg tcacggtgca acactcaacc tatcaacacc tttctcagtt   4860
taccgagagg agttcgcata agttgttccc cgacttccta cgggtcttcc atggggtaac   4920
ataccctaga ctagaccccg gagccacgtg tacgaaatgt acacaaatca gctccaattt   4980
tttgcagatc cggggggctt ggtgcccctg caccaaaagg aaactttttg tgctactatt   5040
atggtactaa cttgttctac ctaacgtgcg tccaagaggc cggcgaaccc acctctccga   5100
taagccgata ctgacccgtg ttgtctgtta gccgacgaga ctacggcggc acaaggccga   5160
cagtcgcgtc cccgcgggcc aagaaaaaca gttctggctg gacaggccac gggacttact   5220
tgacgtcctg ctccgtcgcg ccgatagcac cgaccggtgc tgcccgcaag gaacgcgtcg   5280
acacgagctg caacagtgac ttcgcccttc cctgaccgac gataacccgc ttcacggccc   5340
cgtcctagag gacagtagag tggaacgagg acggctcttt cataggtagt accgactacg   5400
ttacgccgcc gacgtatgcg aactaggccg atggacgggt aagctggtgg ttcgctttgt   5460
agcgtagctc gctcgtgcat gagcctacct tcggccagaa cagctagtcc tactagacct   5520
gcttctcgta gtccccgagc gcggtcggct tgacaagcgg tccgagttcc gcgcgtacgg   5580
gctgccgctc ctagagcagc actgggtacc gctacggacg aacggcttat agtaccacct   5640
tttaccggcg aaaagaccta agtagctgac accggccgac ccacaccgcc tggcgatagt   5700
cctgtatcgc aaccgatggg cactataacg acttctcgaa ccgccgctta cccgactggc   5760
gaaggagcac gaaatgccat agcggcgagg gctaagcgtc gcgtagcgga agatagcgga   5820
agaactgctc aagaagactc gccctgagac cccaagcgta gctattttat tttctaaaat   5880
aaatcagagg tctttttccc cccttacttt ctggggtgga catccaaacc gttcgatcga   5940
attcattgcg gtaaaacgtt ccgtaccttt ttatgtattg actcttatct cttcaagtct   6000
agttccagtc cttgtctacc ttgtcgactt atacccggtt tgtcctatag acaccattcg   6060
tcaaggacgg ggccgagtcc cggttcttgt ctaccttgtc gacttatacc cggtttgtcc   6120
tatagacacc attcgtcaag gacggggccg agtcccggtt cttgtctacc aggggtctac   6180
gccaggtcgg gagtcgtcaa agatctcttg gtagtctaca aaggtcccac ggggttcctg   6240
gactttactg ggacacggaa taaacttgat tggttagtca agcgaagagc gaagacaagc   6300
gcgcgaagac gaggggctcg agttattttc tcgggtgttg gggagtgagc cccgcggtca   6360
ggaggctaac tgactcagcg ggcccatggg cacataggtt atttgggaga acgtcaacgt   6420
aggctgaaca ccagagcgac aaggaaccct cccagaggag actcactaac tgatgggcag   6480
tcgcccccag aaagtaagta cgtcgtacat agttttaatt aaaccaaaaa aaagaattca   6540
taaatgtaat ttaccggtat caacgtaatt acttagccgg ttgcgcgccc ctctccgcca   6600
aacgcataac cgcgagaagg cgaaggagcg agtgactgag cgacgcgagc cagcaagccg   6660
acgccgctcg ccatagtcga gtgagtttcc gccattatgc caataggtgt cttagtcccc   6720
tattgcgtcc tttcttgtac actcgttttc cggtcgtttt ccggtccttg gcatttttcc   6780
ggcgcaacga ccgcaaaaag gtatccgagg cggggggact gctcgtagtg tttttagctg   6840
cgagttcagt ctccaccgct ttgggctgtc ctgatatttc tatggtccgc aaagggggac   6900
cttcgaggga gcacgcgaga ggacaaggct gggacggcga atggcctatg gacaggcgga   6960
aagagggaag cccttcgcac cgcgaaagag tatcgagtgc gacatccata gagtcaagcc   7020
acatccagca agcgaggttc gacccgacac acgtgcttgg ggggcaagtc gggctggcga   7080
cgcggaatag gccattgata gcagaactca ggttgggcca ttctgtgctg aatagcggtg   7140
accgtcgtcg gtgaccattg tcctaatcgt ctcgctccat acatccgcca cgatgtctca   7200
agaacttcac caccggattg atgccgatgt gatcttcttg tcataaacca tagacgcgag   7260
acgacttcgg tcaatggaag cctttttctc aaccatcgag aactaggccg tttgtttggt   7320
ggcgaccatc gccaccaaaa aaacaaacgt tcgtcgtcta atgcgcgtct ttttttccta   7380
gagttcttct aggaaactag aaaagatgcc ccagactgcg agtcaccttg cttttgagtg   7440
caattcccta aaaccagtac tctaatagtt tttcctagaa gtggatctag gaaaacgccg   7500
gcgtttagtt agatttcata tatactcatt tgaaccagac tgtcaatggt tacgaattag   7560
tcactccgtg gatagagtcg ctagacagat aaagcaagta ggtatcaacg gactgagggg   7620
cagcacatct attgatgcta tgccctcccg aatggtagac cggggtcacg acgttactat   7680
ggcgctctgg gtgcgagtgg ccgaggtcta aatagtcgtt atttggtcgg tcggccttcc   7740
cggctcgcgt cttcaccagg acgttgaaat aggcggaggt aggtcagata attaacaacg   7800
gcccttcgat ctcattcatc aagcggtcaa ttatcaaacg cgttgcaaca acggtaacga   7860
tgtccgtagc accacagtgc gagcagcaaa ccataccgaa gtaagtcgag gccaagggtt   7920
gctagttccg ctcaatgtac tagggggtac aacacgtttt ttcgccaatc gaggaagcca   7980
ggaggctagc aacagtcttc attcaaccgg cgtcacaata gtgagtacca ataccgtcgt   8040
gacgtattaa gagaatgaca gtacggtagg cattctacga aaagacactg accactcatg   8100
agttggttca gtaagactct tatcacatac gccgctggct caacgagaac gggccgcagt   8160
tatgccctat tatggcgcgg tgtatcgtct tgaaattttc acgagtagta accttttgca   8220
agaagccccg cttttgagag ttcctagaat ggcgacaact ctaggtcaag ctacattggg   8280
tgagcacgtg ggttgactag aagtcgtaga aaatgaaagt ggtcgcaaag acccactcgt   8340
ttttgtcctt ccgttttacg gcgttttttc ccttattccc gctgtgcctt tacaacttat   8400
gagtatgaga aggaaaaagt tataataact tcgtaaatag tcccaataac agagtactcg   8460
cctatgtata aacttacata aatcttttta tttgtttatc cccaaggcgc gtgtaaag     8518
 
           
             6 
             8175 
             DNA 
             Unknown 
             
               pICAST OMC. 
             
           
            6
ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca     60
gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt    120
tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag    180
tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc    240
ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga    300
gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc    360
gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc    420
tgttccttgg gaggytctcc tctgagtgat tgactacccg tcagcggggg tctttcattt    480
gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg    540
caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta    600
tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa    660
ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc    720
gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg    780
tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt    840
cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt    900
ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt    960
ttgaccttag gtaactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc   1020
aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg   1080
ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca   1140
cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct   1200
tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct   1260
ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt   1320
tatccagccc tcactccttc tctaggcgcc ggccgctcta gcccattaat acgactcact   1380
atagggcgat tcgaatcagg ccttggcgcg ccggatcctt aattaagcgc aattgggagg   1440
tggcggtagc ctcgagatgg gcgtgattac ggattcactg gccgtcgttt tacaacgtcg   1500
tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc   1560
cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tacgcagcct   1620
gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct   1680
ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg   1740
ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt   1800
tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct   1860
acaggaaggc cagacgcgaa ttatttttga tggcgttaac tcggcgtttc atctgtggtg   1920
caacgggcgc tgggtcggtt acggccagga cagtcgtttg ccgtctgaat ttgacctgag   1980
cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct ggagtgacgg   2040
cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg acgtctcgtt   2100
gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct ttaatgatga   2160
tttcagccgc gctgtactgg aggctgaagt tcagatgtgc ggcgagttgc gtgactacct   2220
acgggtaaca gtttctttat ggcagggtga aacgcaggtc gccagcggca ccgcgccttt   2280
cggcggtgaa attatcgatg agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa   2340
cgtcgaaaac ccgaaactgt ggagcgccga aatcccgaat ctctatcgtg cggtggttga   2400
actgcacacc gccgacggca cgctgattga agcagaagcc tgcgatgtcg gtttccgcga   2460
ggtgcggatt gaaaatggtc tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt   2520
taaccgtcac gagcatcatc ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca   2580
ggatatcctg ctgatgaagc agaacaactt taacgccgtg cgctgttcgc attatccgaa   2640
ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa   2700
tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc gctggctacc   2760
ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat   2820
catctggtcg ctggggaatg aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg   2880
gatcaaatct gtcgatcctt cccgcccggt gcagtatgaa ggcggcggag ccgacaccac   2940
ggccaccgat attatttgcc cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc   3000
tgtgccgaaa tggtccatca aaaaatggct ttcgctacct ggagagacgc gcccgctgat   3060
cctttgcgaa tacgcccacg cgatgggtaa cagtcttggc ggtttcgcta aatactggca   3120
ggcgtttcgt cagtatcccc gtttacaggg cggcttcgtc tgggactggg tggatcagtc   3180
gctgattaaa tatgatgaaa acggcaaccc gtggtcggct tacggcggtg attttggcga   3240
tacgccgaac gatcgccagt tctgtatgaa cggtctggtc tttgccgacc gcacgccgca   3300
tccagcgctg acggaagcaa aacaccagca gcagtttttc cagttccgtt tatccgggca   3360
aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc gataacgagc tcctgcactg   3420
gatggtggcg ctggatggta agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc   3480
acaaggtaaa cagttgattg aactgcctga actaccgcag ccggagagcg ccgggcaact   3540
ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat   3600
cagcgcctgg cagcagtggc gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc   3660
ccacgccatc ccgcatctga ccaccagcga aatggatttt tgcatcgagc tgggtaataa   3720
gcgttggcaa tttaaccgcc agtcaggctt tctttcacag atgtggattg gcgataaaaa   3780
acaactgctg acgccgctgc gcgatcagtt cacccgtgtc gatagatctg aacagaaact   3840
catttccgaa gaagacctag tcgaccatca tcatcatcat caccggtaat aataggtaga   3900
taagtgactg attagatgca tttcgactag atccctcgac caattccggt tattttccac   3960
catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag   4020
cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa   4080
ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag   4140
gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga   4200
tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag   4260
agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc   4320
ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag   4380
gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga   4440
tgataatacc atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga   4500
aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt   4560
cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt   4620
ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt   4680
gcttgacatt ggggaattta gcgagagcct gacctattgc atctcccgcc gtgcacaggg   4740
tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga   4800
ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg   4860
accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc   4920
ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc   4980
tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc   5040
ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg   5100
gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc   5160
gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc   5220
aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag   5280
cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt   5340
ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg   5400
gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc   5460
gagggcaaag gaatagagta gatgccgacc gggatctatc gataaaataa aagattttat   5520
ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt   5580
aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat   5640
caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca   5700
gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga   5760
tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc   5820
ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc   5880
tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc   5940
gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc   6000
ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat   6060
ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca   6120
gcgggggtct ttcattcatg cagcatgtat caaaattaat ttggtttttt ttcttaagta   6180
tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt   6240
tgcgtattgg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct   6300
gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga   6360
taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc   6420
cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg   6480
ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg   6540
aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt   6600
tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt   6660
gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg   6720
cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact   6780
ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt   6840
cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct   6900
gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac   6960
cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc   7020
tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg   7080
ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta   7140
aaaatgaagt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca   7200
gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca   7260
tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc   7320
ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa   7380
accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc   7440
agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca   7500
acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat   7560
tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag   7620
cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac   7680
tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt   7740
ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt   7800
gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc   7860
tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat   7920
ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca   7980
gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga   8040
cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg   8100
gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg   8160
ttccgcgcac atttc                                                    8175
 
           
             7 
             8175 
             DNA 
             Unknown 
             
               pICAST OMC. 
             
           
            7
gacgtcggac ttatacccgg tttgtcctat agacaccatt cgtcaaggac ggggccgagt     60
cccggttctt gtctaccttg tcgacttata cccggtttgt cctatagaca ccattcgtca    120
aggacggggc cgagtcccgg ttcttgtcta ccaggggtct acgccaggtc gggagtcgtc    180
aaagatctct tggtagtcta caaaggtccc acggggttcc tggactttac tgggacacgg    240
aataaacttg attggttagt caagcgaaga gcgaagacaa gcgcgcgaag acgaggggct    300
cgagttattt tctcgggtgt tggggagtga gccccgcggt caggaggcta actgactcag    360
cgggcccatg ggcacatagg ttatttggga gaacgtcaac gtaggctgaa caccagagcg    420
acaaggaacc ctcccagagg agactcacta actgatgggc agtcgccccc agaaagtaaa    480
cccccgagca ggccctagcc ctctggggac gggtccctgg tggctgggtg gtggccctcc    540
gttcgaccgg tcgttgaata gacacagaca ggctaacaga tcacagatac tgactaaaat    600
acgcggacgc agccatgatc aatcgattga tcgagacata gaccgcctgg gcaccacctt    660
gactgctcaa gacttgtggg ccggcgttgg gaccctctgc agggtccctg aaacccccgg    720
caaaaacacc gggctggact ccttccctca gctacacctt aggctggggc agtcctatac    780
accaagacca tcctctgctc ttggattttg tcaagggcgg aggcagactt aaaaacgaaa    840
gccaaacctt ggcttcggcg cgcagaacag acgacgtcgt agcaagacac aacagagaca    900
gactgacaca aagacataaa cagactttta atcccggtct gacaatggtg agggaattca    960
aactggaatc cattgacctt tctacagctc gccgagcgag tgttggtcag ccatctacag   1020
ttcttctctg caacccaatg gaagacgaga cgtcttaccg gttggaaatt gcagcctacc   1080
ggcgctctgc cgtggaaatt ggctctggag tagtgggtcc aattctagtt ccagaaaagt   1140
ggaccgggcg tacctgtggg tctggtccag gggatgtagc actggaccct tcggaaccga   1200
aaactggggg gagggaccca gttcgggaaa catgtgggat tcggaggcgg aggagaagga   1260
ggtaggcggg gcagagaggg ggaacttgga ggagcaagct ggggcggagc taggagggaa   1320
ataggtcggg agtgaggaag agatccgcgg ccggcgagat cgggtaatta tgctgagtga   1380
tatcccgcta agcttagtcc ggaaccgcgc ggcctaggaa ttaattcgcg ttaaccctcc   1440
accgccatcg gagctctacc cgcactaatg cctaagtgac cggcagcaaa atgttgcagc   1500
actgaccctt ttgggaccgc aatgggttga attagcggaa cgtcgtgtag ggggaaagcg   1560
gtcgaccgca ttatcgcttc tccgggcgtg gctagcggga agggttgtca atgcgtcgga   1620
cttaccgctt accgcgaaac ggaccaaagg ccgtggtctt cgccacggcc tttcgaccga   1680
cctcacgcta gaaggactcc ggctatgaca gcagcagggg agtttgaccg tctacgtgcc   1740
aatgctacgc gggtagatgt ggttgcactg gatagggtaa tgccagttag gcggcaaaca   1800
agggtgcctc ttaggctgcc caacaatgag cgagtgtaaa ttacaactac tttcgaccga   1860
tgtccttccg gtctgcgctt aataaaaact accgcaattg agccgcaaag tagacaccac   1920
gttgcccgcg acccagccaa tgccggtcct gtcagcaaac ggcagactta aactggactc   1980
gcgtaaaaat gcgcggcctc ttttggcgga gcgccactac cacgacgcga cctcactgcc   2040
gtcaatagac cttctagtcc tatacaccgc ctactcgccg taaaaggcac tgcagagcaa   2100
cgacgtattt ggctgatgtg tttagtcgct aaaggtacaa cggtgagcga aattactact   2160
aaagtcggcg cgacatgacc tccgacttca agtctacacg ccgctcaacg cactgatgga   2220
tgcccattgt caaagaaata ccgtcccact ttgcgtccag cggtcgccgt ggcgcggaaa   2280
gccgccactt taatagctac tcgcaccacc aatacggcta gcgcagtgtg atgcagactt   2340
gcagcttttg ggctttgaca cctcgcggct ttagggctta gagatagcac gccaccaact   2400
tgacgtgtgg cggctgccgt gcgactaact tcgtcttcgg acgctacagc caaaggcgct   2460
ccacgcctaa cttttaccag acgacgacga cttgccgttc ggcaacgact aagctccgca   2520
attggcagtg ctcgtagtag gagacgtacc agtccagtac ctactcgtct gctaccacgt   2580
cctataggac gactacttcg tcttgttgaa attgcggcac gcgacaagcg taataggctt   2640
ggtaggcgac accatgtgcg acacgctggc gatgccggac atacaccacc tacttcggtt   2700
ataactttgg gtgccgtacc acggttactt agcagactgg ctactaggcg cgaccgatgg   2760
ccgctactcg cttgcgcatt gcgcttacca cgtcgcgcta gcattagtgg gctcacacta   2820
gtagaccagc gaccccttac ttagtccggt gccgcgatta gtgctgcgcg acatagcgac   2880
ctagtttaga cagctaggaa gggcgggcca cgtcatactt ccgccgcctc ggctgtggtg   2940
ccggtggcta taataaacgg gctacatgcg cgcgcaccta cttctggtcg ggaagggccg   3000
acacggcttt accaggtagt tttttaccga aagcgatgga cctctctgcg cgggcgacta   3060
ggaaacgctt atgcgggtgc gctacccatt gtcagaaccg ccaaagcgat ttatgaccgt   3120
ccgcaaagca gtcatagggg caaatgtccc gccgaagcag accctgaccc acctagtcag   3180
cgactaattt atactacttt tgccgttggg caccagccga atgccgccac taaaaccgct   3240
atgcggcttg ctagcggtca agacatactt gccagaccag aaacggctgg cgtgcggcgt   3300
aggtcgcgac tgccttcgtt ttgtggtcgt cgtcaaaaag gtcaaggcaa ataggcccgt   3360
ttggtagctt cactggtcgc ttatggacaa ggcagtatcg ctattgctcg aggacgtgac   3420
ctaccaccgc gacctaccat tcggcgaccg ttcgccactt cacggagacc tacagcgagg   3480
tgttccattt gtcaactaac ttgacggact tgatggcgtc ggcctctcgc ggcccgttga   3540
gaccgagtgt catgcgcatc acgttggctt gcgctggcgt accagtcttc ggcccgtgta   3600
gtcgcggacc gtcgtcaccg cagaccgcct tttggagtca cactgcgagg ggcggcgcag   3660
ggtgcggtag ggcgtagact ggtggtcgct ttacctaaaa acgtagctcg acccattatt   3720
cgcaaccgtt aaattggcgg tcagtccgaa agaaagtgtc tacacctaac cgctattttt   3780
tgttgacgac tgcggcgacg cgctagtcaa gtgggcacag ctatctagac ttgtctttga   3840
gtaaaggctt cttctggatc agctggtagt agtagtagta gtggccatta ttatccatct   3900
attcactgac taatctacgt aaagctgatc tagggagctg gttaaggcca ataaaaggtg   3960
gtataacggc agaaaaccgt tacactcccg ggcctttgga ccgggacaga agaactgctc   4020
gtaaggatcc ccagaaaggg gagagcggtt tccttacgtt ccagacaact tacagcactt   4080
ccttcgtcaa ggagaccttc gaagaacttc tgtttgttgc agacatcgct gggaaacgtc   4140
cgtcgccttg gggggtggac cgctgtccac ggagacgccg gttttcggtg cacatattct   4200
atgtggacgt ttccgccgtg ttggggtcac ggtgcaacac tcaacctatc aacacctttc   4260
tcagtttacc gagaggagtt cgcataagtt gttccccgac ttcctacggg tcttccatgg   4320
ggtaacatac cctagactag accccggagc cacgtgtacg aaatgtacac aaatcagctc   4380
caattttttg cagatccggg gggcttggtg cccctgcacc aaaaggaaac tttttgtgct   4440
actattatgg tactttttcg gacttgagtg gcgctgcaga cagctcttca aagactagct   4500
tttcaagctg tcgcagaggc tggactacgt cgagagcctc ccgcttctta gagcacgaaa   4560
gtcgaagcta catcctcccg cacctataca ggacgcccat ttatcgacgc ggctaccaaa   4620
gatgtttcta gcaatacaaa tagccgtgaa acgtagccgg cgcgagggct aaggccttca   4680
cgaactgtaa ccccttaaat cgcrctcgga ctggataacg tagagggcgg cacgtgtccc   4740
acagtgcaac gttctggacg gactttggct tgacgggcga caagacgtcg gccagcgcct   4800
ccggtaccta cgctagcgac gccggctaga atcggtctgc tcgcccaagc cgggtaagcc   4860
tggcgttcct tagccagtta tgtgatgtac cgcactaaag tatacgcgct aacgactagg   4920
ggtacacata gtgaccgttt gacactacct gctgtggcag tcacgcaggc agcgcgtccg   4980
agagctactc gactacgaaa cccggctcct gacggggctt caggccgtgg agcacgtgcg   5040
cctaaagccg aggttgttac aggactgcct gttaccggcg tattgtcgcc agtaactgac   5100
ctcgctccgc tacaagcccc taagggttat gctccagcgg ttgtagaaga agacctccgg   5160
caccaaccga acatacctcg tcgtctgcgc gatgaagctc gcctccgtag gcctcgaacg   5220
tcctagcggc gccgaggccc gcatatacga ggcgtaacca gaactgcttg agatagtctc   5280
gaaccaactg ccgttaaagc tactacgtcg aacccgcgtc ccagctacgc tgcgttagca   5340
ggctaggcct cggccctgac agcccgcatg tgtttagcgg gcgtcttcgc gccggcagac   5400
ctggctaccg acacatcttc atgagcggct atcacctttg gctgcggggt cgtgagcagg   5460
ctcccgtttc cttatctcat ctacggctgg ccctagatag ctattttatt ttctaaaata   5520
aatcagaggt ctttttcccc ccttactttc tggggtggac atccaaaccg ttcgatcgaa   5580
ttcattgcgg taaaacgttc cgtacctttt tatgtattga ctcttatctc ttcaagtcta   5640
gttccagtcc ttgtctacct tgtcgactta tacccggttt gtcctataga caccattcgt   5700
caaggacggg gccgagtccc ggttcttgtc taccttgtcg acttataccc ggtttgtcct   5760
atagacacca ttcgtcaagg acggggccga gtcccggttc ttgtctacca ggggtctacg   5820
ccaggtcggg agtcgtcaaa gatctcttgg tagtctacaa aggtcccacg gggttcctgg   5880
actttactgg gacacggaat aaacttgatt ggttagtcaa gcgaagagcg aagacaagcg   5940
cgcgaagacg aggggctcga gttattttct cgggtgttgg ggagtgagcc ccgcggtcag   6000
gaggctaact gactcagcgg gcccatgggc acataggtta tttgggagaa cgtcaacgta   6060
ggctgaacac cagagcgaca aggaaccctc ccagaggaga ctcactaact gatgggcagt   6120
cgcccccaga aagtaagtac gtcgtacata gttttaatta aaccaaaaaa aagaattcat   6180
aaatgtaatt taccggtatc aacgtaatta cttagccggt tgcgcgcccc tctccgccaa   6240
acgcataacc gcgagaaggc gaaggagcga gtgactgagc gacgcgagcc agcaagccga   6300
cgccgctcgc catagtcgag tgagtttccg ccattatgcc aataggtgtc ttagtcccct   6360
attgcgtcct ttcttgtaca ctcgttttcc ggtcgttttc cggtccttgg catttttccg   6420
gcgcaacgac cgcaaaaagg tatccgaggc ggggggactg ctcgtagtgt ttttagctgc   6480
gagttcagtc tccaccgctt tgggctgtcc tgatatttct atggtccgca aagggggacc   6540
ttcgagggag cacgcgagag gacaaggctg ggacggcgaa tggcctatgg acaggcggaa   6600
agagggaagc ccttcgcacc gcgaaagagt atcgagtgcg acatccatag agtcaagcca   6660
catccagcaa gcgaggttcg acccgacaca cgtgcttggg gggcaagtcg ggctggcgac   6720
gcggaatagg ccattgatag cagaactcag gttgggccat tctgtgctga atagcggtga   6780
ccgtcgtcgg tgaccattgt cctaatcgtc tcgctccata catccgccac gatgtctcaa   6840
gaacttcacc accggattga tgccgatgtg atcttcttgt cataaaccat agacgcgaga   6900
cgacttcggt caatggaagc ctttttctca accatcgaga actaggccgt ttgtttggtg   6960
gcgaccatcg ccaccaaaaa aacaaacgtt cgtcgtctaa tgcgcgtctt tttttcctag   7020
agttcttcta ggaaactaga aaagatgccc cagactgcga gtcaccttgc ttttgagtgc   7080
aattccctaa aaccagtact ctaatagttt ttcctagaag tggatctagg aaaatttaat   7140
ttttacttca aacgccggcg tttagttaga tttcatatat actcatttga accagactgt   7200
caatggttac gaattagtca ctccgtggat agagtcgcta gacagataaa gcaagtaggt   7260
atcaacggac tgaggggcag cacatctatt gatgctatgc cctcccgaat ggtagaccgg   7320
ggtcacgacg ttactatggc gctctgggtg cgagtggccg aggtctaaat agtcgttatt   7380
tggtcggtcg gccttcccgg ctcgcgtctt caccaggacg ttgaaatagg cggaggtagg   7440
tcagataatt aacaacggcc cttcgatctc attcatcaag cggtcaatta tcaaacgcgt   7500
tgcaacaacg gtaacgatgt ccgtagcacc acagtgcgag cagcaaacca taccgaagta   7560
agtcgaggcc aagggttgct agttccgctc aatgtactag ggggtacaac acgttttttc   7620
gccaatcgag gaagccagga ggctagcaac agtcttcatt caaccggcgt cacaatagtg   7680
agtaccaata ccgtcgtgac gtattaagag aatgacagta cggtaggcat tctacgaaaa   7740
gacactgacc actcatgagt tggttcagta agactcttat cacatacgcc gctggctcaa   7800
cgagaacggg ccgcagttat gccctattat ggcgcggtgt atcgtcttga aattttcacg   7860
agtagtaacc ttttgcaaga agccccgctt ttgagagttc ctagaatggc gacaactcta   7920
ggtcaagcta cattgggtga gcacgtgggt tgactagaag tcgtagaaaa tgaaagtggt   7980
cgcaaagacc cactcgtttt tgtccttccg ttttacggcg ttttttccct tattcccgct   8040
gtgcctttac aacttatgag tatgagaagg aaaaagttat aataacttcg taaatagtcc   8100
caataacaga gtactcgcct atgtataaac ttacataaat ctttttattt gtttatcccc   8160
aaggcgcgtg taaag                                                    8175
 
           
             8 
             8161 
             DNA 
             Unknown 
             
               pICAST OMN. 
             
           
            8
ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca     60
gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt    120
tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag    180
tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc    240
ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga    300
gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc    360
gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc    420
tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt    480
gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg    540
caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta    600
tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa    660
ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc    720
gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg    780
tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt    840
cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt    900
ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt    960
ttgaccttag gtaactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc   1020
aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg   1080
ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca   1140
cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct   1200
tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct   1260
ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt   1320
tatccagccc tcactccttc tctaggcgcc ggccgctcta gcccattaat acgactcact   1380
atagggcgat tcgaacacca tgcaccatca tcatcatcac gtcgacgaac agaaactcat   1440
ttccgaagaa gacctactcg agatgggcgt gattacggat tcactggccg tcgttttaca   1500
acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc   1560
tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttacg   1620
cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag   1680
ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat   1740
gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc   1800
gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag   1860
ctggctacag gaaggccaga cgcgaattat ttttgatggc gttaactcgg cgtttcatct   1920
gtggtgcaac gggcgctggg tcggttacgg ccaggacagt cgtttgccgt ctgaatttga   1980
cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg gtgatggtgc tgcgctggag   2040
tgacggcagt tatctggaag atcaggatat gtggcggatg agcggcattt tccgtgacgt   2100
ctcgttgctg cataaaccga ctacacaaat cagcgatttc catgttgcca ctcgctttaa   2160
tgatgatttc agccgcgctg tactggaggc tgaagttcag atgtgcggcg agttgcgtga   2220
ctacctacgg gtaacagttt ctttatggca gggtgaaacg caggtcgcca gcggcaccgc   2280
gcctttcggc ggtgaaatta tcgatgagcg tggtggttat gccgatcgcg tcacactacg   2340
tctgaacgtc gaaaacccga aactgtggag cgccgaaatc ccgaatctct atcgtgcggt   2400
ggttgaactg cacaccgccg acggcacgct gattgaagca gaagcctgcg atgtcggttt   2460
ccgcgaggtg cggattgaaa atggtctgct gctgctgaac ggcaagccgt tgctgattcg   2520
aggcgttaac cgtcacgagc atcatcctct gcatggtcag gtcatggatg agcagacgat   2580
ggtgcaggat atcctgctga tgaagcagaa caactttaac gccgtgcgct gttcgcatta   2640
tccgaaccat ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg tggtggatga   2700
agccaatatt gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg   2760
gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag   2820
tgtgatcatc tggtcgctgg ggaatgaatc aggccacggc gctaatcacg acgcgctgta   2880
tcgctggatc aaatctgtcg atccttcccg cccggtgcag tatgaaggcg gcggagccga   2940
caccacggcc accgatatta tttgcccgat gtacgcgcgc gtggatgaag accagccctt   3000
cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg ctacctggag agacgcgccc   3060
gctgatcctt tgcgaatacg cccacgcgat gggtaacagt cttggcggtt tcgctaaata   3120
ctggcaggcg tttcgtcagt atccccgttt acagggcggc ttcgtctggg actgggtgga   3180
tcagtcgctg attaaatatg atgaaaacgg caacccgtgg tcggcttacg gcggtgattt   3240
tggcgatacg ccgaacgatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac   3300
gccgcatcca gcgctgacgg aagcaaaaca ccagcagcag tttttccagt tccgtttatc   3360
cgggcaaacc atcgaagtga ccagcgaata cctgttccgt catagcgata acgagctcct   3420
gcactggatg gtggcgctgg atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt   3480
cgctccacaa ggtaaacagt tgattgaact gcctgaacta ccgcagccgg agagcgccgg   3540
gcaactctgg ctcacagtac gcgtagtgca accgaacgcg accgcatggt cagaagccgg   3600
gcacatcagc gcctggcagc agtggcgtct ggcggaaaac ctcagtgtga cgctccccgc   3660
cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca tcgagctggg   3720
taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt ggattggcga   3780
taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgtcgata gatctggagg   3840
tggtggcagc aggccttggc gcgccggatc cttaattaac aattgaccgg taataatagg   3900
tagataagtg actgattaga tgcatttcga ctagatccct cgaccaattc cggttatttt   3960
ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga   4020
cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg   4080
tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt   4140
gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat   4200
aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg   4260
aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg   4320
taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt   4380
cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac   4440
acgatgataa taccatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga   4500
tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg   4560
ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg   4620
gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg   4680
aagtgcttga cattggggaa tttagcgaga gcctgaccta ttgcatctcc cgccgtgcac   4740
agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg   4800
cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat   4860
tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg   4920
atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc   4980
aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc   5040
acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg   5100
actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga   5160
ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc   5220
ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc   5280
agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa   5340
tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg   5400
tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc   5460
gtccgagggc aaaggaatag agtagatgcc gaccgggatc tatcgataaa ataaaagatt   5520
ttatttagtc tccagaaaaa ggggggaatg aagaccccaa cctgtaggtt tggcaagcta   5580
gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc   5640
agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta   5700
agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac   5760
aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag   5820
atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag   5880
gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt   5940
tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc   6000
agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt   6060
gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc   6120
gtcagcgggg gtctttcatt catgcagcat gtatcaaaat taatttggtt ttttttctta   6180
agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc   6240
ggtttgcgta ttggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc   6300
ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag   6360
gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa   6420
aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc   6480
gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc   6540
ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg   6600
cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt   6660
cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc   6720
gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc   6780
cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag   6840
agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg   6900
ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa   6960
ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag   7020
gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact   7080
cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc   7140
ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta   7200
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc   7260
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg   7320
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga   7380
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt   7440
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt   7500
gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc   7560
caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc   7620
ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca   7680
gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag   7740
tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg   7800
tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa   7860
cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa   7920
cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga   7980
gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga   8040
atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg   8100
agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt   8160
c                                                                   8161
 
           
             9 
             8161 
             DNA 
             Unknown 
             
               pICAST OMN. 
             
           
            9
gacgtcggac ttatacccgg tttgtcctat agacaccatt cgtcaaggac ggggccgagt     60
cccggttctt gtctaccttg tcgacttata cccggtttgt cctatagaca ccattcgtca    120
aggacggggc cgagtcccgg ttcttgtcta ccaggggtct acgccaggtc gggagtcgtc    180
aaagatctct tggtagtcta caaaggtccc acggggttcc tggactttac tgggacacgg    240
aataaacttg attggttagt caagcgaaga gcgaagacaa gcgcgcgaag acgaggggct    300
cgagttattt tctcgggtgt tggggagtga gccccgcggt caggaggcta actgactcag    360
cgggcccatg ggcacatagg ttatttggga gaacgtcaac gtaggctgaa caccagagcg    420
acaaggaacc ctcccagagg agactcacta actgatgggc agtcgccccc agaaagtaaa    480
cccccgagca ggccctagcc ctctggggac gggtccctgg tggctgggtg gtggccctcc    540
gttcgaccgg tcgttgaata gacacagaca ggctaacaga tcacagatac tgactaaaat    600
acgcggacgc agccatgatc aatcgattga tcgagacata gaccgcctgg gcaccacctt    660
gactgctcaa gacttgtggg ccggcgttgg gaccctctgc agggtccctg aaacccccgg    720
caaaaacacc gggctggact ccttccctca gctacacctt aggctggggc agtcctatac    780
accaagacca tcctctgctc ttggattttg tcaagggcgg aggcagactt aaaaacgaaa    840
gccaaacctt ggcttcggcg cgcagaacag acgacgtcgt agcaagacac aacagagaca    900
gactgacaca aagacataaa cagactttta atcccggtct gacaatggtg agggaattca    960
aactggaatc cattgacctt tctacagctc gccgagcgag tgttggtcag ccatctacag   1020
ttcttctctg caacccaatg gaagacgaga cgtcttaccg gttggaaatt gcagcctacc   1080
ggcgctctgc cgtggaaatt ggctctggag tagtgggtcc aattctagtt ccagaaaagt   1140
ggaccgggcg tacctgtggg tctggtccag gggatgtagc actggaccct tcggaaccga   1200
aaactggggg gagggaccca gttcgggaaa catgtgggat tcggaggcgg aggagaagga   1260
ggtaggcggg gcagagaggg ggaacttgga ggagcaagct ggggcggagc taggagggaa   1320
ataggtcggg agtgaggaag agatccgcgg ccggcgagat cgggtaatta tgctgagtga   1380
tatcccgcta agcttgtggt acgtggtagt agtagtagtg cagctgcttg tctttgagta   1440
aaggcttctt ctggatgagc tctacccgca ctaatgccta agtgaccggc agcaaaatgt   1500
tgcagcactg acccttttgg gaccgcaatg ggttgaatta gcggaacgtc gtgtaggggg   1560
aaagcggtcg accgcattat cgcttctccg ggcgtggcta gcgggaaggg ttgtcaatgc   1620
gtcggactta ccgcttaccg cgaaacggac caaaggccgt ggtcttcgcc acggcctttc   1680
gaccgacctc acgctagaag gactccggct atgacagcag caggggagtt tgaccgtcta   1740
cgtgccaatg ctacgcgggt agatgtggtt gcactggata gggtaatgcc agttaggcgg   1800
caaacaaggg tgcctcttag gctgcccaac aatgagcgag tgtaaattac aactactttc   1860
gaccgatgtc cttccggtct gcgcttaata aaaactaccg caattgagcc gcaaagtaga   1920
caccacgttg cccgcgaccc agccaatgcc ggtcctgtca gcaaacggca gacttaaact   1980
ggactcgcgt aaaaatgcgc ggcctctttt ggcggagcgc cactaccacg acgcgacctc   2040
actgccgtca atagaccttc tagtcctata caccgcctac tcgccgtaaa aggcactgca   2100
gagcaacgac gtatttggct gatgtgttta gtcgctaaag gtacaacggt gagcgaaatt   2160
actactaaag tcggcgcgac atgacctccg acttcaagtc tacacgccgc tcaacgcact   2220
gatggatgcc cattgtcaaa gaaataccgt cccactttgc gtccagcggt cgccgtggcg   2280
cggaaagccg ccactttaat agctactcgc accaccaata cggctagcgc agtgtgatgc   2340
agacttgcag cttttgggct ttgacacctc gcggctttag ggcttagaga tagcacgcca   2400
ccaacttgac gtgtggcggc tgccgtgcga ctaacttcgt cttcggacgc tacagccaaa   2460
ggcgctccac gcctaacttt taccagacga cgacgacttg ccgttcggca acgactaagc   2520
tccgcaattg gcagtgctcg tagtaggaga cgtaccagtc cagtacctac tcgtctgcta   2580
ccacgtccta taggacgact acttcgtctt gttgaaattg cggcacgcga caagcgtaat   2640
aggcttggta ggcgacacca tgtgcgacac gctggcgatg ccggacatac accacctact   2700
tcggttataa ctttgggtgc cgtaccacgg ttacttagca gactggctac taggcgcgac   2760
cgatggccgc tactcgcttg cgcattgcgc ttaccacgtc gcgctagcat tagtgggctc   2820
acactagtag accagcgacc ccttacttag tccggtgccg cgattagtgc tgcgcgacat   2880
agcgacctag tttagacagc taggaagggc gggccacgtc atacttccgc cgcctcggct   2940
gtggtgccgg tggctataat aaacgggcta catgcgcgcg cacctacttc tggtcgggaa   3000
gggccgacac ggctttacca ggtagttttt taccgaaagc gatggacctc tctgcgcggg   3060
cgactaggaa acgcttatgc gggtgcgcta cccattgtca gaaccgccaa agcgatttat   3120
gaccgtccgc aaagcagtca taggggcaaa tgtcccgccg aagcagaccc tgacccacct   3180
agtcagcgac taatttatac tacttttgcc gttgggcacc agccgaatgc cgccactaaa   3240
accgctatgc ggcttgctag cggtcaagac atacttgcca gaccagaaac ggctggcgtg   3300
cggcgtaggt cgcgactgcc ttcgttttgt ggtcgtcgtc aaaaaggtca aggcaaatag   3360
gcccgtttgg tagcttcact ggtcgcttat ggacaaggca gtatcgctat tgctcgagga   3420
cgtgacctac caccgcgacc taccattcgg cgaccgttcg ccacttcacg gagacctaca   3480
gcgaggtgtt ccatttgtca actaacttga cggacttgat ggcgtcggcc tctcgcggcc   3540
cgttgagacc gagtgtcatg cgcatcacgt tggcttgcgc tggcgtacca gtcttcggcc   3600
cgtgtagtcg cggaccgtcg tcaccgcaga ccgccttttg gagtcacact gcgaggggcg   3660
gcgcagggtg cggtagggcg tagactggtg gtcgctttac ctaaaaacgt agctcgaccc   3720
attattcgca accgttaaat tggcggtcag tccgaaagaa agtgtctaca cctaaccgct   3780
attttttgtt gacgactgcg gcgacgcgct agtcaagtgg gcacagctat ctagacctcc   3840
accaccgtcg tccggaaccg cgcggcctag gaattaattg ttaactggcc attattatcc   3900
atctattcac tgactaatct acgtaaagct gatctaggga gctggttaag gccaataaaa   3960
ggtggtataa cggcagaaaa ccgttacact cccgggcctt tggaccggga cagaagaact   4020
gctcgtaagg atccccagaa aggggagagc ggtttcctta cgttccagac aacttacagc   4080
acttccttcg tcaaggagac cttcgaagaa cttctgtttg ttgcagacat cgctgggaaa   4140
cgtccgtcgc cttggggggt ggaccgctgt ccacggagac gccggttttc ggtgcacata   4200
ttctatgtgg acgtttccgc cgtgttgggg tcacggtgca acactcaacc tatcaacacc   4260
tttctcagtt taccgagagg agttcgcata agttgttccc cgacttccta cgggtcttcc   4320
atggggtaac ataccctaga ctagaccccg gagccacgtg tacgaaatgt acacaaatca   4380
gctccaattt tttgcagatc cggggggctt ggtgcccctg caccaaaagg aaactttttg   4440
tgctactatt atggtacttt ttcggacttg agtggcgctg cagacagctc ttcaaagact   4500
agcttttcaa gctgtcgcag aggctggact acgtcgagag cctcccgctt cttagagcac   4560
gaaagtcgaa gctacatcct cccgcaccta tacaggacgc ccatttatcg acgcggctac   4620
caaagatgtt tctagcaata caaatagccg tgaaacgtag ccggcgcgag ggctaaggcc   4680
ttcacgaact gtaacccctt aaatcgctct cggactggat aacgtagagg gcggcacgtg   4740
tcccacagtg caacgttctg gacggacttt ggcttgacgg gcgacaagac gtcggccagc   4800
gcctccggta cctacgctag cgacgccggc tagaatcggt ctgctcgccc aagccgggta   4860
agcctggcgt tccttagcca gttatgtgat gtaccgcact aaagtatacg cgctaacgac   4920
taggggtaca catagtgacc gtttgacact acctgctgtg gcagtcacgc aggcagcgcg   4980
tccgagagct actcgactac gaaacccggc tcctgacggg gcttcaggcc gtggagcacg   5040
tgcgcctaaa gccgaggttg ttacaggact gcctgttacc ggcgtattgt cgccagtaac   5100
tgacctcgct ccgctacaag cccctaaggg ttatgctcca gcggttgtag aagaagacct   5160
ccggcaccaa ccgaacatac ctcgtcgtct gcgcgatgaa gctcgcctcc gtaggcctcg   5220
aacgtcctag cggcgccgag gcccgcatat acgaggcgta accagaactg gttgagatag   5280
tctcgaacca actgccgtta aagctactac gtcgaacccg cgtcccagct acgctgcgtt   5340
agcaggctag gcctcggccc tgacagcccg catgtgttta gcgggcgtct tcgcgccggc   5400
agacctggct accgacacat cttcatgagc ggctatcacc tttggctgcg gggtcgtgag   5460
caggctcccg tttccttatc tcatctacgg ctggccctag atagctattt tattttctaa   5520
aataaatcag aggtcttttt ccccccttac tttctggggt ggacatccaa accgttcgat   5580
cgaattcatt gcggtaaaac gttccgtacc tttttatgta ttgactctta tctcttcaag   5640
tctagttcca gtccttgtct accttgtcga cttatacccg gtttgtccta tagacaccat   5700
tcgtcaagga cggggccgag tcccggttct tgtctacctt gtcgacttat acccggtttg   5760
tcctatagac accattcgtc aaggacgggg ccgagtcccg gttcttgtct accaggggtc   5820
tacgccaggt cgggagtcgt caaagatctc ttggtagtct acaaaggtcc cacggggttc   5880
ctggacttta ctgggacacg gaataaactt gattggttag tcaagcgaag agcgaagaca   5940
agcgcgcgaa gacgaggggc tcgagttatt ttctcgggtg ttggggagtg agccccgcgg   6000
tcaggaggct aactgactca gcgggcccat gggcacatag gttatttggg agaacgtcaa   6060
cgtaggctga acaccagagc gacaaggaac cctcccagag gagactcact aactgatggg   6120
cagtcgcccc cagaaagtaa gtacgtcgta catagtttta attaaaccaa aaaaaagaat   6180
tcataaatgt aatttaccgg tatcaacgta attacttagc cggttgcgcg cccctctccg   6240
ccaaacgcat aaccgcgaga aggcgaagga gcgagtgact gagcgacgcg agccagcaag   6300
ccgacgccgc tcgccatagt cgagtgagtt tccgccatta tgccaatagg tgtcttagtc   6360
ccctattgcg tcctttcttg tacactcgtt ttccggtcgt tttccggtcc ttggcatttt   6420
tccggcgcaa cgaccgcaaa aaggtatccg aggcgggggg actgctcgta gtgtttttag   6480
ctgcgagttc agtctccacc gctttgggct gtcctgatat ttctatggtc cgcaaagggg   6540
gaccttcgag ggagcacgcg agaggacaag gctgggacgg cgaatggcct atggacaggc   6600
ggaaagaggg aagcccttcg caccgcgaaa gagtatcgag tgcgacatcc atagagtcaa   6660
gccacatcca gcaagcgagg ttcgacccga cacacgtgct tggggggcaa gtcgggctgg   6720
cgacgcggaa taggccattg atagcagaac tcaggttggg ccattctgtg ctgaatagcg   6780
gtgaccgtcg tcggtgacca ttgtcctaat cgtctcgctc catacatccg ccacgatgtc   6840
tcaagaactt caccaccgga ttgatgccga tgtgatcttc ttgtcataaa ccatagacgc   6900
gagacgactt cggtcaatgg aagccttttt ctcaaccatc gagaactagg ccgtttgttt   6960
ggtggcgacc atcgccacca aaaaaacaaa cgttcgtcgt ctaatgcgcg tctttttttc   7020
ctagagttct tctaggaaac tagaaaagat gccccagact gcgagtcacc ttgcttttga   7080
gtgcaattcc ctaaaaccag tactctaata gtttttccta gaagtggatc taggaaaacg   7140
ccggcgttta gttagatttc atatatactc atttgaacca gactgtcaat ggttacgaat   7200
tagtcactcc gtggatagag tcgctagaca gataaagcaa gtaggtatca acggactgag   7260
gggcagcaca tctattgatg ctatgccctc ccgaatggta gaccggggtc acgacgttac   7320
tatggcgctc tgggtgcgag tggccgaggt ctaaatagtc gttatttggt cggtcggcct   7380
tcccggctcg cgtcttcacc aggacgttga aataggcgga ggtaggtcag ataattaaca   7440
acggcccttc gatctcattc atcaagcggt caattatcaa acgcgttgca acaacggtaa   7500
cgatgtccgt agcaccacag tgcgagcagc aaaccatacc gaagtaagtc gaggccaagg   7560
gttgctagtt ccgctcaatg tactaggggg tacaacacgt tttttcgcca atcgaggaag   7620
ccaggaggct agcaacagtc ttcattcaac cggcgtcaca atagtgagta ccaataccgt   7680
cgtgacgtat taagagaatg acagtacggt aggcattcta cgaaaagaca ctgaccactc   7740
atgagttggt tcagtaagac tcttatcaca tacgccgctg gctcaacgag aacgggccgc   7800
agttatgccc tattatggcg cggtgtatcg tcttgaaatt ttcacgagta gtaacctttt   7860
gcaagaagcc ccgcttttga gagttcctag aatggcgaca actctaggtc aagctacatt   7920
gggtgagcac gtgggttgac tagaagtcgt agaaaatgaa agtggtcgca aagacccact   7980
cgtttttgtc cttccgtttt acggcgtttt ttcccttatt cccgctgtgc ctttacaact   8040
tatgagtatg agaaggaaaa agttataata acttcgtaaa tagtcccaat aacagagtac   8100
tcgcctatgt ataaacttac ataaatcttt ttatttgttt atccccaagg cgcgtgtaaa   8160
g                                                                   8161
 
           
             10 
             5 
             PRT 
             Artificial Sequence 
             
               GS Linker 
             
           
            10
Gly Gly Gly Gly Ser
1               5